Hydrogeochemical characterization of groundwater and their associated potential health risks.

The present study assessed the human health risk exposure from the consumption of poor quality groundwater in the Lucknow area, a part of Central Ganga alluvial plain in India. Around 27 (n = 27) groundwater samples were collected from the study area. The analytical results of the samples (n = 27) collected indicate silicate and carbonate weathering is the dominant process along with cation exchange, sulfide oxidation, and reverse ion exchange. The type of groundwater is Ca2-Na-HCO3- type having all cations and anions within permissible WHO limits except for iron (Fe2+) and nitrate (NO3-). The high concentrations of Fe2 and NO3- in samples indicate the possibility of a non-geogenic point source for the same in an urban-influenced environment. The ionic concentration of dissolved constituents is used in weighted overlay analysis to generate the water quality index (WQI). WQI indicates that most urban areas (~ 98.52%) have fallen in the good to excellent category except few situated in the highly populated parts of Lucknow. The ionic concentrations of Fe2+ and NO3- have been further used to estimate human health risk by integrating regional urban population density data in Lucknow. The risk map shows alarming risks in the west-central part, where nearly ~ 35% of the total area is at moderate to high health risk.

Nanoporous carbon materials as a sustainable alternative for the remediation of toxic impurities and environmental contaminants: A review.

Due to rapidly deteriorating water resources, the world is looking forward to a sustainable alternative for the remediation of noxious pollutants such as heavy metals and organic and gaseous contaminants. To address this global issue of environmental pollution, nanoporous carbon materials (NPCMs) can be used as a one-stop solution. They are widely applied as adsorbents for many toxic impurities and environmental contaminants. The present review provides a detailed overview of the role of different synthesis factors on the porous characteristics of carbon materials, activating agents, reagent-precursor ratio and their potential application in the remediation. Findings revealed that synthetic parameters result in the formation of microporous NPCMs (SBET: >4000 m3/g; VTotal (cm3/g) ≥ 2; VMicro (cm3/g) ≥ 1), micromesoporous (SBET: >2500 m3/g; VTotal (cm3/g) ≥ 1.5; VMicro (cm3/g) ≥ 0.7) and mesoporous (SBET: >2500 m3/g; VTotal (cm3/g) ≥ 1.5; VMicro (cm3/g) ≥ 0.5) NPCMs. Moreover, it was observed that a narrow pore size distribution (0.5-2.0 nm) yields excellent results in the remediation of noxious contaminants. Further, chemical activating agents such as NaOH, KOH, ZnCl2, and H3PO4 were compared. It was observed that activating agents KОН, H3PO4, and ZnCl2 were generally used and played a significant role in the possible large-scale production and commercialization of NPCMs. Thus, it can be interpreted that with a well-planned strategy for the synthesis, NPCMs with a "tuned" porosity for a specific application, in particular, microporosity for the accumulation and adsorption of energetically important gases (CO2, CH4, H2), micro-mesoporosity and mesoporosity for high adsorption capacity for towards metal ions and a large number of dyes, respectively.

Therapy with Bone Marrow-Derived Autologous Adult Stem Cells in Quadriparesis due to Motor Neuron Disease.

OBJECTIVE: To report the safety and therapeutic effectiveness of application of concentrated bone marrow aspirate in three bedridden patients with weakness in both legs, and monitor potential improvement in neurological outcomes. DESIGN: Case report. Intervention: Five infusions of 3x108 mononuclear cells were administrated with 12 week intervals. Bone marrow (240ML) were obtained from the posterior superior iliac spine and Bone marrow mononuclear cells were enriched by standard manual close method under aseptic condition. RESULTS: During the follow-up study of one year after stem cell implantation, the conditions of all three patients were improved and were confirmed by physical assessment, muscle charting and Electromyography (EMG). One year after stem cell implantation patients who were bedridden before treatment could sit without support and walk with support up to 200 feet at a stretch. CONCLUSION: The local application of a cocktail of regenerative cell population found in an MNC fraction of bone marrow was safe and effective in improving quality of life and muscle strength in ALS patients. This case opens the need for further investigations on Autogenic stem cell transplant therapies for MND disease.

Administration of Autologous Bone Marrow-Derived Stem Cells for Treatment of Cerebral Palsy Patients: A Proof of Concept.

BACKGROUND: Stem cell therapy is a promising treatment for cerebral palsy, which refers to a category of brain diseases that are associated with chronic motor disability in children. Autologous bone marrow stem cells may be a better cell source and have been studied for the treatment of cerebral palsy because of their functions in tissue repair and the regulation of immunological processes. METHODS: To assess autologous marrow stem cells as a novel treatment for patients with moderate-to-severe cerebral palsy, a total of 10 cerebral palsy patients were enrolled in this clinical study with 24 months follow-up. A total of 10 cerebral palsy patients received autologous bone marrow cells transplantation (4.5 × 108 mononuclear cells; 90% viability) into the subarachnoid cavity and rehabilitation. RESULTS: We recorded the gross motor function measurement scores, manual ability function measurement score, and adverse events up to 24 months post-treatment. The gross motor function measurement scores were significantly higher at month 6 post-treatment compared with the baseline scores and were stable up to 24 months follow-up. The increase in manual ability and communication function measurement scores at 6 months were not significant when compared to the baseline score. All the 10 patients survived and none of the patients experienced any serious adverse events or complications. CONCLUSION: Our results indicated that bone marrow derived MNCs are safe and effective for the treatment of motor deficits related to cerebral palsy. Further randomized clinical trials are necessary to establish the efficacy of this procedure.

A quantitative assay for insulin-expressing colony-forming progenitors.

The field of pancreatic stem and progenitor cell biology has been hampered by a lack of in vitro functional and quantitative assays that allow for the analysis of the single cell. Analyses of single progenitors are of critical importance because they provide definitive ways to unequivocally demonstrate the lineage potential of individual progenitors. Although methods have been devised to generate "pancreatospheres" in suspension culture from single cells, several limitations exist. First, it is time-consuming to perform single cell deposition for a large number of cells, which in turn commands large volumes of culture media and space. Second, numeration of the resulting pancreatospheres is labor-intensive, especially when the frequency of the pancreatosphere-initiating progenitors is low. Third, the pancreatosphere assay is not an efficient method to allow both the proliferation and differentiation of pancreatic progenitors in the same culture well, restricting the usefulness of the assay. To overcome these limitations, a semi-solid media based colony assay for pancreatic progenitors has been developed and is presented in this report. This method takes advantage of an existing concept from the hematopoietic colony assay, in which methylcellulose is used to provide viscosity to the media, allowing the progenitor cells to stay in three-dimensional space as they undergo proliferation as well as differentiation. To enrich insulin-expressing colony-forming progenitors from a heterogeneous population, we utilized cells that express neurogenin (Ngn) 3, a pancreatic endocrine progenitor cell marker. Murine embryonic stem (ES) cell-derived Ngn3 expressing cells tagged with the enhanced green fluorescent protein reporter were sorted and as many as 25,000 cells per well were plated into low-attachment 24-well culture dishes. Each well contained 500 µL of semi-solid media with the following major components: methylcellulose, Matrigel, nicotinamide, exendin-4, activin ßB, and conditioned media collected from murine ES cell-derived pancreatic-like cells. After 8 to 12 days of culture, insulin-expressing colonies with distinctive morphology were formed and could be further analyzed for pancreatic gene expression using quantitative RT-PCR and immunoflourescent staining to determine the lineage composition of each colony. In summary, our colony assay allows easy detection and quantification of functional progenitors within a heterogeneous population of cells. In addition, the semi-solid media format allows uniform presentation of extracellular matrix components and growth factors to cells, enabling progenitors to proliferate and differentiate in vitro. This colony assay provides unique opportunities for mechanistic studies of pancreatic progenitor cells at the single cell level.

Characterization of an in vitro differentiation assay for pancreatic-like cell development from murine embryonic stem cells: detailed gene expression analysis.

Embryonic stem (ES) cell technology may serve as a platform for the discovery of drugs to treat diseases such as diabetes. However, because of difficulties in establishing reliable ES cell differentiation methods and in creating cost-effective plating conditions for the high-throughput format, screening for molecules that regulate pancreatic beta cells and their immediate progenitors has been limited. A relatively simple and inexpensive differentiation protocol that allows efficient generation of insulin-expressing cells from murine ES cells was previously established in our laboratories. In this report, this system is characterized in greater detail to map developmental cell stages for future screening experiments. Our results show that sequential activation of multiple gene markers for undifferentiated ES cells, epiblast, definitive endoderm, foregut, and pancreatic lineages was found to follow the sequence of events that mimics pancreatic ontogeny. Cells that expressed enhanced green fluorescent protein, driven by pancreatic and duodenal homeobox 1 or insulin 1 promoter, correctly expressed known beta cell lineage markers. Overexpression of Sox17, an endoderm fate-determining transcription factor, at a very early stage of differentiation (days 2-3) enhanced pancreatic gene expression. Overexpression of neurogenin3, an endocrine progenitor cell marker, induced glucagon expression at stages when pancreatic and duodenal homeobox 1 message was present (days 10-16). Forced expression (between days 16 and 25) of MafA, a pancreatic maturation factor, resulted in enhanced expression of insulin genes, glucose transporter 2 and glucokinase, and glucose-responsive insulin secretion. Day 20 cells implanted in vivo resulted in pancreatic-like cells. Together, our differentiation assay recapitulates the proceedings and behaviors of pancreatic development and will be valuable for future screening of beta cell effectors.

Potential role of CXCL10 in the induction of cell injury and mitochondrial dysfunction.

Chemokines have been known to play a critical role in pathogenesis of chronic pancreatitis and acinar cell death. However, the role played by one of the CXC chemokines: CXCL10 in regulation of acinar cell death has remained unexplored. Hence, this study was designed to assess the role of CXCL10 promoting apoptosis in ex vivo cultured acinar cells. Primary human pancreatic acinar cell cultures were established and exposed to varying doses of CXCL10 for different time intervals. Apoptotic induction was evaluated by both qualitative as well as quantitative analyses. Various mediators of apoptosis were also studied by Western blotting, membrane potential (Psim) and ATP depletion in acinar cells. Analysis of apoptosis via DNA ladder and cell death detection - ELISA demonstrated that CXCL10 induced 3.9-fold apoptosis when administrated at an optimal dose of 0.1 mug of recombinant CXCL10 for 8 h. Quantitative analysis using FACS and dual staining by PI-annexin showed increased apoptosis (48.98 and 53.78% respectively). The involvement of upstream apoptotic regulators like pJNK, p38 and Bax was established on the basis of their increased expression of CXCL10. The change of Psim by 50% was observed in the presence of CXCL10 in treated acinar cells along with enhanced expression of Cytochrome C, apaf-1 and caspase 9/3 activation. In addition, ATP depletion was also noticed in CXCL10 stimulated acinar cells. CXCL10 induces cell death in human cultured pancreatic cells leading to apoptosis and DNA fragmentation via CXCR3 signalling. These signalling mechanisms may play an important role in parenchymal cell loss and injury in pancreatitis.

Mitochondrial dysfunction and apoptosis of acinar cells in chronic pancreatitis.

BACKGROUND: The mechanism of acinar cell death in human chronic pancreatitis (CP) remains largely unexplored. Previous studies have demonstrated the role played by apoptosis and necrosis in experimental pancreatitis; however, their relationship with the progression of CP remains unknown. The present study was carried out to elucidate the role and extent of apoptosis in CP tissues with different histopathological scores and to examine the possible apoptotic pathway involved. METHODS: Pancreatic tissues (25 CP patients) that had been histopathologically graded (I-III) and ten normal pancreatic tissue samples were evaluated for apoptosis by DNA fragmentation and an in situ TUNEL assay. The expression of various apoptotic and antiapoptotic markers in the tissues were studied by immunohistochemistry and Western blotting. To elucidate the role of the mitochondria in acinar cell death, the mitochondrial membrane potential (DeltaPsim) and ATP levels were determined by flow cytometry and a luminometer. RESULTS: The presence of DNA fragmentation and apoptotic nuclei in all CP tissues confirmed the presence of apoptosis. The apoptotic index in CP tissue ranged from 0.09% to 0.86% +/- 0.02% and was highest in grade II (0.7 +/- 0.04%) tissues. Differential upregulation of the apoptotic mediators p53, Bax, cytochrome c, and caspase-3 and -9, and downregulation of antiapoptotic Bcl-2, was observed in CP. DeltaPsim on the order of 1.2-to 2.2-fold and ATP depletion in the range of 23%-84% in CP tissues was observed. CONCLUSIONS: Apoptosis plays an important role both in the initial stages and during the progression of CP, as evident in all tissue grades. Increased DeltaPsim, loss of ATP, and activation of caspases suggests the involvement of intrinsic pathways.

Primary culture of pancreatic (human) acinar cells.

The acinar cell culture plays a very important role in research of pancreatic pathophysiology. The aim of this study was to establish a long-term culture of human (foetal) pancreatic acinar cells in standardized nutrient media with supplements. Acinar cells were prepared from pancreatic tissues obtained from aborted foetus (> or =35 weeks) with no prior pancreatic complications by collagenase digestion and cultured using different media and supplements. The purity and phenotype of acinar cells was confirmed by various staining techniques and FACS. The acinar cell proliferation was determined at different time intervals by Bromo-deoxyuridine (BrdU) incorporation, and metabolic enzyme activity was analysed. The acini could be cultured and maintained in Ham's F-12 K/M199 media in the presence of 5% BSA, 0.1 mg/ml STI, 10 ng/ml EGF, and 10% FCS with the same morphological appearance as that of freshly prepared for 12 days with maximum viability of 80-85% and formation of monolayer without extracellular matrix. A significant BrdU incorporation of acinar cells in primary culture was observed which was maximum (105%) at day four. Higher amylase and lipase activity was seen in freshly isolated acinar cells which decreased with time of the culture. The established human pancreatic acinar cell culture may act as an excellent model to study exocrine dysfunction or pancreatitis in response to acinar cell injury.