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Tourists visiting to endemic zones may acquire Enterotoxigenic Escherichia coli (ETEC) infection resulting into diarrhea due to consumption of contaminated potable waters. In this study, a qPCR assay (SYBR Green), targeting LT1 and ST1 genes was designed to quantify ETEC in potable waters derived from civic water supply. The assay could detect lowest 1CFU/PCR targeting LT1/ST1 gene from ten-fold diluted culture of the reference strain (E. coli MTCC 723) and is ten-fold more sensitive than the conventional PCR. The quantification of the ETEC in potable waters collected from civic supply of a major city of the northern India exhibiting high flow of tourists reveals that all the sites that ran along sewage line were contaminated by the ETEC. Contamination was due to percolation of sewage. The assay could be used for the regular monitoring of potable water in places exhibiting heavy flow of tourists to prevent ETEC induced diarrhea. [Copyright &y& Elsevier]

Abstract: Pseudomonas aeruginosa is an opportunistic pathogen that can, like other bacterial species, exist in antimicrobial resistant sessile biofilms and as free-swimming, planktonic cells. Specific virulence factors are typically associated with each lifestyle and several two component response regulators have been shown to reciprocally regulate transition between biofilm-associated chronic, and free-swimming acute infections. Quorum sensing (QS) signal molecules belonging to the las and rhl systems are known to regulate virulence gene expression by P. aeruginosa. However the impact of a recently described family of novel quorum sensing signals produced by the Pseudomonas Quinolone Signal (PQS) biosynthetic pathway, on the transition between these modes of infection is less clear. Using clonal isolates from a patient developing ventilator-associated pneumonia, we demonstrated that clinical observations were mirrored by an in vitro temporal shift in isolate phenotype from a non-secreting, to a Type III cytotoxin secreting (TTSS) phenotype and further, that this phenotypic change was PQS-dependent. While intracellular type III cytotoxin levels were unaffected by PQS concentration, cytotoxin secretion was dependent on this signal molecule. Elevated PQS concentrations were associated with inhibition of cytotoxin secretion coincident with expression of virulence factors such as elastase and pyoverdin. In contrast, low concentrations or the inability to biosynthesize PQS resulted in a reversal of this phenotype. These data suggest that expression of specific P. aeruginosa virulence factors appears to be reciprocally regulated and that an additional level of PQS-dependent post-translational control, specifically governing type III cytotoxin secretion, exists in this species. [ABSTRACT FROM AUTHOR]

Abstract: An experiment was initiated in 1991 to evaluate crop productivity and define optimum tree density with advancing age of Prosopis cineraria (L.) (Khezri) in an agroforestry system. Plots of P. cineraria at densities of 417 (D 1), 278 (D 2) and 208 trees/ha (D 3) were intercropped with Vigna radiata (L.) (mungbean) in 1995, 1997, 1999 and 2000 with Pennisetum glaucum (L.) R. Br. (pearlmillet) in 1998 and 2001. Tree height and collar diameter increased by 2.5 and 2.2-fold in D 1, 2.2 and 2.4-fold in D 2 and 2.2 and 2.0-fold in D 3 plot, respectively in the 6-year period. The highest crop yields were found in D 2 plots in 1995 and 1996, in D 3 plots in 2000 and in the control plots in 2001. The lowest crop yields were found in D 1 plots throughout the duration of the experiment. Trees produced utilizable biomass of 19.1, 15.8 and 10.3tones/ha and dry leaf weight of 0.85, 0.67 and 0.50tones/ha, respectively in the D 3, D 2 and D 1 plots at the age of 12 years (June 2002). Low soil water content at 1m distance from tree base compared to that at the center of four trees indicated greater utilization of soil water within the tree rooting zone. The yield of the annual crop increased when density of P. cineraria was appropriate (i.e., optimum tree density). But optimum tree density varied with tree size/age due to competition for soil resources. Yield of the annual crops was the highest at optimum tree densities of 278 trees/ha (4m×9m) at 6 and 7 years, 208 trees/ha (8m×6m) at 10 year and <208 trees/ha at 11 years of age. The study indicated greater benefits of P. cineraria tree integrated at optimum density through tree produced and synergistic effects on the annual crops. [Copyright &y& Elsevier]

Abstract: The objective of the study was to fortify calcium in cow milk in order to prepare calcium-enriched heat-stable milk for individuals who may not ingest enough calcium to meet minimum daily requirements. Therefore, cow milk was fortified with calcium at the rate of 50mg/100ml using three salts of calcium, viz. calcium chloride, calcium lactate and calcium gluconate. Upon addition of calcium salts, there was a marked drop in the pH and heat stability. However, restoration of pH to the original value with the addition of disodium phosphate stabilized the fortified milk and enhanced its heat stability over unfortified milk. The maximum in heat stability (HCT) of calcium-fortified cow milk samples remained slightly higher than that of unfortified milk. Metabolic study on mice revealed that calcium bioavailability of cow milk fortified with calcium lactate and calcium gluconate and stabilized with disodium phosphate was slightly higher than unfortified cow milk. Fortification of cow milk with calcium and restoration of its pH resulted in a calcium to phosphorus ratio still greater than one, which is considered ideal for retention of calcium in the body. [Copyright &y& Elsevier]

The performance of the seed-metering device of a pneumatic planter was investigated under laboratory and field conditions to optimise the design and operating parameters for cottonseed planting. The effect of operational speed of the disc, vacuum pressure and shape of the entry of seed hole were evaluated by examining the mean seed spacing, precision in spacing (coefficient of variation), miss index, multiple index, and highest quality of feed index. For picking single seeds, the planter disc had a seed hole of 2·5mm in diameter. The entry cone angle of the hole was varied from 90 to 150°, the speed varied from 0·29 to 0·69m/s, and the vacuum pressure from 1 to 2·5kPa. The metering system of the planter was set to place the seeds at 250mm spacing. It was observed that the planter disc with a 120° entry cone angle gave superior performance at all speeds and operating pressures. However, there was no conclusive statistical evidence to identify a single value of disc speed or vacuum pressure. Lower miss indices were observed at higher pressures and lower speeds, and lower multiple indices at lower pressure and higher speeds. The metering system with a speed of 0·42m/s, and a vacuum pressure of 2kPa produced superior results with a feed index of 94·7% and a coefficient of variation in spacing of 8·6%, recording a mean seed spacing of 251mm. Optimisation of the regression equations incorporating speed of the disc and operating vacuum pressure through iteration further revealed that a disc, operating at speeds from 0·34 to 0·44m/s and a vacuum pressure of 2kPa, yielded similar performance. Based on the optimised operational parameters, performance indices of the pneumatic planter were determined under field condition by measuring the distribution of cotton plants spacing. A mean plant spacing of 298mm was found in the field with a 19·1% precision (coefficient of variation). Within the range of 210–300mm, 49% cotton plants were distributed compared to 88% seeds spacing distribution observed on the laboratory test rig. Displacement of seeds in the field (due to rolling and bouncing) can affect the plant spacing distribution in the field. [Copyright &y& Elsevier]

A field study was carried out on growth and biomass production of Calligonum polygonoides (L.) in different habitats to determine competitive interaction of Dactyloctenium sindicum grass for natural soil resources. Bare dune (BD), bare dune plantation (BDP), semistabilized dune plantation with D. sindicum (SDP), flatland without vegetation (FW), flatland with D. sindicum grass (FG) and flatland plantation with D. sindicum (FGP) were the habitats identified on the basis of micro-topography, presence of D. sindicum grass and plantation. Growth and biomass production of C. polygonoides were observed in relation to changes in biomass from surface vegetation, soil water content and nutrients. D. sindicum grass density and biomass were significantly (P<0.01) high in FG in 1998 whereas, in 1999 and 2000, FGP produced greater biomass. Soil organic matter, NH4–N, NO3–N, PO4–P and K were comparatively greater in the habitats with D. sindicum in 1996. All the nutrients increased in 2000 except NO3–N and the improvement was more in plantation. High (P<0.01) soil water content in FW and BD suggests that D. sindicum had extracted and utilized soil water more efficiently through its extensive root system in the remaining habitats. Soil water content increased in winter and coincided with plant senescence. Plant growth and biomass productions were significantly (P<0.05) less and grass production was greater in FGP. Contrasting result was obtained in BDP habitat in absence of D. sindicum. Difference in soil water content and nutrients seems to be the main cause for productivity differences of C. polygonoides. Lesser plant growth in FGP and SDP habitats was attributed to the competitive effect of D. sindicum grass, which seemed to have a stronger competitive effect on resource utilization. The differences in growth and biomass production accounted for the variation in D. sindicum grass density and root biomass, which was negatively correlated with growth increments and with soil water. The results indicate that surface vegetation affects the productivity of C. polygonoide altering availability of soil resources. Therefore, proper management to ensure water availability will be beneficial to enhance the productivity and the population of this species under afforestation programme. [Copyright &y& Elsevier]

Heterologus transformed Arabidopsis plants improved tolerance against abiotic stress by the modulation of glycosylation of sterols and sterol glycosides. It is due to enhanced activity of Ws SGTL1 enzyme in different stresses. Presently, the effect of WsSGTL1 gene was investigated, focusing on interaction of A. brassicicola fungus on transgenic lines. Overexpressed lines showed restricted lesion, less spore counts and few electrolyte leakages. Biochemical study suggested that the transgenic lines had more glycosylated sterol/phenolic compounds. Relative expression of JA biosynthesis gene and phenylalanine ammonia lyase ( PAL) showed enhanced expression. Photosynthesis and chlorophyll fluorescence imaging revealed that transgenic plants had minimum damage of photosynthetic apparatus because of minimum PSII destruction. [ABSTRACT FROM AUTHOR]

Endothelial dysfunction has been implicated in regional blood flow deficits after resuscitated shock. Whole body hyperthermia generates heat shock proteins (HSP) that attenuate protein denaturation and preserve cellular function. This study was performed to determine the effect of whole body hyperthermia on endothelial function and mesenteric blood flow after resuscitated hemorrhage. Sprague Dawley rats were hemorrhaged to 30% of blood volume and resuscitated with shed blood and an equal of volume of Ringer’s Lactate (Standard Resuscitation, SR). Intravital microscopy was used to determine first order arteriolar (A1) diameters and blood flow (QA1) in the neurovascularly intact ileum at baseline and 90 minutes post resuscitation (T90). Animals treated with SR and whole body hyperthermia (SR-H) were placed into a ventilated heating chamber following hemorrhage and resuscitation to achieve a temperature of 39.5–40°C for 6 hours. Identical microvascular studies were performed 18 hours later. Endothelial function was quantitated by measuring changes in vessel diameter to topical application of the endothelial dependent dilator acetylcholine (Ach, 10−5 M). A1 diameter is reported in microns and QA1 as nanoliters per second (+/− SEM). Tissue was taken from the ileum for HSP72 determination via Western analysis. This study demonstrated that whole body hyperthermia applied after resuscitated hemorrhage results in an upregulation of HSP expression that is associated with preserved endothelial function and restoration of mesenteric blood flow. These results suggest that whole body hyperthermia may serve as an adjunct to standard resuscitation in restoring blood flow and attenuating subsequent organ dysfunction.TABLE—ABSTRACT 19AchQA1HSP72BaselineSR85 ± 5161 ± 15.124 ± .1SR-H90 ± 5226 ± 17.124 ± .1T90SR48 ± 831 ± 7.241 ± .1SR-H88 ± 5&ast;157 ± 20&ast;.924 ± .2&ast;&ast;P = <0.05 vs. T90 SR for ACH, QA1 and HSP72 by Student’s t test. [Copyright &y& Elsevier]

Ghrelin, a hormone predominantly found in the stomach, was recently described as a factor that controls female reproductive function. The aim of our study was to investigate the expression and localization of ghrelin and its active receptor, growth hormone secretagogue receptor type 1a (GHS-R1a) in buffalo ovarian follicles of different follicular size and to investigate role of ghrelin on estradiol (E2) secretion, aromatase (CYP19A1), proliferating cell nuclear antigen (PCNA) and apoptosis regulator Bax gene expression on granulosa cell culture. Using real time PCR and western blot, we measured gene and protein expression of examined factors. Localization was done with immunofluorescence method. Expression of ghrelin increased with follicle size with significantly highest in dominant or pre-ovulatory follicle ( P < 0.05). Expression of GHS-R1a was comparable in medium and large follicle but was higher than small follicles ( P < 0.05). Both the factors were localized in granulosa and theca cells. Pattern of intensity of immunofluorescence was similar with mRNA and protein expression. In the in vitro study granulosa cells (GCs) were cultured and treated with ghrelin each at 1, 10 and 100 ng/ml concentrations for two days after obtaining 75–80 per cent confluence. Ghrelin treatment significantly ( P < 0.05) inhibited E2 secretion, CYP19A1 expression, apoptosis and promoted cell proliferation. In conclusion, this study provides novel evidence for the presence of ghrelin and receptor GHS-R1a in ovarian follilcles and modulatory role of ghrelin on granulosa cell function in buffalo. [ABSTRACT FROM AUTHOR]

The role of growth factors in the modulation of ovarian function is an interesting area of research in reproductive biology. Recently, we have shown the expression and role of IGF, EGF, VEGF and FGF in the follicle and CL. Here, we report the presence of Bone Morphogenetic Proteins (BMPs) and their functional receptors in the corpus luteum (CL) of buffalo. The bubaline CL was classified into four stages according to the morphology and progesterone (P4) concentration. The qPCR, immunoblot and immunohistochemistry studies revealed that BMP2 and BMP Receptors (BMPR1A, BMPR1B and BMPR2) were significantly upregulated during the mid stage whereas BMP4 and BMP7 were upregulated during the early stage of CL (P < 0.05). Studies on primary luteal cell culture (LCC) using mid CL showed a significant time and concentration dependent effect of BMP4 and BMP7 (P < 0.05). At 100 ng ml-1, the BMPs maximally stimulated the transcripts of StAR, CYP11A1 and 3bHSD that paralleled with P4 accretion in the media (P < 0.05). Further, the BMP4 as well as BMP7 upregulated the transcripts of PCNA and downregulated CASPASE3 in the LCC at the same concentration (P < 0.05). Though the combined effect of BMP4 and 7 was significantly higher (P < 0.05) than that of individual one, it was not additive. In conclusion, the expression of BMPs and their receptors were dependent on the stages of CL in the buffalo. Treatment of LCC with BMPs in vitro confirmed the presence of functional receptors that stimulated the P4 production and luteal cell survival. Moreover, the results support the concept that the upregulation of P4 and its biosynthetic pathway enzymes such as CYP11A1, StAR and 3bHSD in the CL is likely due to the autocrine and/or paracrine effects of BMP4 and BMP7 under physiological milieu. [ABSTRACT FROM AUTHOR]

Abstract: Long-term field trials using lignite fly ash (LFA) were carried out in rice crops during the period 1996–2000 at Mine I, Neyveli Lignite Corporation, Tamil Nadu. LFA, being alkaline and endowed with an excellent pozzolanic nature, silt loam texture, and plant nutrients, has the potential to improve the texture, fertility, and crop productivity of mine spoil. The rice crops were the first, third, fifth, and sixth crops in rotation. The other crops, such as green gram (second) and sun hemp (fourth), were grown as green manure. For experimental trials, LFA was applied at various dosages (0, 5, 10, 20, 50, 100, and 200t/ha), with and without press mud (10t/ha), before cultivation of the first crop. Repeat applications of LFA were made at the same dosages in treatments of up to 50t/ha (with and without press mud) before cultivation of the third and fifth crops. Press mud, a lightweight organic waste product from the sugar industry, was used as an organic amendment and source of plant nutrients. Also, a recommended dosage of chemical fertilizer, along with gypsum, humic acid, and biofertilizer as supplementing agents, was applied in all the treatments, including control. With one-time and repeat applications of LFA, from 5 to 20t/ha (with and without press mud), the crop yield (grain and straw) increased significantly (p<0.05), in the range from 3.0 to 42.0% over the corresponding control. The maximum yield was obtained with repeat applications of 20t/ha of LFA with press mud in the third crop. The press mud enhanced the yield in the range of 1.5–10.2% with various dosages of LFA. The optimum dosage of LFA was 20t/ha for both one-time and repeat applications. Repeat applications of LFA at lower dosages of up to 20t/ha were more effective in increasing the yield than the corresponding one-time applications of up to 20t/ha and repeat applications at 50t/ha. One-time and repeat applications of LFA of up to 20t/ha (with and without press mud), apart from increasing the yield, evinced improvement in the texture and fertility of mine spoil and the nutrient content of crop produce. Furthermore, some increase in the content of trace and heavy metals and the level of γ-emitters in the mine spoil and crop produce was observed, but well within the permissible limits. The residual effect of LFA on succeeding crops was also encouraging in terms of eco-friendliness. Beyond 20t/ha of LFA, the crop yield decreased significantly (p<0.05), as a result of the formation of hardpan in the mine spoil and possibly the higher concentration of soluble salts in the LFA. However, the adverse effects of soluble salts were annulled progressively during the cultivation of succeeding crops. A plausible mechanism for the improved fertility of mine spoil and the carryover or uptake of toxic trace and heavy metals and γ-emitters in mine spoil and crop produce is also discussed. [Copyright &y& Elsevier]

Objective. The aim was to determine whether human malignant ascites fluid (MAF) associated with abdominal cancer, including ovarian cancer, contained factors which inhibit angiogenesis as well as others which stimulate this process.Methods. MAF was collected from six patients, four with ovarian cancer, one with gastric cancer, and one with liver metastases. Using the chick chorioallantoic membrane (CAM) the effect of MAF on 7-day-old CAM capillaries was examined for 48 h. Vascular endothelial growth factor (VEGF) was evaluated by ELISA. Five samples of MAF were fractionated by lysine–Sepharose chromatography and the lysine-bound and -unbound fractions were eluted by ∊-amino-n-hexanoic acid. Whole MAF, the lysine-bound and -unbound fractions, and human angiostatin were subjected to SDS–PAGE/Western blot analysis and immunostained after exposure to anti-human plasminogen. Human plasminogen was exposed to conditioned medium from ovarian epithelial cancer (HEY) cells and subjected to similar Western blot analysis.Results. Despite containing VEGF, each MAF sample examined caused a loss of capillaries from the CAM; a similar response was seen using purified human angiostatin. Whole MAF and the lysine-bound fraction contained plasminogen (90 kDa) and a 55-kDa protein which migrated in a similar manner to human angiostatin on Western blot. Both the lysine-bound and -unbound fractions caused a loss of capillaries in the CAM. Human plasminogen exposed to conditioned medium from HEY cells yielded a fragment which was similar in size to angiostatin.Conclusions. MAF from patients with various clinical presentations contains angiostatin and VEGF as well as other factors which are capable of inhibiting angiogenesis. [Copyright &y& Elsevier]

• BMPs were found to be expressed in the placenta during all stages of pregnancy. • BMP4 and BMP7 stimulate trophoblast cell steroidogenesis and angiogensis. • BMP4 and BMP7 possibly act as a cytoprotective factor and promote trophoblast cell survival. The objective of this study was to document the expression and functional role of BMPs in the placental (caruncle; CAR, cotyledon; COT) during different stages of pregnancy in water buffalo. Samples collected from Early pregnancy 1 (EP1); Early pregnancy 2 (EP2), Mid pregnancy (MP), Late pregnancy (LP) while the third stage of oestrus cycle (NP) was taken as control. Also, the synergistic role of BMP4/BMP7 or combination on mRNA expression of vWF, PCNA, StAR, CYP11A1, 3βHSD, and BAX were studied in trophoblast cells cultured (TCC) during an early stage. The qPCR and immunoblotting studies revealed that BMP2, BMPR1A, BMPR1B, and BMPR2 mRNA level was significantly (p < 0.05) upregulated during early pregnancy in COTs while in CARs it was significantly upregulated (p < 0.05) during all the stages of pregnancy.BMP4 mRNA level was significantly upregulated (p < 0.05) during early pregnancy in COTs as well as in CARs. BMP6 expression was significantly upregulated (p < 0.05) during early and late stages of pregnancy. BMP7 mRNA level was upregulated (p < 0.05) during the late stage of pregnancy in COTs. At 100 ng/ml, the BMP4 maximally stimulated the transcripts of StAR, CYP11A1, and 3βHSD while BMP7 maximally stimulated the transcripts of 3βHSD that paralleled with P4 accretion in the media (P < 0.05). BMP4 as well as BMP7 upregulated the transcripts of PCNA, vWF, and downregulated BAX in the TCC (P < 0.05). In conclusion, BMPs are expressed in a regulated manner with stage-specific differences in the placenta and promotes the angiogenesis, proliferation, cell survivability, and steroidogenesis thereby regulating placental function in an autocrine/paracrine manner in water buffalo. [ABSTRACT FROM AUTHOR]