Metabolomic Tools for Secondary Metabolite Discovery from Marine Microbial Symbionts

Marine invertebrate-associated symbiotic bacteria produce a plethora of novel secondary metabolites which may be structurally unique with interesting pharmacological properties. Selection of strains usually relies on literature searching, genetic screening and bioactivity results, often without considering the chemical novelty and abundance of secondary metabolites being produced by the microorganism until the time-consuming bioassay-guided isolation stages. To fast track the selection process, metabolomic tools were used to aid strain selection by investigating differences in the chemical profiles of 77 bacterial extracts isolated from cold water marine invertebrates from Orkney, Scotland using liquid chromatography-high resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance (NMR) spectroscopy. Following mass spectrometric analysis and dereplication using an Excel macro developed in-house, principal component analysis (PCA) was employed to differentiate the bacterial strains based on their chemical profiles. NMR ¹H and correlation spectroscopy (COSY) were also employed to obtain a chemical fingerprint of each bacterial strain and to confirm the presence of functional groups and spin systems. These results were then combined with taxonomic identification and bioassay screening data to identify three bacterial strains, namely Bacillus sp. 4117, Rhodococcus sp. ZS402 and Vibrio splendidus strain LGP32, to prioritize for scale-up based on their chemically interesting secondary metabolomes, established through dereplication and interesting bioactivities, determined from bioassay screening.     

Nonemployment Income and the Economic Base of Michigan Counties: 1959–1986

ABSTRACT Despite the fact that nonemployment income makes up approximately one-third of all personal income, its impact on local area economies has not been closely examined. This study uses Michigan county data to examine the impact of nonemployment income on nonbasic income over a twenty-seven-year period. This impact is compared to the impact of basic income by employing regression analysis to estimate comparative multiplier effects for both types of income. Nonemployment income is found to have a significant impact on nonbasic income, particularly in metropolitan and nonmetropolitan urban counties, where its impact appears to be stronger than that of basic income.     

The nutritional status of asymptomatic HIV-infected Africans: directions for dietary intervention?

OBJECTIVE: To compare the relationships between food (nutrient) intakes and biochemical markers of nutritional status of asymptomatic HIV-infected with HIV-uninfected subjects, to gain more information on the appropriate diet for HIV-infected persons at an early stage of infection. DESIGN: Cross-sectional population-based survey. SETTING: North West Province, South Africa. SUBJECTS: Two hundred and sixteen asymptomatic HIV-infected and 1550 HIV-uninfected men and women volunteers aged 15 years and older, recruited as 'apparently healthy' subjects from 37 randomly selected sites. OUTCOME MEASURES: Food and nutrient intakes, measured with a validated food-frequency questionnaire, and nutritional status indicated by anthropometric and biochemical variables, measured by a standardised methodology. RESULTS: The prevalence of HIV infection in the study population was 11.9%. The anthropometric indices and nutrient intakes of HIV-infected and uninfected subjects did not differ significantly, indicating that these 216 HIV-infected subjects were at an early stage of infection. Of the biochemical nutritional status variables, high-density lipoprotein cholesterol and total cholesterol, haemoglobin, albumin and triglycerides were significantly lower in infected subjects. They also had higher globulin and liver enzyme levels than uninfected subjects. In infected subjects, serum albumin correlated significantly with serum lipids, serum vitamin A, serum vitamin E, serum iron, total iron-binding capacity and haemoglobin. The significant positive correlations of the liver enzymes with serum lipids, albumin, vitamin A and iron, observed in HIV-uninfected subjects, disappeared in the infected subjects. Polyunsaturated fat intake showed significant positive correlations with the increased liver enzymes in infected subjects. A principal components analysis indicated that, in infected subjects, increased liver enzymes correlated with higher consumption of maize meal and lower consumption of meat and vegetables. CONCLUSIONS AND RECOMMENDATIONS: This survey indicated that asymptomatic HIV-infected subjects who followed a diet rich in animal foods had smaller decreases in serum albumin, haemoglobin and lipid variables, and smaller increases in liver enzymes, than those who consumed a diet based on staple foods. This suggests that animal foods are associated with improved nutritional status in HIV-infected persons. These results should be confirmed with intervention studies before dietary recommendations for asymptomatic HIV-infected individuals can be made.     

Carbon-13 in tree rings indicates no record of sea-surface temperature

When the correct values for the temperature coefficient of carbon dioxide solubility in seawater are used, theoretical calculations show that no measurable carbon-isotope redistribution occurs between sea and air for any plausible change in the sea-surface temperature. Although this fact invalidates one possible paleothermometer, it somewhat simplifies the interpretation of carbon-13 data in terrestrial biological samples.     

The determinants of overweight and obesity among 10- to 15-year-old schoolchildren in the North West Province, South Africa - the THUSA BANA (Transition and Health during Urbanisation of South Africans; BANA, children) study

AIM: To investigate the determinants of overweight and obesity among 10- to 15-year-old schoolchildren in a population in the transitional phase in the North West Province of South Africa.METHODS: A cross-sectional survey was used to investigate weight status (anthropometric indicators) and determinants of overweight/obesity including dietary intake, physical activity and socio-economic status. A single, random sample (n=1257), stratified for gender, type of school and ethnic group, was used. Data were collected on demographics, family circumstances, habitual physical activity, dietary intake and anthropometry to evaluate weight status and body fat content. One-way analysis of variance, the generalised linear models procedure of SAS and the Tukey post hoc honest significant difference test were used to analyse the data.RESULTS: Few children were overweight or obese (7.8%) according to International Obesity Task Force (IOTF) standards (body mass index (BMI)-for-age). These standards were compared with other accepted standard values. Both Cole's IOTF/BMI-for-age standard and the sum of skinfold thicknesses standard classified normal-weight status similarly at a level of 92% (P<0.01) and were found to be useful in determining overweight/obesity. The prevalence rate was higher in females and white children, and was more apparent in urban areas, smaller households and children of parents with low- or high-income occupations. Boys and pre-menarcheal girls had mean body fat percentage in the normal/optimal range, whereas that of post-menarcheal girls was moderately high. Few variables showed a significant association with high body fat percentage: in boys, only the number of members in the household and physical activity levels over the weekend; in girls, only age. The overweight/obese boys mostly lived in smaller households, and the overweight/obese post-menarcheal girls were most inactive on both weekdays and weekends, and more overweight with increasing age.CONCLUSION: Smaller households, inactivity and increasing age for girls were found to be determinants that influence the development of overweight/obesity, while female gender and age post-menarche were identified as determinants of higher body fat content. For overweight/obesity prevention, the focus should be on pre-menarcheal girls, aged 10-13 years, using these determinants to identify overweight/obesity risk. Preventive programmes should aim to increase the physical activity of children to improve their current and future weight status.     

Direct observations of structural phase transitions in planar crystallized ion plasmas

Laser-cooled 9Be+ ions confined in two-dimensionally extended lattice planes were directly observed, and the images were used to characterize the structural phases of the ions. Five different stable crystalline phases were observed, and the energetically favored structure could be sensitively tuned by changing the areal density of the confined ions. The experimental results are in good agreement with theoretical predictions for the planar (infinite in two dimensions) one-component plasma. Qualitatively similar structural phase transitions occur, or are predicted to occur, in other experimentally realizable planar systems.     

Changes of Connexin43 Expression in Non‐pregnant Porcine Myometrium Correlate with Progesterone Concentration During Oestrous Cycle

Connexin43 (Cx43) is a major protein of myometrial gap junctions. The number of Cx43 gap junctions increase dramatically with the onset of labour in association with development of synchronized uterine contractions. The formation of myometrial gap junctions follows an increase in the oestrogen to progesterone ratio indicating an important role of steroid hormones in regulating Cx43 expression at term. However, no relationship has been established between the expression of Cx43 in the non‐pregnant myometrium and concentration of steroid hormones during the oestrous cycle. Here, we used immunofluorescence and Western blotting to analyse the expression of Cx43 gap junctions in the myometrium of pre‐pubertal pigs (n = 7) and mature pigs at pre‐ovulatory (n = 7), luteal (n = 5) and late luteal (n = 3) stages of the oestrous cycle. The number of Cx43 gap junctions calculated per 1 mm² of the myometrial section was low in pre‐pubertal pigs and significantly higher (p < 0.022) in pre‐ovulatory animals. In relation to pre‐ovulatory animals the number of myometrial gap junctions was significantly lower (p < 0.019) at the luteal phase and correlated with significantly higher (p < 0.005) concentration of endogenous progesterone. Phosphorylated isoforms of Cx43 protein were expressed in the myometrium of pre‐pubertal pigs and mature animals at pre‐ovulatory and late luteal phases, while they were down regulated at the luteal stage. These results indicate that changes of Cx43 expression in the porcine myometrium during the oestrous cycle may be regulated by progesterone concentration and may contribute to the modulation of uterine motility.     

Localization of Transforming Growth Factor Beta3 in Squamous Metaplasia of the Porcine Oviduct: a Case Report

Squamous metaplasia of the oviduct epithelium is a rare disorder of reproductive organs. We noted squamous metaplasia of the oviduct epithelium in a sow routinely slaughtered at day 2 of the oestrous cycle. Expression of transforming growth factor beta3 (TGF beta3) in the metaplastic epithelia was evaluated by immunohistochemistry, because TGF beta3 appears to play a key role as regulator of a variety of tissue remodelling events. Our results show that TGF beta3 immunostaining was specifically localized to foci of squamous metaplasia of the epithelial linings. Non‐metaplastic epithelial cells of the oviduct were not immunostained with anti‐TGF beta3 antibody. At the subcellular level, TGF beta3‐labelled cells occasionally showed signs of apoptotic cell death. It is concluded that signals produced by TGF beta3 in metaplastic lesions of the oviduct are potentially involved in pathophysiological processes.