Multidrug resistance among uropathogenic clonal group A E. Coli isolates from Pakistani women with uncomplicated urinary tract infections.

OBJECTIVE: Multi-drug resistance (MDR) has notably increased in community acquired uropathogens causing urinary tract infections (UTIs), predominantly Escherichia coli. Uropathogenic E. coli causes 80% of uncomplicated community acquired UTIs, particularly in pre-menopausal women. Considering this high prevalence and the potential to spread antimicrobial resistant genes, the current study was conducted to investigate the presence of clinically important strains of E. coli in Pakistani women having uncomplicated cystitis and pyelonephritis. Women belonging to low-income groups were exclusively included in the study. Seventy-four isolates from urine samples were processed, phylotyped, and screened for the presence of two Single Nucleotide Polymorphisms (SNPs) particularly associated with a clinically important clonal group A of E. coli (CgA) followed by antibiotic susceptibility testing and genome sequence analysis. RESULTS: Phylogroup B2 was most prevalent in patients and 44% of isolates were positive for the presence of CgA specific SNPs in Fumarate hydratase and DNA gyrase subunit B genes. Antibiotic susceptibility testing showed widespread resistance to trimethoprim-sulfamethoxazole and extended-spectrum beta-lactamase production. The infection analysis revealed the phylogroup B2 to be more pathogenic as compared to the other groups. The genome sequence of E. coli strain U17 revealed genes encoding virulence, multidrug resistance, and host colonization mechanisms. CONCLUSIONS: Our research findings not only validate the significant occurrence of multidrug-resistant clonal group A E. coli (CgA) in premenopausal Pakistani women suffering from cystitis and pyelonephritis but also reveal the presence of genes associated withvirulence, and drug efflux pumps. The detection of highly pathogenic, antimicrobial-resistant phylogroup B2 and CgA E. coli strains is likely to help in understanding the epidemiology of the pathogen and may ultimately help to reduce the impact of these strains on human health. Furthermore, the findings of this study will particularly help to reduce the prevalence of uncomplicated UTIs and the cost associated with their treatment in women belonging to low-income groups.

Recurrence mutation in RBBP8 gene causing non-syndromic autosomal recessive primary microcephaly; geometric simulation approach for insight into predicted computational models.

Primary microcephaly is a rare, congenital, and genetically heterogeneous disorder in which occipitofrontal head circumference is reduced by a minimum of three standard deviations (SDs) from average because of the defect in fetal brain development. OBJECTIVE: Mapping of RBBP8 gene mutation that produce autosomal recessive primary microcephaly. Insilco RBBP8 protein models prediction and analysis. METHODS: Consanguineous Pakistani family affected with non-syndromic primary microcephaly was mapped a biallelic sequence variant (c.1807_1808delAT) in the RBBP8 gene via whole-exome sequencing. The deleted variant in the RBBP8 gene in affected siblings (V:4, V:6) of primary microcephaly was confirmed by sanger sequencing. RESULTS: Identified variant c.1807_1808delAT that truncated the protein translation p. Ile603Lysfs*7 and impaired the functioning of RBBP8 protein. This sequence variant was only reported previously in Atypical Seckel syndrome and Jawad syndrome, while we mapped it in the non-syndromic primary microcephaly family. We predicted 3D protein models by using Insilco tools like I TASSER, Swiss model, and phyre2 of wild RBBP8 protein of 897 amino acids and 608 amino acids of the mutant protein. These models were validated through the online SAVES server and Ramachandran plot and refined by using the Galaxy WEB server. A predicted and refined wild protein 3D model was deposited with accession number PM0083523 in Protein Model Database. A normal mode-based geometric simulation approach was used through the NMSim program, to find out the structural diversity of wild and mutant proteins which were evaluated by RMSD and RMSF. Higher RMSD and RMSF in mutant protein reduced the stability of the protein. CONCLUSION: The high possibility of this variant results in nonsense-mediated decay of mRNA, leading to the loss of protein functioning which causes primary microcephaly.

Alleviation of Cadmium Stress by Silicon Supplementation in Peas by the Modulation of Morpho-Physio-Biochemical Variables and Health Risk Assessment.

Agricultural soil quality degradation by potentially toxic elements, specifically cadmium (Cd), poses a significant threat to plant growth and the health of humans. However, the supplementation of various salts of silicon (Si) to mitigate the adverse effect of Cd on the productivity of peas (Pisum sativum L.) is less known. Therefore, the present investigation was designed to evaluate the exogenous application at various levels (0, 0.50, 1.00 and 1.50 mM) of silicate compounds (sodium and potassium silicates) on pea growth, gaseous exchange, antioxidant enzyme activities and the potential health risk of Cd stress (20 mg kg-1 of soil) using CdCl2. The findings of the study showed that Cd stress significantly reduced growth, the fresh and dry biomass of roots and shoots and chlorophyll content. In addition, electrolyte leakage, antioxidant enzymes and the content of Cd in plant tissues were enhanced in Cd-induced stressed plants. An application of Si enhanced the development of stressed plants by modulating the growth of fresh and dry biomass, improving the chlorophyll contents and decreasing leakage from the plasma membrane. Furthermore, Si addition performed a vital function in relieving the effects of Cd stress by stimulating antioxidant potential. Hence, a significant level of metal protection was achieved by 1.00 mM of potassium silicate application under the Cd levels related to stress conditions, pointing to the fact that the Si concentration required for plant growth under Cd stress surpassed that which was required for general growth, enzymatic antioxidants regulation and limiting toxic metal uptake in plant tissues under normal conditions. The findings of this research work provide a feasible approach to reduce Cd toxicity in peas and to manage the entry and accumulation of Cd in food crops.

Effects of dibutyl phthalate and di (2-ethylhexyl) phthalate on hepatic structure and function of adult male mice.

The objective of the present research was to determine if dibutyl phthalate (DBP) and di-(2-ethylhexyl) phthalate (DEHP) alone and combined exposure induced pathological alterations in laboratory reared albino mice. Adult male mice were equally divided (n = 10) into Control, corn oil (CO), DBP, DEHP, and DBP+DEHP treated groups. Dibutyl phthalate (250 mg/kg), DEHP (300 mg/kg), and DBP+DEHP (250+300 mg/kg), respectively, were administered by oral gavage mixed in corn oil (0.2 mL) for 28 days. All animals were sacrificed following 28 days of treatment and blood was collected for serum lipid profiles and liver function tests. Liver samples were also collected for observation of histological changes. Microphotographs of hematoxylin and eosin-stained sections were used for computer-based micrometry. CO, DBP, DEHP, and DBP+DEHP treatment resulted in a significant increase in the mean body and liver weights as compared with the Control group. Histological examination of the livers with DBP and/or DEHP treatment showed marked alterations leading to hepatic hypertrophy. In the treated groups, a significant increase in the mean number of mononucleated, binucleated cells, and oval cells per unit area was noticed with disorganized trabecular arrangement as compared with the Control group. Treatment with DBP and/or DEHP resulted in large regeneration zones in the liver and an increased relative nucleo-cytoplasmic index of mononuclear shepatocytes when compared with the Control group. All treatments caused a significant increases in the liver enzymes and proteins as well as altered serum cholesterol, triglycerides, LDL, and VLDL levels. The histopathological and serological findings confirmed the toxic potentials to hepatic tissue of DBP and DEHP either given alone or in combination.

Tubulin Beta 2C Chain (TBB2C), a Potential Marker of Ovarian Cancer, an Insight from Ovarian Cancer Proteome Profile.

Ovarian cancer (OC) is the most lethal among female reproductive system malignancies. Depending upon the stage at presentation, the five year survival ratio varies from ∼92 to ∼30%. The role of biomarkers in early cancer diagnosis, including OC, is well understood. In our previous study, through an initial screening, we have analyzed eleven proteins that exhibited differential expression in OC using two-dimensional gel electrophoresis (2D-GE) and matrix-assisted laser desorption/ionization-time of flight mass spectrometric (MALDI-TOF MS) analysis. In continuation of our previous study, the present work describes analysis of twenty more proteins that showed aberrant expression in OC. Among these, six showed consistent significant deregulation in the OC false discovery rate [FDR ≤ 0.05]. Upon MS analysis, they were identified as vimentin, tubulin beta 2C chain, tubulin alpha 1C chain, actin cytoplasmic 2, apolipoprotein A-I, and collagen alpha 2(VI) chain [peptide mass fingerprint (PMF) score ≥ 79]. One of the differentially regulated proteins, tubulin beta 2C chain, was found to be significantly (fold change, 2.5) enhanced in OC. Verification by western blot and enzyme-linked immunosorbent assay (ELISA) demonstrated that the tubulin beta 2C chain may serve as a valuable marker for OC (ANOVA p < 0.0001). The assessment of the likely association of TBB2C with OC in a larger population will not only help in developing clinically useful biomarkers in the future but also improve our understanding of the progression of OC disease.

Glucuronyl C5-epimerase is crucial for epithelial cell maturation during embryonic lung development.

Glucuronyl C5-epimerase (Hsepi) is a key enzyme in the biosynthesis of heparan sulfate that is a sulfated polysaccharide expressed on the cell surface and in the extracellular matrix of alveolar walls and blood vessels. Targeted interruption of the Hsepi gene, Glce, in mice resulted in neonatal lethality, which is most likely due to lung atelectasis. In this study, we examined the potential mechanisms behind the defect in lung development. Histological analysis of the lungs from embryos revealed no difference in the morphology between wild-type and mutant animals up to E16.5. This suggests that the initial events leading to formation of the lung primordium and branching morphogenesis are not disturbed. However, the distal lung of E17.5-18.5 mutants is still populated by epithelial tubules, lacking the typical saccular structural characteristic of a normal E17.5 lung. Immunostaining revealed strong signals of surfactant protein-C, but a weaker signal of T1α in the mutant lungs in comparison to WT littermates, suggesting differentiation of type I alveolar epithelial cells (AT1) is impaired. One of the parameters contributed to the failure of AT1 maturation is reduced vascularization in the developing lungs.

Plant growth promoting rhizobacteria alleviates drought stress in potato in response to suppressive oxidative stress and antioxidant enzymes activities.

Maintenance of plant physiological functions under drought stress is normally considered a positive feature as it indicates sustained plant health and growth. This study was conducted to investigate whether plant growth-promoting rhizobacteria (PGPR) Bacillus subtilis HAS31 has potential to maintain potato growth and yield under drought stress. We analyzed trends of chlorophyll concentration, photosynthesis process, relative water content, osmolytes, antioxidants enzymes and oxidative stress, relative growth rate, tuber and aboveground biomass production in two potato varieties, Santae (drought-tolerant) and PRI-Red (drought-sensitive). Plants of both genotypes were treated with 100 g of HAS31 inoculant at 10 days after germination and exposed to different soil relative water contents (SRWC), including 80 ± 5% (well watered), 60 ± 5% (moderate stress) and 40 ± 5% SRWC (severe stress) for 7 days at tuber initiation stage (30 days after germination). The drought stress reduced plant relative growth rate, biomass production, leaf area, number of leaves and tubers, tuber weight, and final yield. The drought-stressed plants showed decline in chlorophyll contents, membrane stability, leaf relative water contents and photosynthetic rate. Under drought stress, enzymatic activity of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD), contents of total soluble sugars, soluble proteins and proline increased. The application of PGPR reduced the impact of drought and maintained higher growth and physio-chemical traits of the plants. The plants with PGPR application showed higher relative growth rate, dry matter production, leaf area, number of tubers, tuber weight and yield as compared to plants without PGPR. The PGPR-HAS31 treated plants maintained higher photosynthetic process, contents of chlorophyll, soluble proteins, total soluble sugars, and enzymatic activities of CAT, POD and SOD as compared to plants without PGPR. The results of the study suggest that plant growth regulators have ability to sustain growth and yield of potato under drought stress by maintaining physiological functions of the plants.

Significance of host heparanase in promoting tumor growth and metastasis.

Heparanase, the sole heparan sulfate degrading endoglycosidase, regulates multiple biological activities that enhance tumor growth, angiogenesis and metastasis. Much of the impact of heparanase on tumor progression is related to its function in mediating tumor-host crosstalk, priming the tumor microenvironment to better support tumor growth and metastasis. We have utilized mice over-expressing (Hpa-tg) heparanase to reveal the role of host heparanase in tumor initiation, growth and metastasis. While in wild type mice tumor development in response to DMBA carcinogenesis was restricted to the mammary gland, Hpa-tg mice developed tumors also in their lungs and liver, associating with reduced survival of the tumor-bearing mice. Consistently, xenograft tumors (lymphoma, melanoma, lung carcinoma, pancreatic carcinoma) transplanted in Hpa-tg mice exhibited accelerated tumor growth and shorter survival of the tumor-bearing mice compared with wild type mice. Hpa-tg mice were also more prone to the development of metastases following intravenous or subcutaneous injection of tumor cells. In some models, the growth advantage was associated with infiltration of heparanase-high host cells into the tumors. However, in other models, heparanase-high host cells were not detected in the primary tumor, implying that the growth advantage in Hpa-tg mice is due to systemic factors. Indeed, we found that plasma from Hpa-tg mice enhanced tumor cell migration and invasion attributed to increased levels of pro-tumorigenic factors (i.e., RANKL, SPARC, MIP-2) in the plasma of Hpa-Tg vs. wild type mice. Furthermore, tumor aggressiveness and short survival time were demonstrated in wild type mice transplanted with bone marrow derived from Hpa-tg but not wild type mice. These results were attributed, among other factors, to upregulation of pro-tumorigenic (i.e., IL35+) and downregulation of anti-tumorigenic (i.e., IFN-γ+) T-cell subpopulations in the spleen, lymph nodes and blood of Hpa-tg vs. wild type mice and their increased infiltration into the primary tumor. Collectively, our results emphasize the significance of host heparanase in mediating the pro-tumorigenic and pro-metastatic interactions between the tumor cells and the host tumor microenvironment, immune cells and systemic factors.

Upregulated BMP-Smad signaling activity in the glucuronyl C5-epimerase knock out MEF cells.

Glucuronyl C5-epimerase (Hsepi) catalyzes the conversion of glucuronic acid to iduronic acid in the process of heparan sulfate biosynthesis. Targeted interruption of the gene, Glce, in mice resulted in neonatal lethality with varied defects in organ development. To understand the underlying molecular mechanisms of the phenotypes, we used mouse embryonic fibroblasts (MEF) as a model to examine selected signaling pathways. Our earlier studies found reduced activities of FGF-2, GDNF, but increased activity of sonic hedgehog in the mutant cells. In this study, we focused on the bone morphogenetic protein (BMP) signaling pathway. Western blotting detected substantially elevated endogenous Smad1/5/8 phosphorylation in the Hsepi mutant (KO) MEF cells, which is reverted by re-expression of the enzyme in the KO cells. The mutant cells displayed an enhanced proliferation and elevated alkaline phosphatase activitywhen cultured in osteogenic medium. Analysis of the genes involved in the BMP signaling pathway revealed upregulation of a number of BMP ligands, but reduced expression of several Smads and BMP antagonist (Grem1) in the KO MEF cells. The high level of Smad1/5/8 phosphorylation was also found in primary calvarial cells isolated from the KO mice. The results suggest that Hsepi expression modulates BMP signaling activity, which, at least partially, is associated with defected molecular structure of heparan sulfate expressed in the cells.

Role of pollination in yield and physicochemical properties of tomatoes (Lycopersicon esculentum).

Very little is known about pollination and its effects on the yield and physicochemical properties of flowering plants in tropical countries. Wind and insect pollinators are among our natural resources because pollination is the most important ecosystem service performed by wind and insects, and is vital to the socio-economic status of human beings. In this experiment, different pollination methods for tomato plants were examined. Self-pollination was encouraged by covering the plants with a plastic sheet. Wind and insects were excluded from these plants, and thus only self-pollination was possible. The experiment occurred during the flowering stage. Wind-pollinated plants were covered with a muslin cloth, which excluded insects, and only wind could pass through the cloth. For insect pollination, plants remained uncovered, allowing free access to insects to pollinate the flowers. At fruit maturity, when fruits were completely red, fruits from each treatment were harvested on the same date and under the same conditions. Results illustrated the substantial importance of insects as pollinators of tomato crops. Open field had greater tomato yield and positive effects on physicochemical properties on fruit than under self and wind pollination.

Design, synthesis, in-vitro thymidine phosphorylase inhibition, in-vivo antiangiogenic and in-silico studies of C-6 substituted dihydropyrimidines.

Thymidine phosphorylase (TP) is an angiogenic enzyme. It plays an important role in angiogenesis, tumour growth, invasion and metastasis. In current research work, we study the effect of structural modification of dihydropyrimidine-2-ones (DHPM-2-ones) on TP inhibition. A series of eighteen new derivatives of 3,4-dihydropyrimidone-2-one were designed and synthesized through the structural modification at C-6 position. All these new derivatives were then assessed for in-vitro inhibition of thymidine phosphorylase (TP) from E. coli. Oxadiazole derivatives 4a-e exhibited excellent TP-inhibition at low micromolar concentration levels better than standard drug 7-deazaxanthine (7-DX). Among all these compounds, 4b was found to be the most potent with IC50 = 1.09 ±â€¯0.004 µM. Anti-angiogenesis potential of representative compounds were also studied in a chorioallantoic membrane (CAM) assay. Here again, compound 4b was found to be the potent anti-angiogenesis compound in a CAM assay. Docking studies were also performed with Molecular Operating Environment (MOE) to further analyse the mode of inhibition of these compounds. Binding mode analysis of the most active inhibitors showed that these are well accommodated into the binding site of enzyme though stable hydrogen bonding and hydrophobic interactions.

Heparin antagonizes cisplatin resistance of A2780 ovarian cancer cells by affecting the Wnt signaling pathway.

Low molecular weight heparin (LMWH), the guideline based drug for prophylaxis and treatment of cancer-associated thrombosis, was recently shown to sensitize cisplatin resistant A2780cis human ovarian cancer cells for cisplatin cytotoxicity upon 24 h pretreatment with 50 µg × mL-1 of the LMWH tinzaparin in vitro, equivalent to a therapeutic dosage. Thereby, LMWH induced sensitization by transcriptional reprogramming of A2780cis cells via not yet elucidated mechanisms that depend on cellular proteoglycans. Here we aim to illuminate the underlying molecular mechanisms of LMWH in sensitizing A2780cis cells for cisplatin. Using TCF/LEF luciferase promotor assay (Top/Flash) we show that resistant A2780cis cells possess a threefold higher Wnt signaling activity compared to A2780 cells. Furthermore, Wnt pathway blockade by FH535 leads to higher cisplatin sensitivity of A2780cis cells. Glypican-3 (GPC3) is upregulated in A2780cis cells in response to LMWH treatment, probably as counter-regulation to sustain the high Wnt activity against LMWH. Hence, LMWH reduces the cisplatin-induced rise in Wnt activity and TCF-4 expression in A2780cis cells, but keeps sensitive A2780 cells unaffected. Consequently, Wnt signaling pathway appears as primary target of LMWH in sensitizing A2780cis cells for cisplatin toxicity. Considering the outstanding role of LMWH in clinical oncology, this finding appears as promising therapeutic option to hamper chemoresistance.

Overexpression of heparanase attenuated TGF-ß-stimulated signaling in tumor cells.

Heparan sulfate (HS) mediates the activity of various growth factors including TGF-ß. Heparanase is an endo-glucuronidase that specifically cleaves and modifies HS structure. In this study, we examined the effect of heparanase expression on TGF-ß1-dependent signaling activities. We found that overexpression of heparanase in human tumor cells (i.e., Fadu pharyngeal carcinoma, MCF7 breast carcinoma) attenuated TGF-ß1-stimulated Smad phosphorylation and led to a slower cell proliferation. TGF-ß1-stimulated Akt and Erk phosphorylation was also affected in the heparanase overexpression cells. This effect involved the enzymatic activity of heparanase, as overexpression of mutant inactive heparanase did not affect TGF-ß1 signaling activity. Analysis of HS isolated from Fadu cells revealed an increase in sulfation of the HS that had a rapid turnover in cells overexpressing heparanase. It appears that the structural alterations of HS affect the ability of TGF-ß1 to signal via its receptors and elicit a growth response. Given that heparanase expression promotes tumor growth in most cancers, this finding highlights a crosstalk between heparanase, HS, and TGF-ß1 function in tumorigenesis.

A Comprehensive Review on L-Asparaginase and Its Applications.

L-asparaginase (LA) catalyzes the degradation of asparagine, an essential amino acid for leukemic cells, into ammonia and aspartate. Owing to its ability to inhibit protein biosynthesis in lymphoblasts, LA is used to treat acute lymphoblastic leukemia (ALL). Different isozymes of this enzyme have been isolated from a wide range of organisms, including plants and terrestrial and marine microorganisms. Pieces of information about the three-dimensional structure of L-asparaginase from Escherichia coli and Erwinia sp. have identified residues that are essential for catalytic activity. This review catalogues the major sources of L-asparaginase, the methods of its production through the solid state (SSF) and submerged (SmF) fermentation, purification, and characterization as well as its biological roles. In the same breath, this article explores both the past and present applications of this important enzyme and discusses its future prospects.

Therapeutic Potential of N-Acetylcysteine for Wound Healing, Acute Bronchiolitis, and Congenital Heart Defects.

BACKGROUND: Wound healing is a composite and vital process in which devitalized tissue layers and cellular structures repair themselves. Bronchiolitis is generally prompted by respiratory syncytial virus or human metapneumovirus; this condition is an acute inflammatory injury of bronchioles. Heart problems that develop before birth are known as congenital heart defects (CHDs), and pregestational diabetes is considered a major predisposing factor of CHDs. N-Acetylcysteine (NAC) is a transformed kind of amino acid cysteine which restores the intracellular levels of the natural antioxidant glutathione when taken internally, thereby assisting the cells' ability to diminish the damaging effects of reactive oxygen species (ROS). OBJECTIVE: In the present communication, NAC's therapeutic potential for wound healing, acute bronchiolitis, and congenital heart defects (CHDs) is critically analyzed by reviewing its effect on the various targets of these diseases. The multifunctional nature of NAC is outlined in a review of evidence from in vitro and in vivo studies. CONCLUSION: In conclusion, NAC could be used as a therapeutic agent in the treatment of wound healing, acute bronchiolitis and congenital heart defects (CHDs). The focus of future research should be the following; (1) to examine NAC clinically to be considered in the treatment of wound healing; (2) to investigate whether NAC could be used alone or with insulin to prevent CHDs in infants with pregestational diabetes; (3) to evaluate the application of NAC as a potential agent for PAH treatment.

Mucinous borderline ovarian tumor with ascites.

Borderline mucinous tumors are epithelial ovarian tumors with low rate of growth and low potential to invade or metastasize and associated with significantly better prognosis and excellent disease-free survival after surgical removal than other epithelial ovarian cancers. The accepted initial treatment is surgical removal of the tumor. Fertility-sparing surgery may suffice in young patients with tumors confined to the ovary. Radical surgery is recommended in patients with advanced disease and advanced age. Long-term surveillance is recommended to document and treat late recurrences. We report a case of a 59 years old postmenopausal patient with complex ovarian mucinous tumor and gross ascites; she had received three lines of chemotherapeutic agents pre-operatively, without any favorable outcome. Then, she went for staging laparotomy and histopathology showed borderline ovarian mucinous tumor required no further treatment and is fine till date.

Serum zinc levels and effects of oral supplementation in pregnant women.

OBJECTIVE: To evaluate the effects of oral zinc supplementation on the serum zinc levels of pregnant women. DESIGN: Experimental (double blinded randomized controlled trial). PLACE AND DURATION OF STUDY: PIMS and KRL Hospital, Islamabad, and community in tehsil Kahuta from April 2003 to April 2004. PATIENTS AND METHODS: Pregnant women of 10 to 16 weeks gestation were invited to enter the study on their booking visit. A sample size of 125 in each group was calculated. After taking an informed consent, they were assigned to control or test group by simple random sampling technique. A detailed questionnaire was filled-up by trained staff and initial evaluation along with serum zinc samples was collected. The subjects were given either zinc sulphate powder, equivalent to 20 mg elemental zinc, or were given placebo treatment along with routine supplements. These patients were followed up throughout the pregnancy by health care providers and their compliance was monitored. At delivery, serum samples were again collected for zinc estimation. The data was entered on computer, cleaned and analyzed. Paired t-test was used for comparison of means. RESULTS: The data of 242 subjects was analyzed at the end of the study. The mean age of the study participants was 25.7 +/- 4.8 years (range 16 to 40). Both the groups were similar in other demographic variables as socioeconomic status, education, BMI, height and weight. One-third of the patients had serum zinc levels below 64 microg/dl at the start of the study. A 128 pairs of pre and post-serum zinc levels were analyzed in the two groups (64 pairs in each group) to compare the means. The zinc supplemented women showed a mean increase of 14.7 microg/dl (95% CI 5-23) (P = 0.002). On the other hand the non-supplemented group showed an actual decrease in the serum zinc level which, however, did not reach statistical significance (P = 0.47). CONCLUSION: Oral zinc supplementation of pregnant women with 20 mg elemental zinc was effective in raising the serum levels of zinc. It is suggested that supplementation trials with larger dose of zinc should be carried out.