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Cytotoxic T‐lymphocyte escape viral variants: how important are they in viral evasion of immune clearance in vivo?

Summary: Although viral variants which are not recognized by epitope‐specific cytotoxic T lymphocytes (CTL) have been shown lo arise during a number of persistent virus infections, in many cases their significance remains controversial: it has been argued that the immune response is sufficiently plastic to contain their replication. In this review, we describe the mechanisms by which amino acid changes in viral proteins may affect epitope recognition by virus‐specific CTL, and discuss the viral and immunological basis for the emergence of viral variants bearing such amino acid changes during infection. We then consider the impact that viral variation may have on the host CTL response and its ability to contain virus replication. We argue that the emergence of a viral variant demonstrates that it must have an in vivo replicative advantage, and that as such, the variant must tip the balance between virus replication and immune control somewhat in favor of the virus. Further, we suggest that although the immune response can evolve to recognize new viral epitopes, the CTL generated following such evolution frequently have a reduced ability to contain virus replication. We conclude that this escape mechanism likely does make a significant contribution to persistence/pathogenesis during a number of different virus infections.

Fungal Speciation Using Quantitative Polymerase Chain Reaction (QPCR) in Patients With and Without Chronic Rhinosinusitis

Objectives/Hypothesis: The objectives of this study were to determine the mycology of the middle meatus using an endoscopically guided brush sampling technique and polymerase chain reaction laboratory processing of nasal mucous; to compare the mycology of the middle meatus in patients with sinus disease with subjects without sinus disease; to compare the responses on two standardized quality‐of‐life survey forms between patients with and without sinusitis; and to determine whether the presence of fungi in the middle meatus correlates with responses on these data sets. Study Design: The authors conducted a single‐blind, prospective, cross‐sectional study. Methods: Patients with sinus disease and a control group without sinus disease were enrolled in the study. A disease‐specific, validated Sinonasal Outcomes Test survey (SNOT‐20) was completed by the subjects and a generalized validated Medical Outcomes Short Form 36 Survey (SF‐36) was also completed. An endoscopically guided brush sampling of nasal mucous was obtained from the middle meatus. Fungal specific quantitative polymerase chain reaction (QPCR) was performed on the obtained sample to identify one of 82 different species of fungus in the laboratory. Statistical analysis was used to categorize the recovered fungal DNA and to crossreference this information with the outcomes surveys. Results: The fungal recovery rate in the study was 45.9% in patients with sinus disease and 45.9% in control subjects. Patients with chronic rhinosinusitis had a mean SNOT‐20 score of 1.80 versus the control group mean score of 0.77 (P < .0001). SF‐36 data similarly showed a statistically significant difference between diseased and control populations with controls scoring a mean of 80.37 and patients with chronic rhinosinusitis scoring a mean of 69.35 for a P value of .02. However, no statistical significance could be ascribed to the presence or absence of fungi recovered, the type of fungi recovered, or the possible impact of fungi on the quality‐of‐life survey results. Conclusion: The recovery rate of fungi from the middle meatus of patients with chronic rhinosinusitis and a control population without chronic rhinosinusitis is 45.9% using QPCR techniques. No direct causation with regard to fungal species or presence was proven; however, a species grouping for future studies is proposed based on trends in this data and other reports. Disease‐specific outcomes surveys revealed a statistically significant difference between the two groups.

Role of aminopeptidase N (CD13) in tumor‐cell invasion and extracellular matrix degradation

We have investigated the effect of monoclonal antibodies (MAbs) specific for aminopeptidase N/CD13 on the invasion of human metastatic tumor cells into reconstituted basement membrane (Matrigel). The invasion of human metastatic tumor cells (SN12M renal‐cell carcinoma, HT1080 fibrosarcoma and A375M melanoma) into Matrigel‐coated filters was inhibited by an anti‐CD 13 MAb, WM15, in a concentration‐dependent manner. However, this MAb did not have any effect on tumor‐cell adhesion and migration to the extracellular matrices, which may be involved in tumor‐cell invasion. MAb WM15 inhibited the degradation of type‐IV collagen by tumor cells in a concentration‐dependent manner. We also found that WM15 inhibited hydrolysing activities towards substrates of aminopeptidases in 3 different tumor cells. Since our previous study indicated that bestatin, an aminopeptidase inhibitor, was able to inhibit tumor‐cell invasion, as well as aminopeptidase activities of murine and human metastatic tumor cells, cell‐surface aminopeptidase N/CD13 may be partly involved in the activation mechanism for type‐IV collagenolysis to achieve tumor‐cell invasion, and anti‐CD13 MAb WM15 may inhibit tumor‐cell invasion through a mechanism involving its inhibitory action on the aminopeptidase N in tumor cells.

Virulent and attenuated canine distemper virus infects multiple dog brain cell types in vitro

Canine Distemper Virus (CDV) produces an encephalitis in dogs that varies with viral strain. We have studied the cell tropisms of two virulent strains (CDV‐SH and CDV A75–17) and an attenuated strain, Rockborn (CDV‐RO), in cultured canine brain cells. Infected cell types were identified by double immunofluorescent labeling of specific cell markers and viral antigens. All viral strains studied produced infection in astrocytes, fibroblasts, and macrophages. Neurons were not infected by CDV A75–17 but were rapidly infected by CDV‐SH and CDV‐RO. Multipolar oligodendrocytes were very rarely infected by any of the virus strains. In contrast, a morphologically distinct subset of bipolar oligodendrocytes were commonly infected by CDV‐SH and CDV‐RO. The kinetics of infection in the astrocytes, oligodendrocytes, neurons, and macrophages varied between strains. Both CDV‐SH and CDV‐RO rapidly infected bipolar oligodendrocytes, astrocytes, neurons, and macrophages by 14 days post infection while infection by CDV A75–17 was delayed until after 28–35 days post infection. The differences in the growth kinetics and cell tropisms for some brain cells, exhibited by the three viral strains examined in this in vitro study, may relate to the different CNS symptoms that these strains produce in vivo.

Mortality factors affecting the leaf‐mining stages of Phyllonorycter (Lepidoptera: Gracillariidae) on oak and birch: 2. Biology of the parasite species

Parasitic Hpenoptera attacking Phyllonorycter species mining leaves of oak and birch were studied at a Cheshire locality during 1974. The host developmental stages killed, and attacked, by each parasite species are analysed. Smaller species and males tend to kill earlier Phyllonorycter instars than do larger species and females, and certain of the endoparasites as well as the ectoparasites must discriminate between small hosts which receive haploid eggs and large hosts which receive diploid eggs. Host‐feeding is very widely practised, and facultative hyperparasitism is engaged in by all except the braconids and a few specialized chalcids. The biology of a parasite species generally allows its allocation to one of three groups according to the size of host larva that it attacks, whether it is endoparasitic or ectoparasitic, whether or not the host continues to develop after parasitization, the extent of its hyperparasitic behaviour, its reproductive capacity, and the width of its host range. Different strategies, together with different specific host and habitat preferences, provide the basis for parasite complexes of high species diversity.

Mycobacteria and the Skin

Prevalence and Risk Factors for Bacterial Food‐Borne Zoonotic Hazards in Slaughter Pigs: A Review

The Hygiene Package and Regulation EC‐2160/2003 require information flow from farm to slaughterhouse to enhance European consumers protection in a ‘farm to fork’ approach. This obligation especially concerns food‐borne zoonotic hazards transmitted to humans through pork consumption, such as thermophilic Campylobacter spp., Listeria monocytogenes, Salmonella enterica and Yersinia enterocolitica. Prevalence estimates of these four hazards are affected by the sampling strategy and diagnostic procedure. Individual prevalence estimates for pig carriage (from digestive contents or lymph nodes collected at slaughterhouse) were higher than individual prevalence estimates for pig shedding (from faeces). Among risk factors described in the literature, poor pen cleaning and disinfection after pig departure to slaughterhouse and poor bio‐security measures are of major significance. Moreover, whereas wet feed increases the risk of pig infection by L. monocytogenes, dry feed is a risk factor for Salm. enterica. Mixing batches of pigs, notably in fattening herds, represents a risk for the transmission of Salm. enterica and Y. enterocolitica. Whereas small herds are more infected by thermophilic campylobacters and Y. enterocolitica, higher prevalence of Salmonella is observed in large herds due to a more frequent mixing of batches. Antibiotic treatment during the finishing period increases the risk of transmission of Salm. enterica. The forenamed elements should be taken into account to characterize farms in a risk assessment approach and to improve zoonotic hazard management in the pork food chain.

Coronavirus y radiología. Consideraciones sobre la crisis

Scalp Verruca from Barber Clippers: an Epidemiologic Assessment of a Common Fomite

Inactivation of interleukin‐8 by aminopeptidase N (CD13)

Aminopeptidase (APN) was found to degrade interleukin‐8 (IL‐8) and inactivate its chemotactic activity. The chemotactic activity of IL‐8 was decreased by APN or neutrophil plasma membranes dose‐ and time‐dependently. The chemotactic activity was not inactivated in the presence of bestatin or WM15 monoclonal antibody. The expression of IL‐8 was measured by flow cytometry. On lipopolysaccharide (LPS) stimulation, IL‐8 expression increased for 60 min and then decreased markedly. In contrast, on treatment with LPS and bestatin, the expression of IL‐8 increased continuously for at least 120 min. These results suggest that the expression and release of IL‐8 from phagocytic cells are regulated by the proteolytic effect of APN on IL‐8. J. Leukoc. Biol. 57: 129–134; 1995.

Rat tracheal organ culture supports replication of parainfluenza 1 (6/94) virus and promotes 6/94 virus rescue from latently infected human brain cells

Rat tracheal organ culture (TOC) supported replication of parainfluenza 1 (6/94) virus. Cell‐associated and cell‐free viruses were found after primary infection of TOC. In contrast to other mammalian systems, rat TOC was capable of maintaining 6/94 virus infectivity after primary infection. Rat TOC may be considered a potential indicator system that could be used to detect virus latent in human tissue.

Acute Respiratory Tract Infection in Daycare Centers for Older Persons

OBJECTIVE: To evaluate the rate of specific pathogens and clinical syndromes associated with acute respiratory tract infections (ARTI) in frail older persons attending daycare. DESIGN: Prospective descriptive study, without intervention. SETTING: Two sites of a senior daycare program providing all‐inclusive care for the older persons in Rochester, New York. PARTICIPANTS: Staff members and participants of the daycare. MEASUREMENTS: Demographic, medical, and physical findings were collected from older subjects at baseline and while ill with respiratory illnesses. Nasopharyngeal specimens for viral and Chlamydia culture and sputum for bacterial culture were obtained from subjects when ill. Acute and convalescent sera were also collected with each illness and examined for viral, chlamydial, and mycoplasma infection. MAIN RESULTS: One hundred sixty‐five illnesses were documented in 165 older daycare participants as well as 113 illnesses among 67 staff members during the 15‐month study. The rate of ARTI in the elderly group was 10.8 per 100 person months. The most common etiologies in both the staff and elderly participants were respiratory syncytial virus (RSV), Influenza A, and coronavirus. The etiologies of illnesses in the staff compared with those in elderly group were similar except that bacterial infections were significantly more common among the elderly (7% vs. 0, P = 0.05). Multiple pathogens were found to cocirculate within centers, and no clear outbreak of a predominant organism was noted. Cough and nasal congestion characterized most illnesses. The elderly experienced significantly more cough, dyspnea, and sputum production than did the staff. There were 10 hospitalizations related to respiratory infections and four deaths during the acute illness among the elderly group and none in staff. CONCLUSIONS: Acute respiratory infections are a common occurrence in both the staff and participants of a senior daycare center and are the cause of substantial morbidity in frail older persons.

Evaluation of a new enzyme‐linked immunosorbent assay (ELISA) in the diagnosis of rhinovirus infection

This study describes the evaluation of a newly developed ELISA for the direct detection of rhinovirus antigens in nasal washings. Of 54 volunteers inoculated with 100 TCID(50) of human rhinovirus type 2 (HRV‐2), 50 (96.6%) and 32 (59%) excreted antigen and virus on at least 1 of 3 days investigated, respectively. Thirty‐three (61%) had significant rises in rhinovirus‐specific IgA by ELISA. Twelve (22%) developed symptoms of colds. Generally the ELISA detected antigen more frequently in volunteers later in the course of infection and provided evidence of infection in a higher proportion of asymptomatic compared with symptomatic volunteers. On the other hand, virus isolation detected virus more frequently earlier in the course of infection and in a higher proportion of symptomatic compared with asymptomatic volunteers. We conclude that rhinovirus antigen detection by ELISA is a simple, rapid, sensitive, and practical test to diagnose a rhinovirus infection and potentially a viable alternative to virus isolation.

Morphometric patterns in Recent and fossil penguins (Aves, Sphenisciformes)

A total of 622 skin specimens, 527 skeletons, and myological data compiled by Schreiweis (1972) were used to investigate morphometric patterns within and among the 18 Recent species of Spheniscidae, and to compare the family with a fighted species, the common diving‐petrel (Pelecanoides urinator), considered by some authorities to be similar to the flighted ancestor of penguins. Fossil penguins also were studied using measurements from 111 skeletal elements representing 18 species. Most external and skeletal measurements follow interspecific rankings in body mass; the latter span a 30‐fold range from 1 kg in Eudyptula minor to 30 kg in Aptenodytes forsteri. Flighted Pelecanoides is only one‐tenth the mass of the smallest spheniscid. Wing areas and body masses of penguins maintain approximate geometric similitude among species; the allometric coefficient is 0.62 ± 0.04. Several skeletal elements are less derived in conformation in the fossil penguins than in modern confamilials. Despite great variation in the lengths of wing and leg skeletons, penguins show relative consistency in proportions of wing elements; pelvic proportions were more variable within the Spheniscidae. Proportions within both limbs are distinctly different from those of Pelecanoides. Analysis of correlation structures within species of penguin revealed three major subsets of skeletal variables that are highly intercorrelated: trunk‐skull lengths, appendicular‐trunk widths, and appendicular lengths. Within these groups, measurements tend to separate by anatomical region and, to a lesser degree, by limb. Canonical analysis of skin and skeletal measurements revealed groupings of spheniscids on the basis of size and relatively complex shape variables. Cluster analyses of taxa on the canonical variates confirmed the similarity of congeners in Eudyptes, Eudyptula and Spheniscus; species of Aptenodytes and, to a lesser degree, Pygoscelis, are more heterogeneous. A cluster analysis of principal components of myological measurements tabulated by Schreiweis (1972) corresponded closely with phenetic groupings using external and skeletal variables. A canonical contrast between spheniscids and Pelecanoides indicated that the families differ in a relatively complex skeletal dimension that only in part reflects overall size. Multivariate assessment of sexual dimorphism in external and skeletal variables indicated that: Eudyptula is least dimorphic; Aptenodytes, Eudyptes, Megadyptes and Spheniscus are moderately dimorphic; and Pygoscelis is most dimorphic. Pelecanoides shows comparatively low dimorphism. Among‐species and within‐species first principal components of skeletal measurements (multivariate axes of skeletal ‘size’) differ from one another in their orientation, and both deviate from isometric size. The first principal component for skeletons of Pelecanoides also deviated from isometric size, but the direction of this allometry is fundamentally different from that in penguins. Estimates of body mass for fossil penguins, based on principal components of available skeletal measurements, indicate that fossil species ranged from 3 kg to 81 kg in total mass; the largest fossil species was approximately 2.5 times as massive as the largest extant spheniscid. Much of the morphometric variation in the Spheniscidae is explainable on locomotory, ecological and thermodynamic grounds, and the associated phenetic groupings conform broadly with traditional generic classifications. The evolutionary significance of mensural correlations, allometric trends and differences between fossil and Recent species are discussed, and the need for a phylogenetic analysis of this highly specialized family of winged‐propelled diving birds is stressed.

Introduction: why don't HIV/AIDS policies work?

21st Annual Symposium 25–27 September 2008 Rhodes, Greece Programme of the 21st annual meeting on: Peripheral Nerve, Neuromuscular and Muscle Disease

Studies of an outbreak of acute hepatitis A: II. Antibody changes to cytomegalovirus and herpesvirus

The acute and convalescent sera from 14 schoolchildren with acute hepatitis A were tested for antibody changes to 70 viral antigens. Marked decreases were noted in the levels of antibody to cytomegalovirus in 5 of the 14 children and in the levels of antibody to herpesvirus type 1 in 3. No such changes were noted in 9 sex‐ and age‐matched healthy control children from the same classes.

European College of Veterinary Internal Medicine‐Companion Animals

Two Nursing Home Outbreaks of Respiratory Infection with Legionella sainthelensi

OBJECTIVE: To describe outbreaks of infection caused by Legionella sainthelensi occurring in older residents of two nursing homes and to determine risk factors for the development of infection. DESIGN: Descriptive epidemiology and a case‐control study. SETTING: Two nursing homes (140 beds and 254 beds in nursing homes A and B, respectively) located in southern Ontario, Canada, experiencing outbreaks of respiratory tract infection in July and August 1994. SUBJECTS: Case‐residents of the two nursing homes who met clinical and laboratory criteria for Legionella infection. Control‐residents were defined as those who were in the homes during the outbreaks and were asymptomatic. MEASUREMENTS: Active surveillance was conducted in both nursing homes to identify symptomatic residents. Residents with fever or respiratory tract symptoms had nasopharyngeal swabs taken for viral antigen detection and culture, urine for Legionella antigen detection, and acute and convalescent serology for viruses, Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella. Chest X‐rays were performed, and an attempt was made to obtain blood and sputum cultures. Water samples from shower heads, faucets, and air conditioning units were collected for Legionella culture and polymerase chain reaction (PCR) assay. A case‐control study was done to assess possible risk factors for legionellosis. RESULTS: Twenty‐nine cases — 17 in nursing home A; 12 in nursing home B — were identified. Four (14%) case‐residents had documented pneumonia and four case‐residents died. Univariate analysis revealed that a history of stroke (odds ratio (OR) 2.3 (95% CI, 1.0–5.3)), eating pureed food (OR 4.6 (95% CI, 1.6–12.7)), and having fluids administered with medication (OR 2.5 (95% CI, 1.0–5.9)) were significant risk factors. Cases were less likely to wear dentures (OR .4 (95% CI, .2‐.9)) or to eat solid food (OR .3, (95% CI, .1‐.6)). Only eating pureed food remained significant in a multivariable analysis (OR 4.6 (95% CI, 1.6–13.0, P = .01)). CONCLUSION: This report describes outbreaks of legionellosis in two nursing homes, representing the first reported outbreaks of infection caused by Legionella sainthelensi. The association with illness of dietary characteristics indicative of swallowing disorders suggests that aspiration was the most likely mode of infection. The diagnosis of legionellosis should be considered during outbreaks of respiratory infection in nursing homes. J Am Geriatr Soc 47:547–552, 1999.

Localization of rhinovirus replication in vitro with in situ hybridization

To facilitate understanding of human rhinovirus (HRV) pathogenesis, methods were developed for detection of HRV infection in vitro using in situ hybridization (ISH). HRV‐14 RNA probes and oligonucleotide probes representing conserved sequences in the 5′‐non‐translated region were labeled with (35)S and used to detect infected HeLa or WI‐38 strain human embryonic lung cells in cytological preparations. ISH was shown to be specific for detection of HRV on a single‐cell basis. Subsequently, in human nasal polyps infected in vitro, both oligonucleotide‐ and riboprobes produced a strong signal in association with ciliated epithelial cells. In human adenoids infected in vitro, a signal was observed in nonciliated epithelial cells. This study shows that HRV replicates in ciliated cells in the epithelium of human nasal polyps infected in vitro, and the presence of viral RNA in non‐ciliated cells of the human adenoid infected in vitro suggests that other cell types may also support rhinovirus replication.

Role of the golgi apparatus in cellular pathology

The Golgi apparatus response to pathological disorders is predominantly as an intermediary component of membrane biogenesis where it is involved in processing, sorting and secretion of materials via secretory granules, and in the formation of lysosomes. A common initial response of the Golgi apparatus to any stress is an alteration or cessation of secretory activity. In the transformed cell, the Golgi apparatus is altered both morphologically and biochemically, suggesting a shift from a secretory to a membrane‐generating mode of functioning. However, since fewer or less well‐developed Golgi apparatus are frequently found in transformed cells, analytical methods of membrane isolation developed for normal tissues may not always yield equivalent results when applied to tumors. Cell surface alterations characteristic of malignant cells may result from modifications occurring at the level of the Golgi apparatus. Some lysosomal dysfunctions may result from underglycosylation of acid hydrolases by the Golgi apparatus. The use of cell‐free systems between endoplasmic reticulum and Golgi apparatus or within Golgi apparatus cisterane is providing a new approach to the elucidation of the role of the Golgi apparatus in normal as well as pathological states.

CHILDHOOD ASTHMA IN BRISBANE: EPIDEMIOLOGICAL OBSERVATIONS

Analysis of hospital admissions for 20 years suggests that there has been an increase in childhood asthma in Brisbane. The characteristic seasonal pattern of asthma with waves in autumn and spring is evident from the second year of age and continues into adult life. It has not been explained, although respiratory infections, allergens and cold changes probably contribute to it. Unlike adults, children shown an increase in asthma in February‐March, ascribed to infections spread at school. Maximal asthma is associated with a mean temperature of 20–21°C. This may be optimal for the production of allergens. Further viral studies of asthmatic attacks are desirable.

IDENTIFICATION OF SEEDS FROM VARIOUS SPECIES OF STROPHANTHUS

In a search for differential specific characters, the outer epidermis of the testa from Strophanthus seeds of 15 species has been examined. A “trichome index” has been defined, and the method of its determination described. It enabled the species studied to be divided into six groups.

A novel method for determination of inorganic oxyanions by electrospray ionization mass spectrometry using dehydration reactions

Novel methods for the determination of inorganic oxyanions by electrospray (ES) ionization mass spectrometry have been developed using dehydration reactions between oxyanions and carboxylic acids at the ES interface. Twelve oxyanions (VO(3) (−), CrO(4) (2−), MoO(4) (2−), WO(4) (2−), BO(3) (3−), SiO(3) (2−), SiO(4) (4−), AsO(4) (4−), AsO(2) (−), SeO(4) (2−), SeO(3) (2−) and NO(2) (−)), out of 16 tested, reacted with at least one of four aminopolycarboxylic acids, i.e. iminodiacetic acid (IDA), nitrilotriacetic acid (NTA), trans‐1,2‐diaminocyclohexane‐N,N,N′,N′‐tetraacetic acid and triethylenetetramine‐N,N,N′,N″,N′″,N′″‐hexaacetic acid, at the ES interface to produce the dehydration products that gave intense mass ion responses, sufficient for trace analysis. As examples, trace determinations of Cr(VI) and silica in water samples were achieved after online ion exchange chromatography, where the dehydration product of CrO(4) (2−) and NTA (m/z 290) and that of SiO(4) (4−) and IDA (m/z 192) were measured. The limits of detection of the respective methods were 17 nM (0.83 ng Cr/ml) for Cr(VI) and 0.17 μM (4.8 ng Si/mL) for SiO(4) (4−). Copyright © 2016 John Wiley & Sons, Ltd.

Viral Respiratory Infections in the Institutionalized Elderly: Clinical and Epidemiologic Findings

OBJECTIVE: To prospectively evaluate the incidence and impact of viral respiratory infection in the institutionalized elderly during a winter season. DESIGN: Prospective descriptive study, without intervention. METHOD: Patients with respiratory illnesses were evaluated by a directed history and physical examination. Nasopharyngeal secretions for viral culture were obtained, and acute and convalescent serum samples were obtained for analysis. Serologic evidence of infection with respiratory syncytial virus (RSV) and parainfluenza were determined by enzyme immunoassay (EIA), and influenza by hemagglutination‐inhibition assay and EIA. SETTING: A 591‐bed nursing home. PARTICIPANTS: Residents with signs or symptoms of acute respiratory illness (nasal congestion, pharyngitis, cough, wheezing, or respiratory difficulty) were eligible for study. RESULTS: A viral etiology was documented in 62 out of 149 illnesses (42%). RSV was the most common virus associated with illness; it was documented in 27% of respiratory illnesses, followed by rhinovirus (9%), parainfluenza (6%), and influenza (1%). RSV was associated with significantly more severe disease when compared with rhinovirus. Clustering of specific viral infections occurred, suggesting nosocomial transmission. CONCLUSIONS: Viruses are an important cause of acute respiratory infections in the institutionalized elderly during the winter months.

A novel real‐time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses

A novel simultaneous detection system for human viruses was developed using a real‐time polymerase chain reaction (PCR) system to identify causes of infection in clinical samples from patients with uncertain diagnoses. This system, designated as the “multivirus real‐time PCR,” has the potential to detect 163 human viruses (47 DNA viruses and 116 RNA viruses) in a 96‐well plate simultaneously. The specificity and sensitivity of each probe–primer set were confirmed with cells or tissues infected with specific viruses. The multivirus real‐time PCR system showed profiles of virus infection in 20 autopsies of acquired immunodeficiency syndrome patients, and detected frequently TT virus, cytomegalovirus, human herpesvirus 6, and Epstein–Barr virus in various organs; however, RNA viruses were detected rarely except for human immunodeficiency virus‐1. Pathology samples from 40 patients with uncertain diagnoses were examined, including cases of encephalitis, hepatitis, and myocarditis. Herpes simplex virus 1, human herpesvirus 6, and parechovirus 3 were identified as causes of diseases in four cases of encephalitis, while no viruses were identified in other cases as causing disease. This multivirus real‐time PCR system can be useful for detecting virus in specimens from patients with uncertain diagnoses. J. Med. Virol. 83:322–330, 2011. © 2010 Wiley‐Liss, Inc.

Nitrogen fixing bacteria in the family Acetobacteraceae and their role in agriculture

For centuries, the Acetobacteraceae is known as a family that harbors many species of organisms of biotechnological importance for industry. Nonetheless, since 1988 representatives of this family have also been described as nitrogen fixing bacteria able to plant growth promotion by a variety of mechanisms. Nitrogen fixation is a biological process that guarantees that the atmospheric N(2) is incorporated into organic matter by several bacterial groups. Most representatives of this group, also known as diazotrophic, are generally associated with soil rhizosphere of many plants and also establishing a more specific association living inside roots, leaves, and others plants tissues as endophyte. Their roles as plant growth‐promoting microorganisms are generally related to increase in plant biomass, phosphate and other mineral solubilization, and plant pathogen control. Here, we report many of these plant growth‐promoting processes related to nitrogen fixing species already described in Acetobacteraceae family, especially Gluconacetobacter diazotrophicus and their importance to agriculture. In addition, a brief review of the state of art of the phylogenetics, main physiological and biochemical characteristics, molecular and functional genomic data of this group of Acetobacteraceae is presented.

Development and evaluation of a real‐time nucleic acid sequence based amplification assay for rapid detection of influenza A

The development and introduction of effective treatment for influenza A in the form of neuraminidase inhibitors have made the rapid diagnosis of infection important especially in high‐risk populations. The aim of this study was to develop a real‐time nucleic acid sequenced based amplification (NASBA) using a molecular beacon that could detect a wide range of influenza A subtypes and strains in a single reaction by targeting a conserved region of the influenza genome, and to evaluate its sensitivity and specificity against traditional laboratory techniques on a range of clinical samples usefulness during the 2003/2004 influenza season. The results demonstrated the assay to be highly sensitive and specific, detecting <0.1 TCID(50) of virus stock. Three hundred eighty‐nine clinical samples were tested in total from two patient groups. Overall, the real‐time NASBA assay detected 64% (66/103) more influenza positive samples than cell culture and direct immunofluorescence (IF) and, therefore, was shown to be more sensitive in detecting influenza A in a wide range of respiratory samples than traditional methods. In conclusion, the real‐time influenza A assay demonstrated clinical usefulness in both hospital and community populations. J. Med. Virol. 74:619–628, 2004. © 2004 Wiley‐Liss, Inc.

Oxidation of N‐heterocyclics: A green approach

[Chemical structure: see text] Environmentally benign oxidation methods satisfy the postulates of green chemistry. Heterocyclic Noxides have applications in synthetic organic chemistry, chemotherapy and agrochemicals. Synthesis of Noxides using green oxidants will be attractive over the conventional methods. The presence of the N‐oxide group in the azine ring makes it more subject to electrophilic and nucleophilic attack and substantially expands the synthetic approaches for the modification of nitrogen‐containing heterocyclics. That is the reason for the increasing interest in the chemistry of heterocyclic N‐oxides. Some reactions adopted for oxidation of N‐heterocyclics have been discussed. Stereochemical and spectroscopic aspects have been mentioned. It will be advantageous if anchored catalysts are employed for industrial exploitation. Several physiochemical aspects of various methods have been discussed.

First detection of spring viraemia of carp virus in common carp (Cyprinus carpio L.) affected by a septicaemic disease in Mexico

Spring viraemia of carp (SVC) is an infectious disease responsible for severe economic losses for various cyprinid species, particularly common carp (Cyprinus carpio carpio). The causative agent is the SVC virus (SVCV), a member of the Sprivivirus genus, Rhabdoviridae family, and a List 1 pathogen notifiable by the World Organization for Animal Health. This study describes the diagnosis of an SVCV pathogen isolated in October 2015 from wild common carp inhabiting a natural lagoon in central Mexico. While neither an epidemic nor fish mortalities were reported, the collected killed specimens exhibited clinical signs of disease (e.g., exopthalmia, moderate abdominal distension and haemorrhaging, as well as internal haemorrhages and adhesions). Histological results of injuries were consistent with the pathology caused by SVCV. This finding was supported by the isolation of a virus in EPC and BF‐2 cells and subsequent RT‐PCR confirmation of SVCV. The phylogenetic analyses of partial SVCV glycoprotein gene sequences classified the isolates into the Ia genogroup. These findings make this the first report of SVCV detection in Mexico, extending the southern geographical range of SVCV within North America. However, since this pathogen was detected in fish inhabiting a natural body of water without tributaries or effluents, it is difficult to estimate the risk of SVCV for other wild/feral cohabitating cyprinid species in the lagoon. The status of this virus is also unknown for other bodies of water within this region.

The role of the combination of IL‐2 and TGF‐β or IL‐10 in the generation and function of CD4(+) CD25(+) and CD8(+)regulatory T cell subsets

Recently, considerable attention has been focused on thymus‐derived CD4(+) regulatory T cells that constitutively express CD25 and have a contact‐dependent, cytokine‐independent mechanism in vitro. However, peripheral CD4(+) and CD8(+) T cells can also be induced to become regulatory T cells. Here we review our studies using the combination of IL‐2 and transforming growth factor β (TGF‐β) to generate regulatory T cell subsets ex vivo, and the work of others using IL‐10 to induce suppressive activity. Under certain conditions, the autocrine effects of TGF‐β and IL‐10 induce peripheral T cells to produce immunosuppressive levels of each of these cytokines. This effect of TGF‐β is IL‐2 dependent. Under other conditions IL‐2 and TGF‐β can induce CD4(+) cells to develop potent contact‐dependent, cytokine‐independent regulatory activity. At present, there is considerable confusion concerning the mechanism of action of CD4(+) CD25(+) cells because cytokine‐producing regulatory T cells generated in the periphery can express CD25 and other markers displayed by naturally occurring, thymus‐derived regulatory T cells. We, therefore, propose a nomenclature that identifies thymus‐derived and peripheral regulatory cells, and that also differentiates T regulatory cells from T helper cells. Because T regulatory cells broadly control T helper cell reactivity, the mechanisms that control regulatory cell function are also reviewed. Finally, the potential use of regulatory T cells generated ex vivo as an adoptive immunotherapy for certain autoimmune diseases, to prevent organ graft rejection, or to prevent pathologic host responses to infectious agents is discussed.

Diseases of pet rodents

The main pet rodent species are introduced with some brief notes on their suitability as pets and normal behaviour. The general clinical features of disease are discussed with some emphasis on species variation. The important diseases that occur in the United Kingdom are described under the headings of respiratory disease, diseases of the alimentary tract, diseases of the central nervous system, skin lesions, musculoskeletal disorders, palpable swellings, urinogenital disorders, diseases of pregnancy and injuries.

Tolerance of one‐month intranasal interferon

Under double‐blind conditions, groups of volunteers (68 in total) were allocated at random to take intranasal solutions of placebo or one of three doses of highly purified leucocyte interferon by intranasal spray twice a day for 28 days. The highest dose would have been expected to protect against experimental colds. Treatment was discontinued because of upper respiratory symptoms as often in each of the interferon groups as in the placebo group. However, it was possible to distinguish clinically between “colds” on placebo and low‐dose interferon and “reactions to treatment” on high‐dose interferon. The features of the reactions to treatment were a protracted build‐up of local symptoms and minor epistaxis. None of the volunteers on the high‐dose interferon were thought to have a definite cold, but viruses were isolated from four out of six volunteers on low‐dose interferon who had definite colds. Previous experiments had also shown this dose to be insufficient to protect against experimental rhinovirus challenge. The dose of interferon that appeared to protect against virus infection caused significant unwanted effects. It is essential to find interferon preparations with less inflammatory activity before interferon can be considered for use as a long‐term prophylactic against the common cold.

Detection and identification of viruses by electron microscopy

Electron microscopy can aid in the rapid diagnosis of viral diseases, as it can be performed in a matter of hours, but on a routine basis it should be used in conjunction with other techniques. Initially, the specimen source and patient symptoms should be ascertained, as these will lend suggestions of possible agents while eliminating others; however, this information should not be allowed to prejudice observation in such a way as to cause oversight of an unlikely pathogen. Second, selection of the method of preparation should be based on sample consistency; extraction, debris clarification, concentration, tissue culture amplification, or embedment may be necessary. Finally, false‐positive results must be avoided by differentiating viruses from cell organelles or debris, mycoplasmal or bacterial contamination, and bacteriophages.

SHELF‐LIFE EXTENSION OF FRESH FISH — A REVIEW PART I — SPOILAGE OF FISH

Giardia lamblia in children and the child care setting: A review of the literature

Ecto‐enzymes: physiology meets pathology

Ecto‐enzymes are catalytic membrane proteins with their active sites outside the cell. They include cholinesterase, which inactivates acetylcholine, and angiotensin‐converting enzyme, which converts angiotensin I to biologically active angiotensin II, and numerous other peptidases, transpeptidases, nucleotidases, phosphodiesterases, and phosphatases. Many CD antigens of leukocytes are ecto‐enzymes; some CD antigens for which no function is currently known are probably ectoenzymes. Expression is highly regulated and correlated with differentiation and activation. Some are highly restricted in distribution; others are ubiquitous. Many are shared between leukocytes and non‐hematogenous cells. Biological functions appear to depend on the type and location of the cell in which expression occurs, and include recycling of nutrients, local control of response to cytokines and hormones, bone formation, cell mobility, invasion, and metastasis. Many novel regulatory functions of ecto‐enzymes remain to be discovered, and may reveal new mechanisms of local extracellular control of cellular function. J. Leukoc. Biol. 67: 285–311; 2000.

Inhibition of Phagocytosis and Interleukin‐1 Production in Pulmonary Macrophages From Rats With Sialodacryoadenitis Virus Infection

To test whether or not sialodacryoadenitis virus (SDAV) infection in rats affects pulmonary macrophage function, we intranasally inoculated pathogen‐free F344 rats with SDAV and collected alveolar and interstitial macrophages 5 d later. We assessed Fc receptor‐mediated attachment and phagocytosis by phase‐contrast microscopic examination of monolayers of alveolar and interstitial macrophages incubated with zymosan, nonopsonized sheep erythrocytes, or erythrocytes opsonized with rabbit antisheeperythrocyte IgG. Alveolar macrophages from virus‐infected rats had significantly (P ≤ .05) lower indices of attachment and phagocytosis of opsonized erythrocytes than control macrophages, but there was no difference in attachment of zymosan particles. Interstitial macrophages were not affected. Alveolar macrophages from SDAV‐infected rats produced significantly less interleukin‐1 than those from control rats, as assessed by testing supematants from lipopolysaccharide‐stimulated macrophage cultures for induction of mouse thymocytes to take up tritiated thymidine. Effects of SDAV infection on lung macrophages could increase host susceptibility to other pathogens or complicate studies of respiratory tract immunity.

Body elongation and decreased reproductive output within a restricted clade of lizards (Reptilia: Scincidae)

Relationships between body shape and relative abdominal size were compared among differentially elongate species within the scincid lizard genus Brachymeles, to investigate how morphological evolution affects the proportion of body volume available to hold eggs and offspring. Relative abdominal size is inversely related to elongation, suggesting that relative clutch mass decreases with addition of abdominal body segments. Shape‐volume relationships contradict trends seen in comparisons among distantly related limbed and limbless squamates (lizards and snakes), in which snakes have relatively more abdominal volume. Comparison within a phylogenetically restricted group allows the identification of functional and ontogenetic factors potentially limiting reproductive output. In Bruchymeles, constraining factors include retention of anterior body segments bearing parasternal ribs, which prevents extension of the clutch anteriorly within the body, and reduction of allometry of abdominal segments, which provides extended series of uniformly‐sized vertebrae for limbless locomotion, but reduces the relative size of the abdomen. The latter trait is associated with overall size reduction, which affects relative egg‐size and packing. Factors constraining abdominal volume in this genus are probably common to other elongate lizards, a morphological group that has been rarely represented in comparative studies of life history.

Non‐Influenza Respiratory Viruses May Overlap and Obscure Influenza Activity

OBJECTIVE: To report the number and timing of influenza A isolates, as well as overlapping respiratory viruses. Co‐circulating respiratory viruses may obscure the determination of influenza activity. DESIGN: Prospective clinical surveillance for the new onset of respiratory illness followed by viral cultures during seven separate influenza seasons. SETTING: The Wisconsin Veterans Home, a skilled nursing facility for veterans and their spouses. RESULTS: Influenza A isolates were encountered in greater numbers than non‐influenza A isolates during three seasons. Seasonal variability is striking. In December 1992, we identified a large outbreak of respiratory illness. Influenza type B was cultured from 102 residents. In December 1995, influenza A was cultured from 285 people in Wisconsin. At that time, we identified outbreaks of respiratory illness in two of our four buildings. Based on statewide data, we suspected an influenza outbreak; however, 26 isolates of parainfluenza virus type 1 were cultured with no influenza. The potential importance of culturing at the end of the season was demonstrated in 1991–1992 when an outbreak of respiratory syn‐cytial virus (RSV) overlapped and extended beyond influenza A activity. CONCLUSIONS: When interpreting new clinical respiratory illnesses as a basis for declaring an outbreak of influenza A, clinicians should realize that co‐circulating respiratory viruses can account for clinical illnesses. Clinicians might utilize healthcare dollars efficiently by performing cultures to focus the timing of influenza A chemoprophylaxis. Cultures could be performed when clinical outbreak criteria are approached to confirm an outbreak. Culturing of new respiratory illness could begin again before the anticipated discontinuation of prophylaxis (approximately 2 weeks). J Am Geriatr Soc 47:1087–1093,1999.

Astrocytic reaction predominance in chronic encephalitis of junin virus‐infected rats

Junin virus antigen distribution and astrocytic reaction to prolonged infection were characterized in rat brain by the PAP technique. During the acute stage of neurologic disease following intracerebral inoculation, Junin antigen was detected in 100% of animals, strongly in most neurons but also to a much lesser degree in scattered astrocytes, dropping to 20% of rats at 540 days postinfection. Initially labeled in all brain areas, viral antigen gradually disappeared from hippocampus but persisted irregularly in cerebral cortex, basal ganglia, Purkinje cells, pons, and medulla oblongata. Such a pattern suggests that specific neuronal subpopulations, in spite of apparently unaltered cell morphology, may persistently harbor the virus, leading on occasion to a delayed neurologic syndrome. During both the acute and chronic stages of disease, a mild inflammatory exudate was observed, characterized by the presence of T and B lymphocytes, as well as macrophages and unidentified round cells. GFAP immunostaining showed increased astrocytic reaction as infection lapsed into chronicity. Corpus callosum, hippocampus, and cerebellum exhibited the sharpest reactive astrocytosis, followed by basal ganglia, pons, and medulla oblongata, whereas in cerebral cortex it was considerably less. Astrocyte activation, which failed to correlate with viral antigen presence in neurons, seems to result from a generalized condition, possibly including diffusible brain factors triggered by viral infection. Such widespread astroglial reaction may thus contribute to the outcome of the late neurologic syndrome.

Evaluation of an enzyme‐linked immunosorbent assay that measures rhinovirus‐specific antibodies in human sera and nasal secretions

Rhinovirus‐specific antibodies have traditionally been detected by their ability to neutralise the homologous rhinovirus serotype in tissue culture. Recently, however, we have described an enzyme‐linked immunosorbent assay that detects rhinovirus‐specific antibodies in sera and nasal secretions [Barclay and Al‐Nakib, 1987]. Here we describe an evaluation of the ELISA in a study involving 71 adult volunteers inoculated intranasally with human rhinovirus type 2 (HRV‐2). Pre‐and post‐inoculation serum samples and pre‐inoculation nasal washings were tested for the presence of HRV‐2‐specific antibodies by ELISA. Such antibodies were associated with protection against infection when present locally in nasal secretions, but when also present in the serum they were associated with protection against both infection and the development of illness. The antibody concentrations showed strong correlation with each other and with that of antibodies detected by the neutralisation test. Following HRV‐2 infection, rises in HRV‐2‐specific IgA in sera detected by ELISA occurred more frequently than rises in neutralizing antibody. These results suggest that the ELISA is a sensitive and reliable indicator of recent infection, as well as a predictor of homologous immune status.

Microstructure, evolution, and ontogeny of scale surfaces in cordylid and gerrhosaurid lizards

The distal oberhautchen and proximal beta layer of 41 species and every genus of cordylid and gerrhosaurid lizard were examined with a scanning electron microscope. Scale organs in these families are concentrated on the face, and scale organs occur singly or in pairs below the keel on the dorsals of cordylids but not gerrhosaurids. Most species examined possess primitive, lenticular scale organs. Unique and previously unknown, projecting scale organs occur in two species of Cordylus. Scale organs appear to be absent in species of Chamaesaura. The scale surfaces of cordylids are covered in macrohoneycomb. Individual compartments of cordylid macrohoneycomb are not intracellular structures but may be overlain by apparent oberhautchen cells. In some cordylid species, part of each cell is free and projects away from the scale surface as a pointed flap. Gerrhosaurids lack macrohoneycomb. Instead, their scale surfaces are covered in lamellae without dentate borders, 50–100 times as long as wide. The epidermis of Platysaurus is unique among cordylids in possessing lamellae 1–8 times as wide as long that overlap macrohoneycomb in some regions of the body. The scale surfaces of Cordylus cordylus undergo an ontogenetic change similar to that recently described for the xenosaurid genus Shinisaurus. Phylogenetic analysis of scale surface features allows the partitioning of Cordylus into three species groups and Chamaesaura into two species groups. Synapomorphies of the species of Chamaesaura, the species of Platysaurus, and the genera of the gerrhosaurids are also described. © 1995 Wiley‐Liss, Inc.

IN VITRO EFFECTS OF MONOLAURIN COMPOUNDS ON ENVELOPED RNA AND DNA VIRUSES

Monolaurin alone and monolaurin with tert‐butylhydroxyanisole (BHA), methylparaben, or sorbic acid were tested for in vitro virucidal activity against 14 human RNA and DNA enveloped viruses in cell culture. At concentrations of 1% additive in the reaction mixture for 1 h at 23°C, all viruses were reduced in infectivity by >99.9%. Monolaurin with BHA was the most effective virucidal agent in that it removed all measurable infectivity from all of the viruses tested. The compounds acted similarly on all the viruses and reduced infectivity by disintegrating the virus envelope.

Evolution of the middle ear apparatus in talpid moles

The middle ear structures of eight species of mole in the family Talpidae (Mammalia: Eulipotyphla) were studied under light and electron microscopy. Neurotrichus, Parascalops, and Condylura have a simple middle ear cavity with a loose ectotympanic bone, ossicles of a “microtype” morphology, and they retain a small tensor tympani muscle. These characteristics are ancestral for talpid moles. Talpa, Scalopus, Scapanus, and Parascaptor species, on the other hand, have a looser articulation between malleus and ectotympanic bone and a reduced or absent orbicular apophysis. These species lack a tensor tympani muscle, possess complete bullae, and extensions of the middle ear cavity pneumatize the surrounding basicranial bones. The two middle ear cavities communicate in Talpa, Scapanus, and Parascaptor species. Parascaptor has a hypertrophied malleus, a feature shared with Scaptochirus but not found in any other talpid genus. Differences in middle ear morphology within members of the Talpidae are correlated with lifestyle. The species with middle ears closer to the ancestral type spend more time above ground, where they will be exposed to high‐frequency sound: their middle ears appear suited for transmission of high frequencies. The species with derived middle ear morphologies are more exclusively subterranean. Some of the derived features of their middle ears potentially improve low‐frequency hearing, while others may reduce the transmission of bone‐conducted noise. By contrast, the unusual middle ear apparatus of Parascaptor, which exhibits striking similarities to that of golden moles, probably augments seismic sensitivity by inertial bone conduction. J. Morphol. © 2006 Wiley‐Liss, Inc.

Interactions of Pseudorabies Virus With Swine Alveolar Macrophages: Effects of Virus Infection on Cell Functions

In order to assess the effect of Pseudorabies virus (PRV) infection on the function of swine alveolar macrophages (AM), lung lavage cells were cultured, infected with one of six strains of PRV, and various activities were measured. Activity measurement included viability, phagocytosis of yeast, phagosome‐lysosome fusion, phagocytosis of opsonized particles, and superoxide release. AM were infected with 5 × 10(‐3) PFU/cell, and the comparative assessment of functions was performed at 18‐20 h postinfection. Cell viability in PRV‐infected cultures ranged from 79 to 94% of the viability in noninfected cultures. Phagocytosis of yeast was significantly reduced only in the AM cultures infected with the strain S‐62. Phagosome‐lysosome fusion was depressed in cultures infected with the strains S‐62, 4892, 3816, and BUK. The phagocytosis of opsonized sheep red blood cells showed significant differences between noninfected and PRV‐infected cultures in all cases except cultures infected with the strain PRV‐C. The O(2) release after stimulation with opsonized zymosan was significantly reduced in all the PRV‐infected cultures. The effect of PRV infection on AM functions that are related to the bacterial activity of such cells suggests that PRV‐induced AM dysfunction might have a role in the increased susceptibility of PRV‐infected pigs to bacterial pneumonia.

Appearance of complement components and immunoglobulins on nasopharyngeal epithelial cells following naturally acquired infection with respiratory syncytial virus

Nasopharyngeal epithelial cells (NPEC) were collected from 144 infants and children with respiratory syncytial virus (RSV) infection, and were analyzed by fluorescent antibody techniques for the presence of cell‐bound complement (C(/) (3)), IgA, IgC, and IgM class of immunoglobulins (Ig), and respiratory syncytial virus antigen. Viral antigen was present on the surface of NPEC in 100% of samples obtained in the first 3 days of illness. The percentage of patients positive for RSV antigen declined steadily, so that no patient still expressed viral antigen on NPEC by 57 days after the onset of illness. Cell‐bound IgA, IgC, and IgM could be detected in most of the samples tested in the first 13 days after the onset of illness. Subsequently, the frequency of detection of cell‐bound Ig gradually declined. Only 8–3 3 % of patients tested 57 days after the onset of illness expressed IgA, IgG, or IgM on NPEC. About 45% of samples tested in the first 8 weeks after the onset of illness exhibited complement binding to NPEC. The percentage of subjects showing cell‐bound C(/) (3) reached a maximum at 8–13 says after the onset of illness, while cell–bound C(/)(3) could not be detected in any of the samples collected 57–90 days after the onset of illness. Although cell‐bound C(/) (3) was generally present in association with cell‐bound Ig on NPEC, in a small percentage (4.6%) of patients cell‐bound C(/)(3) could be detected in the absence of any cell–bound Ig. Cell‐bound C(/)(3) and IgA, IgG, and IgM were present with equal frequency in patients with all forms of clinical disease caused by RSV, and in patients less than or greater than 6 months of age at the onset of illness.

SCIENTIFIC INFORMATION DOCUMENT: Campylobacter infection

Innovative Drug Treatments for Viral and Autoimmune Myocarditis

Myocarditis has been shown to be a common cause of cardiomyopathy and is believed to account for 25% of all cases in human beings. Unfortunately, the disease is difficult to detect before a myopathic process ensues. Treatment of myocarditis‐induced heart failure includes the standard regimen of diuretics, digoxin, angiotensin‐converting enzyme inhibitors, and currently, β‐adrenergic blockers. Treatment of myocarditis itself is dependent on the etiology of the illness. Treatments under investigation include immunosuppressants, nonsteroidal antiinflammatory agents, immunoglobulins, immunomodulation, antiadrenergics, calcium‐channel blockers, angiotensinconverting enzyme inhibitors, nitric oxide inhibition (e.g., aminoguanidine), and antiviral agents. Despite advances in treatment, more work needs to be done in the early detection of myocarditis. Additionally, better means need to be established for distinguishing between viral and autoimmune forms of the disease, so that appropriate treatment can be instituted.

Program and Abstracts of the 11th INTERNATIONAL RES CONGRESS and 24th NATIONAL MEETING OF THE RETICULOENDOTHELIAL SOCIETY: Westin Kauai, Kauai, Hawaii, October 17–21, 1987

Enzyme immunoassay for respiratory syncytial virus: Rapid detection in nasopharyngeal secretions and evaluation of isolates representing different RSV subgroups

The presence of respiratory syncytial virus (RSV) was investigated by immunofluorescent antibody (IFA) technique and by an enzyme immunoassay (EIA) in 169 samples of nasopharyngeal secretions of infants and children with acute respiratory infections. Of 31 samples positive by EIA, 25 were positive by IFA. In 24 samples from a retrospective study, RSV positive by IFA and/or tissue culture isolation (TCI), 22 were also positive by EIA. The EIA was also evaluated with 111 RSV isolates in Hep2 cell cultures representing different RSV subgroups. All were positive by EIA. © 1993 Wiley‐Liss, Inc.

defensive publications in an r&d race

This paper considers a multistage dynamic R&D race in which the competitors strategically publish research findings. Publications change the prior art, thus affecting patentability. Firms publish when they are behind in the race and their rival is close to winning it. Publication sets back both competitors and gives the follower a chance to catch up. Publications prolong the race. Firms are more likely to publish the more patient they are, and the higher their probability of success. Asymmetry between the firms generates additional incentives to publish such as protecting profits from a previous patent and increasing a strong firm's probability of winning. When firms face a joint decision on publications and the intensity of research, publications substitute investment.

Molecular biology of human pathogenic viruses

Refining rare weed trait syndromes along arable intensification gradients

QUESTION: How does the conservation (rarity) value of arable weed communities differ along intensification gradients? Which functional traits best distinguish the weed communities of more and less extensively managed fields? Can the same traits predict the rarity of individual weed species? LOCATION: Hungary. METHODS: Using relevé data from 60 cereal and 70 stubble fields, together with weed trait data, we characterized community responses to arable intensification using functional trait analyses based on trait‐convergence and trait‐divergence assembly patterns. We also examined how well the broad‐scale rarity status of species predicts their occurrence along intensification gradients, and how it maps onto our functional classifications describing intensification responses. RESULTS: The response of weeds to intensification in cereal fields was best described by a functional classification based on species' flowering duration, maximum height and seed weight: weeds of extensively managed fields have short flowering seasons (2–5 mo) and particularly large or small seeds. The highest proportions of rare species also happen to be found in these groups. The rarest weeds among these species tend to be late‐winter and early‐summer annuals, while the rare species of stubble fields tend to be broad‐leaved with low nitrogen requirements, small seeds and short height. Stubble fields showed a decline in weed cover with increasing application of fertilizer and distance from the field edge, but we could detect no strong associations of management factors with trait composition, perhaps because the intensification gradient across these fields was shorter. CONCLUSIONS: Many rare Hungarian weeds are associated with traditional extensive farming practices. They are particularly characterized by short, midsummer flowering periods and by preference for low nitrogen levels, but a range of trait syndromes must be considered to understand their ecology and conservation. Analyses based on trait‐divergence patterns, rather than trait‐convergence patterns, provide better insights into the functional composition of weed communities, emphasizing the importance of disruptive filters in weed community assembly and the need for improved methods to detect such effects.

The effect of plant species on soil nitrogen mineralization

1.. To ascertain the influence of different plant species on nitrogen (N) cycling, we performed a long‐term garden experiment with six grasses and five dicots with different potential growth rates, that are adapted to habitats with different nutrient supplies. We measured in situ N mineralization and nitrification of the soil under monocultures of each species during the fourth year of the experiment. 2.. We focused on the effects of the different species on (i) annual net N mineralization; (ii) the seasonal pattern of N mineralization; and (iii) the fraction of the total N mineralization that is nitrified. Our hypothesis was that plant species of nutrient‐rich habitats would enhance the N mineralization compared with species of nutrient‐poor habitats. 3.. The results demonstrate a strong influence of the species on net N mineralization and net nitrification, both of which fluctuated strongly during the year. Overall, species from high fertility habitats increased N mineralization and nitrification more than species from low fertility habitats. About 90% of the mineralized ammonium was oxidized to nitrate. There was no significant difference in this proportion in the plots of species from nutrient‐rich, moderate and nutrient‐poor habitats.

Current issues and forthcoming events

Enzyme‐Linked lmmunosorbent assay: A rapid reproducible test for the measurement of rabies antibody

An enzyme‐linked immunosorbent assay (ELISA) was developed for detecting IgG antibodies to the Pitman Moore strain of rabies virus in sera from subjects immunised with HDCS vaccine. End‐point titres of antibody were determined using a pocket calculator preprogrammed to analyse absorbence values of test sera and negative controls. The assay was highly reproducible, and very close agreement was achieved when the results were compared with those of the mouse neutralization test. Rabies enzyme‐linked immunosorbent assay is easy to perform, the results are obtained within 48 hours, and because it employs inactivated virus, the test can be used safely without special containment facilities.

Polymerase chain reaction and sequencing for typing rhinovirus RNA

Primers were designed and tested for their ability to distinguish rhinoviruses from enterovi‐ruses. A primer set derived from the 5′‐UTR/VP coding region junction was able to amplify all the rhinovirus serotypes tested. Enteroviruses were either not amplified by these primer pairs or produced a band of larger size that could easily be discriminated from the rhinovirus‐specific product. In contrast, primers embedded in the 5′‐UTR region alone were able to amplify both rhinovirus and enterovirus RNA. It is shown that rhino‐viruses could be specifically typed by sequencing the amplicon derived from this 5′‐UTR set. The sequences of the 5′‐UTR region often previously unsequenced rhinoviruses were derived. The sequences obtained cluster into two groups: 18,41, 15, 30, 63, 31,56, and 44; and 17, 69, and 70. Ampliconsfrom serotypes 17, 69, and 70 also group by sequence with the equivalent region of HRV14 from the genetic database, while the others group with 2 and 89. © 1994 Wiley‐Liss, inc.

Detection of enteric adenoviruses with synthetic oligonucleotide probes

The abilities of hybridization probes to detect all human adenovirus types and to identify enteric adenovirus types were evaluated. The efficiency of hybridization was compared to other tests currently in routine laboratory use on clinical specimens from young children with gastroenteritis, Probes were derived from various regions of the adenovirus types 2 and 41 genomes, and were evaluated by hybridization with a series of DNA quantities from 1 μg to 10 pg of one adenovirus type from each human subgenus, lambda phage, and HEp 2 cells. The sensitivity of hybridization with the HPII probe (92.7%), containing the conserved hexon gene, compared well with EM (54.6%), culture and neutralization (45.5%), and enzyme immunoassay (61.8%). The sensitivity of detection of enteric adenovirus isolates by the cloned Bg/II D fragment probe (92.9%) and by a synthetic probe (85.7%), manufactured from type‐specific sequences of the Ad41 hexon gene were comparable to Ad40/Ad41 specific enzyme immunoassay (84.6%). Hybridization was found to be a sensitive method of adenovirus detection in comparison to traditional methods of laboratory diagnosis. Synthetic oligonucleotides enable specific detection of individual enteric adenovirus types. Hybridization had additional advantages over other tests in identifying cases of infection with more than one adenovirus type and in allowing an estimate of the concentration of adenovirus in the specimen.

Mitotane (o, p‘‐DDD) treatment in a cat with hyperadrenocorticism

An 11‐year‐old male castrated Persian cat with spontaneous hyperadrenocorticism was presented. Both adrenals were grossly enlarged and calcified. A diagnosis of pituitary‐dependent hyperadrenocorticism was made. Signs of hyperadrenocorticism resolved with long‐term mitotane treatment. Concurrent diabetes mellitus resolved after 220 days of therapy. No severe adverse drug reactions were noted.

Detection of cytomegalovirus and Epstein‐Barr virus in labial salivary glands in Sjogren's syndrome and non‐specific sialadenitis

To investigate the role of herpes viruses in Sjogren's syndrome, minor (labial) salivary gland tissues from Sjogren's syndrome and from non‐specific sialadenitis were examined for Epstein‐Barr virus (EBV) and human cytomegalovirus (HCMV) DNA by the polymerase chain reaction. Almost half of all salivary glands studied contained EBV and/or HCMV. There was, however, no significant difference between the detection of EBV or HCMV in salivary glands from patients with Sjogren's syndrome or non‐specific sialadenitis. The findings are consistent with the persistence of EBV and HCMV in minor salivary glands following primary infection, but do not indicate a direct role for either virus in the aetiology of Sjogren's syndrome, and do not exclude reactivation of the viruses in this disease.

Prospective Hematologic and Clinicopathologic Study of Asymptomatic Cats With Naturally Acquired Feline Immunodeficiency Virus Infection

Prospective studies were performed over a 28‐ to 77‐month period (median, 66 months) on 5 cats with naturally acquired feline immunodeficiency virus (FIV) infection in an attempt to correlate hematologic and Clinicopathologic changes with the emergence of clinical disease. On presentation, all cats were asymptomatic; free of opportunistic infections; and had normal complete blood counts, bone marrow morphologies, marrow progenitor frequencies, and progenitor in vitro growth characteristics. During study, 2 cats remained healthy, 2 cats showed mild clinical signs, and 1 cat developed a malignant neoplasm (ie, bronchiolar‐alveolar adenocarcinoma). Although persistent hematologic abnormalities were not observed, intermittent peripheral leukopenias were common. In 3 of 5 FlV‐seropositive cats, lymphopenia (< 1,500 lymphs/μL; normal reference range, 1,500 to 7,000 lymphs/μL) was a frequent finding and the absolute lymphocyte counts had a tendency to progressively decline. One of the other 2 cats had consistently low to low‐normal absolute neutrophil counts (1,300 to 4,800 segs/μL; mean, 2,730 segs/μL; normal reference range, 2,500 to 12,500 segs/μL), and the remaining cat had consistently normal leukograms, except for a transient period (ie, 11 months) of benign lymphocytosis (7,200 to 13,430 lymphs/μL) early in the study. Periodic examinations of bone marrow aspirates revealed normal to slightly depressed myeloid‐to‐erythroid ratios with normal cellular morphology and maturation. Bone marrow abnormalities observed late in the study included mild dysmor‐phic changes (ie, megaloblastic features) in 2 cats, and a significant decrease (60% of controls, P < .001) in the frequencies of burst‐forming units erythroid (BFU‐E) in marrow cultures of FIV‐seropositive cats compared with uninfected control cats. Serum biochemical profiles were unremarkable throughout the study, with the exception of hyperglobulinemia (ie, polyclonal gammopathy) in 2 of 5 cats. Peripheral blood and bone marrow findings were of no apparent prognostic value. These results confirm the long latency between natural FIV infection and the development of life‐threatening clinical disease. Chronic FIV infection, like infection with human immunodeficiency virus, can be associated with derangements in peripheral blood cell counts, as well as pertubations in marrow cell morphologies and hematopoietic progenitor frequencies before the terminal symptomatic stages of retroviral disease, when persistent cytopenias are prominent.

The accuracy of nurses' estimates of their absenteeism

gaudine a & gregory c (2010) Journal of Nursing Management 18, 599–605 The accuracy of nurses' estimates of their absenteeism Aim The purpose of the present study was to determine the accuracy of nurses’ self‐reports of absence by examining: (1) the correlation, intra‐class correlation, and Cronbach’s alpha for self‐reported absence and absence as reported in organizational records, (2) difference in central tendency for the two measures of absence and (3) the percentage of nurses who underestimate their absence. Background Research on nurses’ absenteeism has often relied on self‐reports of absence. However, nurses may not be aware of their actual absenteeism, or they may underestimate it. Method Self‐reported absence from questionnaires completed by 215 Canadian nurses was compared with their absence from organizational records. Results There is a strong positive correlation, a strong intra‐class correlation and Cronbach’s alpha for the two measures of absence. However, there is a difference in central tendency that is related to the majority of nurses in this study (51.1%) underestimating their days absent from work. Conclusions Research examining the predictors of absence may consider measuring absence with self‐reports. Nevertheless, nurses demonstrated a bias to underestimate their absence. Implications for nursing management Feedback interventions to reduce absenteeism can be developed to include providing nurses with accurate information about their absence.

Characterization of precipitates size distribution: validation of low‐voltage STEM

The size distribution of second phase precipitates is frequently determined using conventional transmission electron microscopy (CTEM). However, other techniques, which present different advantages, can also be used for this purpose. In this paper, we focus on high angle annular dark field (HAADF) in TEM and scanning TEM (STEM) in scanning electron microscopy (SEM) imaging modes. The mentioned techniques will be first described, then compared to more conventional ones for the measurement of carbides size distribution in two FeCV and FeCVNb model alloys. This comparative study shows that STEM in SEM, a technique much easier to undertake compared to TEM, is perfectly adapted for size distribution measurements of second phase particles, with sizes ranging between 5 and 200 nm in these systems.

Can plant litter affect net primary production of a typical steppe in Inner Mongolia?

Question: Litter (dead leaves or stems) affects production by conserving soil moisture. However, that role is not clear for grasslands where most precipitation falls during the growing season when the demand for water is high. Our question was: Does litter affect forage production in such an environment? Location: Typical steppe, Inner Mongolia. Methods: We examined the role of plant litter in two experiments where litter was either removed or added in a protected or heavily grazed site, respectively, in autumn and in spring in a split plot design. The treatments (control, moderate and heavy litter application) were applied once in five replications but repeated at new locations in each of 3 years. This was done to examine only the direct effect of litter on annual net primary production and selected plant characteristics and not potential secondary effects. We also measured soil moisture and soil temperature. Results: Removing litter caused a reduction in the amount of grass (Leymus chinensis) that was produced, but litter addition caused an inconsistent effect among years, with moderate applications producing the most positive effects. Litter removal resulted in shorter and less dense plants of L. chinensis and Carex duriuscula, while heavy litter addition in autumn reduced plant height of both Cleistogenes squarrosa and C. duriuscula. Conclusions: Litter was effective for enhancing soil moisture status and reducing soil heat units in the typical steppe of Inner Mongolia. Therefore, litter mass may serve as an index of grassland health in such environments.

Functional traits explain amphibian distribution in the Brazilian Atlantic Forest

AIM: Species distributions are one of the most important ways to understand how communities interact through macroecological relationships. The functional abilities of a species, such as its plasticity in various environments, can determine its distribution, species richness and beta diversity patterns. In this study, we evaluate how functional traits influence the distribution of amphibians, and hypothesize which functional traits explain the current pattern of amphibian species composition. LOCATION: Atlantic Forest, Brazil. TAXON: Amphibia (Anura and Gymnophiona) METHODS: Using potential distributions of Brazilian amphibians from Atlantic Forest based on their functional traits, we analysed the influence of biotic and abiotic factors on species richness, endemism (with permutation multivariate analysis) and beta diversity components (i.e. total, turnover and nestedness dissimilarities). RESULTS: Environmental variables explained 59.5% of species richness, whereas functional traits explained 15.8% of species distribution (geographical species range) for Anuran and 88.8% for Gymnophiona. Body size had the strongest correlation with species distribution. Results showed that species with medium to large body size, and species that are adapted to living in open areas tended to disperse from west to east direction. Current forest changes directly affected beta diversity patterns (i.e. most species adapted to novel environments increase their ranges). Beta diversity partitioning between humid and dry forests showed decreased nestedness and increased turnover by increasing altitude in the south‐eastern region of the Atlantic Forest. MAIN CONCLUSIONS: Our study shows that functional traits directly influence the ability of the species to disperse. With the alterations of the natural environment, species more apt to these alterations have dispersed or increased their distribution, which consequently changes community structure. As a result, there are nested species distribution patterns and homogenization of amphibian species composition throughout the Brazilian Atlantic Forest.

Probable Vitamin K‐Deficient Bleeding in Two Cats With Malabsorption Syndrome Secondary to Lymphocytic Plasmacytic Enteritis

Two cats with intestinal malabsorption developed a hemorrhagic diathesis. Although unsubstantiated, the probable cause of bleeding was a chronic malabsorption of fat and the fat‐soluble vitamin K. When treated_with vitamin Ki per os, one cat's clotting times were only partially corrected. Since vitamin K, is actively absorbed in the proximal small intestine, the incomplete response of this case to orally administered vitamin K, was predictable. The infrequent occurrence of bleeding in animals with malabsorption is, in part, attributable to the ileal and colonic absorption of bacterially derived vitamin K(2). For this reason, nonspecific use of antibiotics in these animals is contraindicated. Since long‐chain, polyunsaturated fats impair vitamin K absorption, dietary fat given to animals with malasorption should be restricted to medium‐ and short‐chain, saturated fats. Vitamin K should be administered subcutaneously to these animals if prolonged clotting times or active bleeding is present, and routinely prior to surgery. Oral supplementation with vitamin K(3), which is absorbed in the colon and less lipid soluble than vitamin Ki, should be given to animals with malabsorption that are maintained as outpatients. Adequate dosage levels of vitamin K(3), however, are yet to be established for the cat, and dose‐dependent hemolytic anemia is a probable toxic manifestation.

Ultrastructure of Euspermatozoa of cerithiacean gastropods (Prosobranchia: Mesogastropoda)

Euspermatozoa of selected cerithiacean gastropods have been studied using transmission electron microscopy and the results compared (primarily) with previous studies of mesogastropod and neogastropod euspermatozoa. Cerithiacean euspermatozoa each possess a well‐defined acrosome (extremely varied in shape), a short (2.25—3 μm), very electron‐dense nucleus, an elongate midpiece, and an elongate glycogen piece. A dense ring structure associated with the plasma membrane occurs at the junction of the midpiece and glycogen piece. While features such as the dense ring structure and the systematic periaxonemal arrangement of “glycogen” granules can be understood from a purely functional standpoint, it is suggested that euspermatozoon features also provide information of taxonomic and phylogenetic relevance. On the basis of euspermatozoon midpiece structure, true cerithiaceans can be easily distinguished from other mesogastropods and from neogastropods and are divided tentatively into two groups: Group 1 (Turritellidae, Cerithiidae, Australaba (family uncertain), Planaxidae, Potamididae (subfamily Batillariinae)), and Group 2 (Potamididae (subfamily Potamidinae), Modulidae, Obtortio (family uncertain)). Using midpiece and acrosomal features, group 1 can be further subdivided into two subgroups: Subgroup 1(i) (Turritellidae, Cerithiidae, Australaba) and Subgroup 1(ii) (Planaxidae, Potamididae (subfamily Batillariinae)). It is suggested that the pronounced differences existing between the two subfamilies of the Potamididae may indicate the necessity for a separate family for the Batillariinae.

Gastrointestinal Motility and Disease in Large Animals

An understanding of the relationship between gastrointestinal (GI) motility and disease is imperative for the proper treatment of large animal patients, especially as new therapeutic agents become available. However, the abundance of information that has become available in the last 2 decades makes gaining this understanding a formidable task. This article summarizes the changes in GI motility caused by some common diseases and conditions encountered in large animal practice, such as GI obstruction, postoperative ileus, resection and anastomosis, diarrhea, endotoxemia, GI parasitism, hypocalcemia, and pregnancy. J Vet Intern Med 1996;10:51–59. Copyright © 1996 by the American College of Veterinary Internal Medicine.

Lymphocyte activation

Aetiology and outcome in 90 cats presenting with dyspnoea in a referral population

Objectives: Dyspnoea is an unspecific severe presenting sign, which can be life threatening and requires prompt treatment. Dyspnoeic cats often have significant underlying disease. Underlying aetiologies in cats that presented with dyspnoea were reviewed, and associations with patient signalment and outcome were investigated. Methods: The case records of 90 dyspnoeic cats were retrospectively reviewed and separated into different groups depending on aetiology (cardiac, respiratory, neoplastic and trauma). Duration of clinical signs, presentation, hospitalisation length and survival were analysed. Results: Cardiac (38 per cent), respiratory (32 per cent) and neoplastic (20 per cent) diseases were common causes of feline dyspnoea. Cats with respiratory causes had longer duration of clinical signs (P<0·001) before presentation. Cats with neoplasia were significantly older (P<0·001). No significant difference in respiratory rates was present between the groups (P = 0·154). High heart rates (P<0·001) and abnormalities on cardiac auscultation were more likely in the cardiac group. Clinical Significance: Frequent causes of dyspnoea in cats were cardiac disease followed by respiratory causes and neoplastic conditions. Heart rate on presentation and presence of heart murmurs or gallops are useful to identify cardiac causes. Improved proportion surviving was found in the respiratory group (P = 0·027), whereas cats with neoplasia had the worst outcome.

Minireview The influence of season, photoperiod, and pineal melatonin on immune function

Abstract: In addition to the well‐documented seasonal cycles of mating and birth, there are also significant seasonal cycles of illness and death among many animal populations. Challenging winter conditions (i.e., low ambient temperature and decreased food availability) can directly induce death via hypothermia, starvation, or shock. Coping with these challenges can also indirectly increase morbidity and mortality by increasing glucocorticoid secretion, which can compromise immune function. Many environmental challenges are recurrent and thus predictable; animals could enhance survival, and presumably increase fitness, if they could anticipate immunologically challenging conditions in order to cope with these seasonal threats to health. The annual cycle of changing photoperiod provides an accurate indicator of time of year and thus allows immunological adjustments prior to the deterioration of conditions. Pineal melatonin codes day length information. Short day lengths enhance several aspects of immune function in laboratory studies, and melatonin appears to mediate many of the enhanced immunological effects of photoperiod. Generally, field studies report compromised immune function during the short days of autumn and winter. The conflict between laboratory and field data is addressed with a multifactor approach. The evidence for seasonal fluctuations in lymphatic tissue size and structure, as well as immune function and disease processes, is reviewed. The role of pineal melatonin and the hormones regulated by melatonin is discussed from an evolutionary and adaptive functional perspective. Finally, the clinical significance of seasonal fluctuations in immune function is presented. Taken together, it appears that seasonal fluctuations in immune parameters, mediated by melatonin, could have profound effects on the etiology and progression of diseases in humans and nonhuman animals. An adaptive functional perspective is critical to gain insights into the interaction among melatonin, immune function, and disease processes.

A New Disease Emerges: Severe Acute Respiratory Syndrome (SARS)

Effect of Gd‐DTPA‐BMA on blood and myocardial T(1) at 1.5T and 3T in humans

PURPOSE: To compare T(1) values of blood and myocardium at 1.5T and 3T before and after administration of Gd‐DTPA‐BMA in normal volunteers, and to evaluate the distribution of contrast media between myocardium and blood during steady state. MATERIALS AND METHODS: Ten normal subjects were imaged with either 0.1 mmol/kg (N = 5) or 0.2 mmol/kg (N = 5) of Gd‐DTPA‐BMA contrast agent at 1.5T and 3T. T(1) measurements of blood and myocardium were performed prior to contrast injection and every five minutes for 35 minutes following contrast injection at both field strengths. Measurements of biodistribution were calculated from the ratio of ΔR(1) (ΔR (1myo)/ΔR (1blood)). RESULTS: Precontrast blood T(1) values (mean ± SD, N = 10) did not significantly differ between 1.5T and 3T (1.58 ± .13 sec, and 1.66 ± .06 sec, respectively; P > 0.05), but myocardium T(1) values were significantly different (1.07 ± .03 sec and 1.22 ± .07 sec, respectively; P < 0.05). The field‐dependent difference in myocardium T(1) postinjection (T(1)@3T – T(1)@1.5T) decreased by approximately 72% relative to precontrast T(1) values, while the field‐dependent difference of blood T(1) decreased only 30% postcontrast. Measurements of ΔR (1myo)/ΔR (1blood) were constant for 35 minutes postcontrast, but changed between 1.5T and 3T (0.46 ± .06 vs. 0.54 ± .06, P < 0.10). CONCLUSION: T(1) is significantly longer for myocardium (but not blood) at 3T compared to 1.5T. The differences in T(1) due to field strength are reduced following contrast administration, which may be attributed to changes in ΔR (1myo)/ΔR (1blood) with field strength. J. Magn. Reson. Imaging 2006. © 2006 Wiley‐Liss, Inc.

Meeting report on the 12th International Congress of Histochemistry and Cytochemistry (ICHC)

The International Congress of Histochemistry and Cytochemistry (ICHC) promoted in San Diego La Jolla (CA, USA), the 12th meeting where researchers of all over the world presented their work and the most innovative methods in histochemical disciplines. A summary of the last meeting is reported. © 2005 Wiley‐Liss, Inc.

Chikungunya disease and chloroquine treatment

Diversity and distribution of ferns in sub‐Saharan Africa, Madagascar and some islands of the South Atlantic

Aim This paper reports the diversity and endemism patterns of African ferns, and explores the potential role of diversity refuges and environmental and historical factors in the shaping of these patterns. Material and locations The extant fern taxa occupying Africa south of the Sahara, Madagascar and some islands of the South Atlantic. Methods The number of taxa in each area or operational geographical unit (OGU) was scored, and the correlation between this number and physical and climatic variables analysed by standard pairwise and stepwise multiple regression analysis (SPR and SMR). The effects of biological factors such as dispersal capacity, reproductive biology, genetic features and certain physiological adaptations were evaluated by comparing the number of species in each OGU. Floral affinities among OGUs were analysed using non‐metric multi‐dimensional scaling (NMS) and parsimonic analysis of dispersion (PAD), and compared with β‐turnover and inter‐OGU distances. Results OGU area, elevation and the distance between refuges determined the composition of local floras, but only greater OGU area and the existence of higher maximum elevations increased species richness. The distance between refuges also affected the number of endemic species, especially on islands. The biological features studied only slightly influenced fern distribution. The main climatic predictor of species number was humidity. SPR and SMR revealed three main groups of ferns with different ecological trends. NMS and PAD analyses separated the four areas of highest diversity in Africa, three of which are inhabited by ferns with distinct ecological requirements. The fourth area was Madagascar, which shows an accumulation of endemic and relict diversity that is not easy to explain. Main conclusions The distribution of ferns in Africa has been influenced by refuges. These probably allowed many species to recolonize the neighbouring areas after the extinctions of the Pleistocene. Three major components were detected in the African flora: Guinea‐Congolian thermophilous, cold‐tolerant Afro‐montane, and Southern drought‐tolerant elements. These are related to the three main refuge areas, i.e. the Gulf of Guinea area, the eastern tropical region, and the Cape region. Endemicity in ferns was found to be lower than that of seed plants due to the higher dispersability of fern spores. The distance between OGUs seems to be the main predictor of the number of endemic fern species these areas contain.

Plasmapheresis combined with interferon: An effective therapy for multiple sclerosis

The rationale for the use of interferon (IFN) in the treatment of multiple sclerosis (MS) is based on its recognized antiviral and immunomodulating actions. The pathogenesis of MS is believed to be due to an immunologic response in a genetically predisposed individual, localized within the central nervous system white matter, and triggered by exposure to an environmental agent such as a virus. Based on our personal experience we find that the efficacy of IFN therapy is hampered in MS patients by the presence of an interferon inhibitor factor (IIF) in the patients' sera which we have isolated and characterized. When plasmapheresis (PP) was done on 24 MS patients with intermittent 3‐day administration of IFN‐a and human leukocyte IFN, marked increase of IFN in 18 patients and modest increase in three patients correlated with clinical improvement. Three clinical nonresponders showed no increase in IFN levels following therapy. The ability to remove IIF and lymphokine inhibitor factor (LIF) by PP may explain the successful treatment of our patients. We describe the evaluation of helper T cells, suppressor T cells, HLADR antigen, natural killer cells, and monocyte/macrophage cell populations by flow cytometry before and after PP. A significant increase in these immune‐competent cells correlated with marked improvement in Kurtzke disability status scale in 13 patients, while eight stabilized. Patients showing progression of the disease either showed decrease or no change in these parameters after therapy. Encouraging results from this pilot study suggest that PP combined with immunomodulatory regimens of IFN may be an effective therapy for MS.

Rhinovirus C is associated with wheezing and rhinovirus A is associated with pneumonia in hospitalized children in Morocco

Human rhinovirus (RV) is commonly associated with severe acute lower respiratory infections (ALRI) in children. We aimed to describe the distribution of RV species and associations between RV species and clinical features in children hospitalized with clinically severe pneumonia (CSP) in Morocco. Nasopharyngeal aspirates (NPAs) were collected from 700 children, 2–59 months of age, admitted with CSP to the Hôpital d'Enfants de Rabat in Morocco. At least one respiratory virus was identified in 92% of children, of which RV was the most common (53%). PCR assays, sequencing, and phylogenetic tree analyses were carried out on 183 RV‐positive NPAs to determine RV species and genotypes. Of 157 successfully genotyped NPAs, 60 (38.2%) were RV‐A, 8 (5.1%) were RV‐B, and 89 (56.7%) were RV‐C. Wheezing and cyanosis were more common in RV‐C‐positive than RV‐A‐positive children (80.9% vs. 56.7%; P = 0.001 for wheezing and 10.1% vs. 0%; P = 0.011 for cyanosis). Physician's discharge diagnosis of pneumonia was more frequent among RV‐A‐positive (40.0%) than RV‐C‐positive children (20.2%; P = 0.009). RV‐A and RV‐C showed distinct seasonal patterns. Our findings suggest that RV‐C is associated with wheezing illness while RV‐A is associated with pneumonia. J. Med. Virol. 89:582–588, 2017. © 2016 Wiley Periodicals, Inc.

Protective effect of OK‐432 (streptococcal preparation) on murine fulminant hepatitis following mouse hepatitis virus infection

The effects of OK‐432 (streptococcal preparation) on murine fulminant hepatitis were investigated. Hepatitis was induced by injection of mouse hepatitis virus type 2 (MHV‐2) at a strength of either 1 times 10(3) or 1 times 10(4) plaque‐forming units (PFU). Mice without OK‐432 treatment died within 5 days, whereas mice pre‐inoculated with OK‐432 showed survival rates of 50% (1 times 10(3)PFU) or 10% (1 times 10(4)PFU) after 60 days. Survival time was not prolonged if OK‐432 was injected after MHV‐2. Examined histologically, mice not treated with OK‐432 showed severe haemorrhagic necrosis of the liver, often panlobular. Treated mice showed less necrosis; the least necrosis was observed in those injected with OK‐432 before MHV‐2. In those mice injected first with OK‐432 and then with 1 times 10(3)PFU of MHV‐2 that survived 7 days, autopsy showed a very slight and focal hepatic necrosis, with follicular infiltration by lymphocytes and macrophages. Mitogenic reaction of spleen cells was remarkably less than normal in mice with MHV‐2 injection. However, mice injected with OK‐432 before MHV‐2 (same treatment as mice showing high survival rates) showed relatively high reactivity in comparison with mice not treated with OK‐432.

WSAVA Guidelines for the vaccination of dogs and cats

Expression and function of chemokines during viral infections: from molecular mechanisms to in vivo function

Recruitment and activation of leukocytes are important for elimination of microbes, including viruses, from infected areas. Chemokines constitute a group of bioactive peptides that regulate leukocyte migration and also contribute to activation of these cells. Chemokines are essential mediators of inflammation and important for control of viral infections. The profile of chemokine expression contributes to shaping the immune response during viral infection, whereas viral subversion of the chemokine system allows the virus to evade antiviral activities of the host. In this review, we discuss the role of chemokines in host‐defense against virus infections, and we also look deeper into the virus‐cell interactions that trigger chemokine expression as well as the cellular signaling cascades involved.

Clinical Practice Guideline for the Evaluation of Fever and Infection in Older Adult Residents of Long‐Term Care Facilities: 2008 Update by the Infectious Diseases Society of America

Residents of long‐term care facilities (LTCFs) are at great risk for infection. Most residents are older and have multiple comorbidities that complicate recognition of infection; for example, typically defined fever is absent in more than one‐half of LTCF residents with serious infection. Furthermore, LTCFs often do not have the on‐site equipment or personnel to evaluate suspected infection in the fashion typically performed in acute care hospitals. In recognition of the differences between LTCFs and hospitals with regard to hosts and resources present, the Infectious Diseases Society of America first provided guidelines for evaluation of fever and infection in LTCF residents in 2000. The guideline presented here represents the second edition, updated by data generated over the intervening 8 years. It focuses on the typical elderly person institutionalized with multiple chronic comorbidities and functional disabilities (e.g., a nursing home resident). Specific topic reviews and recommendations are provided with regard to what resources are typically available to evaluate suspected infection, what symptoms and signs suggest infection in a resident of an LTCF, who should initially evaluate the resident with suspected infection, what clinical evaluation should be performed, how LTCF staff can effectively communicate about possible infection with clinicians, and what laboratory tests should be ordered. Finally, a general outline of how a suspected outbreak of a specific infectious disease should be investigated in an LTCF is provided.

Systemic Vasculitis with Severe Cutaneous Manifestation as a Suspected Idiosyncratic Hypersensitivity Reaction to Fenbendazole in a Cat

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Feline Hybridoma Growth Factor/lnterleukin‐6 Activity

An assay system was developed to measure feline hybridoma growth factor (HGF)/interleukin‐6 (IL‐6) activity in biological samples containing many kinds of cytokines by using the proliferation of the newly established mouse‐rat hybridoma clone. B3B1. The proliferative response of this B3B1 clone was IL‐6‐specific, and could not be promoted by other cytokines including IL‐1, IL‐2, IL‐3, and granulocyte‐colony‐stimulating factor (G‐CSF), The anti‐human B‐cell stimulatory factor 2 (BSF‐2)/IL‐6 antiserum did not neutralize feline HGF/IL‐6 activity in conditioned media prepared from feline con A‐stimulated splenocytes and unstimulated alveolar macrophages, indicating antigenic differences between species. Feline HGF/IL‐6 was eluted into the fractions corresponding to a molecular weight of 30,000–40,000 in gel filtration, and into the fractions at a salt concentration of 0.2–0.3 M NaCI in anion exchange chromatography. The physicochemical properties of feline HGF/IL‐6 were slightly different from those of murine and human IL‐6.

Direct detection of rhinoviruses by an enzyme‐linked immunosorbent assay

This paper describes the first enzyme‐linked immunosorbent assay for the detection of rhinovirus antigens in clinical specimens (nasal washings), either directly or following overnight cell culture amplification. The assay takes approximately 48 hours to perform and utilizes the same rabbit antirhinovirus hyperimmune serum as both the capture and detecting antibody. The latter has been biotin‐labelled and is detected via a streptavidin 3‐galactosidase preformed complex. This new assay has been found to be very sensitive, detecting human rhinovirus (HRV)‐EL and HRV‐2 at titres as low as 10(1.8) TCID(50) 100 μl(−1) and < 10(1) TCID(50) 100 μl(−1), respectively. Furthermore, when 57 different human rhinovirus serotypes were tested in both the HRV‐EL and HRV‐2 ELISA systems a total of 49 (86%) were found to be cross‐reactive. Of 36 clinical specimens tested by virus isolation, cell‐culture‐amplified (CCA) ELISA, and direct ELISA, 15 were positive by isolation, 11 by CCA‐ELISA, and 11 by direct ELISA. The overall correlation of the CCA and direct ELISA techniques with virus isolation was found to be 88.9% and 66.7%, respectively. The present study demonstrates that the ELISA system developed is a sensitive technique for the diagnosis of rhinovirus infections.

Preliminary Characterisation of Torovirus‐Like Particles of Humans: Comparison With Berne Virus of Horses and Breda Virus of Calves

Pleomorphic virus‐like particles have been observed by electron microscopy in the faeces of children and adults with diarrhoea. Some of these particles were approximately 100 nm in diameter and had a “fringe” of closely applied peplomers approximately 10 nm long; they closely resembled Berne virus of horses and Breda virus of calves, the two representatives of a newly proposed family called the Toroviridae. In one sample a toroidal nucleoprotein‐like structure was observed within the particles. For two samples a buoyant density of 1.14 g/ml was determined by centrifugation through a sucrose density gradient. One sample possessed a haemagglutination for rat erythrocytes. The serological relationship between these different viruses was observed by immune electron microscopy, haemagglutination inhibition, and serum neutralisation. The role of these viruslike particles as candidate pathogens of humans is discussed.

Two models of multiple sclerosis: Experimental allergic encephalomyelitis (EAE) and theiler's murine encephalomyelitis virus (TMEV) infection. A pathological and immunological comparison

Theiler's murine encephalomyelitis virus (TMEV) infection and experimental allergic encephalomyelitis (EAE) are considered among the best models of human multiple sclerosis (MS). In both models, clinical disease is characterized by paralysis, while pathological changes consist of inflammatory demyelination. In both models there is a genetic influence on susceptibility/resistance to the development of disease. This has been thoroughly studied in TMEV infection, and it has been found to depend on both major histocompatibility complex (MHC) and non‐MHC genes. At least four genes have been so far identified. Because of this genetic influence, some strains of mice are more susceptible to both clinical and pathological changes than others, and susceptibility appears to best correlate with the ability of a certain murine strain to develop a delayed‐type hypersensitivity (DTH) response to viral antigens. We have also observed that even among mice which are equally susceptible clinically, striking differences may be seen under pathological examination. These consist of different gradients of severity of inflammation, particularly in regards to the macrophage component. There is an inverse relationship between the number of macrophages, and their length of stay in the CNS, and the ability of mice to remyelinate their lesions. The most severe lesions are in SJL/J mice, and remyelination in this strain is extremely poor. The least severe lesions in terms of macrophage invasion are in strains such as NZW and RIIIS/J, and these are able to remyelinate lesions very successfully. Murine chronic relapsing EAE (CR‐EAE) shows pathological changes in many ways similar to those in TMEV‐infected SJL/J mice, although less severe in terms of degrees of macrophage infiltration and tissue destruction. Mice with CR‐EAE have a correspondingly limited ability to remyelinate their lesions. In both models the pathology appears to be mediated through a DTH response. However, while in EAE the DTH response is clearly against neuroantigens, the response in TMEV infection is against the virus itself. The end result in both models would be that of myelin destruction through a lymphotoxincytokine‐mediated mechanism. The importance of the DTH response in both models is well illustrated by the effects of tolerance induction in EAE and TMEV infection to neuroantigens and virus, respectively. These are important models of human MS, since the current hypothesis is that a viral infection early in life, on the appropriate genetic background, may trigger a secondary misdirected immune response which could be directed either against myelin antigens and/or possible persistent virus(es). © 1995 Wiley‐Liss, Inc.

Genetic variability of alcohol dehydrogenase among Australian Drosophila species: Correlation of ADH biochemical phenotype with ethanol resource utilization

Alcohol dehydrogenase (ADH) activities, electrophoretic phenotypes, and the extent of ethanol resource utilization are compared for three groups of species distinguishable on ecological criteria: 1) the cosmopolitan species D. melanogaster, a frequent inhabitant of wineries; 2) fruit‐baited species of the typically Australian subgenus Scaptodrosophila: D. lativittata, D. nitidithorax and D. howensis; and 3) Scaptodrosophila species not attracted to fermented‐fruit baits being collected by sweeping in temperate rain forests (D. inornata, D. collessi) or from Hibiscus flowers (D. hibisci). D. melanogaster showed the highest levels of ADH activity and an electrophoretic polymorphism with two active allelic forms, while group 2) species showed intermediate ADH activities and polymorphisms, which were consistent with “high activity” and “low activity” allelic forms in natural populations of these species, and group 3) species showed only “low activity” forms. Ethanol resource utilization follows the same sequence, being 1 > 2 > 3(D. howensis and D. collessi were not tested). Therefore the species considered shown an association of ADH biochemical phenotype, laboratory ethanol utilization, and resources utilized.

High protection of animals lethally infected with influenza virus by aprotinin‐rimantadine combination

The successful therapeutic synergism of aprotinin and rimantadine, which are known to attack different viral targets, was demonstrated in influenza‐virus infected animals. Combined treatment with these drugs of mice infected with a highly lethal does of mouse‐adapted influenza virus prevented the development of separate administration of aprotinin and rimantadine induced 35% and 15% protection, respectively.

WSAVA Guidelines for the Vaccination of Dogs and Cats

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Therapeutic spectrum of interferon‐β in ischemic stroke

Ischemic stroke is devastating and a major cause of morbidity and mortality worldwide. To date, only clot retrieval devices and/or intravenous tissue plasminogen activators (tPA) have been approved by the US‐FDA for the treatment of acute ischemic stroke. Therefore, there is an urgent need to develop an effective treatment for stroke that can have limited shortcomings and broad spectrum of applications. Interferon‐beta (IFN‐β), an endogenous cytokine and a key anti‐inflammatory agent, contributes toward obviating deleterious stroke outcomes. Therefore, exploring the role of IFN‐β may be a promising alternative approach for stroke intervention in the future. In the present review, we have discussed about IFN‐β along with its different mechanistic roles in ischemic stroke. Furthermore, therapeutic approaches targeting the inflammatory cascade with IFN‐β therapy that may be helpful in improving stroke outcome are also discussed.

RSV‐hRV co‐infection is a risk factor for recurrent bronchial obstruction and early sensitization 3 years after bronchiolitis

To assess risk factors of recurrent bronchial obstruction and allergic sensitization 3 years after an episode of acute bronchiolitis, whether after ambulatory care treatment or hospitalization. A monocentric prospective longitudinal study including infants aged under 1 year with acute bronchiolitis was performed, with clinical (severity score), biological (serum Krebs von den Lungen 6 antigen), and viral (14 virus by naso‐pharyngeal suction detection) assessments. Follow‐up included a quaterly telephone interview, and a final clinical examination at 3 years. Biological markers of atopy were also measured in peripheral blood, including specific IgEs towards aero‐ and food allergens. Complete data were available for 154 children. 46.8% of them had recurrent wheezing (RW). No difference was found according to initial severity, care at home or in the hospital, respiratory virus involved, or existence of co‐infection. A familial history of atopy was identified as a risk factor for recurrent bronchial obstruction (60% for RW infants versus 39%, P = 0.02), as living in an apartment (35% versus 15%, P = 0.002). 18.6% of the infants were sensitized, with 48.1% of them sensitized to aeroallergens and 81.5% to food allergens. Multivariate analysis confirmed that a familial history of atopy (P = 0.02) and initial co‐infection RSV‐hRV (P = 0.02) were correlated with the risk of sensitization to aeroallergens at 3 years. Familial history of atopy and RSV‐hRV co‐infection are risk factors for recurrent bronchial obstruction and sensitization.

Toxicologic effects of ribavirin in cats

Ribavirin, a broad‐spectrum antiviral agent active in vitro against a number of RNA and DNA viruses, has been associated with moderate toxicity in laboratory animals and humans. Clinically, ribavirin has been used effectively in persons primarily to treat life‐threatening viral diseases such as acute haemorrhagic fever or viral pneumonia of infants. In order to evaluate the feasibility of using this antiviral agent in cats, the effects of oral (p.o.), intramuscular (i.m.) and intravenous (i.v.) doses of ribavirin in 27 9‐month‐old specific‐pathogen‐free cats were evaluated by haematology, clinical chemistries, bone marrow biopsies and histopathology. Ribavirin was administered once daily for 10 consecutive days at a dose of either 11, 22, or 44 mg/kg after which all cats were euthanatized and necropsied. Most cats receiving 22 or 44 mg of ribavirin/kg became anorectic and suffered some degree of weight loss (0.2 to 0.6 kg), and about one‐third of the cats developed diarrhoea and/or mucous membrane pallor. Icterus or haemorrhage was not observed. The most profound and consistent haemato‐logic change, particularly among the moderate and high dosage groups regardless of route of administration, was a significant and severe thrombocytopenia (range, 33–78% reduction in mean platelet counts vs. baseline). Other changes, particularly reductions in total WBC and neutrophils and reductions in RBC and PCV, tended to occur at lower ribavirin dosages, but generally they were not statistically significant. Cats given 44 mg of ribavirin/kg i.v. showed significant decreases in leukocyte variables, including total WBC (P = 0.016), neutrophils (P= 0.026) and lymphocytes (P= 0.047). Mild‐to‐moderate increases in serum alanine aminotransferase and alkaline phosphatase activities occurred at doses of 22 and 44 mg/kg. Evaluation of bone marrow biopsies before and after treatment revealed that cats given 11 mg of ribavirin/kg had mild megakaryocytic (MK) hypoplasia, whereas cats receiving 22 or 44 mg/kg had progressively severe degrees of MK hypoplasia and dysplasia, asynchronous MK maturation, and increased myeloidrerythroid ratio. Pathologic changes in ribavirin‐treated cats generally were mild and included primarily enteritis (seven cats) and hepatocellular vacuolation and/or centrilobular necrosis (seven cats). Results of this study in cats indicated that daily administration of ribavirin at a dose range of 11 to 44 mg/kg induced a dose‐related toxic effect on bone marrow, primarily on megakaryocytes and erythroid precursors, and at the higher dosages it suppressed numbers of circulating leukocytes.

Occurrence in water of viruses of public health significance

Detection of human bocavirus in respiratory, fecal, and blood samples by real‐time PCR

Human bocavirus (HBoV) has been detected worldwide in respiratory samples. Two real‐time PCR assays, targeting the non‐structural protein (NP‐1) and viral protein (VP‐1) genes, were designed and validated to detect HBoV in patients with respiratory disease, gastroenteritis, or systemic illness. Sensitivity of the NP‐1 and VP‐1 assays were equal to the conventional PCR assay previously described by Allander et al. [2005: Proc Natl Acad Sci USA 102: 12891–12896] being 100%, and giving specificity of 94% and 93%, respectively. There was no cross‐reaction identified with unrelated respiratory agents, or to human DNA. The limits of detection were 10 copies of genomic DNA equivalents per reaction for both assays. The assays were used to screen three different sample populations, combined nose, and throat swabs (n = 96) from children with acute respiratory disease, fecal samples (n = 375) from adults, and children with gastroenteritis and whole blood (n = 229) collected from 31 immunocompromised children taken over an 18‐month period. In total 17 (18%) respiratory samples and 18 (4.8%) fecal samples were identified as having HBoV present. Of the pediatric whole blood specimens investigated, HBoV was detected in six (2.6%) samples from four patients. In summary, two real‐time PCR assays targeting different genes were designed and validated for use as screening methods for the detection of HBoV. HBoV was found in three different specimen types: parent‐collected combined nose–throat swabs, fecal samples collected from symptomatic individuals and whole blood from immunocompromised children. J. Med. Virol. 81:488–493, 2009. © 2009 Wiley‐Liss, Inc.

ORIGINAL ARTICLE: Taxonomy and diversity of Afroalpine Chironomidae (Insecta: Diptera) on Mount Kenya and the Rwenzori Mountains, East Africa

Aim Anthropogenic climate change is expected to result in the complete loss of glaciers from the high mountains of tropical Africa, with profound impacts on the hydrology and ecology of unique tropical cold‐water lakes located downstream from them. This study examines the biodiversity of Chironomidae (Insecta: Diptera) communities in these scarce Afroalpine lake systems, in order to determine their uniqueness in relation to lowland African lakes and alpine lakes in temperate regions, and to evaluate the potential of Afroalpine Chironomidae as biological indicators to monitor future changes in the ecological integrity of their habitat. Location Mount Kenya (Kenya) and Rwenzori Mountains (Uganda). Methods The species composition of Afroalpine chironomid communities was assessed using recent larval death assemblages extracted from the surface sediments of 11 high‐mountain lakes between 2900 and 4575 m. Results were compared with similar faunal data from 68 East African lakes at low and middle elevation (750–2760 m), and with literature records of Chironomidae species distribution in sub‐Saharan Africa, the Palaearctic region and elsewhere. All recovered taxa were fully described and illustrated. Results The 11‐lake analysis yielded 1744 subfossil chironomid larvae belonging to 16 distinct taxa of full‐grown larvae, and three taxa of less differentiated juveniles. Eleven of these 16 are not known to occur in African lakes at lower elevation, and eight taxa (or 50% of total species richness) appear restricted to the specific habitat of cold lakes above 3900 m, where night‐time freezing is frequent year‐round. The faunal transition zone coincides broadly with the Ericaceous zone of terrestrial vegetation (c. 3000–4000 m). Snowline depression during the Quaternary ice ages must have facilitated dispersion of cold‐stenothermous species among the high mountains of equatorial East Africa, but less so from or to the Palaearctic region via the Ethiopian highlands. Main conclusions Chironomid communities in glacier‐fed lakes on Africa's highest mountains are highly distinct from those of lowland African lakes, and potentially unique on a continental scale. By virtue of excellent preservation and their spatial and temporal integration of local community dynamics, chironomid larval death assemblages extracted from surface sediments are powerful biological indicators for monitoring the hydrological and ecological changes associated with the current retreat and loss of Africa's glaciers.

Isolation and Identification of Enteric Adenoviruses

Thirty‐four out of 64 faecal samples with adenovirus particles, as seen by electron microscopy, were found to contain adenovirus 40 or 41 by direct isolation and neutralization in Chang's conjunctival cells, mostly within one week. (Ad40 and 41 candidate viruses are serologically related.) 6 other adenovirus specics were isolated; 6 samples gave equivocal results, and 18 were negative. A genus‐specific ELISA with an antihexon coat yielded positive results in 40 out of 55 samples; the test failed to identify adenovirus antigen in 10 out of 17 specimens, which were found negative by culture. All of them were negative by immunfluorescence of inoculated Chang cell cultures. Hence the failures are probably due to insufficient amount of virus in the samples. The predominance of only two adenovirus species associated with gastroenteritis in children and the ease of cultivating and identifying them should help to elucidate their etiological significance.