Occurrence of Antibiotic Resistance Genes and Bacterial Markers in a Tropical River Receiving Hospital and Urban Wastewaters

The occurrence of emerging biological contaminants including antibiotic resistance genes (ARGs) and Faecal Indicator Bacteria (FIB) is still little investigated in developing countries under tropical conditions. In this study, the total bacterial load, the abundance of FIB (E. coli and Enterococcus spp. (ENT)), Pseudomonas spp. and ARGs (bla_TEM, bla_CTX-M, bla_SHV, bla_NDM and aadA) were quantified using quantitative PCR in the total DNA extracted from the sediments recovered from hospital outlet pipes (HOP) and the Cauvery River Basin (CRB), Tiruchirappalli, Tamil Nadu, India. The abundance of bacterial marker genes were 120, 104 and 89 fold higher for the E. coli, Enterococcus spp. and Pseudomonas spp., respectively at HOP when compared with CRB. The ARGs aadA and bla_TEM were most frequently detected in higher concentration than other ARGs at all the sampling sites. The ARGs bla_SHV and bla_NDM were identified in CRB sediments contaminated by hospital and urban wastewaters. The ARGs abundance strongly correlated (r ≥ 0.36, p < 0.05, n = 45) with total bacterial load and E. coli in the sediments, indicating a common origin and extant source of contamination. Tropical aquatic ecosystems receiving wastewaters can act as reservoir of ARGs, which could potentially be transferred to susceptible bacterial pathogens at these sites.     

Mortality in sickle cell anemia in Africa: a prospective cohort study in Tanzania

Background

The World Health Organization has declared Sickle Cell Anemia (SCA) a public health priority. There are 300,000 births/year, over 75% in Africa, with estimates suggesting that 6 million Africans will be living with SCA if average survival reaches half the African norm. Countries such as United States of America and United Kingdom have reduced SCA mortality from 3 to 0.13 per 100 person years of observation (PYO), with interventions such as newborn screening, prevention of infections and comprehensive care, but implementation of interventions in African countries has been hindered by lack of locally appropriate information. The objective of this study was to determine the incidence and factors associated with death from SCA in Dar-es-Salaam.

Methods and Findings

A hospital-based cohort study was conducted, with prospective surveillance of 1,725 SCA patients recruited from 2004 to 2009, with 209 (12%) lost to follow up, while 86 died. The mortality rate was 1.9 (95%CI 1.5, 2.9) per 100 PYO, highest under 5-years old [7.3 (4.8–11.0)], adjusting for dates of birth and study enrollment. Independent risk factors, at enrollment to the cohort, predicting death were low hemoglobin (<5 g/dL) [3.8 (1.8–8.2); p = 0.001] and high total bilirubin (≥102 µmol/L) [1.7 (1.0–2.9); p = 0.044] as determined by logistic regression.

Conclusions

Mortality in SCA in Africa is high, with the most vulnerable period being under 5-years old. This is most likely an underestimate, as this was a hospital cohort and may not have captured SCA individuals with severe disease who died in early childhood, those with mild disease who are undiagnosed or do not utilize services at health facilities. Prompt and effective treatment for anemia in SCA is recommended as it is likely to improve survival. Further research is required to determine the etiology, pathophysiology and the most appropriate strategies for management of anemia in SCA.     

Human exposures and potential health hazard assessment of Hg and Pb in some major imported frozen fish species in Nigeria

The levels of mercury and lead in three major species of imported frozen fish—Atlantic mackerel (Scomber scombrus), sardine (Sardinella sindesis), and scumbera (Scumbria spp)—were determined and potential health hazard resulting from their ingestion by Nigerians was assessed. Sample digestion was carried out as prescribed in the Food and Agricultural Organisation/Swedish International Development Agency wet digestion method. Quantitation of the Hg and Pb content of the digested samples and blanks was carried out spectrophotometrically using thiozone as the ligand. The ranges of level of Hg in the samples in mg/Kg were: 0.03–0.42 for Scomber scombrus and 0.03–0.37 and 0.020–0.31 for Scumbria and Sardinella sindesis, respectively. Also, Pb had value ranges in mg/Kg of 0.20–0.28, 0.24–0.32, and 0.15–0.32 in Scomber scombrus, Scumbria, and Sardinella sindesis, respectively. Potential health hazard assessment that was carried out showed no risk status for children and adults consuming Sardinella sindesis and scumbera and adults consuming Scomber scombrus (HQ < 1). The results further showed a risk (HQ = 1.022 and THI = 1.036) for children taking Scomber scombrus.     

The discovery and unique pharmacological profile of RO4938581 and RO4882224 as potent and selective GABAA α5 inverse agonists for the treatment of cognitive dysfunction

Lead optimisation of the imidazo[1,5-a][1,2,4]-triazolo[1,5-d][1,4]benzodiazepine class led to the identification of two clinical leads [RO4882224 (11) and RO4938581 (44)] functioning as novel potent and selective GABA_A α5 inverse agonists. The unique pharmacological profiles and optimal pharmacokinetic profiles resulted in in vivo activity in selected cognition models.     

Isolation of sodium chloride tolerant cell lines and plants in finger millet

Sodium chloride tolerant cell lines of finger millet were isolated from embryogenic cultures growing on MS medium supplemented with picloram (2 mg I^-1), kinetin (0.1 mg l^-1) and sodium chloride (1 %) at the end of 6 passages. The sodium chloride tolerant cell lines showed better growth in comparison with control at all concentrations of sodium chloride tested, with optimum growth at 0.25 % NaCl. When the tolerant lines were grown for 3 passages in absence of NaCl, the growth was lower than that of the tolerant lines tested immediately at the end of 6 passages of selection. NaCl tolerant calli had more Na¹ in comparison with control and they regenerated plants in presence of 1 % NaCl, while the control lines failed to differentiate. When screened in a hydroponics system with 1 % NaCl, the tolerant plants grew to maturity while the control plants failed to grow.     

Desulfonation of Linear Alkylbenzenesulfonate Surfactants and Related Compounds by Bacteria

Pseudomonas putida S-313 (= DSM 6884) grew in sulfate-free medium when the sole sulfur source supplied was one of several arylsulfonates involved in the synthesis, application, or biodegradation of linear alkyl-benzenesulfonate (LAS) surfactants. 2-(4-Sulfophenyl)butyric acid, 4-n-butyl-1-methyl-6-sulfotetralin, and 4-toluenesulfonic acid were each completely utilized during growth, as were the model LAS 1-(4-sulfophenyl) octane and the arylsulfonate dyestuff Orange II. The product in each case was the corresponding phenol, which was identified by gas chromatography-mass spectrometry or ¹H nuclear magnetic resonance. Stoichiometric conversion of 4-toluenesulfonic acid to 4-cresol was observed. The molar growth yields observed were 2.4 to 2.8 kg of protein per mol of S, which were comparable to the yield for sulfate. Commercial LAS disappeared from growth medium inoculated with strain S-313, but negligible growth occurred; digestion of cells in alkali led to recovery of the LAS mixture, which seemingly sorbed to the cells. However, mixed culture L6 was readily obtained from batch enrichment cultures containing commercial LAS as a sole sulfur source and an inoculum from domestic sewage. Culture L6 desulfonated components of the LAS surfactant to the corresponding phenols, which were identified by gas chromatography-mass spectrometry. Compounds with shorter alkyl chains were desulfonated preferentially, as were the centrally substituted isomers. In the presence of 200 μM sulfate, culture L6 grew well and LAS disappeared, although this was due purely to sorption, as shown by digestion of the cells in alkali. Thus, under sulfate-limited conditions, LAS can be desulfonated directly.     

Effects of nutrient (N,P, C) enrichment upon the littoral diatom community of an oligotrophic high-mountain lake

The effects of nutrient additions upon the epilithic diatom communities and the algal standing crop were investigated in the oligotrophic, softwater Lake Piccolo Naret, situated in the Swiss alps. Nutrient-diffusing flower pot substrates were filled with either N (0.15 mol NaNO₃), P (0.015 mol Na₂HPO₄) or C (0.15 mol NaHCO₃) or combinations of them. Twenty-five pots representing eight treatments were placed into the lake in July 1991 and sampled after 42 days of exposure.On the surfaces of all pots containing P we measured higher algal biomasses as on the control pots. The chlorophyll-a maximum of 12.9 µg cm^{– 2} was obtained on NPC pots (0.47 µg cm^{– 2} on control pots). On pots with P, NP or NPC supply high amounts of green algae were detected, also reflected in an increased chl-b/chl-c ratio related to the controls which showed algal communities dominated by diatoms.The diatom communities on the control pots as well as on the pots with N, P and NP had a structure similar to the epilithic community in Lake Piccolo Naret (dominance identity > 58 %). However, the community structures of the diatoms from pots with C addition (C, NC, PC and NPC) differed considerably. This is discussed in view of the cell densities of dominant diatom species. For further comparisons the results of two additional high-mountain lakes are used. By means of a cluster analysis it could be shown that epilithic diatom communities were considerably influenced by C addition, while N and P supply were of minor importance.     

Novel CCR1 antagonists with oral activity in the mouse collagen induced arthritis

Cinnamides as novel CCR1 antagonist chemotypes are described with high affinity to human and rodent receptors. A1B1 and A4B7 showed oral activity in the mouse collagen induced arthritis.     

Effect of pyridoxamine on chemical modification of proteins by carbonyls in diabetic rats: characterization of a major product from the reaction of pyridoxamine and methylglyoxal

Advanced glycation end products (AGEs) from the Maillard reaction contribute to protein aging and the pathogenesis of age- and diabetes-associated complications. The alpha-dicarbonyl compound methylglyoxal (MG) is an important intermediate in AGE synthesis. Recent studies suggest that pyridoxamine inhibits formation of advanced glycation and lipoxidation products. We wanted to determine if pyridoxamine could inhibit MG-mediated Maillard reactions and thereby prevent AGE formation. When lens proteins were incubated with MG at 37 degrees C, pH 7.4, we found that pyridoxamine inhibits formation of methylglyoxal-derived AGEs concentration dependently. Pyridoxamine reduces MG levels in red blood cells and plasma and blocks formation of methylglyoxal-lysine dimer in plasma proteins from diabetic rats and it prevents pentosidine (an AGE derived from sugars) from forming in plasma proteins. Pyridoxamine also decreases formation of protein carbonyls and thiobarbituric-acid-reactive substances in plasma proteins from diabetic rats. Pyridoxamine treatment did not restore erythrocyte glutathione (which was reduced by almost half) in diabetic animals, but it enhanced erythrocyte glyoxalase I activity. We isolated a major product of the reaction between MG and pyridoxamine and identified it as methylglyoxal-pyridoxamine dimer. Our studies show that pyridoxamine reduces oxidative stress and AGE formation. We suspect that a direct interaction of pyridoxamine with MG partly accounts for AGE inhibition.     

Oxaloacetate decarboxylase of <Emphasis Type="Italic">Archaeoglobus fulgidus</Emphasis>: cloning of genes and expression in <Emphasis Type="Italic">Escherichia coli</Emphasis>

Archaeoglobus fulgidus harbors three consecutive and one distantly located gene with similarity to the oxaloacetate decarboxylase Na⁺ pump of Klebsiella pneumoniae (KpOadGAB). The water-soluble carboxyltransferase (AfOadA) and the biotin protein (AfOadC) were readily synthesized in Escherichia coli, but the membrane-bound subunits AfOadB and AfOadG were not. AfOadA was affinity purified from inclusion bodies after refolding and AfOadC was affinity purified from the cytosol. Isolated AfOadA catalyzed the carboxyltransfer from [4-¹⁴C]-oxaloacetate to the prosthetic biotin group of AfOadC or the corresponding biotin domain of KpOadA. Conversely, the carboxyltransferase domain of KpOadA exhibited catalytic activity not only with its pertinent biotin domain but also with AfOadC.