Water Quality and Associated Human Health Risk Assessment Related to Some Ions and Trace Elements in a Series of Rural Roma Communities in Transylvania, Romania.

This research aims to assess the content of some ions and trace elements in water sources in 24 rural Roma communities in Transylvania in order to assess the human health risk associated with exposure to such elements and ions. To this end, eight ions (F-, Cl-, Br-, NO2-, NO3-, SO42-, PO43-, NH4+) and ten trace elements (Cr, Ni, As, Pb, Cd, Mn, Cu, Zn, Fe, and Hg) were determined in 71 water samples by ion chromatography coupled with a conductivity detector for ions and atomic absorption spectrophotometry for all trace elements. General parameters were also determined. Non-conformity (as number of samples), according to the EU Drinking Water Directive, was observed as follows: pH (7), EC (7), hardness (1), oxidizability (15), Cl- (4), NO3- (30), SO42- (6), Fe (16), Mn (14), As (3), and Ni (1 sample). The incidence of ions was Cl- (71), SO42- (70), F- (67), NO3- (65), NH4+ (21), Br- (10), PO43-, and NO2- (1 sample) and for trace elements, Mn (59), Fe (50), As (38), Ni (32), Cu (29), Zn (28), Cd (12), Cr (11), and Pb (3 samples). Hg was not detected. Non-carcinogenic (HI) values exceeded one for As in 13 Roma communities, with higher values for children than for adults. For NO3-, the HI values were >1 in 12 for adults and 14 communities for children. The carcinogenic risk (CR) for As through ingestion ranged from 0.795 to 3.50 × 10-4 for adults and from 1.215 to 5.30 × 10-4 for children. CR by dermal contact was in the range of ×10-6 both for adults and children.

LC-MS Approach to Decipher a Light Chain Chromatographic Peak Splitting of a Monoclonal Antibody.

PURPOSE: Monoclonal antibodies (mAbs), like other protein therapeutics, are prone to various forms of degradation, some of which are difficult to distinguish from the native form yet may alter potency. A generalizable LC-MS approach was developed to enable quantitative analysis of isoAsp. In-depth understanding of product quality attributes (PQAs) enables optimization of the manufacturing process, better formulation selection, and decreases risk associated with product handling in the clinic or during shipment. METHODS: Reversed-phase chromatographic peak splitting was observed when a mAb was exposed to elevated temperatures. Multiple LC-MS based methods were applied to identify the reason for peak splitting. The approach involved the use of complementary HPLC columns, multiple enzymatic digestions and different MS/MS ion dissociation methods. In addition, mAb potency was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The split peaks had identical masses, and the root cause of the peak splitting was identified as isomerization of an aspartic acid located in the complementarity-determining region (CDR) of the light chain. And the early eluting and late eluting peaks were collected and performed enzymatic digestion to confirm the isoAsp enrichment in the early eluting peak. In addition, decreased potency was observed in the same heat-stressed sample, and the increased isoAsp levels in the CDR correlate well with a decrease of potency. CONCLUSION: Liquid chromatography-mass spectrometry (LC-MS) has been utilized extensively to assess PQAs of biological therapeutics. In this study, a generalizable LC-MS-based approach was developed to enable identification and quantitation of the isoAsp-containing peptides.

G.I.S. Surveillance of Chronic Non-occupational Exposure to Heavy Metals as Oncogenic Risk.

INTRODUCTION: The potential oncogenic effect of some heavy metals in people occupationally and non-occupationally exposed to such heavy metals is already well demonstrated. This study seeks to clarify the potential role of these heavy metals in the living environment, in this case in non-occupational multifactorial aetiology of malignancies in the inhabitants of areas with increased prevalent environmental levels of heavy metals. METHODS: Using a multidisciplinary approach throughout a complex epidemiological study, we investigated the potential oncogenic role of non-occupational environmental exposure to some heavy metals [chrome (Cr), nickel (Ni), copper (Cu), zinc (Zn), cadmium (Cd), lead (Pb) and arsenic (As)-in soil, drinking water, and food, as significant components of the environment] in populations living in areas with different environmental levels (high vs. low) of the above-mentioned heavy metals. The exposures were evaluated by identifying the exposed populations, the critical elements of the ecosystems, and as according to the means of identifying the types of exposure. The results were interpreted both epidemiologically (causal inference, statistical significance, mathematical modelling) and by using a GIS approach, which enabled indirect surveillance of oncogenic risks in each population. RESULTS: The exposure to the investigated heavy metals provides significant risk factors of cancer in exposed populations, in both urban and rural areas [ χ2 test (p < 0.05)]. The GIS approach enables indirect surveillance of oncogenic risk in populations. CONCLUSIONS: The role of non-occupational environmental exposure to some heavy metals in daily life is among the more significant oncogenic risk factors in exposed populations. The statistically significant associations between environmental exposure to such heavy metals and frequency of neoplasia in exposed populations become obvious when demonstrated on maps using the GIS system. Environmental surveillance of heavy metals pollution using GIS should be identified as an important element of surveillance, early detection, and control of neoplastic risks in populations, at the level of a single locality, but even on a wider geographical scale.

Lack of association of delta-aminolevulinate dehydratase polymorphisms with blood lead levels and hemoglobin in Romanian women from a lead-contaminated region.

As part of a project on environmental pollution, this study aimed to evaluate associations between blood lead (BPb) levels, hemoglobin (Hb) content, and single-nucleotide polymorphisms (SNPs) of delta-aminolevulinic acid dehydratase (ALAD) gene in 129 unrelated women from Romania. Five SNPs (rs1805313, rs2228083, rs1805312, rs1800435, rs1139488) were analyzed with respect to haplotype structure and impact on BPb levels and Hb content with proportional odds and analysis of covariance models. Combinations of SNPs were rare (16%). Low haplotype diversity was found with seven haplotypes. One rare haplotype implied the C allele of rs1800435, often referred to as the ALAD2 allele (frequency 8.6%). The putative risk genotype (CC) occurred in only one woman with BPb below 0.5 microg/dl. Median BPb was 4.8 microg/dl and differed markedly by community with a level of 12.5 microg/dl near a mining-spill region. Hb was regular (interquartile range 12.3-13.7 g/dl) and not correlated with BPb, although quantitatively lower in women living near the spill region. No significant associations were found for BPb or Hb with SNPs, haplotypes, or diplotypes. BPb levels were higher in this region than in populations from industrialized countries but without hematotoxic effects. An impact of ALAD2 on BPb or Hb was not seen in these women.

Variation of the N-acetyltransferase 2 gene in a Romanian and a Kyrgyz population.

As part of a project on environmental disasters in minority populations, this study aimed to evaluate differences in the sequence of N-acetyltransferase 2 (NAT2) as a metabolic susceptibility gene in yet unexplored ethnicities. Eight single nucleotide polymorphisms (SNP) in the NAT2 coding region and a variant in the 3' flanking region were analyzed in 290 unrelated Kyrgyz and 140 unrelated Romanians by SNP-specific PCR analysis. The variants 341C, 481T, and 803G were less and 857A more prevalent in Kyrgyz (P < 0.0001). The variant at site 857 indicates Asian descent. 282C>T and 590G>A showed no significant variation by ethnicity. 364G>A and 411A>T turned out to be monomorphic. Database comparisons of the NAT2 minor allele frequencies support that Romanians belong to Caucasians and Kyrgyz are in between Caucasians and East Asians. The distributions of predicted haplotypes differed significantly between the two ethnicities where the Kyrgyz showed a higher genetic diversity. The haplotype without mutations was more common in Kyrgyz (40.1% in Kyrgyz, 29.3% in Romanians). Accordingly, the imputed slow acetylator phenotype was less prevalent in Kyrgyz (35.2% versus 51.4% in Romanians). We found pronounced ethnic differences in NAT2 genotypes with yet unknown effect on the health risks for environmental or occupational exposures in minority populations.

Dietary imbalance in some communities of Transylvania.

The relationship between diet intake and nutritional needs may be the most important factor which influences the health status from a nutritional point of view. Essential for maintaining health status is to keep this two constituents in balance. To evaluate the diet and nutritional status, we enrolled in an observational cross-sectional study, a population of 312 people, 202 females and 110 males, categorized by age (20-40 years, 41-65 years and over 65 years) and by physical exertion (low, medium and high). The food intake was under reference values in the case of vegetable proteins (-9.99%), carbohydrates (-24.17%) and energy (-10.29%) and over the reference values of animal fats (+24.15%) in the female population and under the reference values of carbohydrates (-24.44%) and over these values in the case of animal fats (+29.83%), total fats (+13.78%) and animal proteins (+15.02%) in the male population of the study. The investigation of the arterial pressure shows that 18.5% of females and 13.5% of males have arterial hypertension which means that for the entire study population the incidence was 16%. This data correlated with the values of body mass index which were over the normal values in 35.5% of females and 23.5% of males, and in 32.38% of the whole study population, indicated a high percent of overweight and together with the deficiency from the lipids metabolism shows the significance of the effect of the over-intake of fats, especially from animal origin, on the health status and in particular, on the cardiovascular system. On the other hand, the body mass indices were found under the low limits in 23.5% of females and in 15.3% of males, and in 21.3% of the study population, thus the correlation with the low carbohydrate and energy intake is obvious.