Isolation of bacteriophages from commercial sera

Summary Commercially available sera contain bacterial viruses. The possible origin of serum bacteriophages and the implication of this observation for tissue culture studies are discussed. Research supported by a Postdoctoral Fellowship (1 FO2 HD 50,527-1) from the National Institute of Child Health and Human Development.     

α‐MSH activates immediate defense responses to UV‐induced oxidative stress in human melanocytes

Exposure of cultured human melanocytes to ultraviolet radiation (UV) results in DNA damage. In melanoma, UV‐signature mutations resulting from unrepaired photoproducts are rare, suggesting the possible involvement of oxidative DNA damage in melanocyte malignant transformation. Here we present data demonstrating immediate dose‐dependent generation of hydrogen peroxide in UV‐irradiated melanocytes, which correlated directly with a decrease in catalase activity. Pretreatment of melanocytes with α‐melanocortin (α‐MSH) reduced the UV‐induced generation of 7,8‐dihydro‐8‐oxyguanine (8‐oxodG), a major form of oxidative DNA damage. Pretreatment with α‐MSH also increased the protein levels of catalase and ferritin. The effect of α‐MSH on 8‐oxodG induction was mediated by activation of the melanocortin 1 receptor (MC1R), as it was absent in melanocytes expressing loss‐of‐function MC1R, and blocked by concomitant treatment with an analog of agouti signaling protein (ASIP), ASIP‐YY. This study provides unequivocal evidence for induction of oxidative DNA damage by UV in human melanocytes and reduction of this damage by α‐MSH. Our data unravel some mechanisms by which α‐MSH protects melanocytes from oxidative DNA damage, which partially explain the strong association of loss‐of‐function MC1R with melanoma.     

α‐MSH tripeptide analogs activate the melanocortin 1 receptor and reduce UV‐induced DNA damage in human melanocytes

One skin cancer prevention strategy that we are developing is based on synthesizing and testing melanocortin analogs that reduce and repair DNA damage resulting from exposure to solar ultraviolet (UV) radiation, in addition to stimulating pigmentation. Previously, we reported the effects of tetrapeptide analogs of α‐melanocortin (α‐MSH) that were more potent and stable than the physiological α‐MSH, and mimicked its photoprotective effects against UV‐induced DNA damage in human melanocytes. Here, we report on a panel of tripeptide analogs consisting of a modified α‐MSH core His⁶‐d ‐Phe⁷‐Arg⁸, which contained different N‐capping groups, C‐terminal modifications, or arginine mimics. The most potent tripeptides in activating cAMP formation and tyrosinase of human melanocytes were three analogs with C‐terminal modifications. The most effective C‐terminal tripeptide mimicked α‐MSH in reducing hydrogen peroxide generation and enhancing nucleotide excision repair following UV irradiation. The effects of these three analogs required functional MC1R, as they were absent in human melanocytes that expressed non‐functional receptor. These results demonstrate activation of the MC1R by tripeptide melanocortin analogs. Designing small analogs for topical delivery should prove practical and efficacious for skin cancer prevention.     

Sequence heterogeneity of theenvelope-like domain in cultivated allotetraploidGossypium species and their diploid progenitors

Retroviralenvelope (env)-like sequences in 2 cultivated allotetraploid cottons and their diploid progenitors have been identified and characterized in this study. DNA sequence analysis reveals that these sequences are heterogeneous. The observed sequence diversity, however, seems to preserve coding information. This is evidenced by the detection of the transmembrane domain (TM), which is the most conserved feature of the divergent retroviral env genes. The high ratio of synonymous to nonsynonymous changes suggests that these sequences are evolving under purifying selection. Phylogenetic analysis shows thatGossypium sequences closely cluster with a lineage of plant endogenous retroviruses that have anenv-like gene. These results provide evidence for the antiquity and the wide diversity ofenv-like sequences in theGossypium genome.     

Growth assessment of diabetic rat fetuses under the influence of insulin and melatonin: a morphologic study

Hyperglycemia-induced oxidative stress makes an important contribution to the etiology of diabetic teratogenicity namely fetal growth and congenital dysmorphogenesis. The aim of this study is to evaluate the protective roles of melatonin and insulin against diabetic's embryolethality and teratogenicity. Diabetes was induced to virgin Sprague Dawley albino rats by a single peritoneal injection of alloxan. Thirty pregnant rats were divided equally into 5 groups: 1) Control 2) Diabetic 3) Diabetic insulin 4) Diabetic melatonin 5) Diabetic melatonin-insulin. Insulin and melatonin were administered daily throughout the whole gestational period. Fetuses were collected on day 20 of gestation and were examined for malformations and growth disorders. A significant increase in fetal growth parameters (Macrosomia) were noticed in the diabetic group compared to the control. Melatonin prevents the appearance of soft tissue anomalies, but it leads to fetal growth restriction of diabetic rats (Microsomia). No significant changes were noticed in fetal growth parameters in diabetic insulin or in diabetic melatonin-insulin groups compared to the control. Congenital anomalies were not seen in diabetic insulin and in diabetic melatonin-insulin groups while the rate of resorption was reduced in both groups when compared to the diabetic group. In conclusion, co-administration of melatonin with insulin leads to a slight non significant improvement of the protective role of insulin against diabetic embryolethality, teratogenicity and fetal growth changes.     

A virulence and antimicrobial resistance DNA microarray detects a high frequency of virulence genes in Escherichia coli isolates from Great Lakes recreational waters

Escherichia coli is generally described as a commensal species with occasional pathogenic strains. Due to technological limitations, there is currently little information concerning the prevalence of pathogenic E. coli strains in the environment. For the first time, using a DNA microarray capable of detecting all currently described virulence genes and commonly found antimicrobial resistance genes, a survey of environmental E. coli isolates from recreational waters was carried out. A high proportion (29%) of 308 isolates from a beach site in the Great Lakes carried a pathotype set of virulence-related genes, and 14% carried antimicrobial resistance genes, findings consistent with a potential risk for public health. The results also showed that another 8% of the isolates had unusual virulence gene combinations that would be missed by conventional screening. This new application of a DNA microarray to environmental waters will likely have an important impact on public health, epidemiology, and microbial ecology in the future.     

A time series analysis of the relationship of ambient temperature and common bacterial enteric infections in two Canadian provinces

The incidence of enteric infections in the Canadian population varies seasonally, and may be expected to be change in response to global climate changes. To better understand any potential impact of warmer temperature on enteric infections in Canada, we investigated the relationship between ambient temperature and weekly reports of confirmed cases of three pathogens in Canada: Salmonella, pathogenic Escherichia coli and Campylobacter, between 1992 and 2000 in two Canadian provinces. We used generalized linear models (GLMs) and generalized additive models (GAMs) to estimate the effect of seasonal adjustments on the estimated models. We found a strong non-linear association between ambient temperature and the occurrence of all three enteric pathogens in Alberta, Canada, and of Campylobacter in Newfoundland-Labrador. Threshold models were used to quantify the relationship of disease and temperature with thresholds chosen from 0 to −10°C depending on the pathogen modeled. For Alberta, the log relative risk of Salmonella weekly case counts increased by 1.2%, Campylobacter weekly case counts increased by 2.2%, and E. coli weekly case counts increased by 6.0% for every degree increase in weekly mean temperature. For Newfoundland-Labrador the log relative risk increased by 4.5% for Campylobacter for every degree increase in weekly mean temperature.     

The Unitary Association method in biochronology and its potential stratigraphic power between benthic and planktic organisms: A case study on foraminifers from Paleocene–Eocene strata of southern Egypt

Although foraminifers have been extensively used for biochronology, no quantitative evaluation of the quality of their application is available. In this work we apply a quantitative approach – the Unitary Association (UA) method – to evaluate the relative quality of planktic and benthic foraminifers in biostratigraphy. Based on 12,321 specimens belonging to 65 planktic and 132 benthic foraminifer species in 117 samples from the Paleocene–Eocene strata of the Kharga-Baris Oases (Western Desert, Egypt), the potential stratigraphic resolution power of benthic foraminifers is quantitatively assessed and compared to planktic foraminifers. The UA method accounts for superpositional contradictions between sets of coexisting taxa and generates unitary associations, which are conceptually similar to Oppel zones. The analysis produced fifteen unitary associations for the Paleocene–Eocene benthic Foraminifera of the studied sections, instead of three classical zones; most of these unitary associations are easily identified by the restricted occurrence of a single species. Although planktic Foraminifera have a higher seriation reality leading to a more robust biozonation, benthic Foraminifera have a higher stratigraphic resolution, at least at such regional scale. Furthermore, there was a significant positive correlation between the number of unitary associations and the number of species (R = 0.78 for planktic and 0.70 for benthic foraminifers; both with p < 0.001). These results corroborate that quantitative techniques may lead to a higher resolution in biochronology, even when applied to poor biochronological markers. Moreover, in contrast to simple spatiotemporal analyses, the Unitary Association method can accurately assess the stratigraphic potential of a given taxonomic group.