Effect of 5-(p-hydroxyphenyl)-5-phenylhydantoin (p-HPPH) enantiomers,major metabolites of phenytoin,on the occurrence of chronic-gingival hyperplasia: in vivo and in vitro study

The purpose of this study was to assess the possible role of the (R)- and (S)- enantiomers of the phenytoin metabolite p-HPPH in the pathogenesis of gingival hyperplasia (GH). About 98% of circulating p-HPPH is in the (S)-form. There were significant differences between patients with and without GH in (R)-p-HPPH level (0.055 vs 0.042 g·ml^–1), both enantiomer/racemate level ratios, and R/S enantiomeric ratio (0.0313 vs 0.0232); an increase in serum (R)-p-HPPH level was observed in patients with GH. In separate experiments, the effect of p-HPPH enantiomers on the proliferation of the normal human dermal fibroblast was studied. The in vitro study showed that (R)-p-HPPH selectively stimulated fibroblast growth. The results suggest that the least abundant metabolite, (R)-p-HPPH, is the most toxic with respect to gingival hyperplasia.     

Interleukin-1Β, interleukin-6, and growth hormone levels in human follicular fluid

Purpose To investigate possible relationships of interleukin-1 (IL-1, interleukin-6 (IL-6), and growth hormone (GH) with biochemical variables in human follicular fluid (FF) and selected in vitro fertilization (IVF) parameters.Methods A total of 67 FF samples (n=67 patients undergoing oocyte retrieval for IVF) was evaluated. IL-1, IL-6, GH, hLH, FSH, PRL, hCG, testosterone, total protein, fibrinogen, sialic acid, ₁-antitrypsin, plasminogen levels, and spectrophotometric absorbance at 458 nm were analyzed for selected FF. IL-6 and GH levels of serum and FF samples were also compared (n=23).Results Immunoreactive levels of IL-1, IL-6, and GH were detected in all FF samples. A positive correlation existed for IL-6 (r=0.5069, P=0.0161 when serum-to-FF levels were compared (concentration ratio, 11.857). Smaller-volume follicles (<4 ml) were associated with high IL-1 levels (P=0.0229, and an additional tendency of IL-1 to decrease with increasing embryo cleavage and scoring was observed. With the exception of a weak positive correlation between follicular IL-1 and testosterone levels (r=0.3128, P=0.025, no other relationship with biochemical variables or IVF parameters (etiology, e.g., endometriosis) could be implicated.Conclusions Substantially higher IL-6 levels occurred in FF compared to serum, thus supporting intrafollicular production. Interleukin- 1,IL-6, and GH levels in FF are, however, unsuitable markers for in vitro fertilization outcome.     

Oxytocin mobilizes calcium from a unique heparin-sensitive and thapsigargin-sensitive store in single myometrial cells from pregnant rats

Intracellular free Ca²⁺ concentration ([Ca²⁺]_i) was monitored using the fluorescence from the dye Fura-2-AM in single myometrial cells from pregnant rats. Oxytocin and acetylcholine applied to the cell evoked an initial peak in [Ca²⁺]_i followed by a smaller sustained rise which was rapidly terminated upon removal of acetylcholine or persisted after oxytocin removal. A Ca²⁺ channel blocker (oxodipine) and external Ca²⁺ removal decreased both the transient and sustained rises in [Ca²⁺]_i suggesting that Ca²⁺ influx through L-type Ca²⁺ channels participated in the global Ca²⁺ response induced by oxytocin. However, the initial peak in [Ca²⁺]_i produced by oxytocin was mainly due to Ca²⁺ store release: it was abolished by inclusion of heparin [which blocks inositol 1,4,5-trisphosphate (InsP ₃) receptors] in the pipette (whole-cell recording mode of patch-clamp) and external application of thapsigargin (which blocks sarcoplasmic reticulum Ca²⁺-ATPases). In contrast, the transient Ca²⁺ response induced by oxytocin was unaffected by ryanodine. Moreover, caffeine failed to induce a rise in [Ca²⁺]_i but reduced the oxytocin-induced transient Ca²⁺ response. The later sustained rise in [Ca²⁺]_i produced by oxytocin was due to the entry of Ca²⁺ into the cell as it was suppressed in external Ca²⁺-free solution. The Ca²⁺ entry pathway is permeable to Mn²⁺ ions, in contrast to that described in various vascular and visceral smooth muscle cells. Oxytocin-induced Ca²⁺ release is blocked by the oxytocin antagonist d(CH₂)₅[Tyr(Me)²,Thr⁴,Tyr-NH ₂ ⁹ ]OVT. The prolonged increase in [Ca²⁺]_i after oxytocin removal is rapidly terminated by addition of the oxytocin antagonist suggesting that oxytocin dissociation from its receptor is very slow. The oxytocin stimulation of [Ca²⁺]_i was insensitive to incubation with pertussis toxin, and blocked by a pipette solution containing anti-q/₁₁ antibody. These data show that myometrial cells possess an unique heparin-sensitive and thapsigargin-sensitive store that can be mobilized by activation of oxytocin receptors which couples with a Gq/G₁₁-protein to activate phospholipase C.     

High prevalence of type 2 diabetes in an urban settlement in Kerala, India

Background. Prevalence of type 2 or non insulin dependent diabetes mellitus is high among Indians living in India as well as abroad. Prevalence among persons of Indian origin in many countries is greater than that of people of other ethnic extraction. The Indian state of Kerala is distinguished by a high level of achievement in the health sector, characterised by both lower mortality rates and greater density of health care institutions that ensure access to most people. These attributes make the prevalence of diabetes and the pattern of its management in Kerala worth studying. Objective. To estimate the prevalence of diabetes among persons 20 years or older in an urban housing settlement in Trivandrum city, the capital of Kerala, as well as study the management of the disease in subjects affected. Design. Cross sectional survey for detecting diabetes and other chronic diseases in all willing residents of an urban housing settlement in Trivandrum, the capital city of Kerala, as part of a preventive campaign against lifestyle diseases. Fasting plasma glucose, serum triglycerides, cholesterol, height, weight and blood pressure were measured, and a detailed questionnaire administered to ascertain previous diabetic status and management. Results. Overall prevalence of type 2 diabetes is 16.3%. In the 30-64 age group, age standardised prevalence is 13.7%. Gender differences in prevalence are negligible. Greater prevalence is associated with advancing age, body mass index above 24.99, sedentary habits, serum total cholesterol > 239, serum triglycerides > 149, hypertension and smoking. Compared to non-diabetics, diabetics have greater mean and range of fasting plasma glucose values (8.87 + /-3.6 mM/l as against 4.34 + /-0.53 mM/l). 32 out of 38 diabetics among the subjects (82.4%) were already diagnosed even before the survey; of them, 89% were on medication. 3% of subjects had impaired fasting glucose, or FPG level between 110-125 mg/dl. Conclusion. Prevalence of type 2 diabetes among a group of urban residents in Trivandrum city in Kerala is very high. This is associated also with a high detection rate and compliance to treatment.     

Apolipoprotein E promotes the binding and uptake of β-amyloid into Chinese hamster ovary cells in an isoform-specific manner

The 4 allele of apolipoprotein E gene is a major risk factor for Alzheimer's disease. However, the mechanism by which the E4 isoform of apolipoprotein E increases the risk of Alzheimer's disease is poorly understood. To determine whether the isoform-specific effects of apolipoprotein E may be mediated via clearance of bound β-amyloid, we examined the uptake of β-amyloid 1–40 into Chinese hamster ovary cells in the presence or absence of the apolipoprotein E isoforms E2, E3 and E4. Apolipoprotein E2 and E3 treatments were associated with higher association of β-amyloid with cells as compared to treatment with E4. Heparin blocked the association of β-amyloid with cells, as did an antibody to one of the apolipoprotein E receptors (the low-density lipoprotein receptor-related protein).

Thus, the apolipoproteins E2 and E3, but not E4, may play important roles in the clearance of β-amyloid from the extracellular space via the low-density lipoprotein receptor-related protein.     

Immunolocalization of CD1d in human intestinal epithelial cells and identification of a beta2-microglobulin-associated form

In order to better understand the role of intestinal CD1d, we sought to define the cellular localization and further characterize the biochemical structure of CD1d in human intestinal epithelial cells (IEC). Using a CD1d-specific rabbit anti-gst-CD1d antibody, immunoprecipitation of radiolabeled cell surface proteins detected a previously identified 37 kDa protein as well as a 48-50 kDa protein which were confirmed by Western blotting with a CD1d-specific mAb, D5. Immunoprecipitation of protein lysates with the CD1d-specific mAb, D5 and 51.1.3, and the beta2-microglobulin (beta2m)-specific mAb, BBM.1, followed by N-glycanase digestion and Western blotting with the D5 mAb showed that the 48-50 kDa protein was a beta2m-associated, CD1d glycoprotein. CD1d was immunolocalized to the apical and lateral regions of native small and large intestinal IEC as defined by confocal laser microscopy using the D5 mAb and the rabbit anti-gst-CD1d antibody. In addition, a large apical intracellular pool of CD1d was identified. Identical observations were made with polarized T84 cells. Selective biotin labeling of apical and basolateral cell surfaces followed by immunoprecipitation with the D5 mAb, N-glycanase digestion and avidin blotting confirmed the presence of glycosylated CD1d on both cell surfaces and immunolocalization of the 37 kDa non-glycosylated form of CD1d to the apical cell surface. These studies show that CD1d is located in an ideal position for luminal antigen sampling and presentation to subjacent intraepithelial lymphocytes.     

Pediatric intestinal transplantation

Advancements in donor management, organ preservation and operative techniques, as well as immunosuppressive therapies, have provided children with intestinal failure and its complications a chance not only for enteral autonomy but also long-term survival through intestinal transplantation (ITx). First described in the 1960’s, experience has grown in managing these complex patients both pre- and post-transplant. The goals of this review are to provide a brief history of intestinal transplantation and intestinal rehabilitation in pediatric patients, followed by focused discussions of the indications for ITx, induction and maintenance immunosuppression therapies, common post-operative complications, and outcomes/quality of life post-transplant.     

Different pattern of risk factors for atopic and nonatopic asthma among children – report from the Obstructive Lung Disease in Northern Sweden Study

BACKGROUND: A cross-sectional study was performed among 78-year-old schoolchildren during the winter of 1996 in three municipalities in the most northern province of Sweden, Norrbotten. The study was the starting point of a longitudinal study of asthma, rhinitis, eczema, and type-1 allergy, and provided data on prevalence and risk factors for these conditions. The aim of the present study was to validate the classification of asthma based on a parental questionnaire, and to examine risk factors for atopic and nonatopic asthma. METHODS: The ISAAC questionnaire with additional questions was distributed by the schools to the parents. The response rate was 97%, and 3431 completed questionnaires were returned. The children in Kiruna and Lule? were also invited to be skin tested, and 2149 (88%) were tested with 10 common airborne allergens. A structured interview was administered by pediatricians in stratified samples of the children to test the validity of the diagnosis of asthma based on the questionnaire. RESULTS: After the validation study, the prevalence of "ever asthma" was estimated to be 8.0%. The specificity of the question, "Has your child been diagnosed as having asthma by a physician?", was high, >99%, while the sensitivity was around 70%. The strongest risk factor for "ever asthma" was a positive skin test (OR 3.9). Risk factors for asthma in the asthmatics who were not sensitized were family history of asthma, OR 3.6; breast-feeding less than 3 months, OR 1.8; past or present dampness at home, OR 1.8; smoking mother, OR 1.7; and male sex, OR 1.6. Among the sensitized asthmatics, only a family history of asthma was a significant risk factor (OR 3.0), while breast-feeding less than 3 months was not associated with an increased risk (OR 1.0). A synergistic effect between genetic and environmental factors was found especially in the nonatopic asthmatics; the children with a family history of asthma who had a smoking mother and past or present dampness at home had an OR for "ever asthma" of 13. CONCLUSIONS: Different risk-factor patterns were found for asthma and type-1 allergy. In addition, the risk factors for atopic or allergic asthma diverged from those for nonatopic asthma.