Dynamic article: composite antropyloric valve and gracilis muscle transposition for total anorectal reconstruction: a preliminary report.

BACKGROUND: Technique and functional outcomes of anorectal reconstruction using an antropyloric graft have been reported previously. This technique had reasonable initial outcomes but lacked voluntary function. OBJECTIVE: We hereby report the initial results of patients who underwent gracilis muscle wrapping around the perineally transposed antropyloric valve in an attempt to improve voluntary fecal control. SETTING: This study was conducted at a single tertiary care institution. PATIENTS: Eight adult patients (7 men and 1 woman) with a median age of 38 years (range, 19-51 years) underwent this procedure. Seven patients already had anorectal reconstruction with a transposed antropyloric valve, and 1 patient with severely damaged anal sphincter complex underwent single-stage composite antropylorus transposition with a gracilis muscle wrap. MAIN OUTCOME MEASURES: The primary outcome measures were anatomical integrity and functional status of the composite graft in the perineum. RESULTS: No operative mortality or serious procedure-related morbidity occurred in any patient. The median postoperative resting pressure was 29 mmHg (range, 22-38 mmHg) and squeeze pressure was 72.5 mmHg (range, 45-267 mmHg). There was a significant improvement in the squeeze pressure following surgery (p = 0.039). Also, the St. Mark's incontinence scores significantly improved in all patients and varied between 7 and 9 (p = 0.003). The ability to defer defecation and the reduced frequency of leakage accidents were the prime reasons for improved postgraciloplasty outcomes in these patients. On personal interviews, all patients who underwent this procedure were satisfied with the results of their surgery. LIMITATIONS: A longer follow-up with a larger sample size is required. Quality-of-life data have not been evaluated in this study. CONCLUSIONS: Gracilis muscle wrapping around a perineally transposed antropyloric valve is possible and improves the voluntary control and overall functional outcomes in a select group of patients with end-stage fecal incontinence requiring anal replacement (Supplemental Digital Content 1, http://links.lww.com/DCR/A173).

Soil fungi for mycoremediation of arsenic pollution in agriculture soils.

AIMS: Soil arsenic (As) contamination of food-chains and public health can be mitigated through fungal bioremediation. To enumerate culturable soil fungi, soils were collected from the As-contaminated paddy fields (3-35 mg kg(-1) ) of the middle Indo-Gangetic Plains. METHODS AND RESULTS: Total 54 fungal strains were obtained and identified at their molecular level. All strains were tested for As tolerance (from 100 to 10,000 mg l(-1) arsenate). Fifteen fungal strains, tolerant to 10,000 mg l(-1) arsenate, were studied for As removal in-vivo for 21 days by cultivating them individually in potato dextrose broth enriched with 10 mg l(-1) As. The bioaccumulation of As in fungal biomass ranged from 0·023 to 0·259 g kg(-1). The biovolatilized As ranged from 0·23 to 6·4 mg kg(-1). CONCLUSIONS: Higher As bioaccumulation and biovolatilization observed in the seven fungal strains, Aspergillus oryzae FNBR_L35; Fusarium sp. FNBR_B7, FNBR_LK5 and FNBR_B3; Aspergillus nidulans FNBR_LK1; Rhizomucor variabilis sp. FNBR_B9; and Emericella sp. FNBR_BA5. These fungal strains were also tested and found suitable for significant plant growth promotion in the calendula, withania and oat plants in a greenhouse based pot experiment. SIGNIFICANCE AND IMPACT OF STUDY: These fungal strains can be used for As remediation in As-contaminated agricultural soils.

Mathematical Modeling of Oxygen Diffusion from Capillary to Tissues during Hypoxia through Multiple Points Using Fractional Balance Equations with Memory.

The diffusion of oxygen through capillary to surrounding tissues through multiple points along the length has been addressed in many clinical studies, largely motivated by disorders including hypoxia. However relatively few analytical or numerical studies have been communicated. In this paper, as a compliment to physiological investigations, a novel mathematical model is developed which incorporates the multiple point diffusion of oxygen from different locations in the capillary to tissues, in the form of a fractional dynamical system of equations using the concept of system of balance equations with memory. Stability analysis of the model has been conducted using the well known Routh-Hurwitz stability criterion. Comprehensive analytical solutions for the differntial equation problem in the new proposed model are obtained using Henkel transformations. Both spatial and temporal variation of concentration of oxygen is visualized graphically for different control parameters. Close correlation with simpler models is achieved. Diffusion is shown to arise from different points of the capillary in decreasing order along the length of the capillary i.e. for the different values of z. The concentration magnitudes at low capillary length far exceed those further along the capillary. Furthermore with progrssive distance along the capillary, the radial distance of diffusion decreases, such that oxygen diffuses only effectively in very close proximity to tissues. The simulations provide a useful benchmark for more generalized mass diffusion computations with commercial finite element and finite volume software including ANSYS FLUENT.

Production and characterization of sustainable vermimanure derived from poultry litter and rice straw using tiger worm Eisenia fetida.

Poultry litter (PL) and rice straw (RS), commonly available waste materials, pose severe threat to environment, if not properly managed. As viable waste treatment method, vermi-transformation of PL into enriched vermimanure was done using RS and cow dung (CD) with different feedstocks (FS) combinations like FS0(CD without earthworm), FS1(CD), FS2(1CD: 1RS), FS3(1CD: 1PL) and FS4(1CD: 1RS: 1PL) for 110 days. Increased growth performance (P < 0.05) of Eisenia fetida, macronutrient levels, and a consistently lower carbon-to-nitrogen ratio (C/N) emphasize the importance of RS and PL in the vermimanuring process. Several analytical techniques have revealed the presence of functional groups, nitrate (NO3-), phosphate (PO43-), and potassium ions (K+) as well as the high porosity of the matured vermimanures. Therefore, using earthworms, the feedstock FS4(1CD: 1RS: 1PL) could be successfully biotransformed into sustainable manure lowering the usage of chemical fertilizers and rice straw burning.

Membrane-damaging potential of natural L-(-)-usnic acid in Staphylococcus aureus.

The purpose of this investigation was to try to understand the antibacterial mechanism of L-(-)-usnic acid isolated for the first time from fruticose lichen Usnea subfloridana using clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC) of L-(-)-usnic acid against the clinical isolates of MRSA and reference strain S. aureus MTCC-96 (SA-96) was in the range 25-50 µg/ml. Treatment of both reference and clinical strains (MRSA-ST 2071) with four-fold MIC concentrations (100-200 µg/ml) of L-(-)-usnic acid reduced the viability of cells without damaging the cell wall. However, the loss of 260 nm absorbing material and increase in propidium iodide uptake was observed in both of the strains. Similarly, a combined effect of L-(-)-usnic acid (25-50 µg/ml) and 7.5 % NaCl resulted in a reduced number of viable cells within 24 h in comparison to the control. These observations clearly indicate that L-(-)-usnic acid exerts its action by disruption of the bacterial membrane. Further, in vivo efficacy showed that L-(-)-usnic acid significantly (p < 0.001) lowered the microbial load of spleen at doses ranging from 1 to 5 mg/kg. Further, toxicity studies in infected mice at doses 20 times higher than the efficacious dose indicated L-(-)usnic acid to be safe. Paradoxically, L-(-)usnic acid exhibited changes in serum triglycerides, alkaline phosphatase (ALKP) and liver organ weight in the healthy mice administered with only 25 mg/kg body weight. The results obtained in this study showed that natural L-(-)-usnic acid exerts its antibacterial activity against MRSA by disruption of the cell membrane. Further, the natural L-(-)-usnic acid was found to be safe up to 100 mg/kg body weight, thereby, making it a probable candidate for treating S. aureus infections.

UV-B induced differential effect on growth and nitrogen metabolism in two cyanobacteria under copper toxicity.

In the present study, impact of low (UV-B(L): 0.1 µmol m(-2) s(-1)) and high (UV-BH: 1.0 µmol m(-2) s(-1)) fluence rates of ultraviolet-B on growth and nitrogen metabolism in two cyanobacteria: Phormidium foveolarum and Nostoc muscorum under copper toxicity (2 and 5 µM) was investigated after 24 and 72 h of experiments. Copper and UV-BH treatment suppressed growth but more in N. muscorum which was accompanied by significant accumulation of Cu. Nitrate and nitrite uptake rates and activities of nitrogen assimilating enzymes i.e. nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS) and glutamate synthase (GOGAT) except glutamate dehydrogenase activity (GDH; aminating) were decreased following treatments of Cu and UV-BH, and under combined treatments the effect was greater. On contrary, UV-BL declined Cu toxicity significantly. The study concludes that Cu and UV-BH suppressed the activity of NR, NiR, GS and GOGAT (except GDH) hence decreased growth. However, UV-BL showed cross tolerance in test organisms against Cu toxicity up to certain extent. Phormidium foveolarum is comparatively less sensitive against UV-BH and excess Cu, a situation likely exists in nature, hence it may be used as a biofertilizer for sustainable agriculture.

Impact of mass drug administration on elimination of lymphatic filariasis in Surat city, India.

Lymphatic filariasis (LF) is one of the major public health problems in some of the endemic districts in India including Surat city of Gujarat province. Historical data reveals that in 1960s, Surat city had infection rate of about 23% and infectivity rate of 1.6%. Since then, Surat city has been reporting the cases of Lymphoedema and hydrocele. Filaria Control Unit was established under National Filaria Control Programme to detect and provide treatment to the cases. Based on the reports of NFCP, Surat City has been considered as LF endemic. During 2004, the country launched campaign of Elimination of Lymphatic Filariasis through Mass Drug Administration (MDA) with annual single dose of 6 mg/kg body weight of DEC tablets in all LF endemic districts including Surat city. Four rounds of MDA (2004-2007) had shown 41% reduction in mf rate, with drastic reduction in infection rate of 88% and 100% in infectivity rate. Serious adverse effect (SAE) after 4th round of MDA was insignificant (< 0.5%) during 2007. An assessment by surveying 5058 people in different parts of Surat city revealed the drug distribution coverage of more than 95% but actual drug compliance between 70-90%. Analysis of the data revealed that though the overall Microfilaria rate has been reduced due to MDA, higher Microfilaria rate was noticed in North zone of city where the migrant populations influx is higher. The observation and analysis of the data in Surat city towards elimination of Lymphatic filariasis has been discussed in this paper.

Large area multi-filamentary plasma source for large volume plasma device-upgrade.

This paper discusses the salient features and plasma performance of the newly installed Large Area Multi-Filamentary Plasma Source (LAMPS) in large volume plasma device-upgrade. The plasma source is designed to exhibit a plasma electron density of ∼1018 m-3, low electron temperature (∼eV), and a uniform plasma cross section of 2.54 m2. The directly heated LAMPS emits accelerated primary energetic electrons when it is biased with a negative discharge voltage with respect to the anode. The hairpin shaped tungsten (W) filaments, each of diameter 0.5 mm and length 180 mm, are heated to a temperature of 2700 K by feeding ∼19.5A to each filament. The LAMPS consists of 162 numbers of filaments, and it has been successfully operated with a total investment of 50 kW of electrical power. The LAMPS as a laboratory plasma source is characterized by large operational life, ease of handling, better compatibility to high pressure conditions, and advantages over other contemporary plasma sources, viz., oxide coated cathodes, RF based sources, and helicon sources, when producing plasma over large cross sections and fill volumes. Pulsed argon plasma is produced with quiescence (δnene≪1%) using LAMPS for the duration of 50 ms and a reasonably good radial uniformity (Ln = 210 cm) is achieved. Good axial uniformity is also observed over the entire length of the device. Initial measurements on plasma parameters have yielded plasma density of ∼2×1017m-3 with existing set of filaments. A plasma density of ∼1018 m-3 is envisaged for larger thickness of filaments, such as 0.75 and 1.0 mm, with the existing plasma source assembly setup.

Dissecting the transcriptome in cardiovascular disease.

The human transcriptome comprises a complex network of coding and non-coding RNAs implicated in a myriad of biological functions. Non-coding RNAs exhibit highly organized spatial and temporal expression patterns and are emerging as critical regulators of differentiation, homeostasis, and pathological states, including in the cardiovascular system. This review defines the current knowledge gaps, unmet methodological needs, and describes the challenges in dissecting and understanding the role and regulation of the non-coding transcriptome in cardiovascular disease. These challenges include poor annotation of the non-coding genome, determination of the cellular distribution of transcripts, assessment of the role of RNA processing and identification of cell-type specific changes in cardiovascular physiology and disease. We highlight similarities and differences in the hurdles associated with the analysis of the non-coding and protein-coding transcriptomes. In addition, we discuss how the lack of consensus and absence of standardized methods affect reproducibility of data. These shortcomings should be defeated in order to make significant scientific progress and foster the development of clinically applicable non-coding RNA-based therapeutic strategies to lessen the burden of cardiovascular disease.

The Forkhead factor FoxQ1 influences epithelial differentiation.

The Forkhead family of transcription factors comprises numerous members and is implicated in various cellular functions, including cell growth, apoptosis, migration, and differentiation. In this study, we identified the Forkhead factor FoxQ1 as increased in expression during TGF-ß1 induced changes in epithelial differentiation, suggesting functional roles of FoxQ1 for epithelial plasticity. The repression of FoxQ1 in mammary epithelial cells led to a change in cell morphology characterized by an increase in cell size, pronounced cell-cell contacts, and an increased expression of several junction proteins (e.g., E-cadherin). In addition, FoxQ1 knock-down cells revealed rearrangements in the actin-cytoskeleton and slowed down cell cycle G1-phase progression. Furthermore, repression of FoxQ1 enhanced the migratory capacity of coherent mammary epithelial cells. Gene expression profiling of NM18 cells indicated that FoxQ1 is a relevant downstream mediator of TGF-ß1-induced gene expression changes. This included the differential expression of transcription factors involved in epithelial plasticity, for example, Ets-1, Zeb1, and Zeb2. In summary, this study has elucidated the functional impact of FoxQ1 on epithelial differentiation.

Nucleotide variation and physical mapping of ribosomal genes using FISH in genus Tor (Pisces, Cyprinidae).

Molecular cytogenetic studies were carried out for localization of 18S and 5S ribosomal DNAs on chromosomes of three cyprinid fish species viz., T. khudree, T. mussullah and T. mosal mahanadicus using two color fluorescence in situ hybridization (FISH). All the species typically possessed 100 diploid chromosomes with minor variation in karyo-morphology. The 18S rDNA signals were observed on two pair of chromosomes in T. khudree and T. mussullah, and three pairs in T. mosal mahanadicus. The location of 18S signals also showed affinity to silver nitrate and chromomycin A3 staining. Similarly, variation in localization of 5S rDNA among the three species has been detected with the presence of FISH signals on one pair of chromosome in T. khudree and T. mussullah, and on two pairs in T. mosal mahanadicus. These molecular markers could be used as species specific markers for taxonomic identification and can further add in understanding the dynamics of genome organization and karyotypic evolution of these species. The 18S rDNA region was sequenced that generated 1811, 1810 and 1776 bp long 18S sequence in T. khudree, T. mussullah and T. mosal mahanadicus, respectively. The 18S rDNA sequence showed 95-98% identity among the subject species. Similarly, 5S sequencing generated 203 bp long fragments in these species with 100% identity in coding and 9.63% variability in non-transcribed spacer regions. The nucleotide sequence variations could be used for understanding the genetic diversity and will add new informative characters in comparative genomics. These results, in general, would enhance the value and interpretation of ecological assessment data for conservation of Tor species.

Development and quality evaluation of honey based carrot candy.

Candy was prepared with 3 different combinations of honey and carrot by using 750 g honey + 1,000 g carrot (T1), 1,000 g honey + 1,000 g carrot (T2) and 1,250 g honey + 1,000 g carrot (T3). To establish the best product, sensory evaluation was done on 9-point Hedonic scale. T1 was found to be most preferred candy. Further the T1 candy was assessed for overall quality during storage at room temperature (25-30 °C) for 6 months. Candy can be preserved safely for 6 months in both glass and LDPE packaging materials.

Heat shock protein 70-associated peptides elicit specific cancer immunity.

Vaccination of mice with heat shock protein 70 (hsp70) preparations derived from the Meth A sarcoma, but not from normal tissues, renders the mice immune to a substantial challenge with Meth A sarcoma. The immunogenicity is dose dependent and tumor specific. Treatment of an antigenically active hsp70 preparation with ATP followed by removal of low-molecular weight material leaves hsp70 intact, as judged by SDS-PAGE but results in loss of antigenicity, as judged by tumor rejection assays. Separation of this low-molecular weight material on a C18 reverse-phase column shows a diverse array of peptides with molecular mass between 1,000 and 5,000 daltons. Our data indicate that antigenicity of hsp70 preparations derives, not from hsp70 per se, but from associated peptides. These observations may suggest a novel method of using the peptide-binding property of hsp70 for specific vaccination against cancer and infectious diseases.

Calreticulin, a peptide-binding chaperone of the endoplasmic reticulum, elicits tumor- and peptide-specific immunity.

Calreticulin (CRT), a peptide-binding heat shock protein (HSP) of the endoplasmic reticulum (ER), has been shown previously to associate with peptides transported into the ER by transporter associated with antigen processing (Spee, P., and J. Neefjes. 1997. Eur. J. Immunol. 27: 2441-2449). Our studies show that CRT preparations purified from tumors elicit specific immunity to the tumor used as the source of CRT but not to an antigenically distinct tumor. The immunogenicity is attributed to the peptides associated with the CRT molecule and not to the CRT molecule per se. It is further shown that CRT molecules can be complexed in vitro to unglycosylated peptides and used to elicit peptide-specific CD8(+) T cell response in spite of exogenous administration. These characteristics of CRT closely resemble those of HSPs gp96, hsp90, and hsp70, although CRT has no apparent structural homologies to them.

T cells from late tumor-bearing mice express normal levels of p56lck, p59fyn, ZAP-70, and CD3 zeta despite suppressed cytolytic activity.

Loss of T cell-associated signal transduction molecules has recently been implicated in immune suppression in tumor-bearing hosts. In the present study, we have examined this and related phenomenon extensively in a large number of tumor-bearing mice, analyzed individually. Splenic T cells from tumor-bearing mice were isolated and characterized with respect to the following: (a) levels of three tyrosine kinases, p56lck, p59fyn, and ZAP-70; (b) expression of CD3-zeta; (c) alloreactive responses; and (d) antigen-specific responses. Contrary to recent reports, T cells from tumor-bearing mice were observed to express normal levels of lck, fyn, ZAP-70, and CD3-zeta. Further, T cells showed healthy alloreactive and antigen-specific responses until approximately 3 wk after post tumor challenge, when the tumors constituted approximately 20% of the body weight. Alterations with respect to some parameters were observed only in mice that had been bearing larger tumors for a considerably longer period. As human tumors are unlikely to grow to such large sizes (e.g., > 20% of the total body weight), the significance of the alterations in T cell expression of lck, fyn, ZAP-70, or CD3-zeta in the immune status of cancer patients is unclear. Altogether, these results indicate that alterations in T cell signal transduction molecules do not account for the profound tumor-specific suppression observed during tumor growth.

The dual nature of specific immunological activity of tumor-derived gp96 preparations.

Mice immunized with optimal doses of autologous tumor-derived gp96 resist a challenge with the tumor that was the source of gp96. Immunization with quantities of gp96 5-10 times larger than the optimal dose does not elicit tumor immunity. This lack of effect is shown to be an active, antigen-specific effect, in that immunization with high doses of tumor-derived gp96, but not normal tissue-derived gp96, downregulates the antitumor immune response. Furthermore, immunization with fractionated doses of gp96 elicits the same kind and level of response as elicited by a single dose equivalent to the total of the fractionated doses. This is true of the tumor-protective doses as well as the high downregulatory doses of gp96. The downregulatory activity can be adoptively transferred by CD4(+) but not CD8(+) T lymphocytes from mice immunized with high doses of gp96. These observations indicate that immunization with gp96 induces a highly regulated immune response that, depending upon the conditions of immunization, results in tumor immunity or downregulation.

The endoplasmic reticular heat shock protein gp96 is transcriptionally upregulated in interferon-treated cells.

A cDNA clone complementary to an interferon (IFN)-induced mRNA approximately 3 kb in length was identified and sequenced revealing homology with the endoplasmic reticular heat shock protein/ATPase gp96. Both IFN-alpha and -gamma transcriptionally upregulate expression of this gene. gp96 transcripts, protein, and ATPase activity are shown to be enhanced as a result of IFN treatment in two human cell lines and this effect requires de novo protein synthesis. gp96 molecules have recently been implicated in the presentation of endogenous antigens. A number of the key elements in this pathway, the transporter proteins, the major histocompatibility complex (MHC)-linked units of the proteasomes and the MHC class I molecules are known to be IFN inducible. Our results show that yet another molecule suggested to play an accessory role in the endogenous presentation pathway is IFN inducible. Further, our studies represent the first demonstration of modulation of expression of a heat shock protein by a cytokine and identify a new enzymatic activity upregulated in IFN-treated cells.

Heat shock protein-peptide complexes, reconstituted in vitro, elicit peptide-specific cytotoxic T lymphocyte response and tumor immunity.

Heat shock protein (HSP) preparations derived from cancer cells and virus-infected cells have been shown previously to elicit cancer-specific or virus-specific immunity. The immunogenicity of HSP preparations has been attributed to peptides associated with the HSPs. The studies reported here demonstrate that immunogenic HSP-peptide complexes can also be reconstituted in vitro. The studies show that (a) complexes of hsp70 or gp96 HSP molecules with a variety of synthetic peptides can be generated in vitro; (b) the binding of HSPs with peptides is specific in that a number of other proteins tested do not bind synthetic peptides under the conditions in which gp96 molecules do; (c) HSP-peptide complexes reconstituted in vitro are immunologically active, as tested by their ability to elicit antitumor immunity and specific CD8+ cytolytic T lymphocyte response; and (d) synthetic peptides reconstituted in vitro with gp96 are capable of being taken up and re-presented by macrophage in the same manner as gp96- peptides complexes generated in vivo. These observations demonstrate that HSPs are CD8+ T cell response-eliciting adjuvants.

Effect of soy protein isolate incorporation on quality characteristics and shelf-life of buffalo meat emulsion sausage.

Incorporation of soy protein isolate (SPI) at 0, 15, and 25% levels in buffalo meat was investigated for production, quality and shelf life evaluation of emulsion sausage (ES). Quality of ES was evaluated by pH, moisture content, thiobarbituric acid (TBA) number, total plate count (TPC), and Yeast and mold count, sensory, characteristics and instrumental colour and texture measurements. It was found that pH and moisture content were slightly affected, TBA number remained unaffected. TPC of ES fresh sample was found in the range 3.7-4.3 log cfu/g. ES was acceptable to the panelists and incorporation of SPI did not affect the acceptability. SPI incorporation increased Hunter L and b values but decreased a value and instrumental hardness. During storage (0°C), L, a, b values fl uctuated irregularly. It was concluded that incorporation of SPI slightly improved texture, juiciness and colour of emulsion sausage.

In vitro release kinetics of graft matrices from Lannea coromandelica (Houtt) gum for treatment of colonic diseases by 5-ASA.

Polyacrylamide graft Jhingan gum (Jh-g-PAMs) was synthesized adopting microwave assisted graft co-polymerization technique. The synthesized graft copolymer was characterized by various analytical techniques such as Elemental analysis, FTIR, TGA, XRD and NMR. Following standard protocol, drug matrix tablets using 5-Aminosalicylic Acid (5-ASA) were prepared and swelling and erosion studies were carried out in different pH dissolution media. The result revealed that maximum swelling and erosion took place in pH 7.4 while the lowest was recorded in pH 1.2. The 'in vitro' drug release studies revealed that grades with higher grafting % exhibited more sustained release. The highest sustained release was observed in Jh-g-PAM 3 (%G 1231) in pH 1.2 while the least was observed in native gum in pH 7.4. Furthermore, the kinetic studies revealed that 'n' values in all dissolution media lies within 0.5-1.0 which suggested non-Fickian diffusion mode of release. From the above results, it can be said that controlled release of 5-ASA using graft material was successful and hence it can be explored for treatment of colon related diseases.