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Necrotizing enterocolitis (NEC) is a devastating neonatal intestinal inflammatory disease, occurring primarily in premature infants, causing significant morbidity and mortality. The pathogenesis of NEC is associated with an excessive inflammatory IL-8 response. In this study, we hypothesized that this excessive inflammatory response is related to an immature expression of innate immune response genes. To address this hypothesis, intestinal RNA expression analysis of innate immune response genes was performed after laser capture microdissection of resected ileal epithelium from fetuses, NEC patients and children and confirmed in ex vivo human intestinal xenografts. Changes in mRNA levels of toll-like receptors (TLR)-2 and -4, their signaling molecules and transcription factors (MyD88, TRAF-6 and NFκB1) and negative regulators (SIGIRR, IRAK-M, A-20 and TOLLIP) and the effector IL-8 were characterized by qRT-PCR. The expression of TLR2, TLR4, MyD88, TRAF-6, NFκB1 and IL-8 mRNA was increased while SIGIRR, IRAK-M, A-20 and TOLLIP mRNA were decreased in fetal vs. mature human enterocytes and further altered in NEC enterocytes. Similar changes in mRNA expression were observed in immature, but not mature, human intestinal xenografts. Confirmation of gene expression was also validated with selective protein measurements and with suggested evidence that immature TRL4 enterocyte surface expression was internalized in mature enterocytes. Cortisone, an intestinal maturation factor, treatment corrected the mRNA differences only in the immature intestinal xenograft. Using specific siRNA to attenuate expression of primary fetal enterocyte cultures, both TOLLIP and A-20 were confirmed to be important when knocked down by exhibiting the same excessive inflammatory response seen in the NEC intestine. We conclude that the excessive inflammatory response of the immature intestine, a hallmark of NEC, is due to a developmental immaturity in innate immune response genes.

Anopheles culicifacies sensu lato collected from Rameshwaram island, Tamil Nadu state, India was identified as species B based on the diagnostic inversion karyotype Xab 2g1+h1 as observed in polytene chromosomes. Among male mitotic karyotypes made from larval neurogonial cells, two types were observed: one with an acrocentric Y-chromosome and the other with a sub-metacentric Y-chromosome, both had sub-metacentric X and metacentric autosomes. The Rameshwaram population is identical to species B in its genetic relationship with species A and C as determined by experimental hybridizations (sterile and fertile male hybrids, respectively).

IPEX Syndrome is a rare disorder presenting in the first few months of life and characterized by a triad of severe autoimmune enteropathy (AIE), endocrinopathy (usually Type I diabetes or thyroiditis), and dermatitis. In addition, patients frequently present with other autoimmune phenomena including hemolytic anemia, thrombocytopenia, neutropenia, hepatitis, and glomerulonephropathy. The disease has been linked to mutations in the FOXP3 gene, located on the X-chromosome (Xp11.23-Xp13.3). FOXP3 encodes a 48-kD protein that is a member of the forkhead/winged-helix family of transcription factors and is essential for development of CD4+CD25+ regulatory T-cells. We have evaluated a large cohort of patients with IPEX symptoms who were referred to our lab for FOXP3 gene sequencing. In this cohort, we have identified 26 different pathogenic FOXP3 mutations in 39 families. In addition, we have identified 2 mutations that are likely non-pathogenic, as they have been found in unaffected family members, or have been identified in the general population. We have also collected a large cohort of patients with the IPEX phenotype, who do not have a FOXP3 mutation and are therefore “IPEX-like”. Combining our mutations with those reported by others in the literature, there are now a total of 55 FOXP3 mutations identified in 84 families with IPEX syndrome. Of these, 16 are splicingmutations, 26missense, 8 frameshift (including 1 premature ATG, 2 alternative ATG, and 1 premature termination), 2 in-frame deletions, 1 complex(insertion/deletion/frameshift), and 2 polyadenylation site mutations. The majority of mutations are located in the forkhead DNA-binding domain of the protein (26/55 exons 9-11). The two most common mutations are a missense mutation in exon 11 (c.1150G>A) resulting in an Ala to Thr substitution at residue 384 (p.A384T) and a 3 nucleotide inframe deletion in Exon 7 leading to a single amino acid deletion (p.251del) in the leucine zipper. These mutations were found in 11 and 6 families respectively. The remainder of the mutations are equally distributed throughout the gene and affect all exons and include mutations in other important functional domains including the upstream proline-rich region (exon 1-4) and the leucine-zipper (exons 6-7). We also describe some basic genotypephenotype correlations that are beginning to emerge as a result of the identification of more FOXP3 genemutations in IPEX.We believe that by increasing our knowledge of the molecular basis of IPEX syndrome we will better understand how specific FOXP3 mutations impact regulatory T cell development which will aid our understanding of autoimmune diseases in general.

Adult colonic mucosa is highly fucosylated relative to immature mucosa, and this fucosylation is dependent upon bacterial colonization. Toll-like receptor-4 (TLR4) was hypothesized to mediate this colonization-dependent fucosylation. Germ-free (GF) and bacteria-depleted (BD) mice were compared with conventional and mutant mice with regard to induction of epithelial fut2 mRNA levels and α1,2-fucosyltransferase (FUT2) activity. Induction of these activities by recolonization were compared with induction by LPS (TLR4 ligand) andmonocolonization by bacteriodes and E. coli strains. Cultured human erythroleukemia (Hel) cells were used to study fucosylated TLR4 In Vitro. Colonization-dependent induction of fucosylation occurs only in TLR2-/and wild-type mice but not in TLR4-/and MyD88-/mice, suggesting TLR4-MyD88 dependent signaling is important for fucosylation. TLR4 is fucosylated in the colonocytes of only GF and BD mice. LPS activate fut2 mRNA and FUT2 activity in the absence of bacterial colonization. Fucose-utilizing Bacteroides fragilis induce colonic fucosylation, but not a fucose-non-utilizing mutant. In Hel cells, which express fucosylated TLR4 activates fut2 mRNA and fucosylation. TLR4-specific siRNA inhibits this activation, and fucosidase treatment abolished TLR4-dependent activation of fut2 mRNA, confirming the central role of fucosylated-TLR4 in activating fut2 and cell surface fucosylation. Fucosylated-TLR4 expressed on uncolonized colonocytes mediates bacterially-induced activation of fut2 mRNA and FUT2 activity without activating inflammation, and is a necessary initial step in the communication between the colonizing bacteria and the host epithelium.

Forty-seven proved cases of intestinal tuberculosis admitted to the surgical ward of MKCG Medical College, Berhampur from 1985 to 1987 were subjected to laparotomy. The common pathology found were tubercles over the peritoneum, multiple strictures of intestine, ileocaecal mass, perforation of the intestines, bands and adhesions and mesenteric node involvement. The patients with acute abdomen were operated in emergency and rest as an elective procedure. Conservative surgeries like stricturoplasty, local intestinal resection, perforation closure, by-pass procedures and local ileocaecal resection were done in most of the cases and only in 2 cases right hemicolectomy was done. Biopsy was taken from the viscera, peritoneum and mesenteric nodes. Postoperative mortality was 6.4%, mostly due to toxaemia and fluid and electrolyte imbalance. Postoperative complications in most of the cases were wound infection. All were given a short course of antituberculosis regimen containing INH, rifampicin and ethambutol. Patients were followed up to one year and definite improvement was noted.

Fine structural analysis of the trophic and sexual stages of Plasmodium vivax obtained from naturally infected humans revealed that in general, the structural features as well as certain specialized functions such as haemoglobin ingestion and utilization etc., are similar to those described for other malaria parasites. Young trophozoite is characterised by less condense cytoplasm with scattering of free ribosomes and minimum amount of endoplasmic reticulum. The trophozoite grows by feeding on the host cell cytoplasm with its cytostome. Older trophozoite gets considerably enlarged and its cytoplasm appears coarsely granular due to an increase in endoplasmic reticulum and ribosomal content. The two sexual forms, the macro- and microgametocyte, can be distinguished on the basis of their fine structure. The macrogametocyte has high ribosomal density and more osmiophilic bodies than microgametocyte and possesses a compact dense nucleus with nucleolus-like region whereas the microgametocyte has large diffuse nucleus without nucleolus. Caveola-vesicle complexes and cytoplasmic clefts are observed in all erythrocytes infected with P. vivax and the incidence of these host cell alterations increases as the intraerythrocytic parasite matures.

Erythrocytic schizogony is initiated by repeated division of the nucleus in rapid sequence. Another important feature of the developing schizont is the reappearance of segments of thick inner membrane and formation of lobes around these segments. These bulbous protuberances signal the start of budding process which ends in complete segmentation of the cell. The cytoplasmic organelles which de-differentiate at the beginning of trophozoite stage are formed again. Asynchrony during merozoite formation is observed in some schizonts where fully mature merozoites are seen lying in the parasitophorous vacuole while the mother cell is still in the process of giving rise to new merozoites.

The neonatal Fc receptor FcRn functions in rodents to transfer maternal gammaglobulins (IgG) from mother to young via the neonatal intestine. In humans most of this transfer is presumed to occur in utero via the placenta. The fetus swallows amniotic fluid that contains immunogiobulins but it is unknown whether this transfer also occurs via the fetal intestine. Human FcRn has been identified in the human syncitiotrophoblast and plays a role in the materno-fetal transfer of antibody. It is also expressed in adult intestine and may be important in IgG catabolism. Objective: We hypothesize that the FcRn receptor is expressed in the human fetal intestine and may play a role in IgG transport from the amniotic fluid/colostrum into the fetal circulation. We wish to demonstrate expression of the FcRn 1) along different segments of the human fetal intestine and 2) in a human non-malignant fetal intestinal epithelial cell line (H4). Methods: Lysates were prepared from human fetal stomach, small intestine and colon from a 22 week gestation fetus and from H4 cells. Caco-2 cells and T-84 cells were used as positive controls. Prepared lysates were run on Western blots and were probed with anti-FcRn antibodies. Bound antibody was detected by HRP-conjugated secondary antibody and enhanced chemiluminescence. FcRn expression was normalized against GAPDH and quantitated by densitometry. Immunostaining of human fetal intestine, neonatal mouse intestine (+ contro[), H4 cells and T-B4 ceils (+ control), was done with a rabbit anti-human FcRn antibody and detected with the appropriate conjugated secondary antibody for microscopy. Results: The FcRn heavy chain was detected from the fetal intestine and H4 cells on Western blot analysis. On immunostaining, a subapical staining pattern for FcRn was seen on human fetal intestine. The H4 cells also stained positive for FcRn. Conclusion: FcRn expression is demonstrated in the human fetal intestine and in the human non-malignant fetal intestinal epithelial cell line (H4) suggesting that FcRn may play a role in the uptake and transport of IgG in the fetus.

Region-Specific Regulation and Cortisone Induction of Glycosyltransferases Expression in the Developing Intestine

Molecular geometries, binding energies, ionization potentials and dipole moments are calculated by the SINDO1 method for a large number of molecules containing C, N and O atoms. Comparison is made with MINDO/3, MNDO and where possible with STO-3G results. The explicit data and an error statistics show the relative merits of SINDO1.

Calcium arsenate dihydrate is precipitated at pH 7.0 and its dissolution in aqueous solutions at temperatures of 35, 40, 45 and 50°C and a pH of between 3.0 and 8.0 is investigated. The thermodynamic parameters ΔG0, ΔS0 and ΔH0 for the process are evaluated. Temperature dependence of solubility is obtained by the equations −log KTCA = 9078.138 T − 34.0468 + 0.0821421 T

Glucose tolerance tests have been performed on five patients with Huntington's chorea and no difference in response has been observed compared with seven controls. Insulin tolerance tests have been performed on 12 patients with Huntington's chorea and 10 controls. Blood samples were taken at regular intervals for 75 minutes and analysed for blood glucose, insulin, and growth hormone (HGH). There was no difference between the groups in the hypoglycaemia which developed. The patients, however, had an earlier elevation of HGH than the controls. The difference was highly significant (P less than 0.001, P less than 0.02) 30 and 35 minutes after the intravenous injection of insulin. The patients, although awake, ceased to have choreiform movements for at least the last 60 minutes of the insulin tolerance tests. Our observations of HGH release imply that hypothalamic activity is altered in Huntington's chorea. Further observations of HGH release may therefore be of value in its diagnosis.