title: Chapter 7 Use of Recombinant Vaccinia Virus Vectors for Cell Biology

text: This chapter describes the use of several of the recombinant vaccinia expression systems, focuses on the systems that are most useful for cell biologists, and discusses their advantages and limitations. Vaccinia-mediated expression can be used for assessing cellular localization, posttranslational modifications, oligomerization, and transport and turnover rates. The system provides a rapid method for screening mutant proteins for expression and targeting. It is an excellent way of quickly deciding which mutant proteins might be worth further studying using stable expression systems. Expression of foreign genes using Vaccinia virus is based on recombinant viruses constructed by insertion of complementary DNA (cDNA) into the nonessential thymidine kinase (TK) gene. Both direct and indirect methods of expression are possible. The foreign gene can be inserted into the vaccinia genome by homologous recombination using a plasmid with flanking regions of vaccinia DNA. The recombinant virus is selected, expanded, and used to infect cells, which then express high levels of the foreign protein. Recombinant vaccinia viruses are generated by subcloning the foreign gene into a plasmid transfer vector so it is flanked by DNA from the vaccinia (TK) gene, which is nonessential for growth of the virus in tissue culture. This plasmid is then transfected into vaccinia-infected cells. Homologous recombination of the plasmid and the vaccinia genome generates a recombinant virus with an inactive TK gene.


Recommendations for the prevention of transmission of SARS during GI endoscopy


title: Determinants of Diarrhea in Viral Enteritis: The Role of Ion Transport and Epithelial Changes in the Ileum in Transmissible Gastroenteritis in Piglets

text: To understand mechanisms of viral diarrhea further, we studied ileal ion transport in vitro in relation to mucosal changes and epithelial differentiation in transmissible gastroenteritis in piglets, an invasive viral enteritis thought to involve mainly proximal intestine. In infected pigs, at the height of diarrhea, short-circuited ileal epithelium failed actively to transport Na(+) and Cl(−), and there was a defect of glucose-mediated Na(+) transport. The Cl(−) secretory response to theophylline remained intact. Conductance measurements indicate that paracellular permeability may be reduced and transcellular transport may be altered. A mucosal lesion was observed at the time of the transport changes, characterized by villus blunting, crypt hyperplasia, and immature crypt-type enterocytes on the villus epithelium, deficient in disaccharidase and (Na(+), K(+))ATPase activity but rich in thymidine kinase. Consideration of the major determinants of diarrhea in this invasive enteritis must take into account not only altered mucosal function and differentiation but also the extent of intestinal involvement, including the ileum, a major site of fluid absorption in the intestine.


El enterovirus D68 sí que existe en España


title: Pathogenesis of infectious bronchitis nephritis 1. Morphometric analysis of kidney proximal tubular epithelium in chickens

text: The ultrastructure of the proximal tubular epithelial cells in chicken kidneys was examined throughout the course of an experimental infection with infectious bronchitis virus. A quantitative assessment of the structural changes in the cells was related to these in normal cells. Significant alterations were detected in the membrane structures and in the mitochondria. There was a reduction in surface area of the microvillus membrane, the basolateral membrane and the apical tubular membrane. There were alterations in the shape of mitochondrial profiles and a decrease in the volume density of mitochondria. Vesicular structures, which are a possible site of viral release, were observed in the lateral surface of cells. These changes in the functional components of the cells indicate impaired transport of ions and water. The results demonstrate the value of stereological methods for the study of viral-host cell interactions in the pathogenesis of viral disease.


title: Ruminant pestiviruses

text: The ruminant pestiviruses, bovine virus diarrhoea virus (BVDV) and border disease virus (BDV) are highly successful and important pathogens which infect ruminant species worldwide. Although the serological relationships among ruminant pestiviruses require further clarification, there is growing evidence for two antigenic groups, one of which predominates in cattle and one in sheep. The success of pestiviruses stems from the ability of the non-cytopathic (NCP) biotype of the virus to cross the placenta and establish a persistent infection (PI) in the developing foetus. This biotype should be regarded as the ‘normal’ biotype with the cytopathic (CP) biotype being an abnormal virus that is usually isolated only from PI animals dying from mucosal disease. Recent molecular evidence points to CP viruses arising from their NCP counterparts by recombination events that include the insertion of host RNA and/or the duplication of viral RNA sequences. However, the biological mechanism through which CP viruses kill cells remains unknown. Virtually all CP and NCP viruses cause only mild, transient clinical symptoms in healthy adult animals and stimulate a protective immune response. Despite the urgent requirement for a safe, effective vaccine, there is still no commercial vaccine that has been shown to immunize dams so that foetal infection is prevented. In the absence of an effective vaccine, reliable diagnostic techniques are essential to implement effective control measures. There is now a range of monoclonal antibody-based enzyme-linked immunosorbent assays for identifying PI or convalescent animals. These tests are specific, rapid, sensitive and reliable but may themselves become redundant as they are superceded by ever-increasing molecular biology-based techniques.


title: Pathogenesis of Aujeszky's disease virus infection in swine tracheal organ culture

text: Two different strains of Aujeszky's disease virus (ADV) were inoculated into swine tracheal organ culture. Both viruses replicated in and destroyed the tracheal epithelium and epithelial cells. ADV antigen was first localized in ciliated epithelial cells by fluorescent antibody and immunoperoxidase examinations. Corresponding to the distribution of ADV antigen, many ADV particles were observed in ciliated epithelial cells. Results demonstrated that the tracheal epithelium infected with ADV is reduced in its ciliary activity.


title: Pathological features of lymphoid tissues in cats with natural feline immunodeficiency virus infection

text: A range of tissues from a total of 17 cats naturally infected with the feline immunodeficiency virus was examined histologically. In 11 cases, chronic inflammatory lesions were present in various tissues including, most commonly, the intestine, brain and lung. Extensive inflammation in the intestinal wall was present in seven of the cats. No particular bacterial organisms were demonstrated in these inflammatory lesions. A range of changes was present in the lymph nodes, including hyperplasia, atrophy or a mixed pattern. Erythrophagocytosis was a consistent feature. The changes resembled those reported in human acquired immunodeficiency syndrome as a result of infection with human immunodeficiency virus.


title: Immunohistochemical demonstration of spread of Aujeszky's disease virus to the porcine central nervous system after intestinal inoculation

text: Aujeszky's disease virus (ADV) was injected into the duodenal lumen of eight specific pathogen-free pigs aged 5 weeks. The infected pigs did not show any diarrhoea or nervous symptoms, but they developed characteristic necrotizing enteritis and myenteric plexitis, accompanied by follicular necrosis in the Peyer's patches. ADV antigen was detected in the submucosa of the dome area of Peyer's patches, lymphatic follicles, Meissner's and Auerbach's plexuses, solar ganglia and thoracic spinal ganglia. These findings suggest that ADV spreads from the intestinal mucosa to the central nervous system via the autonomic nerves.


Viajar por trabajo. ¿Un nuevo perfil de viajero de alto riesgo?


title: Porcine reproductive and respiratory syndrome (PRRS): A review, with emphasis on pathological, virological and diagnostic aspects

text: Despite early attempts to control the spread of the disease, porcine reproductiveand respiratory syndrome (PRRS) has now become endemic in many countries including Britain. The occurrence of subclinical herd infections, the prolonged circulation of virus within herds and probable aerogenic virus spread all mitigated against the success of control measures. The origin of the disease is unknown but the causative agent has been shown to be an arterivirus with shared features to lactate dehydrogenase virus of mice. There is evidence of extreme genetic and antigenic variability between American and European isolates. PRRS virus has a predilection for alveolar macrophages and does notgrow in most cell lines. In infected pigs, viraemia can persist for many weeks in the face of circulating antibodies and little is known about the mechanisms by which immunity to infection develops. A wide spectrum of disease has been reported from the field, accompanied in some cases by heavy economic losses. Reproductive and perinatal losses were most prominent when the disease first appeared. In the endemic phase, PRRS may be more significant as a contributory factor to a post-weaning respiratory syndrome of young pigs of 3–8 weeks. On-farm techniques have been developed to reduce the recycling of PRRS virus from older infected nursery pigs to the younger newly weaned pig. Vaccines are now marketed for the control of PRRS, but are not licensed for use in Britain. Improvements in knowledge of virion composition and antigenic stability and in the nature of the immune response of the pig should result in genetically engineered subunit vaccines becoming available. Diagnosis of PRRS is still difficult as many animals do not show clinicalsigns and may only be detected by serology and often only when other respiratory diseases are being investigated. Now that the infection is widespread, serological testing must be properly targeted and interpreted to give meaningful results about virus circulation. An increasing arsenal of diagnostic methods are becoming available to detect virus in both fresh and fixed specimens. The pathogenic mechanisms of PRRS remain poorly defined and more work is needed to reveal the nature of the interaction between PRRS virus and other factors in disease.


title: l-glutamine stimulates jejunal sodium and chloride absorption in pig rotavirus enteritis

text: Rotavirus enteritis is the leading cause of diarrhea in infants worldwide. A research priority of the World Health Organization is to develop oral rehydration solutions containing amino acids or other additives that will stimulate intestinal absorption more efficiently than the current glucose-based oral rehydration solutions. Glutamine is the principal metabolic fuel of the small bowel and a putative stimulator of mucosal repair. This report describes the transport response to mucosal l-glutamine following intestinal injury caused by porcine rotavirus. Peak symptoms and mucosal damage were observed 2–7 days after oral rotavirus inoculation. In vitro transport studies of the maximally injured region, the midjejunum (80% reduction in lactase), surprisingly, showed transport responses to l-glutamine (30 mmol/L) Sand l-alanine (30 mmol/L) that were similar qualitatively and quantitatively to those observed in control tissue. Subsequent application of mucosal d-glucose (30 mmol/L) caused additional stimulation of electrogenic Na(+) transport, but the response to glucose was blunted (P < 0.05) in the infected tissues. Glutamine and alanine enhanced Na(+) absorption to a similar degree (2–2.5 μEq · cm(−2) · h(−1)), but glutamine stimulated equal amounts of electrogenic and electroneutral NaC1 absorption, whereas alanine had no significant effect on net Cl(−) flux. Glutamine is a potentially useful substrate for investigation in oral rehydration solutions for infant diarrhea.


Severe Acute Respiratory Syndrome, Pulmonary Function Tests, and Quality of Life: Lessons Learned


title: Colonic Compensation in Transmissible Gastroenteritis of Swine

text: Absorption of water and electrolytes by the small and large intestine was examined using a nonabsorbable marker technique in 3-day-old and 3-wkold pigs. One-half of the pigs in each group were orally infected with transmissible gastroenteritis virus; the remaining pigs served as controls. Threeday-old control pigs concentrated the nonabsorbable fluid marker twelve f old along the small and large intestine, indicating an efficiency of about 95% in absorption of the exogenous daily fluid load presented to the intestine. In contrast, the marker concentration in infected pigs showed no change whatsoever along either the small or large intestine, indicating a complete absence of net fluid absorption or secretion in these animals. Three-week-old control pigs concentrated the marker similarly to the 3-day-old group, with the bulk of the fluid absorption occurring in the small intestine. Infected pigs in the 3-wk-old group had marked net fluid secretion in the proximal small intestine, so that about twice the fluid load was presented to the large intestine of the 3-wk-old infected pigs as compared to the 3-day-old infected group. However, in contrast to the 3-day-old infected group, the large intestine of the 3-wk-old infected pigs increased fluid absorption some six times over the control, and this compensatory response prevented diarrhea in these older animals. Analysis of luminal contents indicated that in the older pigs, unabsorbed carbohydrate was almost completely fermented to short-chain fatty acids in the colon, whereas in the younger pigs the carbohydrate passed through the colon unchanged. These results demonstrate that development of microbial digestion, together with rapid short-chain fatty acid absorption, is a primary feature responsible for the colonic compensation in the older pigs with transmissible gastroenteritis.


Human cancer vaccines()


Who's boss


The timing is right: Evolution of AIDS-causing HIV strains is consistent with history of chloroquine use


Síndrome respiratorio agudo grave: ¿final o paréntesis de una epidemia?


Viral Gastroenteritis


docid: wsbxrssg

title: A unifying hypothesis for pathogenesis and prevention of necrotizing enterocolitis


Tendencias de la salud internacional: retos y oportunidades para nuestra sociedad


Was a neurovirulent influenza virus the cause of amyotrophic lateral sclerosis and parkinsonism-dementia on Guam?


title: Diagnosis of bovine cryptosporidiosis by an enzyme-linked immunosorbent assay

text: This paper describes an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of cryptosporidiosis. A monoclonal antibody with a high affinity against an oocyst antigen was used to set up the test. The efficiency of this assay was compared with that of the flotation test; 275 calf faecal samples were examined by the two methods. There was 96% agreement between the two tests. For the 11 conflicting samples, the two tests were repeated and a modified Ziehl-Neelsen staining was performed on faecal smears. All these 11 samples contained few oocysts, but only five and six of them were shown to be positive by the ELISA and flotation tests, respectively. The degree of sensitivity of the ELISA and flotation tests is comparable; samples heavily or moderately contaminated with oocysts are detected by both methods. This ELISA is reliable and never gives rise to false positive results. Nevertheless, as with the flotation test, the occasional case containing very few oocysts will not always be detected by this test. If necessary, very accurate diagnosis can be made by a staining technique or by a direct immunofluorescent assay. In veterinary medicine, the ELISA seems to be a method of choice; it appears to be a fast and reliable technique which could be used as a routine test for the detection of Cryptosporidium oocysts. Nevertheless the degree of sensitivity must always be borne in mind. There is no need for a microscopic examination, which is an additional advantage.


title: Monoclonal antibodies from Epstein-Barr virus-transformed lymphocytes of common marmosets (Callithrix jacchus) immune to malaria

text: The B lymphocytes of the common marmoset Callithrix jacchus can be immortalised by infection with Epstein-Barr virus (EBV) in vitro (Desranges et al., 1976). C. jacchus is susceptible to infection with the blood stages of several species of malaria parasite including the line designated MVF1 (Mitchell et al., 1988) from which it recovers and shows immunity to reinfection. By exploiting these two phenomena, EBV-transformed, marmoset lymphoblastoid cell lines secreting antibodies to malaria parasite antigens have been generated and cloned. We believe this to be the first time that monoclonal antibodies (MAbs) have been raised from common marmosets. Since numerous and diverse human pathogens can infect this small primate in the laboratory, these methods may prove generally applicable for the generation of MAbs whose specificities derive from immune responses to infection.


title: A new conjugate for the ELISA quantitation of porcine IgA

text: The preparation of a new soluble immune complex conjugate (anti-IgA-IgA-peroxidase), suitable for use in the ELISA technique is described. This conjugate requires pure IgA and specific anti-IgA, but the enzyme (peroxidase) was easily conjugated to the IgA.


Proceedings Fifth ESWI Influenza Conference


Growth and phototherapy


Rational hand hygiene during COVID-19 pandemic


title: Measurement of the concentration of murine IgG monoclonal antibody in hybridoma supernatants and ascites in absolute units by sensitive and reliable enzyme-linked immunosorbent assays (ELISA)

text: We have investigated an enzyme-linked immunosorbent assay (ELISA) for mouse IgG using affinity-purified goat anti-mouse antibodies for capture and detection. This assay was used to measure the absolute or weight/volume concentration of murine monoclonal antibody in hybridoma supernatants. Bovine or horse serum did not interfere with the assay, which permitted reliable measurement of all murine IgG subclasses except IgG3 in the 1–20 ng/ml range. Antibody capture was essentially complete in the optimized assay. In combination with an antigen-dependent ELISA, the assay allowed estimation of the absolute concentration of specific monoclonal antibody in ascites. These rapid and relatively simple assays may be applicable in many situations in which a practical means of measuring murine monoclonal antibodies in weight/volume units is needed.


title: Cystatin C-Properties and use as diagnostic marker

text: This chapter focuses on the most well characterized inhibitors—cystatin C—and provide some information on its structure, biochemical properties, its role in normal and abnormal physiological processes, as well as on its use as a diagnostic marker. A major part of the cysteine proteases are evolutionary related to the structurally well–defined cysteine protease papain and are called papain–like cysteine proteases. The biological roles and the cystatin superfamily inhibitors of papain–like cystein proteases are also discussed. The aminoacid sequence and schematic structure of human cystatin C is also presented. The evolutionary relationships among all known inhibitory active human cystatins and kininogen cystatin domains are diagrammatically represented. The distribution of cystatins in body fluids and additional functions attributed to cystatin C are described. The serum or plasma cystatin C is used as a marker for glomerular filtration rate (GFR). The urine cystatin C is used as a marker for proximal tubular damage. The two types of brain hemorrhage associated with Cystatin C amyloid deposits are also demonstrated. The conditions connected with deposition of amyloid β–protein in cystatin C and cerebral hemorrhage is also provided.


Subject index volumes 146–155 1992


Proceedings of the 2nd seminar on emerging infectious diseases, December 7, 2012 – Current trends and proposals


title: N-linked protein glycosylation in the rat parotid gland during aging

text: N-Linked protein glycosylation was examined in vitro in dispersed rat parotid acinar cells from young adult (3–6 months) and aged (22–24 months) rats. A small decrease in general protein production was observed with cells from aged animals (∼20% lower incorporation of [(14)C]leucine into 10% CCI(3) COOH insoluble protein during continuous pulse labeling). Incorporation of [(3)H]mannose into N-linked glycoproteins by aged cells was further reduced (∼35%). Similarly microsomal membranes from parotid glands of aged animals showed ∼50% reduction in the synthesis of mannosylphosphoryl dolichol, a key intermediate in the dolichol pathway of protein N-glycosylation. Man-P-Dol synthase, the microsomal enzyme responsible for production of this saccharide-lipid, displayed no change in apparent K(m) for GDP-mannose when preparations from aged animals were utilized, but did show ∼50% reduction in V(max). Following β-adrenoreceptor activation, cells from both young adult and aged glands showed increased N-linked protein glycosylation almost to the same extent (∼2-fold). The data suggested that in aged rat parotid cells there is a basal reduction of activity in the pathway responsible for asparagine-linked protein glycosylation, but that following exocytotic stimuli this pathway responds in a manner comparable to cells from young adult glands.


title: Methods for studying naturally occurring human pain and their analogues

text: Methods for investigating human pain have been developed over the last 100 years. Typically, researchers focus on people with clinical pain, or on healthy participants undergoing laboratory-controlled pain-induction techniques focussed mostly on exogenously generated skin nociception. Less commonly investigated are acute pain experiences that emerge naturally. Six common painful complaints were identified: headache, muscular pain, visceral pain, menstrual pain, dental pain, and pain associated with upper respiratory tract infection. Methods used to recruit participants with the natural occurrence of each pain complaint were identified, and features of their use reviewed. Also reviewed were experimental analogues designed to mimic these pains, with the exception of menstrual pain. Headache and menstrual pain appear to be most effectively researched in their naturally occurring form, whereas muscle and dental pain may be more easily induced. Upper respiratory tract infection and abdominal pain provide further challenges for researchers. Summary guidance is offered, and directions for methods development outlined.


Author index volumes 146–155 1992


title: Pathogenesis of Theiler's Murine Encephalomyelitis Virus

text: Theiler's murine encephalomyelitis virus belongs to the family of picornaviridae. Picornaviruses are small ( “pico”), phylogenetically related RNA viruses. Based on different biochemical and biophysical characteristics picornaviruses are subdivided into four groups: enteroaphthovirus (foot-and-mouth disease virus), cardiovirus [encephalomyocarditis virus (EMCV), Mengo virus], and rhinovirus (human rhinovirus). Theiler's murine encephalomyelitis virus was originally classified among the picornaviridae as an enterovirus because of its biological similarities with poliovirus. Further comparison of the complete genome of TMEV BeAn 8386 strain identifies remarkable similarities at the level of nucleotides and predicted amino acids between BeAn and the cardioviruses EMCV and Mengo virus. Theiler's murine encephalomyelitis virus is a single-stranded nonenveloped RNA virus. The viral RNA is of positive sense, having the same polarity as mRNA. Viral mRNA lacks the cap structure found at the 5’ end of almost all eukaryotic mRNAs.


The conference on vaccines protective against enteric diseases


Requiem pour l’autopsie


title: The 5′-Untranslated Region of Picornaviral Genomes

text: Picornaviruses are small naked icosahedral viruses with a single-stranded RNA genome of positive polarity. According to current taxonomy, the family includes four genera: Enterouirus (polioviruses, coxsackieviruses, echoviruses, and other enteroviruses), Rhinovirus, Curdiouirus [encephalomyocarditis virus (EMCV), mengovirus, Theiler's murine encephalomyelitis virus (TMEV)], and Aphthouirus [foot-and-mouth disease viruses (FMDV)]. There are also some, as yet, unclassified picornaviruses [e.g., hepatitis A virus (HAW] that should certainly be assessed as a separate genus. Studies on the molecular biology of picornaviruses might be divided into two periods: those before and after the first sequencing of the poliovirus genome. The 5'-untranslated region (5-UTR) of the viral genome was one of the unexpected problems. This segment proved to be immensely long: about 750 nucleotides or ∼10% of the genome length. There were also other unusual features (e.g., multiple AUG triplets preceding the single open reading frame (ORF) that encodes the viral polyprotein). This chapter shows that the picornaviral 5-UTRs are not only involved in such essential events as the synthesis of viral proteins and RNAs that could be expected to some extent, although some of the underlying mechanisms appeared to be quite a surprise, but also may determine diverse biological phenotypes from the plaque size or thermosensitivity of reproduction to attenuation of neurovirulence. Furthermore, a close inspection of the 5-UTR structure unravels certain hidden facets of the evolution of the picornaviral genome. Finally, the conclusions drawn from the experiments with the picornaviral5-UTRs provide important clues for understanding the functional capabilities of the eukaryotic ribosomes.


title: A core competency model for Chinese baccalaureate nursing graduates: A descriptive correlational study in Beijing

text: BACKGROUND: A review of the literature showed that the core competencies needed by newly graduated Chinese nurses were not as of yet undocumented. OBJECTIVE: To develop a psychometrically sound instrument for identifying and measuring the core competencies needed by Chinese nursing baccalaureate graduates. DESIGN: Descriptive correlational and multicentre study. SETTING: Seven major tertiary teaching hospitals and three major medical universities in Beijing. PARTICIPANTS: 790 subjects, including patients, nursing faculty members, doctors and nurses. METHOD: A reliable and valid self-report instrument, consisting of 58 items, was developed using multiple methods. It was then distributed to 790 subjects to measure nursing competency in a broader Chinese context. The psychometric characteristics of reliability and validity were supported by descriptive and inferential analyses. RESULTS: The final instrument consists of six dimensions with 47 items. The content validity index was 0.90. The overall scale reliability was 0.97 with dimensions range from 0.87 to 0.94. Six domains of core competencies were identified: professionalism; direct care; support and communication; application of professional knowledge; personal traits; and critical thinking and innovation. CONCLUSION: The findings of this study provide valuable evidence for a psychometrically sound measurement tool, as well as for competency-based nursing curriculum reform.


title: DNA immunization and central nervous system viral infection

text: This chapter discusses the virus infections of the central nervous system (CNS) and DNA vaccines. Mild central nervous system (CNS) symptoms, such as headache and drowsiness, can result from systemically elevated cytokine levels and therefore are common in many virus infections, even in the absence of the infection of the CNS. CNS infection is quite unusual and is initiated either as a result of the viremia or, more rarely, as a result of neural spread. The poliovirus infects the anterior horn motor neurons of the spinal cord, causing poliomyelitis, the disease for which the virus is named. DNA vaccination is a relatively new entrant in the vaccine sweepstakes, but is viewed with optimism, for a number of reasons. DNA vaccines encoding the nucleoprotein from lymphocytic choriomeningitis virus can confer protection against the normally lethal intracranial challenge. In rabies, in a mouse model, immunization with plasmids encoding the rabies glycoprotein conferred complete protection against subsequent viral challenge. Several virus-induced CNS diseases may be explained by their triggering of autoimmunity. Experimental autoimmune encephalomyelitis is a well-characterized CNS disease induced by the administration of certain CNS proteins.


SARS: Solución mediante Acciones y Respuestas Sensatas


title: Characterization of the Sendai virus V protein with an anti-peptide antiserum

text: The Sendai virus V protein, which is a fusion of the P and V ORFs of the P gene, was characterized with antisera to a portion of the V ORF and compared to the P protein. The only property found in common with P is that V is also highly phosphorylated, and this is so even when these proteins are expressed independently of the other viral proteins. Otherwise, V was not found in virions, was not strongly associated with viral nucleocapsids like P, and anti-V had no effect on viral RNA synthesis in vitro under conditions where anti-P was highly inhibitory. The available evidence suggests that V may playa role in RNA synthesis, but it is not an essential one like that of the P protein.


docid: ghzwqbhu

title: Infections in day-care centers


title: Characterization of proteins encoded by ORFs 2 to 7 of Lelystad virus

text: The genome of Lelystad virus (LV), a positive-strand RNA virus, is 15 kb in length and contains 8 open reading frames (ORFs) that encode putative viral proteins. ORFs 2 to 7 were cloned in plasmids downstream of the Sp6 RNA polymerase promoter, and the translation of transcripts generated in vitro yielded proteins that could be immunoprecipitated with porcine anti-LV serum. Synthetic polypeptides of 15 to 17 amino acids were selected from the amino acid sequences of ORFs 2 to 7 and antipeptide sera were raised in rabbits. Antisera that immunoprecipitated the in vitro translation products of ORFs 2 to 5 and 7 were obtained. Sera containing antibodies directed against peptides from ORFs 3 to 7 reacted positively with LV-infected alveolar lung macrophages in the immunoperoxidase monolayer assay. Using these antipeptide sera and porcine anti-LV serum, we identified three structural proteins and assigned their corresponding genes. Virions were found to contain a nucleocapsid protein of 15 kDa (N), an unglycosylated membrane protein of 18 kDa (M), and a glycosylated membrane protein of 25 kDa (E). The N protein is encoded by ORF7, the M protein is encoded by ORF6, and the E protein is encoded by ORF5. The E protein in virus particles contains one or two N-glycans that are resistant to endo-β-N-acetyl-d-glucosaminidase H. This finding indicates that the high-mannose glycans are processed into complex glycans in the Golgi compartment. The protein composition of the LV virions further confirms that LV is evolutionarily related to equine arteritis virus, simian hemorrhagic fever virus, and lactate dehydrogenase-elevating virus.


Antibiothérapie par voie générale en pratique courante dans les infections respiratoires hautes de l'adulte et l'enfant. Argumentaire


Los animales como vectores de las enfermedades emergentes


title: A prominent antigenic surface polypeptide involved in the biogenesis and function of the vaccinia virus envelope()

text: Polypeptides of the vaccinia virus envelope exposed on the surface were identified by means of sulfo-N-hydroxysuccinimidobiotin as a surface tag. Among surface expressed polypeptides is the 35-kDa antigen, previously designated Ag35. Both monoclonal (mAb) and monospecific affinity pure antibodies directed against Ag35 neutralized vaccinia infectiousness, indicating that this prominent surface antigen has a function during early virus-host cell interactions. The binding of several monoclonal antibodies to various regions of Ag35 was tested by reacting CNBr fragments, derived from the polypeptide, employing Western blotting. All mAbs tested reacted with the same region of Ag35. Estimation of the molecular weights (MW), based on migration of the CNBr peptides in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed that those partial digestion products which contained a proline-rich 99 amino acid limit digest fragment were present at a position approximately 12.5 kDa larger than that predicted from the DNA sequence. By contrast, partial and limit digest products lacking the proline-rich fragment migrated to the MW position expected from the length of the DNA sequence. This observation demonstrates that departure from a predicted 22.3 kDa to an anomalous MW of Ag35 is conferred by the proline-rich peptide. The surface location of Ag35 was confirmed by immune electron microscopy. In a competition test the binding specificity of mAb and affinity-purified antibodies at the surface of virions could be demonstrated. Evidence for an association of Ag35 with the virus envelope at various stages during biogenesis of vaccinia was obtained by immune electron microscopy of whole mounts and thin sections. Presence of Ag35 as an early component of immature and mature virions, probably residing in the bilayer membrane structure was detected. A distinction can, therefore, be made between Ag35 and several other vaccinia envelope polypeptides which are synthesized as late functions and added during late stages of envelope assembly.


Síndrome respiratorio agudo grave. Una nueva enfermedad infecciosa emergente


title: Virus Infection of Polarized Epithelial Cells

text: This chapter focuses on the interaction of viruses with epithelial cells. The role of specific pathways of virus entry and release in the pathogenesis of viral infection is examined together with the mechanisms utilized by viruses to circumvent the epithelial barrier. Polarized epithelial cells in culture, which can be grown on permeable supports, provide excellent systems for investigating the events in virus entry and release at the cellular level, and much information is being obtained using such systems. Much remains to be learned about the precise routes by which many viruses traverse the epithelial barrier to initiate their natural infection processes, although important information has been obtained in some systems. Another area of great interest for future investigation is the process of virus entry and release from other polarized cell types, including neuronal cells.


title: Central oxytocin increases food intake and daily weight gain in rats

text: The present study was performed to investigate the effects of centrally administered oxytocin on weight gain and food intake in rats. Two substrains of Sprague-Dawley rats (A and B) differing in average daily weight gain were used. Female rats of substrain A gained 2 g per day and males gained 7 g. Female rats of substrain B gained 5 g per day and males gained 8 g. Animals were implanted with a stainless steel guide cannula, allowing ICV injections into the lateral ventricle. ICV injections of 1, 5, or 10 μg of oxytocin or isotonic saline in a volume of 5 μl were given. In females, ICV treatment with either saline or 5 μg of oxytocin caused a transient loss of weight within 24 h of treatment. However, in the more slowly growing females of substrain A depression in body weight was observed after a single treatment with saline, whereas the body weight of oxytocin-treated females showed less marked depression and rapidly returned to the pretreatment weight. After a 3-day treatment period an even greater difference in daily weight gain was seen between oxytocin-treated and saline-treated female rats of substrain A. In contrast, no difference in daily weight gain or food intake was observed between oxytocin- and saline-treated male rats of substrain A, nor in females or males of the more rapidly growing substrain B. Intraperitoneal injections of 5 μg of oxytocin did not influence food intake or daily weight gain in female rats of substrain A. These data suggest that oxytocin may act centrally to influence food intake and daily weight gain in slowly growing female Sprague-Dawley rats.


Cumulative Chapter Titles Keyword Index, Volume 1 – 50


title: AIDS-Myelopathy: A Neuropathological Study

text: Vacuolar myelopathy belongs to the AIDS-associated diseases. It is characterized by vacuolation and infiltration of the long tracts of the spinal cord by macrophages. The clinical and morphological findings of 8 AIDS-patients with vacuolar myelopathy are reported here. The syndrome developed during the final stages of AIDS and was associated with HIV-encephalopathy in 5 cases. The vacuoles were mainly due to intramyelinic swelling and vacuolation. Vacuolated macrophages and axons contributed only to a minor degree. In one case only, HIV-antigens were detected immunohistochemically. The results are discussed in the light of modern pathogenetical concepts of HIV-related diseases.


title: Human brain evolution and the “Neuroevolutionary Time-depth Principle:” Implications for the Reclassification of fear-circuitry-related traits in DSM-V and for studying resilience to warzone-related posttraumatic stress disorder

text: The DSM-III, DSM-IV, DSM-IV-TR and ICD-10 have judiciously minimized discussion of etiologies to distance clinical psychiatry from Freudian psychoanalysis. With this goal mostly achieved, discussion of etiological factors should be reintroduced into the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-V). A research agenda for the DSM-V advocated the “development of a pathophysiologically based classification system”. The author critically reviews the neuroevolutionary literature on stress-induced and fear circuitry disorders and related amygdala-driven, species-atypical fear behaviors of clinical severity in adult humans. Over 30 empirically testable/falsifiable predictions are presented. It is noted that in DSM-IV-TR and ICD-10, the classification of stress and fear circuitry disorders is neither mode-of-acquisition-based nor brain-evolution-based. For example, snake phobia (innate) and dog phobia (overconsolidational) are clustered together. Similarly, research on blood-injection-injury-type-specific phobia clusters two fears different in their innateness: 1) an arguably ontogenetic memory-trace-overconsolidation-based fear (hospital phobia) and 2) a hardwired (innate) fear of the sight of one's blood or a sharp object penetrating one's skin. Genetic architecture-charting of fear-circuitry-related traits has been challenging. Various, non-phenotype-based architectures can serve as targets for research. In this article, the author will propose one such alternative genetic architecture. This article was inspired by the following: A) Nesse's “Smoke-Detector Principle”, B) the increasing suspicion that the “smooth” rather than “lumpy” distribution of complex psychiatric phenotypes (including fear-circuitry disorders) may in some cases be accounted for by oligogenic (and not necessarily polygenic) transmission, and C) insights from the initial sequence of the chimpanzee genome and comparison with the human genome by the Chimpanzee Sequencing and Analysis Consortium published in late 2005. Neuroevolutionary insights relevant to fear circuitry symptoms that primarily emerge overconsolidationally (especially Combat related Posttraumatic Stress Disorder) are presented. Also introduced is a human-evolution-based principle for clustering innate fear traits. The “Neuroevolutionary Time-depth Principle” of innate fears proposed in this article may be useful in the development of a neuroevolution-based taxonomic re-clustering of stress-triggered and fear-circuitry disorders in DSM-V. Four broad clusters of evolved fear circuits are proposed based on their time-depths: 1) Mesozoic (mammalian-wide) circuits hardwired by wild-type alleles driven to fixation by Mesozoic selective sweeps; 2) Cenozoic (simian-wide) circuits relevant to many specific phobias; 3) mid Paleolithic and upper Paleolithic (Homo sapiens-specific) circuits (arguably resulting mostly from mate-choice-driven stabilizing selection); 4) Neolithic circuits (arguably mostly related to stabilizing selection driven by gene–culture co-evolution). More importantly, the author presents evolutionary perspectives on warzone-related PTSD, Combat-Stress Reaction, Combat-related Stress, Operational-Stress, and other deployment-stress-induced symptoms. The Neuroevolutionary Time-depth Principle presented in this article may help explain the dissimilar stress-resilience levels following different types of acute threat to survival of oneself or one's progency (aka DSM-III and DSM-V PTSD Criterion-A events). PTSD rates following exposure to lethal inter-group violence (combat, warzone exposure or intentionally caused disasters such as terrorism) are usually 5–10 times higher than rates following large-scale natural disasters such as forest fires, floods, hurricanes, volcanic eruptions, and earthquakes. The author predicts that both intentionally-caused large-scale bioevent-disasters, as well as natural bioevents such as SARS and avian flu pandemics will be an exception and are likely to be followed by PTSD rates approaching those that follow warzone exposure. During bioevents, Amygdala-driven and locus-coeruleus-driven epidemic pseudosomatic symptoms may be an order of magnitude more common than infection-caused cytokine-driven symptoms. Implications for the red cross and FEMA are discussed. It is also argued that hospital phobia as well as dog phobia, bird phobia and bat phobia require re-taxonomization in DSM-V in a new “overconsolidational disorders” category anchored around PTSD. The overconsolidational spectrum category may be conceptualized as straddling the fear circuitry spectrum disorders and the affective spectrum disorders categories, and may be a category for which Pitman's secondary prevention propranolol regimen may be specifically indicated as a “morning after pill” intervention. Predictions are presented regarding obsessive-compulsive disorder (OCD) (e.g., female-pattern hoarding vs. male-pattern hoarding) and “culture-bound” acute anxiety symptoms (taijin-kyofusho, koro, shuk yang, shook yong, suo yang, rok-joo, jinjinia-bemar, karoshi, gwarosa, Voodoo death). Also discussed are insights relevant to pseudoneurological symptoms and to the forthcoming Dissociative-Conversive disorders category in DSM-V, including what the author terms fright-triggered acute pseudo-localized symptoms (i.e., pseudoparalysis, pseudocerebellar imbalance, psychogenic blindness, pseudoseizures, and epidemic sociogenic illness). Speculations based on studies of the human abnormal-spindle-like, microcephaly-associated (ASPM) gene, the microcephaly primary autosomal recessive (MCPH) gene, and the forkhead box p2 (FOXP2) gene are made and incorporated into what is termed “The pre-FOXP2 Hypothesis of Blood-Injection-Injury Phobia.” Finally, the author argues for a non-reductionistic fusion of “distal (evolutionary) neurobiology” with clinical “proximal neurobiology,” utilizing neurological heuristics. It is noted that the value of re-clustering fear traits based on behavioral ethology, human-phylogenomics-derived endophenotypes and on ontogenomics (gene–environment interactions) can be confirmed or disconfirmed using epidemiological or twin studies and psychiatric genomics.


SARS: una nueva enfermedad desenmascarada


title: Enterotoxigenic Escherichia coli Infections in Newborn Calves: A Review

text: Diarrhea caused by enterotoxigenic Escherichia coli is an infectious bacterial disease of calves that occurs during the first few days of life. The Escherichia coli that cause the disease possess special attributes of virulence that allow them to colonize the small intestine and produce an enterotoxin that causes hypersecretion of fluid into the intestinal lumen. These enterotoxigenic Escherichia coli are shed into the environment by infected animals in the herd and are ingested by newborn calves soon after birth. There is some natural immunity to enterotoxigenic Escherichia coli, however, it often fails to protect calves born and raised under modern husbandry conditions. Hence, methods have been developed to stimulate protective immunity by vaccination of the dam. The protective antibodies are transferred passively to calves through the colostrum.


title: Respiratory Viruses and Mycoplasma Pneumoniae Infections at the Time of the Acute Exacerbation of Chronic Otitis Media

text: The present study was undertaken to ascertain whether or not patients with chronic otitis media are infected with viruses or Mycoplasma at the time of sudden increase in otorrhea. From 26 patients with acute exacerbation of chronic otitis media, sera were collected at the time of sudden increase in otorrhea and three to four weeks later. These paired sera were examined for antibody titer to respiratory viruses (21 species) and Mycoplasma pneumoniae. Of them, influenza B virus and RSV infections were demonstrated in four and two cases, respectively. Examinations showed no infection in 10 control cases without acute exacerbation. In 36 cases of acute exacerbation of chronic otitis media, attempts were made to isolate viruses and Mycoplasma pneumoniae from the pharynx and otorrhea. Consequently, influenza B virus was detected in pharyngeal mucous scrapings in two cases and RSV in one. The probability of respiratory virus infection leading to acute exacerbation of chronic otitis media appears to be lower than that provoking acute otitis media in children and infants. However, the present data suggest that the development of respiratory virus infection in patients with chronic otitis media may cause an increase in the otorrhea, eventually resulting in an acute exacerbation of inflammation.


Hybridoma technology: new developments of practical interest


Compound name, code numcer and subject index, vol. 38


IChemE Special Issue on Climate Change


title: Treatment and Control of Neonatal Diarrhea in Calves

text: Treatment and control of acute neonatal diarrhea in calves are outlined and discussed. The difficulty in making a definitive etiological diagnosis makes effective treatment and control also difficult and largely empirical. Physiological events in calves with diarrhea are known, and fluid therapy is directed toward treating dehydration and acidosis. Whether affected calves should receive antibacterial agents orally is an open question. Principles of control of diarrhea in calves are outlined and discussed.


title: Biochemical and biophysical characterization of the reovirus cell attachment protein σ1: Evidence that it is a homotrimer

text: The oligomerization state of the reovirus cell attachment protein σ1 (49K monomeric molecular weight) was determined by biochemical and biophysical means. Full-length (protein product designated A) and C-terminal truncated (protein product designated B) serotype 3 reovirus Sl mRNA transcripts synthesizedin vitro were cotranslated in a rabbit reticulocyte lysate, and the products were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under conditions which allowed for the identification of oligomeric forms of σl. A total of four oligomeric protein bands (corresponding to A(3), A(2)B(1), A(1)B(2), and B3, respectively) was consistently observed, which suggests that the protein is made up of three monomeric subunits. Biophysical characterization of purified σ1 using column filtration and sucrose gradient sedimentation analysis confirmed the highly asymmetric shape of σ1 and allowed us to determine the molecular weight of the native protein to be ∼132K (a trimer). Similar biophysical analysis on the two tryptic fragments of the σ1 [N-terminal fibrous tail (26K monomeric molecular weight) and the C-terminal globular head (23K monomeric molecular weight)] yielded molecular weights of 77K and 64K, respectively, both again corresponding to trimers. We therefore conclude that protein σ1 is a homotrimer and provide, with supportive experimental evidence, a rationale for the anomalous behavior of the oligomeric protein in SDS-polyacrylamide gels, which, coupled with chemical cross-linking studies, has in part led to the previous suggestion that σ1 might be a higher order oligomer.


title: Rotavirus in infant-toddler day care centers: Epidemiology relevant to disease control strategies()

text: A 15-month prospective longitudinal study of diarrhea and rotavirus (RV) infection was conducted concurrently in infants and toddlers in day care centers (DCCs) and in a large pediatric clinic in Houston. The mean number of children in the DCCs was 223; the diarrhea rate during the first 12 months was 2.62 episodes per child-year. Rotavirus accounted for approximately 10% of the total episodes of diarrhea in the pediatric clinic and DCC populations, but 50% during the winter months. The occurrence of RV in the DCCs paralleled that seen in the pediatric clinic. The annual rate of RV infection in DCCs was 0.55 episodes per child-year, with diarrhea occurring in only 40% of the episodes (0.22 episodes per child-year). There were 45 diarrhea outbreaks in DCCs, for a mean of 3.8 per center per year; nine of these outbreaks were associated with RV. Polyacrylamide gel electrophoresis of RNA genome patterns of RV strains from eight of these outbreaks showed that in seven outbreaks a single strain was identified in children in that DCC, whereas multiple strains were identified simultaneously in the community. The age distributions of symptomatic and asymptomatic RV infections in DCC study children were not significantly different. In symptomatic RV-infected children in DCCs, 42% had RV identified in stool specimens within 2 days before diarrhea occurred. Thirty-eight DCC children had more than one episode of RV infection, but only five had two symptomatic RV infections. Diarrhea caused by RV is common in children in DCCs, often occurs in outbreaks due to the same strain, and parallels disease in the community; asymptomatic RV infection is also common in children in DCCs.


title: The TYMV tRNA-like structure

text: The genomic RNA from turnip yellow mosaic virus presents a 3′-end functionally and structurally related to tRNAs. This report summarizes our knowledge about the peculiar structure of the tRNA-like domain and its interaction with tRNA specific proteins, like RNAse P, tRNA nucleotidyl-transferase, aminoacyl-tRNA synthetases, and elongation factors. It discusses also the biological role of this structure in the viral life cycle. A brief survey of our knowledge of other tRNA mimicries in biological systems, as well as their relevance for understanding canonical tRNA, will also be presented.


Seasonal meanderings and novel virus strains


Epidemiología del daño pulmonar agudo y síndrome de distrés respiratorio agudo()


title: Antibiotics in feed, with special reference to pigs: a veterinary viewpoint

text: The historical background of the present UK legislation is briefly reviewed, and the current legal situation as it pertains to the UK and European Community is summarised. The reasons for the inclusion of antibiotics in feed are outlined and discussed. In the first case, feedstuffs may have to be medicated for reasons of disease prevention and control. In large herds, especially in loose housing, mass medication by injection is not possible and even minimal disease herds will become infected with airborne pathogens from time to time. In the second case, continuous low-level feeding of antibiotics to growing pigs (growth promoters) suppresses the growth of harmful bacteria and allows healthier pigs to grow faster and more efficiently, thus producing cheaper food at the same time as improving welfare. The legal requirements regarding medicines available only on prescription are outlined and discussed. The fears and worries of the general public are noted and found to be overstated because of misconceptions and misunderstanding. It is concluded that food and milk produced today will remain safe provided the safety rules are adhered to.


Neonatal necrotizing enterocolitis: Pathogenesis, classification, and spectrum of illness


Special edition editorial celebrating 25 years of publication


Volume 24, 1979 author and subject index


title: Hospital admission due to respiratory viral infections in moderate preterm, late preterm and term infants during their first year of life

text: BACKGROUND: Respiratory viral infections are a major cause of hospitalisation in infants <1 year and might cause severe symptoms in preterm infants. Our aim was to analyse admissions due to respiratory infections in moderate, late and term infants, and to identify risk factors for hospitalisation in preterm versus term. METHODS: Prospective study in a cohort of moderate and late preterm, and term infants born between October/2011 and December/2012. Admissions due to respiratory infections during the first year of life were analysed and compared among moderate (32–33), late (34–36) and term infants. Sixteen respiratory viruses were detected by RT-PCR. Clinical data were collected. RESULTS: 30 (20.9%) out of 143 preterm infants required admission for respiratory infection, versus 129 (6.9%) of 1858 term infants born in the same period (p < 0.0001, OR: 3.6 CI 2.0 to 5.0). Hospitalised children had a higher prevalence of hyaline membrane disease (HMD) at birth (p < 0.001, OR: 7.7 CI: 2.121 to 27.954) and needed more mechanical ventilation (p < 0.001, OR: 5.7 CI: 1.813 to 18.396). Virus was identified in 25/30 (83%) preterm babies, and in 110/129 (85%) term infants. The most frequent viruses in preterm infants were RSV (76%) rhinovirus (20%). Clinical and epidemiological characteristics among term and preterm infants were similar. CONCLUSIONS: The risk of respiratory admissions during the first year of life is up to 3.6 times higher in moderate and late preterm. Once admitted, clinical features of respiratory episodes requiring hospitalisation are similar among term and preterm infants. Hyaline membrane disease and mechanical ventilation were also risk factors for respiratory admissions.


Human Retrovirus and Multiple Sclerosis


title: CD4 rat × rat and mouse × rat T cell hybridomas produced by fusion of established T cell lines and clones to W/Fu (C58NT)D

text: Previously, fusion of established T cell lines or clones has been claimed to be difficult. We now report our experiences in the fusion of both long term cultures of rat T cell clones and mouse T cell lines to rat W/Fu (C58NT)D. Upon fusion of rat T cell clones the hybrids obtained expressed antigen specificities identical to those of the parent clones. In addition, C58 was used for interspecies hybridisation of murine T cell lines. The specificity of intra- and inter-species hybrids was maintained by subcloning. We conclude that the C58 cell line can be used to generate continuously growing monoclonal T-cell reagents of sufficient stability using both intra- and inter-species hybri disation.


title: Clinical manifestations of exacerbations of cystic fibrosis associated with nonbacterial infections

text: The purpose of this study was to determine whether acute pulmonary exacerbations of cystic fibrosis associated with nonbacterial infections are clinically distinguishable from other exacerbations. Eighty exacerbations in 54 patients were studied. Exacerbations associated with influenza (n=8) were compared with those associated with other nonbacterial infections (n=15) and those in which no nonbacterial infection was detected (n=57). Patients with influenza had lower Shwachman scores and were more likely to be seroposifive for C-reactive protein than patients in the other two groups. Patients with influenza had lower Shwachman scores and were more likely to be seropositive for C-reactive protein than patients in the other two groups. Patients with influenza had a mean decrease in forced expiratory volume per second of 26%, compared with test results obtained before the exacerbation. In contrast, the mean decrease in forced expiratory volume per second was 6% for other nonbacterial infections and 12% for the group without nonbacterial infection (p<0.05 for both comparisons). The forced expiratory flow in first 25% of vital capacity decreased 44% in the influenza group compared with 13% and 17% in the other two groups, respectively (p<0.01 for both comparisons). The influenza group also had a higher proportion of patients with at least a 20% decrease in forced expiratory volume per second and forced expiratory flow in first 25% of vital capacity than the other two groups had (p<0.05 for all comparisons). These data suggest that influenza is associated with severe exacerbations in patients with cystic fibrosis and support recommendations for efforts to prevent influenza in this population.


title: Characterization of defective-interfering RNAs of rubella virusgenerated during serial undiluted passage

text: During serial undiluted passage of rubella virus (RUB) in Vero cells, two species of defective-interfering (DI) RNAs of approximately 7000 and 800 nucleotides (nts) in length were generated (Frey, T. K., and Hemphill, M. L., Virology 164, 22–29, 1988). In this study, these DI RNAs were characterized by molecular cloning, hybridization with probes of defined sequence, and primer extension. The 7000-nt DI RNA species were found to be authentic DI RNAs which contain a single 2500- to 2700-nt deletion in the structural protein open reading frame (ORF) region of the genome. The 800-nt RNAs were found to be subgenomic DI RNAs synthesized from the large DI RNA templates. Analysis of the extent of the deletions using a reverse-transcription-PCR protocol revealed that the 3′ end of the deletions did not extend beyond the 3′ terminal 244 nts of the genome. The 5′ end of the deletions did not extend into the nonstructural protein ORF; however, DI RNAs in which the subgenomic start site was deleted were present. Following serial undiluted passage of seven independent stocks of RUB, this was the only pattern of DI RNAs generated. DI RNAs of 2000 to 3000 nt in length were the majority DI RNA species in a persistently infected line of Vero cells, showing that other types of RUB DI RNAs can be generated and selected. However, when supernatant from the persistently infected cells was passaged, the only DI RNAs present after two passages were 7000 nts in length, indicating that this species has a selective advantage over other types of DI RNAs during serial passage.


title: Recognition of a 170 kD protein in mammalian Golgi complexes by an antibody against malarial intraerythrocytic lamellae

text: Human erythrocytes infected with the malarial parasite Plasmodium falciparum contain flattened membrane lamellae. It has been suggested that the lamellae may be involved in the sorting of malarial proteins to the cytoplasm and the cell membrane of the host erythrocyte. We have previously shown that the lamellae accumulate sphingolipids by virtue of their lipid composition in a manner similar to the trans-Golgi and the trans-Golgi network in mammalian cells. In this paper, we show by immunofluorescence microscopy that a monoclonal antibody to the lamellae labelled a perinuclear organelle that colocalized with WGA and the mannose-6-phosphate receptor in cultured mammalian cells. Immunoelectron microscopy experiments revealed that LWLI labels cisternae of the trans-face and the trans-Golgi network. Western blot analysis of subcellular fractions using LWLI detected a 170 kD protein which is associated with the luminal side of Golgi membranes of rat liver and is conserved in all cell lines studied. Our results indicate that (i) the 170 kD protein is a novel marker of the mammalian trans-Golgi and the trans-Golgi network and (ii) in addition to similarities in their morphological and lipid characteristics, the lamellae induced by P. falciparum in erythrocytes share proteinaceous determinants with the Golgi apparatus of mammalian cells.


title: The Effect of Calfhood Diseases on Growth of Female Dairy Calves During the First 3 Months of Life in New York State

text: Our objective was to study the effects of pneumonia (cumulative incidence, 25%), diarrhea (29%), umbilical infection (14%), and umbilical hernia (15%) on BW and height gains during the first 3 mo of life. Female dairy calves (n = 410) born from January to December 1990 in 18 commercial herds in New York state were used. Average daily gains during the 1st, 2nd, and 3rd mo were 374, 596, and 719 g, respectively; average gain was 565 g during the 3-mo period. Average monthly height gains during the 1st, 2nd, and 3rd mo were 4.4, 5.6, and 5.7 cm, respectively. Use of multiple linear regression, with farms treated as random effects, indicated that treated, verified pneumonia was associated with a reduction in average daily gain of 66 g and that failure of passive transfer reduced average daily gain by 48 g during the 1st mo. During the 2nd mo, neither disease nor failure of passive transfer affected average daily gain. During the 3rd mo, each additional week of pneumonia reduced average daily gain by 14 g, and umbilical infection reduced average daily gain by 96 g. Each additional week of diagnosed pneumonia reduced total BW gain during the first 3 mo by 0.8 kg. Similarly, each week of pneumonia reduced total height gain by 0.2 cm and failure of passive transfer by 0.9 cm. Prevention of chronic pneumonia and umbilical infection may improve average daily gain of calves.


Pandemic influenza: learning from the present


title: Interferon induction in swine lymphocyte antigen-defined miniature pigs

text: Interferon was induced in two groups of swine lymphocyte antigen (SLA)-defined miniature pigs with polyinosinic: polycytidylic acid complexed with poly-L-lysine and carboxymethylcellulose. The group 1 pigs were low antibody-response phenotypes (SLA(a/a), SLA(a/c), SLA(c/c)), and the group 2 pigs were high antibody-response phenotypes (SLA(d/d), SLA(d/g), SLA(g/g)). Six hours after induction the antiviral titres were not influenced by the SLA group, but higher titres were observed in females. Higher antiviral titres were found in group 2 pigs before treatment and 24 hours after treatment, and higher titres were found in female pigs. The antiviral titres before and after treatment were also influenced by the sire. Group 2 pigs had lower total leucocyte counts before treatment, and there was a significant reduction in leucocyte numbers in both groups six hours after induction, due mainly to a large reduction in lymphocyte counts.


Re: Letter to the Editor of Public Health in response to 'Nipah virus infection: gaps in evidence and its public health importance'


title: Three-generation toxicity study of rats ingesting Brown HT in the diet

text: Brown HT was fed to rats of both sexes over three generations at dietary concentrations designed to provide daily intakes of 0, 50, 250 and 500 mg Brown HT/kg body weight/day. During the study a number of females died or failed to nurse their litters. This was so severe following the first mating of F(1) adults that the animals were remated to provide the next generation. None of these effects were related to treatment. Body weight and food and water intakes were not adversely affected by treatment. No effects of treatment were seen on reproductive performance or foetal and pup development, apart from slight evidence of a treatment-related retarded ossification of the third sternebrae. Organ weights at autopsy showed two changes, one of which was increased kidney weights which, although not present in every generation, seemed to be related to treatment. The other, increased caecum weights, occurred in adult high-dose females of early generations, but not in males or later generations of the study. Apart from brown coloration of tissues, macroscopic and microscopic examination revealed no treatment-related changes. It was concluded that the no-untoward-effect level in the present study was 250 mg Brown HT/kg/day.


title: Vaccines for preventing enterotoxigenic Escherichia coli infections in farm animals

text: Fimbrial vaccines are routinely given parenterally to pregnant cattle, sheep and swine to protect suckling newborn calves, lambs and pigs against enterotoxigenic Escherichia coli (ETEC) infections. Such vaccines are practical and effective because (1) most fatal ETEC infections in farm animals occur in the early neonatal period when the antibody titres in colostrum and milk are highest; (2) more than 90% of the ETEC in farm animals belong to a small family of fimbrial antigen types: (3) fimbriae consist of good protein antigens on the bacterial surface where they are readily accessible to antibody; (4) fimbriae are required for a critical step (adhesion-colonization) early in the pathogenesis of the disease. ETEC infections continue to be a significant clinical problem in farm animals in spite of extensive use of fimbriae-based vaccines. Definitive data on the efficacy of the commercial vaccines in field use are not available. The prevailing perception among animal health professionals is that the vaccines are effective, that the problem occurs chiefly among non-vaccinated animals, and that in some herds vaccination moves peak prevalence of disease from the first to the second or third week after birth, when mortality is lower. It has been suggested that extensive use of vaccines will rapidly select for the emergence of novel or previously low prevalence fimbrial antigen types. There is no evidence that this has happened after a decade of routine vaccine use in the United States. However, there is no active direct surveillance for such emergence. In contrast to the rational development of vaccines to provide passive lacteal protection against ETEC in suckling neonates, comparatively little progress has been made in providing the knowledge required for development of vaccines to protect against postweaning ETEC infections in swine.


title: Manure and Microbes: Public and Animal Health Problem?

text: Most environmental concerns about waste management either have focused on the effects of nutrients, especially N and P, on water quality or have emphasized odor problems and air quality. Microbes from manure are often low on the priority list for control and remediation, despite the fact that several out-breaks of gastroenteritis have been traced to livestock operations. The pathogens discussed in this paper include protozoans (Cryptosporidium parvum, Giardia spp.), bacteria (Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., and Mycobacterium paratuberculosis), and some enteric viruses. Clinical symptoms, prospects for zoonotic infection, and control methods other than the use of antimicrobials are considered. Recommendations to avoid disease transmission include taking steps to ensure the provision of clean, unstressful environments to reduce disease susceptibility and the careful handling and spreading of manure from animals at high risk for infection, especially young calves. Composting and drying of manure decrease the number of viable pathogens. Environmental controls, such as filter strips, also reduce the risk of water contamination.


title: Regulation of t cell responses during central nervous system viral infection

text: This chapter focuses on the contribution of T cells to the pathogenesis of neurologic disease and discusses specific examples of how individual T cell effector functions can be regulated during central nervous system's (CNS) viral infections. T cells can serve a variety of functions as part of the host immune response during CNS viral infection. They can participate directly in viral clearance from the brain, or they can promote the survival of the host without exerting any direct effect on virus replication. Only a small number of T cells infiltrate the brain under normal circumstances. This paucity of immune surveillance of baseline is one of several reasons why the CNS has often been characterized as an “immunologically privileged” site. T cell-mediated lysis of infected cells has been demonstrated to be an important mechanism of viral clearance from tissues other than the CNS. In several well-characterized animal models of CNS viral infection, part of the elicited T cell response actually contributes to the pathology and adverse outcome of disease. Neurotropic lymphocytic choriomeningitis virus infection of adult mice is the premier example of this phenomenon.


title: Diagnosis of Cryptosporidium on a sheep farm with neonatal diarrhea by immunofluorescence assays

text: An outbreak of diarrhea in neonatal lambs occurred on a a sheep farm in northern Ohio. Diarrhea commenced as early as 1 week of age and lasted for about 3–4 days. Although 100% of the newborn lambs were affected, most had recovered by 3 weeks of age. Cryptosporidium infection appeared to be the cause of diarrhea. Fecal examination of nine diarrheic newborn lambs (5–10 days old), 23 older lambs (2–3 weeks old, six with diarrhea) and 23 clinically normal ewes by immunofluorescence assays revealed infection rates of 100%, 78.3% and 17.4%, respectively. Most newborn lambs had high oocyst counts. Ewes were considered to be an important source of infection for lambs.


docid: waacoaeq

title: Influence of Sodium Bicarbonate on Growth and Health of Young Calves

text: Fifty-four Holstein and Jersey calves were assigned at 4 days of age within breed and sex to one of four treatments: control consisting of colostrum, milk replacer, and starter; buffered colostrum and replacer (.6% sodium bicarbonate) and starter (2% sodium bicarbonate); acidified colostrum (1% propionic), untreated replacer, and starter; and acidified, buffered colostrum (1% propionic, .6% sodium bicarbonate), buffered replacer (.6% sodium bicarbonate), and starter (2% sodium bicarbonate). The feeding regimen was colostrum once daily, day 4 to 14; milk replacer once daily, day 15 to 28; and calf starter ad libitum, day 4 to 84. Bull calves were fed for 42 days and heifers for 84 days. Calves fed acidified colostrum refused more feed and were less efficient from day 4 to 14 than calves fed buffered colostrum. Bulls were more sensitive to acidified colostrum than heifers. Starter intake, total dry matter intake, and average daily gains were similar for all calves during days 4 to 84. Rumen fluid from calves fed diets with sodium bicarbonate was higher in acetate and lower in propionate and lactate than that from calves fed diets without sodium bicarbonate. Sodium bicarbonate improved intake of acidified colostrum during the first 2 or 3 days of feeding but had no other effect on gain or feed intake.


title: Virus-Induced Immunopathology

text: Several viruses cause damage to the tissue by immunopathological mechanisms. This chapter presents the principal examples of immunopathogenesis caused by the viruses, accompanied by speculations about its management. The most common mechanism of lesion development in virus induced immunopathology involves T cells. Usually, it seems that when CD8(+) T cells act as the controlling cell type, lesions are acute and the outcome is decided quickly. The classic example is provided by LCM in mice. The newest candidate may turn out to be SNV infection in humans. Lesions orchestrated primarily by CD4(+) T cells can be either acute or long-lasting. Curiously, in the LCMV example, if CD8(+) T cells are removed from the scene, immunopathological responses may still occur and these involve CD4(+) T cells. Such responses are far more chronic and of lower grade than those mediated by CD8(+) T lymphocytes. One possible sequel to chronic inflammatory responses to viruses is that autoreactive inflammatory reactions are initiated and an autoimmune disease occurs. The adage that an ounce of prevention is worth a pound of cure is certainly true in the field of viral pathogenesis. Preventing viral infection or manipulating immune processes during the initial phases of infection is far more successful than attempting to counteract pathological events once underway.


Cardiopulmonary Resuscitation for Ebola patients: Ethical considerations


title: Proteolysis and antigen presentation by MHC class II molecules

text: Proteolysis is the primary mechanism used by all cells not only to dispose of unwanted proteins but also to regulate protein function and maintain cellular homeostasis. Proteases that reside in the endocytic pathway are the principal actors of terminal protein degradation. The proteases contained in the endocytic pathway are classified into four major groups based on the active-site amino acid used by the enzyme to hydrolyze amide bonds of proteins: cysteine, aspartyl, serine, and metalloproteases. The presentation of peptide antigens by major histocompatibility complex (MHC) class II molecules is strictly dependent on the action of proteases. Class II molecules scour the endocytic pathway for antigenic peptides to bind and present at the cell surface for recognition by CD4(+) T cells. The specialized cell types that support antigen presentation by class II molecules are commonly referred to as professional antigen presenting cells (APCs), which include bone marrow-derived B lymphocytes, dendritic cells (DCs), and macrophages. In addition, the expression of certain endocytic proteases is regulated either at the level of gene transcription or enzyme maturation and their activity is controlled by the presence of endogenous protease inhibitors.


title: Virus Entry into Animal Cells

text: In addition to its many other functions, the plasma membrane of eukaryotic cells serves as a barrier against invading parasites and viruses. It is not permeable to ions and to low molecular weight solutes, let alone to proteins and polynucleotides. Yet it is clear that viruses are capable of transferring their genome and accessory proteins into the cytosol or into the nucleus, and thus infect the cell. While the detailed mechanisms remain unclear for most animal viruses, a general theme is apparent like other stages in the replication cycle; their entry depends on the activities of the host cell. In order to take up nutrients, to communicate with other cells, to control the intracellular ion balance, and to secrete substances, cells have a variety of mechanisms for bypassing and modifying the barrier properties imposed by their plasma membrane. It is these mechanisms, and the molecules involved in them, that viruses exploit.


Lenguaje médico: el rayo neológico que no cesa


title: Monoclonal antibodies against bovine immunoglobulins and their use in isotype-specific ELISAs for rotavirus antibody

text: Monoclonal antibodies (MCA) against bovine immunoglobulin (BIg) isotypes were produced and characterized. MCAs were obtained which react specifically with IgG, IgG(1), IgG(2) or IgA while MCAs against IgM showed a partial cross-reaction with affinity purified IgA. MCAs with optimal characteristics for application in ELISA were selected and used as conjugates in an indirect double antibody sandwich assay (IDAS) and as the capturing antibody in an antibody capture assay (ACA) for the isotype-specific detection of antibodies against rotavirus. Based on theoretical grounds, experimental analysis of inter- and intra-isotype competition in IDAS and ACA, respectively, and direct comparison of both tests, the IDAS was selected for the detection of IgG(1) and IgG(2) anti-rotavirus antibodies. The ACA was the test of choice for the detection of IgM and IgA anti-rotavirus antibodies. The isotype specificity of these tests relies on the specificity of the MCAs and was confirmed for each test by the observation that samples containing rotavirus antibodies of only 1 particular isotypee reacted only in the homologous assay. The MCAs against bovine Ig isotypes and isotype-specific ELISAs were found to be very useful in the study of humoral mucosal immunity in calves infected with rotavirus.


Penicillium marneffei: penicilliosis and the red peril in the east


title: Immunosorbent Electron Microscopy For Detection Of Viruses

text: This chapter discusses the newer modifications of immunosorbent electron microscopy (ISEM) methods in both plant and animal virology. ISEM methods presented in the chapter include all the techniques where the “solid phase principle” is essential in a way similar to other solid phase immunoassays. These methods include (1) the antibody-coated grid technique (AB-CGT); (2) the protein A-coated grid technique (PA-CGT); (3) the protein A-coated bacteria technique (PA-CBT); and (4) the antigen-coated grid technique (AG-CGT). In all ISEM methods, one of the components of the system is adsorbed to a solid phase. In AG-CGT, PA-CGT, and AB-CGT, one of the reagents is adsorbed to an electron microscopic grid, while in PA-CBT protein A is naturally present on the surface of a bacterium that serves as a solid support. In ISEM methods, the viruses can be statistically evaluated and numerically expressed as number of virions per unit of area, and can, therefore, be statistically evaluated. Thus, these methods optimize the results of a test by quantifying the effects of the quality of the supporting grid, the time of adsorption, the pH, the presence of salts, and the type of staining. The ISEM also permits a detailed study of antigenic variations in the same genus of virus, and thus would visually pinpoint the type or strain differences.


title: Nucleotide sequence of the bean strain of southern bean mosaic virus()

text: The genome of the bean strain of southern bean mosaic virus (SBMV-B) comprises 4109 nucleotides and thus is slightlyshorter than those of the two other sequenced sobemoviruses (southern bean mosaic virus, cowpea strain (SBMV-C) and rice yellow mottle virus (RYMV)). SBMV-B has an overall sequence similarity with SBMV-C of 55% and with RYMV of 45%. Three potential open reading frames (ORFs) were recognized in SBMV-B which were in similar positions in the genomes of SBMV-C and RYMV. However, there was no analog of SBMV-C and RYMV ORF 3. From a comparison of the predicted sequences of the ORFs of these three sobemoviruses and of the noncoding regions, it is suggested that the two SBMV strains differ from one another as much as they do from RYMV and that they should be considered as different viruses.


Mini‐Symposium Severe acute respiratory syndrome (SARS): Reshaping global public health


title: Mouse hepatitis virus type 3 infection provokes a decrease in the number of sinusoidal endothelial cell fenestrae both in vivo and in vitro

text: Fenestrations of hepatic endothelial cells play an active role as a sieving barrier allowing extensive exchange between the blood and liver parenchyma. Alteration of these structures may be induced in the course of various pathological events and provoke important perturbations of liver function. We demonstrate here that sinusoidal endothelial cells are permissive for mouse hepatitis virus 3 (MHV3) in vivo and in vitro and that this infection leads to a striking decrease in the number of fenestrae. The disappearance of these structures observed under scanning electron microscopy or in cryofracture preparations in vivo and in vitro cannot be reversed by the action of cytochalasin B on the microfilament network. The decrease in the porosity seems to be related directly to the productive infection of the endothelial cells, because it was not observed in A/J mice resistant to the virus and in susceptible BALB/c mice immunized with a thermosensitive mutant in which no viral replication occurs. In conclusion, a viral infection of liver endothelial cells may cause extensive loss of the fenestrations and thus lead to important functional pertubations.


title: Pathology and toxicology of beluga whales from the St. Lawrence Estuary, Quebec, Canada. Past, present and future

text: An indigenous population of 450–500 beluga whales (Delphinapterus leucas) inhabiting the St. Lawrence Estuary has been exposed chronically for more than 50 years to a complex mixture of industrial pollutants including organochlorinated compounds (OC), polycyclic aromatic hydrocarbons (PAH) and heavy metals. From 1983 to 1990, we have necropsied 45 well preserved carcasses out of a total of 120 beluga whales reported dead over this period. Of these 45 animals, nine were affected by 10 malignant neoplasms. Fifteen animals (33%) were affected by pneumonia. Milk production was compromised in eight of 17 mature females (41%), by inflammatory changes (seven animals) and cancer (one animal) which affected the mammary glands. Opportunistic bacteria were found in pure culture, and/or in significant amounts in at least two organs in 20 belugas (44%). The concentrations of both total PCBs and highly chlorinated PCB congeners were much higher in St. Lawrence animals than in Arctic beluga whales. OC-induced immunosuppression has been repeatedly demonstrated in a wide variety of animal species. Therefore, it is probable that the immune functions of St. Lawrence beluga whales are impaired. Benzo[α]pyrene adducts were detected in 10 of the 11 St. Lawrence beluga whales of which tissues (six livers, 10/11 brains) were analyzed by a method based on HPLC. No such adducts were found in four Arctic animals. Since benzo[α]pyrene is one of the most potent chemical carcinogens known to man, these compounds might be responsible for some of the cancers observed in that population. Overall, our findings contrast vividly with those of others who found that cancers are exceedingly rare in free-ranging odontocete populations and that the major causes for mortalities in these populations are bacteria, parasites, and trauma.


title: Prevalence of Cryptosporidium and Giardia infections in cattle in Aragón (northeastern Spain)

text: Faecal samples from 554 bovines randomly selected at 30 farms in Aragón were examined to investigate the prevalence of Cryptosporidium and Giardia infections. C. parvum oocysts were identified by using the Ziehl-Neelsen modified technique in 109 (19.7%) bovines ranging from 3 days old to adults. Positive animals were found in 19 (63.3%) farms. As much as 44.4% of calves aged 3–4 days were infected, but infection rates peaked at 6–15 days of age (76.7%). Nevertheless, prevalence was also high in weaning calves aged 1.5–4 months (14%), fattening calves and heifers 4–24 months old (7.7%) and adults (17.8%). Diarrhoea was recorded in 78.6% of suckling and 29.4% of weanling calves infected by C. parvum, but it was only found to be statistically associated with infection in suckling calves (P < 0.01). All calves shedding moderate or many oocysts had diarrhoea, whereas asymptomatic infection was always correlated with few oocysts in faeces. Cryptosporidial infections were always asymptomatic in bovines older than 4 months. Giardia cysts were identified in 65 bovines (11.7%) from 16 (53.3%) of the farms surveyed. Infection rates were significantly higher in suckling (14.1%) and weanling calves (38%) than in bovines older than 4 months (2.2%) (P < 0.001). Diarrhoea was recorded in 45.5% of suckling and 10.9% of weanling calves infected by Giardia, but it was not found to be statistically associated with infection. In fact, infection rates were higher in non-diarrhoeic than in diarrhoeic calves.


Síndrome de distrés respiratorio agudo. Conclusiones y perspectivas de futuro()


title: Mouse hepatitis virus ORF 2a is expressed in the cytosol of infected mouse fibroblasts

text: A 266-bp fragment of cDNA from within gene B, ORF 2a, of MHV-A59 was used to construct a vector encoding a bacterial/viral fusion protein. Antiserum raised against this fusion protein specifically immunoprecipitates a 30K protein from infected 17CI-1 mouse fibroblasts. The protein is localized primarily in the cytosol and not in the membranes. This is consistent with its predicted sequence and potential role as an RNA binding protein.


title: Syndemics, sex and the city: Understanding sexually transmitted diseases in social and cultural context

text: This paper employs syndemics theory to explain high rates of sexually transmitted disease among inner city African American and Puerto Rican heterosexual young adults in Hartford, CT, USA. Syndemic theory helps to elucidate the tendency for multiple co-terminus and interacting epidemics to develop under conditions of health and social disparity. Based on enhanced focus group and in-depth interview data, the paper argues that respondents employed a cultural logic of risk assessment which put them at high risk for STD infection. This cultural logic was shaped by their experiences of growing up in the inner city which included: coming of age in an impoverished family, living in a broken home, experiencing domestic violence, limited expectations of the future, limited exposure to positive role models, lack of expectation of the dependency of others, and fear of intimacy.


Global health is our health


title: Mapping of Linear Antigenic Sites on the S Glycoprotein of a Neurotropic Murine Coronavirus with Synthetic Peptides: A Combination of Nine Prediction Algorithms Fails To Identify Relevant Epitopes and Peptide Immunogenicity Is Drastically Influenced by the Nature of the Protein Carrier

text: The elucidation of the antigenic structure of the S glycoprotein of murine coronaviruses will provide further understanding of the Complex pathogenicity of these viruses. In order to identify linear antigenic determinants, the primary structure of the S glycoprotein of murine hepatitis virus strain A59 was analyzed with a combination of nine epitope prediction algorithms. Fifteen potential epitopes were synthesized chemically and injected into BALB/c mice to study their biological relevance. This approach failed to identify novel important epitopes. Furthermore, the algorithms were unable to identify as antigenic the previously mapped immunodominant epitope A [C Daniel, R. Anderson, M. J. Buchmeier, J. O. Fleming, W. J. M. Spaan, H. Wege, and Talbot, P. J. (1993) J. Virol. 67, 1185-1194]. Interestingly, peptide A coupled to KLH induced an immune response that stimulated the immune response induced by the corresponding region of the protein much more accurately than when the same peptide was coupled to BSA. This included drastically enhanced competition with monoclonal antibodies and protection from virus challenge. These findings emphasize the shortcomings of amino acid sequence-based epitope prediction algorithms and demonstrate the critical importance of the carrier when synthetic peptides are considered as potential vaccines.


title: An outbreak of rotavirus-associated neonatal necrotizing enterocolitis()

text: An outbreak of necrotizing enterocolitis and hemorrhagic gastroenteritis occurred in two nurseries during 25 days in August 1982. Eleven of the 40 patients in these nurseries during that time developed disease (attack rate 27.5%). In seven of the 10 patients with gastrointestinal disease, stool samples tested for human rotavirus were positive by ELISA, whereas in 20 unaffected infants, no stools tested demonstrated HRV (P=0.0001). Eleven staff members had serologic evidence of recent HRV infection. Comparison of risk factors traditionally associated with the development of NEC between the affected and unaffected infants revealed no significant differences. Rotavirus infection was the only finding that was highly correlated with this epidemic.


title: A simplified procedure for studies of intestinal immunity in rabbits

text: As interest in the development of oral vaccines continues to rise, alternative animal models for studies of mucosal immunity are needed. The present study examines a simplified procedure for delivering antigen to rabbit Peyer's patches via an indwelling cannula. The cannula was placed 3–4 cm proximal to the Peyer's patch, and was used to deliver four weekly doses of the potent mucosal immunogen, cholera toxin (CT). Anti-CT specific fecal secretory IgA (S-IgA), serum IgG and serum IgA were found in essentially equal amounts in rabbits with cannulas and in rabbits fitted with Thiry-Vella (T-V) isolated ileal loops. In contrast to animals with T-V loops, the intestinal flora of animals with cannulas contained less bacterial overgrowth with Pseudomonas sp. Further, the villus architecture remained histologically normal in appearance and there were fewer post-surgical complications associated with this technique than with T-V loops. This simplified technique should allow wider use of rabbits in studies of mucosal immunity.


title: A nose-brain pathway for psychotropic peptides: evidence from a brain evoked potential study with cholecystokinin

text: The access of substances to the brain is of particular relevance for the etiology and treatment of psychiatric and neurologic diseases. This study provides functional evidence for a direct access of peptides to the human brain after intranasal administration. Effects were compared of intranasal (IN, 10 μg) and intravenous (IV, 0.25 and 2.5 μg) administered cholecystokinin-8 (CCK) on the auditory event related potential (AERP) in 20 healthy subjects. Also, plasma concentration of cortisol and ACTH were monitored. The study was designed as a placebo-controlled, double-blind within-subject cross-over comparison. AERPs were recorded while the subject performed on an attention task (oddball task). Plasma CCK concentrations after IN administration of CCK were comparable to those after IV administration of 0.25 μg CCK, but were substantially lower than those after 2.5 μg CCK. The P3 complex of the AERP was markedly increased following the IN administration of CCK (p < .01) compared to placebo and to the IV administration of 0.25 μg This pattern was more obvious in women than men. Increases in plasma ACTH concentrations after CCK reached significance selectively following the IN mode of administration (p < .01).


title: Early interactions between animal viruses and the host cell: relevance to viral vaccines

text: Viral recognition of specific receptors in the host cell plasma membrane is the first step in virus infection. Attachment is followed by a redistribution or capping of virus particles on the cell surface which may play a role in the uptake process. Certain viruses penetrate the plasma membrane directly but many, both enveloped and non-enveloped viruses, are endocytosed at coated pits and subsequently pass into endosomes. The low pH environment of the endosome facilitates passage of the viral genome into the cytoplasm. For some viruses the mechanism of membrane penetration is now known to be linked to a pH-mediated conformational change in external virion proteins. As a consequence of infection there are alterations in the permeability of the plasma membrane which may contribute to cellular damage. Recent advances in the understanding of these processes are reviewed and their relevance to the development of new strategies for vaccines emphasised.


Index des mots-clés


Compound Name, Code Number and Subject Index. Vol. 42


title: Passive immunity to bovine rotavirus in newborn calves fed colostrum supplements from cows immunized with recombinant SA11 rotavirus core-like particle (CLP) or virus-like particle (VLP) vaccines

text: Heterotypic passive immunity to IND (P[5]G6) bovine rotavirus (BRV) was evaluated. Three groups of calves (n = 5 per group) were fed 1% pooled colostrum supplements (birth to 7 days of age) from BRV seropositive cows vaccinated with recombinant SA11(P[2]G3) rotavirus-like particles (VLPs), recombinant SA11 rotavirus core-like particles (CLPs), or inactivated SA11 rotavirus (SA11). Control calves (n = 5 per group) received either pooled colostrum from unvaccinated (BRV field exposure seropositive) control cows, or no colostrum. IgG1 antibody titers to IND BRV for the pooled colostrum were: 1048576 (VLP); 1048576 (CLP); 262144 (SA11); and 16384 (control colostrum). Elevated titers of BRV neutralizing (VN) antibodies were present in VLP colostrum (98 000), and SA11 colostrum (25 000), but not in CLP colostrum (1400), compared to colostrum from nonvaccinates (2081). Calves were orally inoculated with virulent IND BRV at 2 days of age and challenged at post-inoculation day (PID) 21. Calves were monitored daily for diarrhea and faecal BRV shedding through PID 10 and post-challenge day (PCD) 10. After colostrum feeding, the IgG1 antibody titers were highest in serum and faeces of calves fed VLP and CLP colostrum, but VN and IgA antibodies were highest in calves fed VLP colostrum. After BRV inoculation, calves fed colostrum from vaccinated cows had significantly fewer days of BRV-associated diarrhea and BRV shedding than control calves. All calves fed VLP colostrum were protected from diarrhea after BRV inoculation; two calves shed BRV. In the CLP colostrum group, one calf developed BRV-associated diarrhea and all calves shed virus. In the SA11 colostrum group, three calves developed BRV-associated diarrhea and four calves shed virus. BRV-associated diarrhea and shedding occurred in 9 of 10 control calves. Active IgM antibody responses occurred in faeces and/or serum of most calves after BRV inoculation. However, the highest active antibody responses (IgM and IgG1 in serum, and IgM, IgG1 or IgA in faeces) after BRV inoculation were in calves fed control or no colostrum, in association with clinical diarrhea in most of these calves. After challenge at PID 21, BRV-associated diarrhea and shedding were of short duration or absent, in all groups. These results demonstrate the efficacy of colostrum from VLP vaccinated cows to provide heterologous, passive protection against BRV diarrhea and shedding in calves. In comparison, calves fed CLP or SA11 colostrum were only partially protected against BRV diarrhea or shedding.


Cumulative Chapter Titles Keyword Index, Volume 1–45


title: Evaluation of free or liposome-encapsulated ribavirin for antiviral therapy of experimentally induced feline infectious peritonitis

text: Ribavirin, either free in aqueous solution or incorporated into liposomes, was evaluated in 50 specific-pathogen-free kittens after experimental challenge exposure with feline infectious peritonitis virus (fipv). Ribavirin was administered daily for 10 to 14 days at 16·5 mg kg(−1) bodyweight given per os, intramuscularly or intravenously beginning 18 hours after kittens were challenge-exposed with fipv. All kittens, including ribavirin-treated and untreated kittens, succumbed to FIP. Clinical signs of disease were more severe in the ribavirin-treated kittens and their mean survival times were shortened. The clinical efficacy of free ribavirin given intravenously at a reduced dosage (5·5 mg kg(−1) bodyweight) was compared to that of ribavirin incorporated into lecithin-containing liposomes (5 mg kg(−1)) intravenously. Drugs were given once daily for three consecutive days of each week for three weeks, beginning 18 hours after virus challenge exposure. There was no significant difference either in survival rate or severity of disease between kittens given free ribavirin, liposomal ribavirin or saline only. Because of its intrinsic toxicity and low therapeutic index against fipv and its marginal antiviral activities in vivo at maximal doses, ribavirin cannot presently be recommended as primary antiviral chemotherapy against fip.


title: Identification of a sorting signal for the regulated secretory pathway at the N-terminus of pro-opiomelanocortin

text: The N-terminal 26 amino acids of the prohormone pro-opiomelanocortin (POMC) were investigated to determine whether this region has the capacity to act as a sorting signal for the regulated secretory pathway. Constructs were made using the N-terminal 101, 50, 26 or 10 amino acids of POMC fused to the chloramphenicol acetyltransferase (CAT) reporter protein and expressed in AtT20 cells to show that at least the first 26 amino acids were required to sort CAT to the regulated secretory pathway. Full length POMC was mutated by deleting amino acids 2–26 from the N-terminal region. Analysis of Neuro-2a cells expressing this mutation compared to wild type POMC indicated that these 26 amino acids contain information essential for sorting POMC to the regulated secretory pathway. The results presented here suggest the presence of a conformation-dependent signal in the N-terminal 26 amino acids of POMC responsible for sorting POMC to the regulated secretory pathway.


title: A new pharyngitis model using capsaicin in rats

text: 1. 1. Application of capsaicin solution onto the rat pharyngeal mucosa caused a well-reproducible increase in vascular permeability in the pharynx. 2. 2. Capsaicin-induced pharyngeal inflammation was unaffected by a histamine H(1) blocker and non-steroidal anti-inflammatory agents, whereas dexamethasone was effective in its inhibition. 3. 3. FK224, a dual antagonist of tachykinin NK(1) and NK(2) receptors, and FK888, a selective antagonist of NK(1) receptor, significantly inhibited capsaicin-induced plasma exudation in the pharynx. 4. 4. In capsaicinized animals, the application of capsaicin solution in the pharyngeal mucosa did not induce pharyngitis. 5. 5. These results suggest that the mechanism of the capsaicin-induced pharyngitis primarily involves tachykinins.


Introduction to issue of highlighted research presented at the 2015 National Foundation for Infectious Diseases Annual Conference on Vaccine Research


title: Necrotizing Enterocolitis: Treatment Based on Staging Criteria

text: Neonatal necrotizing enterocolitis is the most important cause of acquired gastrointestinal morbidity or mortality among low birthweight infants. Prematurity alone is probably the only identifiable risk factor. Although the etiology is unknown NEC has many similarities to an infectious disease. Proper staging helps improve reporting and the management of NEC.