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PubMed1999

Impact of the introduction of A/Sydney/5/97 H3N2 influenza virus into South Africa.

In 1998 South Africa experienced a major influenza epidemic that was characterized by extensive illness and an unusually early season. The impact of the epidemic was charted by measuring proxy indexes of influenza activity such as school absenteeism and excess mortality in persons older than 65 years. Viruses isolated from patients of all age groups were analyzed both antigenically and at the molecular level to determine the characteristics of the influenza strain responsible for the outbreaks. The study revealed that influenza activity was detected as early as the middle of April and peaked toward the end of May and early June. School absenteeism correlated with a sharp rise in virus isolation during this period. Consumption of influenza-related pharmaceuticals, as well as mortality figures, also corresponded to the increased absenteeism and virus isolation. Characterization of the viruses isolated during 1997 and 1998 showed clearly that the epidemic was caused by the introduction of the A/Sydney/5/97-like H3N2 influenza strain into South Africa in 1998. With no prior exposure to this virus strain, which is antigenically distinct from the viruses that had been present in this country in 1997, the population was highly susceptible, resulting in an early, rapid spread of influenza. This epidemic has highlighted the importance of having an influenza vaccine specifically formulated for the Southern Hemisphere. If the 1998 vaccine had not contained the A/Sydney/5/97 strain, the widespread outbreaks in South Africa would have been far worse in terms of morbidity, mortality, and economic loss. This, in turn, emphasizes the need for increased influenza surveillance and international cooperation.

Absenteeism↗

PubMed2006

Automated screening using microfluidic chip-based PCR and product detection to assess risk of BK virus-associated nephropathy in renal transplant recipients.

The cost-effective detection of viral particles in bodily fluids could enable more effective responses to viral outbreaks, whether isolated clinical cases, or influenza epidemics. In renal transplant recipients, complications arising from high levels of BK virus can lead to graft dysfunction, graft loss, and/or reduced patient survival. We describe a microfluidic system for the sensitive analysis of BK virus (viral load) in unprocessed urine samples that are applied directly onto the chip, thus avoiding labor-intensive processing and sources of inter-assay variability. Integration of small volume genetic amplification (PCR) and electrophoretic analysis detects as few as 1-2 viral copies, distinguishes between high, medium and low levels of virus and reliably identifies viral loads requiring clinical intervention. As a first step to wider application in the clinic and in the field, the present work presents an entirely microchip-based system, validated against conventional clinical methods using clinical samples.

BK Virus↗

PubMed1994

Occurrence of domoic acid in Washington state razor clams (Siliqua patula) during 1991-1993.

The presence of domoic acid in aquatic species was reported for the first time in the United States in the late summer of 1991 in Monterey Bay, California. By October of 1991, domoic acid was found in razor clams (Siliqua patula) and in the viscera of Dungeness crab (Cancer magister) along the coasts of Washington and Oregon. In response to this outbreak, the National Marine Fisheries Service, in cooperation with the Washington State Department of Fisheries began analysis of Washington State razor clams for the period from November 1991 to June 1993. This survey indicated that domoic acid levels in the edible portion of the razor clams peaked in December of 1991 (average of all Washington state coastal sites: 106 ppm) and followed a slow decline to the present day low levels (< 5 ppm). Sixteen months after the maximum level, domoic acid has not completely disappeared from the razor clams from the Washington State beaches. Unlike mussels (Mytilus edulis), where the toxin is found only in the viscera, domoic acid distributes itself throughout the various body parts of the razor clam. The highest concentration occurs in the foot or "digger" and the lowest in the siphon or "neck." The concentration of domoic acid in the razor clam foot reached a high of 230 ppm.

Animals↗

PubMed1993

Baculovirus expression of the ADRV gene 5 encoded protein produces an oligomerized, antigenic, and immunogenic VP6 protein.

Adult diarrheal rotavirus (ADRV) is a currently noncultivatable group B human rotavirus responsible for epidemic outbreaks of gastroenteritis in China. Gene segment 5 of ADRV encodes the major inner capsid protein, VP6. ADRV gene 5 was inserted into a recombinant baculovirus by homologous recombination between baculovirus shuttle plasmid pACYM1-AD5 and AcNPV genomic DNA. Baculovirus recombinants were selected visually and plaque purified and VP6 expression was detected by Coomassie staining of PAGE-separated proteins. The baculovirus-expressed gene 5 polypeptide is 44 kDa, the same as for the major inner capsid protein present on EDTA-treated ADRV virions and in vitro-expressed VP6 protein. The expressed protein is oligomeric and in the absence of reducing agents multimerizes to apparent trimer, hexamer, and greater molecular mass as assayed by SDS-PAGE. The VP6 protein is immunoprecipitable by hyperimmune serum to ADRV, human ADRV convalescent serum, by a group B-specific monoclonal antibody and by porcine group B rotavirus infection serum. The baculovirus-expressed protein is immunogenic and antibodies to the expressed protein recognize ADRV virions. The ADRV VP6 protein should be useful for developing diagnostic assays for serum antibodies to group B rotavirus as well as for generating hyperimmune serum and monoclonal antibodies for detecting viral antigen from ADRV and other group B rotaviruses.

Antibodies, Monoclonal↗

PubMed2002

Bidirectional infection and release of Rift Valley fever virus in polarized epithelial cells.

Rift Valley Fever (RVF) virus is an arbovirus and is responsible for large outbreaks of disease predominantly in sub-Saharan Africa. However, several aspects of RVF virus transmission, such as high viremia, multiple vector species, and broad host range, result in a pathogen with high likelihood of geographic spread. RVF virus infection in humans and livestock is characterized by broad dissemination of RVF virus antigens throughout the body. We sought insight into the high pathogenicity and broad tropism of this virus through a characterization of its interaction with polarized epithelial cells. Our results indicate that infection and release of RVF virus in polarized epithelial cells occurs at both apical and basolateral membranes and hence is bidirectional. Furthermore, our results indicate that RVF virus causes disruptions in both the microfilament and the microtubule networks. These disruptions may provide a mechanism for bidirectional release of RVF virions.

Actin Cytoskeleton↗

PubMed1999

Synthesis and formation of an EMS correlated contaminant in biotechnologically manufactured L-tryptophan.

Contaminants in biotechnologically manufactured L-Tryptophan (Trp) are suspected to be responsible for the outbreak of an unknown autoimmune disease in 1989. The contaminants, found in Trp-lots of a Japanese manufacturer, are classified in EMS-correlated and non EMS-correlated substances. Up to now six EMS-correlated substances are known. One of these compounds is 2-(3'-indolylmethyl)-indole (IMT). IMT was detected as a major contaminant in two investigated EMS-associated trp-samples. In a seven step chemical synthesis IMT was obtained for use as a reference substance. A model system to investigate the formation of IMT was created using Trp and 3-indolylmethanol (IM). IMT formation was observed at acidic and alkaline pH-values and the optimal molar ratio of Trp to IM is 100:1. In addition an IMT formation was observed from indole, formaldehyde and Trp as well as from Trp and 3-indolylacetaldehyde.

Biotechnology↗

PubMed2004

Emerging encephalitogenic viruses: lyssaviruses and henipaviruses transmitted by frugivorous bats.

Three newly recognized encephalitogenic zoonotic viruses spread from fruit bats of the genus Pteropus (order Chiroptera, suborder Megachiroptera) have been recognised over the past decade. These are: Hendra virus, formerly named equine morbillivirus, which was responsible for an outbreak of disease in horses and humans in Brisbane, Australia, in 1994; Australian bat lyssavirus, the cause of a severe acute encephalitis, in 1996; and Nipah virus, the cause of a major outbreak of encephalitis and pulmonary disease in domestic pigs and people in peninsula Malaysia in 1999. Hendra and Nipah viruses have been shown to be the first two members of a new genus, Henipavirus, in the family Paramyxoviridae, subfamily Paramyxovirinae, whereas Australian bat lyssavirus is closely related antigenically to classical rabies virus in the genus Lyssavirus, family Rhabdoviridae, although it can be distinguished on genetic grounds. Hendra and Nipah viruses have neurological and pneumonic tropisms. The first humans and equids with Hendra virus infections died from acute respiratory disease, whereas the second human patient died from an encephalitis. With Nipah virus, the predominant clinical syndrome in humans was encephalitic rather than respiratory, whereas in pigs, the infection was characterised by acute fever with respiratory involvement with or without neurological signs. Two human infections with Australian bat lyssavirus have been reported, the clinical signs of which were consistent with classical rabies infection and included a diffuse, non-suppurative encephalitis. Many important questions remain to be answered regarding modes of transmission, pathogenesis, and geographic range of these viruses.

Animals↗

PubMed1988

Recent trends in human leptospirosis in Italy.

The recent epidemiological trends of human leptospirosis in Italy were investigated using data collected for the years 1981-1985. A total of 626 hospitalized patients with clinical diagnoses of suspected leptospirosis were reported by hospital centers from several Italian regions. Epidemiological, clinical and seroimmunological data were collected in 517 of these cases and examined by the National Center for Leptospirosis. Serological findings in 33.5% of these subjects met the criteria for confirmation of the disease. In 21.8% of the subjects, low titer antibodies were detected, which possibly reflected previous leptospiral infections. An early antibiotic treatment of the current infection may also have lowered the seroimmunological response in some of these patients. In 59.3% of the confirmed cases, modes of transmission were allotted equally between accidental events and recreational or occupational activities. Drinking water from an open air fountain emerged as an uncommon mode of transmission; it was responsible for an outbreak of 33 cases of leptospirosis. In another 37.07% of the subjects, it was impossible to establish the mode of transmission. Respiratory or influenza-like symptoms were the only clinical signs of illness in 21.2% of the patients with confirmed leptospirosis. In comparison to the sixties and seventies, the prevalence of infecting serovars showed increasing incidence of infections due to serovars of the Javanica (11.0%) and Australis (11.0%) serogroups and an important decrease in the Bataviae serogroup infections (from 58.8% in rice-field workers in the forties to 0.6% in the years 1981-1985). Sejroe serogroup infections accounted for 4.5 per cent of confirmed cases of leptospirosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMedDate not supplied

Linkage and independence of AIDS and Kaposi disease: the interaction of human immunodeficiency virus and some coagents.

Through epidemiological considerations we conclude that full-blown AIDS may occur only if the index patient is infected by the human immunodeficiency virus (HIV) and, in addition, by some other infectious coagent. Since the dynamical behavior of the spread of AIDS cases with manifestation of Kaposi's sarcoma differs fundamentally from that of the non-Kaposi cases, we conjecture that two independent coagents (together with HIV) are responsible for the outbreak of full-blown AIDS with or without manifestation of Kaposi's sarcoma, respectively. Our formal epidemiological considerations appear to be supported by recent microbiological findings.

Acquired Immunodeficiency Syndrome↗

PubMed1985

Sanguinarine: its potential as a liver toxic alkaloid present in the seeds of Argemone mexicana.

The alkaloid sanguinarine reported to be responsible for several outbreaks of epidemic dropsy in the tropics was examined for its hepatotoxic potential in rats. The studies showed that a single i.p. dose (10 mg/kg) of sanguinarine not only increased the activity of SGPT and SGOT substantially but also caused a significant loss of microsomal cytochrome P-450 and benzphetamine N-demethylase activity. Furthermore, the treated rats exhibited considerable loss of body and liver weight, peritoneal edema and slightly enlarged livers with fibrinous material. Microscopic examination of the liver tissue showed progressive cellular degeneration and necrosis further substantiating that sanguinarine is a potential hepatotoxic alkaloid.

Alanine Transaminase↗

PubMed2005

Molecular epidemiology of enterovirus 71 over two decades in an Australian urban community.

Enterovirus 71 (EV71), first isolated in 1969, has been responsible for numerous outbreaks of hand, foot and mouth disease (HFMD) with a small proportion of cases associated with neurological disease. Since 1997 there has been a significant increase in both the prevalence and virulence of EV71 in the Asia-Pacific region. We have examined the genetic diversity of EV71 in a large Australian city (Sydney N.S.W.) over a nineteen-year period. We determined the VP1 gene sequence of forty-eight EV71 strains isolated between 1983 and 2001. Analysis by molecular phylogeny revealed the presence of four subgenogroups B2, B4, C1 and C2. The results indicate that the major lineage circulating in Sydney N.S.W. was subgenogroup C1 with a recent switch in dominance to B4 in 2000 and 2001.

Australia↗

PubMed1997

Human calicivirus genogroup II capsid sequence diversity revealed by analyses of the prototype Snow Mountain agent.

The Snow Mountain agent (SMA) is the prototype genogroup II and serotype 3 human calicivirus responsible for epidemic outbreaks of acute gastroenteritis. We have cloned the region of the SMA genome that encodes the single capsid protein. The predicted amino acid sequence of the capsid protein is distinct from other calicivirus strains that have been termed SMA-like based on sequence similarity between the RNA polymerase regions and IEM reactivity. In a previous report, a high sequence similarity in a small region of the RNA polymerase between SMA and another strain, OTH-25, suggested that the capsid proteins of OTH-25 and SMA would be very similar. In this report, we show that the capsid proteins of OTH-25 and SMA are more distinct than was predicted by similarity in the RNA polymerase. In addition, phylogenetic analysis of a region of the RNA polymerase and of the N-terminal conserved domain of the capsid protein of 12 human caliciviruses resulted in trees with different topologies, suggesting that recombination has occurred within this group of viruses. Molecular characterization of the prototype calicivirus strains is important in determining the relationships between capsid similarity at the amino acid level, genetic grouping by sequence comparison, and antigenic reactivity.

Amino Acid Sequence↗

PubMed1988

Genetic and molecular characterization of an epidemic plasmid coding for multidrug resistance in Salmonella typhimurium of human origin.

All 201 multidrug resistant Salmonella typhimurium strains isolated from epidemics in India contained nonconjugative (157 strains) or conjugative (44 strains) Inc F1me multiresistance plasmids. Two small R-plasmids of 7 MDa which coded for resistance to either ampicillin or streptomycin and sulfamethoxazole were also detected along with other plasmids. The small plasmids were members of group 1 and group 2 incompatibility groups. Restriction endonuclease analysis of conjugative (96 MDa) and nonconjugative (88 MDa) Inc F1me plasmids showed considerable similarity except for the presence of unique fragments among both the groups and the loss of fragments corresponding to the smaller size of the nonconjugative plasmid. A single Inc F1me plasmid appears responsible for various outbreaks of multiresistant S. typhimurium in different parts of India.

Conjugation, Genetic↗

PubMed1993

Concentration of fish enveloped viruses from large volumes of water.

The validity of several concentration procedures for the detection of fish enveloped viruses present in large volumes of water was determined. Viral haemorrhagic septicaemia virus (VHSV) was used to evaluate adsorption/elution to positively-charged MK filter cartridges for the concentration of enveloped viruses. For fresh water, the efficiency of the procedure ranged from 12 to 100%, with a mean recovery of 57%. In seawater samples, the recoveries varied from 15 to 100%, with a mean recovery of 59%. The same virus was used in methods such as organic flocculation and ammonium sulphate flocculation with very poor recoveries of infectious virus, caused by the inactivation of VHSV in both procedures. Concentration of seawater samples from tanks housing sea bass or gilthead affected by viral erythrocytic infection and lymphocystis, respectively, were carried out. In both cases, the viruses responsible for the outbreaks were detected by electron microscopy in the concentrated water samples.

Adsorption↗

PubMed1991

Manufacturing processes at two French rapeseed oil companies: possible relationships to toxic oil syndrome in Spain.

The toxic oil syndrome (TOS) epidemic that occurred in Spain in spring 1981 has been associated with the consumption of rapeseed oil that was denatured with aniline for industrial use but diverted for human consumption. The precise aetiologic agent in the oil responsible for the outbreak has not been identified. To learn more about possible contaminants and how the contamination might have occurred, we visited two French companies that process rapeseed oil and that were identified in Spanish administrative and judicial records as the ones exporting aniline-denatured rapeseed oil to Spain in 1981. With the apparently full and voluntary co-operation of personnel at both companies, we reviewed the processes involved in manufacturing, treating and transporting rapeseed oil, and we have summarized the information provided to us. Of particular importance is the finding that oil exported to Spain was taken from stock, the rest of which was sold for human consumption in the French domestic market, apparently without any adverse health effects. The differences between the oil exported to Spain and the oil sold as food in France were that aniline equivalent to 2% of the weight of the oil was added to most of the Spanish oil but not to that sold in France, and that contamination of the Spanish oil may have occurred in the tank trucks used for transportation to Spain, which had previously carried industrial chemicals. There is no assurance that the trucks were cleaned appropriately for transporting a food product before the oil was loaded for the journey to Spain. Since the clinical manifestations of TOS are not those of aniline toxicity, we conclude that the aetiological agent of TOS is likely to be one of the following: (1) a contaminant in the aniline, (2) a contaminant introduced during transportation, (3) a reaction product of normal oil components or materials used in refining with either aniline or the potential contaminants mentioned under (1) or (2) above.

Aniline Compounds↗

PubMed1993

Production of polyclonal antibodies against 1,1'-ethyliden bis[L-tryptophan] (EBT), a potential contaminant causing eosinophilia-myalgia syndrome (EMS).

Eosinophilia-myalgia syndrome (EMS) is thought to be caused by the intake of contaminated L-tryptophan products. 1,1'-Ethyliden bis[L-tryptophan] (EBT) is one of the contaminants in the L-tryptophan products and is thought to be a possible cause of EMS. In the present study we report that anti-EBT antibodies are generated by immunizing rabbits with the low molecular weight substance, EBT (molecular weight 434). These antibodies may be available tools for elucidating the mechanisms responsible for the outbreak and/or development of EMS.

Animals↗

PubMed2004

Survival of Escherichia coli O157:H7, O111:H- and O26:H11 in artificially contaminated chocolate and confectionery products.

To date, the survival of Escherichia coli O157:H7 and other verocytotoxin-producing E. coli (VTEC) in chocolate and other confectionery products has not been fully established, unlike Salmonella, which have been responsible for occasional outbreaks of infection linked to contaminated chocolate and related products, although none of these outbreaks have been related to products produced in the United Kingdom. The United Kingdom Biscuit, Cake, Chocolate and Confectionery Alliance commissioned this study to obtain information on the decline and potential survival of E. coli, particularly verocytotoxin-producing strains, in reduced aw confectionery products chocolate, biscuit cream and mallow. These products were artificially contaminated with high (4 log10 cfu/g) and low (2 log10 cfu/g) levels of E. coli O157:H7, O111:H- and O26:H11 and their survival, as affected by storage temperature (10, 22 and 38 degrees C), was monitored over 12 months. Preliminary studies to establish the best inoculation and recovery procedures indicated that differences between counts on selective and non-selective media used were not sufficiently different to influence the outcome of this study. Irrespective of sample type, rapid decline was observed in products stored at 38 degrees C and increased survival occurred in products stored at 10 degrees C. In chocolate (average aw 0.40), these bacteria were detected for up to 43 days in samples stored at 38 degrees C. At 22 degrees C they survived for up to 90 days and in product stored at 10 degrees C they could still be detected after 366 days storage. In biscuit cream (average aw 0.75) they survived for 2 days at 38 degrees C, 42 days at 22 degrees C and 58 days at 10 degrees C. Whilst mallow (aw ca. 0.73) was not stored at 38 degrees C, these bacteria could still be detected in samples stored for up to 113 and 273 days at 22 and 10 degrees C, respectively. The observed prolonged survival of these bacteria under conditions of reduced aw and lowered storage temperature in this study is supported by previous studies with Salmonella and E. coli O157:H7 in other foods. In the same way that Salmonella bacteria can survive for long periods, in excess of 12 months, in chocolate, this study provides evidence that E. coli, including pathogenic strains, can also survive for similar periods of time. Assuming the routes of transmission are similar, controls currently used by the confectionery industry to prevent contamination by Salmonella should also be effective against E. coli, including VT-producing strains, providing that all raw materials have been suitably processed, stored and handled before and during manufacture.

Cacao↗

PubMed2006

Production of authentic SARS-CoV M(pro) with enhanced activity: application as a novel tag-cleavage endopeptidase for protein overproduction.

The viral proteases have proven to be the most selective and useful for removing the fusion tags in fusion protein expression systems. As a key enzyme in the viral life-cycle, the main protease (M(pro)) is most attractive for drug design targeting the SARS coronavirus (SARS-CoV), the etiological agent responsible for the outbreak of severe acute respiratory syndrome (SARS) in 2003. In this study, SARS-CoV M(pro) was used to specifically remove the GST tag in a new fusion protein expression system. We report a new method to produce wild-type (WT) SARS-CoV M(pro) with authentic N and C termini, and compare the activity of WT protease with those of three different types of SARS-CoV M(pro) with additional residues at the N or C terminus. Our results show that additional residues at the N terminus, but not at the C terminus, of M(pro) are detrimental to enzyme activity. To explain this, the crystal structures of WT SARS-CoV M(pro) and its complex with a Michael acceptor inhibitor were determined to 1.6 Angstroms and 1.95 Angstroms resolution respectively. These crystal structures reveal that the first residue of this protease is important for sustaining the substrate-binding pocket and inhibitor binding. This study suggests that SARS-CoV M(pro) could serve as a new tag-cleavage endopeptidase for protein overproduction, and the WT SARS-CoV M(pro) is more appropriate for mechanistic characterization and inhibitor design.

Calbindins↗