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Biomedical subjects

J A Marshall

Publications and source records attributed to J A Marshall.

At least 19 recordsLinked to original sources
PubMed2007

Molecular features of astrovirus associated with a gastroenteritis outbreak in an aged-care centre.

The study presented here was conducted in order to gain a better understanding of the role of astroviruses (AsVs) in outbreaks of gastroenteritis among the elderly. This report is the first to provide detailed information on the molecular characteristics of an AsV causing an outbreak in an aged-care centre and is the first to clearly establish that individuals infected in such an outbreak were, in fact, elderly. The outbreak under investigation took place in Victoria, Australia, in October 2005. Twelve individuals (mean age +/- standard deviation [SD] 85.5 +/- 12.3 years) became ill during the outbreak from a total population of 86 susceptible residents. The mean duration (+/-SD) of illness was 2.3 +/- 1.6 days; symptoms included diarrhoea, abdominal pain, nausea and headache. No bacterial pathogens were detected. AsV was identified in five faecal specimens using electron microscopy and reverse transcription-polymerase chain reaction methodologies; no other gastroenteritis virus was detected. Nucleotide sequence analysis indicated the AsV identified could be assigned to the 1d lineage of AsV serotype 1 and that the AsV was not a recombinant form. The findings, taken together with previous work, indicate the AsV serotype most commonly associated with gastroenteritis outbreaks among the elderly is serotype 1 AsV.

Aged, 80 and over↗
PubMed2005

Evaluation of a commercial enzyme immunoassay for detection of norovirus in outbreak specimens.

The aim of the study presented here was to use faeces from 41 gastroenteritis outbreaks (130 specimens) in Victoria, Australia, to evaluate the sensitivity and specificity of the RIDASCREEN norovirus enzyme immunoassay (EIA) kit relative to reverse transcription-polymerase chain reaction and/or electron microscopy. Seven specimens known to contain sapovirus, adenovirus, astrovirus and rotavirus were also tested. For single-specimen diagnosis the kit gave a specificity and sensitivity of 47% and 71%, respectively; altering the positivity cut-off to give a specificity of 73% reduced the sensitivity to 44%. Thus, the kit cannot be recommended for single-specimen diagnosis. One specimen containing adenovirus but not norovirus was identified as non-specifically positive by the EIA kit. If the criterion used for outbreak positivity was at least one EIA-positive specimen per outbreak, the kit's outbreak sensitivity was 94% but the outbreak specificity was only 60%.

Caliciviridae Infections↗
PubMed2005

The SPINK1 N34S mutation is not associated with Type 2 diabetes mellitus in a population of the USA.

AIMS: Mutations in the serine protease inhibitor (SPINK1) gene have been associated with all forms of chronic pancreatitis. Recently, an association of SPINK1 mutations with early-onset Type 2 diabetes mellitus has been reported in patients from Bangladesh. Therefore, we determined the frequency of SPINK1 N34S mutations in patients with Type 2 diabetes mellitus from the USA. METHODS: The study population of Hispanic and non-Hispanic white people consisted of 387 patients with Type 2 diabetes and familial clustering of the disease, 232 family members without diabetes, 259 patients with Type 2 diabetes without a family history, and 302 ethnically matched healthy controls as part of the San Luis Valley Diabetes Study. We performed linkage- and association-analysis in 82 multiplex families with Type 2 diabetes mellitus. RESULTS: No significant linkage or allele sharing was detected between Type 2 diabetes mellitus and the SPINK1 locus. The frequency of the N34S mutation was determined by fluorescence polarization and was similar between patients (n = 14/387 patients with familial clustering; n = 2/259 patients without family history) and controls (n = 5/232 family members without diabetes; n = 10/302 individuals). Variables such as ethnicity, age of diabetes onset and percentage of individuals with impaired glucose tolerance did not differ significantly between carriers and homozygous normal individuals. CONCLUSION: The SPINK1 N34S mutation appears not to predispose Hispanic or non-Hispanic white people from the USA to the development of Type 2 diabetes mellitus.

Chronic Disease↗
PubMed2004

Long-term features of norovirus gastroenteritis in the elderly.

Noroviruses are important pathogens in both sporadic cases and outbreaks of gastroenteritis in humans. Noroviruses can affect individuals of all ages in a variety of settings, but are a particularly important cause of gastroenteritis in aged-care facilities. The relationship between clinical symptoms and norovirus excretion and the possible role of asymptomatic carriage of norovirus in the elderly are poorly understood. This study examined symptoms and norovirus excretion in elderly individuals associated with a norovirus outbreak in an aged-care facility. Ten individuals aged 79-94 years were recruited for the study. Nine were symptomatic and one was an asymptomatic contact who subsequently developed gastroenteritis. The 10 participants were interviewed regarding their clinical symptoms between two and six times over a three-week study period. One or more sequential faecal samples were collected from all participants over the same period and tested by reverse transcription-polymerase chain reaction for the presence of norovirus. Norovirus was detected in faecal samples from all 10 study participants and was commonly detected in formed stools. In the nine symptomatic participants, acute symptoms such as diarrhoea and vomiting had largely resolved by the third or fourth day of illness, but non-specific symptoms such as headache, thirst and vertigo could persist for as long as 19 days. Both acute and non-specific symptoms appeared to resolve and recur in some participants. The median excretion time for norovirus was 8.6 days (range 2-15 days) in symptomatic participants (N=5). There was no general relationship between the duration of norovirus excretion and the duration of either acute or non-specific symptoms. A faecal sample collected from the asymptomatic contact the day before gastroenteritis symptoms began was positive for norovirus, demonstrating prodromal excretion of norovirus. The results of this study indicate that infection control guidelines should consider both long-term excretion and prodromal excretion of norovirus, and the possibility that formed stools can contain norovirus. Furthermore, the care of elderly individuals recovering from a norovirus infection should take long-term non-specific clinical symptoms into account.

Aged↗
PubMed2004

Failure to detect norovirus in a large group of asymptomatic individuals.

Noroviruses are a major cause of both sporadic and epidemic gastroenteritis in humans, but the mechanisms by which norovirus circulates within the community are poorly understood. In this study, we examined the hypothesis that asymptomatic people act as a reservoir for norovirus. Faecal specimens from 399 asymptomatic individuals were tested for norovirus by reverse transcription polymerase chain reaction (RT-PCR) methodology, and no norovirus was detected. The failure to detect norovirus was not apparently due to the test sample being resistant to norovirus infection, nor to the presence of PCR inhibitors in the test sample. The findings suggest that, if norovirus is carried by asymptomatic people, the carriage rate is very low; the upper bound (95% confidence interval, binomial distribution) of the carriage rate was only 0.8%. Thus, it is unlikely that asymptomatic people are an important reservoir for norovirus.

Adolescent↗
PubMed2004

Insulin resistance as a modifier of the relationship between dietary fat intake and weight gain.

OBJECTIVE: To investigate whether insulin resistance modifies the rate of weight gain associated with a high percent of energy intake from dietary fat. DESIGN: Longitudinal, observational population study. SUBJECTS: A total of 782 nondiabetic Hispanic and non-Hispanic white free-living adult residents of the San Luis Valley in Colorado. MEASUREMENTS: Subjects were seen up to three times over a 14-y period. Weight, height, fasting insulin and glucose, diet by 24 h recall, and self-reported physical activity were collected at each visit. RESULTS: Percentage of energy intake from dietary fat was positively associated with weight gain over time (P=0.0103). High intake of dietary fat was more strongly related to weight gain in women than in men, and in those with lower total energy intake levels. The relationship between weight change and relative macronutrient intake also varied by baseline insulin sensitivity (P=0.0025). Weight gain over time in individuals with relative insulin resistance at baseline, as measured by QUICKI, was the greatest among those who consumed a higher percent of energy from fat. CONCLUSION: Percentage of total intake from dietary fat predicts weight change independent of total energy intake. Nondiabetic, insulin-resistant individuals are particularly susceptible to the weight gain associated with high levels of dietary fat intake. Further investigation into the relationship between insulin resistance, diet, and weight gain is warranted.

Adult↗
PubMed2004

Admixture in the Hispanics of the San Luis Valley, Colorado, and its implications for complex trait gene mapping.

Hispanic populations are a valuable resource that can and should facilitate the identification of complex trait genes by means of admixture mapping (AM). In this paper we focus on a particular Hispanic population living in the San Luis Valley (SLV) in Southern Colorado. We used a set of 22 Ancestry Informative Markers (AIMs) to describe the admixture process and dynamics in this population. AIMs are defined as genetic markers that exhibit allele frequency differences between parental populations >or=30%, and are more informative for studying admixed populations than random markers. The ancestral proportions of the SLV Hispanic population are estimated as 62.7 +/- 2.1% European, 34.1 +/- 1.9% Native American and 3.2 +/- 1.5% West African. We also estimated the ancestral proportions of individuals using these AIMs. Population structure was demonstrated by the excess association of unlinked markers, the correlation between estimates of admixture based on unlinked marker sets, and by a highly significant correlation between individual Native American ancestry and skin pigmentation (R2= 0.082, p < 0.001). We discuss the implications of these findings in disease gene mapping efforts.

Adult↗
PubMed2002

A multiplex allele specific polymerase chain reaction (MAS-PCR) on the dihydrofolate reductase gene for the detection of Cryptosporidium parvum genotypes 1 and 2.

A multiplex allele specific polymerase chain reaction (MAS-PCR) based on the Cryptosporidium parvum dihydrofolate reductase (dhfr) gene sequence differentiates genotype 1 ('Human') from 2 ('Cattle') in a 1-step reaction. The MAS-PCR was validated on a panel of 34 microscopically positive C. parvum faecal samples of human and animal origin in comparison with 2 published PCR-restriction fragment length polymorphism (RFLP) methods targeting dhfr and the oocyst wall protein (cowp) genes. A validation panel of 37 negative faecal samples of human and animal origin was also tested in comparison with the cowp PCR-RFLP. MAS-PCR was found to be as sensitive for species detection as the most sensitive of the other tests, and detected more mixed genotype infections than the two other tests combined. In addition the MAS-PCR showed equivalent detection sensitivity in comparison with a published nested RFLP targeting the SSU rRNA gene, on a panel of prepared mixed genotype samples. The 1-step reaction is simpler and less expensive to perform than the RFLP methods, while the C. parvum specific amplicons and those for genotypes 1 and 2 (575, 357 and 190 bp respectively) can be easily distinguished on agarose gel.

Alleles↗
PubMed2002

The conformation of the mature dimeric human immunodeficiency virus type 1 RNA genome requires packaging of pol protein.

The packaging of a mature dimeric RNA genome is an essential step in human immunodeficiency virus type 1 (HIV-1) replication. We have previously shown that overexpression of a protease (PR)-inactive HIV-1 Gag-Pro-Pol precursor protein generates noninfectious virions that contain mainly monomeric RNA (M. Shehu-Xhilaga, S. M. Crowe, and J. Mak, J. Virol. 75:1834-1841, 2001). To further define the contribution of HIV-1 Gag and Gag-Pro-Pol to RNA maturation, we analyzed virion RNA dimers derived from Gag particles in the absence of Gag-Pro-Pol. Compared to wild-type (WT) dimeric RNAs, these RNA dimers have altered mobility and low stability under electrophoresis conditions, suggesting that the HIV-1 Gag precursor protein alone is not sufficient to stabilize the dimeric virion RNA structure. The inclusion of an active viral PR, without reverse transcriptase (RT) and integrase (IN), rescued the stability of the virion RNA dimers in the Gag particles but did not restore the mobility of the RNAs, suggesting that RT and IN are also required for virion RNA dimer maturation. Thin-section electron microscopy showed that viral particles deficient in RT and IN contain empty cone-shaped cores. The abnormal core structure indicates a requirement for Gag-Pro-Pol packaging during core maturation. Supplementing viral particles with either RT or IN via Vpr-RT or Vpr-IN alone did not correct the conformation of the dimer RNAs, whereas expression of both RT and IN in trans as a Vpr-RT-IN fusion restored RNA dimer conformation to that of the WT virus and also restored the electron-dense, cone-shaped virion core characteristic of WT virus. Our data suggest a role for RT-IN in RNA dimer conformation and the formation of the electron-dense viral core.

Cell Line↗
PubMed2001

Synthesis of (-)-deoxypukalide, the enantiomer of a degradation product of the furanocembranolide pukalide.

A convergent stereoselective synthesis of (-)-deoxypukalide is described. This substance has not yet been found in Nature but is obtained through deoxygenation of pukalide, the first naturally occurring furanocembrane to be structurally elucidated. The route features a new intraannular furan synthesis that entails treatment of a 4-oxopropargylic beta-keto ester with silica gel. The product of this novel reaction, a 3-carboxy 2,5-bridged furan, is formed in 96% yield. The synthetic strategy was strongly directed by molecular mechanics calculations, which provided valuable insight into stereodefining steps including double bond stereochemistry and butenolide configuration.

Animals↗
PubMed2001

Synthesis of stereopentad analogues of the C14-C22 segment of callystatin A through additions of chiral allenylzinc reagents to stereotriads.

The addition of (P)- and (M)-allenylzinc reagents, prepared in situ through Pd-catalyzed metalation of (R)- and (S)-3-butyn-2-ol mesylates, to diastereomeric stereotriad aldehydes 8, 13, 18, and 23 of syn,syn, syn,anti, anti,anti, and anti,syn stereochemistry was examined. Additions to the former two aldehydes afforded the four anti adducts with high diastereoselectivity and negligible mismatching. Significant mismatching was observed with the latter two aldehydes and the (M)-allenylzinc reagent. An evaluation of possible transition states is presented in consideration of steric and dipolar control elements.

Aldehydes↗
PubMed2001

Synthesis of a C1-C21 subunit of the protein phosphatase inhibitor tautomycin: a formal total synthesis.

The synthesis of a C1-C21 subunit of tautomycin is described. The convergent route employs enantioenriched allenylstannane and zinc reagents derived from (S)-3-butyn-2-ol methanesulfonate. These reagents react with appropriate aldehyde segments to yield syn and anti adducts with high diastereoselectivity. The derived lithioalkynes are joined stepwise to a CO equivalent, (MeONMe)2C=O, to afford an intermediate ketone which is converted to the core spiroketal moiety of tautomycin upon acid treatment. Chain elongation by another addition of the aforementioned allenylzinc reagent to a spiroketal aldehyde proceeds with high diastereoselectivity to install the remaining stereocenters. The resulting homopropargylic alcohol adduct is converted to a methyl ketone through intramolecular hydrosilylation of the alkyne and Tamao oxidation of the derived five-membered siloxane. This ketone proved identical to an intermediate employed by Chamberlin in a prior total synthesis of tautomycin.

Antifungal Agents↗
PubMed2001

Soybean sterol composition and utilization by Phytophthora sojae.

The sterol fraction of Glycine max (soybean) was found to contain a mixture of 13 major sterols which differed dramatically in composition between seeds and shoots. Typical C4-desmethyl Delta(5)-sterols, including sitosterol, predominate the sterol mixture of shoots, whereas C4-methyl sterol intermediates, cycloartenol and 24(28)-methylene cycloartanol, accumulate in seeds. The significance of modified sterol profile of shoot compared to seed was relevant to the physiology of Phytophthora sojae, a phytopathogen of soybean shown to be auxotrophic for sterol. Sterols native to the host plant containing a C4-methyl group, such as cycloartenol, were not utilized by the fungus. Alternatively, all Delta(5)-sterols added to the culture media of P. sojae supported normal growth and promoted viable oospore production. The results demonstrate the importance of sterols in plant-fungal interactions and offer the possibility of bioengineering the phytosterol pathway for resistance to phytopathogens which scavenge specific sterols of the host plant to complete the life cycle.

Molecular Structure↗
PubMed2001

Simultaneous isolation of verotoxin-producing strains of Escherichia coli O128:H2 and viruses in gastroenteritis outbreaks.

Three outbreaks of gastroenteritis from which the Verotoxin producing Escherichia coli serotype O128:H2 was isolated are reported. In addition Norwalk-like viruses were detected in patients from two of the outbreaks and astrovirus in the third outbreak. While it cannot be specifically determined which of these agents played the major role in these outbreaks, the findings suggest that the viral agents need to be considered in investigations of gastroenteritis outbreaks, regardless of whether bacterial enteropathogens have also been isolated. This study points to a strong need to investigate gastroenteritis outbreaks for both bacterial and viral agents and to review in detail the asymptomatic carriage rate of Verotoxin-producing bacteria and gastroenteritis-associated viral agents; these areas of public health significance have been largely neglected.

Australia↗