FUT2 non-secretor status is associated with Type 1 diabetes susceptibility in Japanese children.

AIM: To examine the contribution of the FUT2 gene and ABO blood type to the development of Type 1 diabetes in Japanese children. METHODS: We analysed FUT2 variants and ABO genotypes in a total of 531 Japanese children diagnosed with Type 1 diabetes and 448 control subjects. The possible association of FUT2 variants and ABO genotypes with the onset of Type 1 diabetes was statistically examined. RESULTS: The se2 genotype (c.385A>T) of the FUT2 gene was found to confer susceptibility to Type 1A diabetes in a recessive effects model [odds ratio for se2/se2, 1.68 (95% CI 1.20-2.35); corrected P value = 0.0075]. CONCLUSIONS: The FUT2 gene contributed to the development of Type 1 diabetes in the present cohort of Japanese children.

Variants associated with autoimmune Type 1 diabetes in Japanese children: implications for age-specific effects of cis-regulatory haplotypes at 17q12-q21.

AIMS: The aim of this study was to clarify the significance of previously reported susceptibility variants in the development of autoimmune Type 1 diabetes in non-white children. Tested variants included rs2290400, which has been linked to Type 1 diabetes only in one study on white people. Haplotypes at 17q12-q21 encompassing rs2290400 are known to determine the susceptibility of early-onset asthma by affecting the expression of flanking genes. METHODS: We genotyped 63 variants in 428 Japanese people with childhood-onset autoimmune Type 1 diabetes and 457 individuals without diabetes. Possible association between variants and age at diabetes onset was examined using age-specific quantitative trait locus analysis and ordered-subset regression analysis. RESULTS: Ten variants, including rs2290400 in GSDMB, were more frequent among the people with Type 1 diabetes than those without diabetes. Of these, rs689 in INS and rs231775 in CTLA4 yielded particularly high odds ratios of 5.58 (corrected P value 0.001; 95% CI 2.15-14.47) and 1.64 (corrected P value 5.3 × 10-5 ; 95% CI 1.34-2.01), respectively. Age-specific effects on diabetes susceptibility were suggested for rs2290400; heterozygosity of the risk alleles was associated with relatively early onset of diabetes, and the allele was linked to the phenotype exclusively in the subgroup of age at onset ≤ 5.0 years. CONCLUSIONS: The results indicate that rs2290400 in GSDMB and polymorphisms in INS and CTLA4 are associated with the risk of Type 1 diabetes in Japanese children. Importantly, cis-regulatory haplotypes at 17q12-q21 encompassing rs2290400 probably determine the risk of autoimmune Type 1 diabetes predominantly in early childhood.

Consumption of a single serving of red raspberries per day reduces metabolic syndrome parameters in high-fat fed mice.

Using an animal model for diet-induced metabolic disease, we have shown previously that the addition of raspberry juice concentrate (RJC) and raspberry puree concentrate (RPC) at a level of 10% of kcal, equivalent to four servings per day, to an obesogenic high-fat, western-style diet (HF) significantly reduced body weight gain, serum resistin levels, and altered the expression of hepatic genes related to lipid metabolism and oxidative stress. This study was designed to examine the effect of a lower level of RJC or RPC consumption, at a level representing a single serving of food per day (2.5% of kcal). For ten weeks, four groups of C57BL/6J mice (n = 8 ea.) were fed: low fat (LF), HF, HF + RJC, or HF + RPC diets. Intake of RJC and RPC decreased final body weight. Hepatic lipid accumulation was significantly decreased in HF + RPC- and HF + RJC-fed mice, compared to HF-fed mice. Further, the relative expression of hepatic genes including Heme oxygenase 1 (Hmox1) and Hormone sensitive lipase (Lipe), were altered by RPC or RJC consumption. In this mouse model of diet-induced metabolic disease, consumption of the equivalent of a single daily serving of either RPC or RJC improved metabolism in mice fed HF diet. We hypothesize that the phytochemicals contained in raspberries, and/or their subsequent metabolites, may be acting to influence gene expression and other regulatory pathways, to produce the metabolic improvements observed in this study.

Estrogen-prolactin dependency in 7,12-dimethylbenz(a)anthracene-induced tumors.

Hormonal influences on dimethylbenz(a)anthracene-induced tumor growth were investigated in detail by endocrine ablation and replacement of hormones. The majority of tumors regressed following ablation and most of them were reactivated by subsequent administrations of estrogen (0.1 to 5 mug) or prolactin (2 mg). Increasing numbers of tumors, however, were not stimulated by prolactin when administration was delayed, and a basal level of estradiol (0.01 mug) in addition to prolactin was required for reactivation of tumors. Nafoxidine hydrochloride, a competitor of estrogen at the receptor sites, arrested growth of a large portion of dimethylbenz(a)anthracene-induced tumors in intact animals but failed to retard growth of prolactin-stimulated tumors. On withdrawal of prolactin-nafoxidine, rapid regression of tumor occurred and readministration of prolactin failed to activate most of the tumors for as long as 28 days. Our results give good supporting evidence that estrogen plays a primary role in tumor growth. The interactions of prolactin and estrogen at tumor sites are necessary for regulatory events related to tumor growth.

On the mechanism of prolactin and estrogen action in 7,12 dimethylbenz(A)anthracene-induced mammary carcinoma in the rat. II. In vivo tumor responses and estrogen receptor.

In order to test the in vivo effect of prolactin on estrogen receptor (ER) binding capacity in tumors induced by 7,12 dimethylbenz(a)anthrancene (DMBA-tumor), growth of the tumors from changes in prolactin and estrogen levels was compared retrospectively with cytoplasmic ER levels. It was demonstrated that some tumors required prolactin, some needed prolactin-estrogen during their growth period anda small number were not influenced by hormonal milieu. ER was present in hormonally dependent tumors but was low or absent in hormonaly-independent tumors. Deletion of hormones by endocrine ablation in the host rat resulted in tumor regression loss of ER. Replenishment of ER and subsequent tumor growth were accomplished by injection of prolactin or prolactin-estrogen in endocrine ablated rats but were not achieved in rats bearing tumors exposed to prolactin-nafoxidine. Our results demonstrate that both estrogen and prolactin were essential for growth of hormonally dependent DMBA-tumors. Tumor growth was also prevented when cytoplasmic ER was not replenished , indicating that ER may be an indispensable prerequisite for growth. Prolactin, independently of or cooperatively with estrogen, stimulated ER binding capacity. These results support the hypothesis that there may exist a prolactin regulatory mechanism of estrogen action at the tumor site. The interactions of estrogen and prolactin in situ in modulating hormonal receptor binding capacities may contribute to the overall stimulatory effect of these two hormones on DMBA-tumors.

Micropenis and the AR Gene: mutation and CAG repeat-length analysis.

Various mutations of the AR gene and expanded CAG repeats at exon 1 of that gene have been reported in patients with hypospadias or genital ambiguity. However, the role of the AR gene has not been systemically studied in those with isolated micropenis lacking hypospadias or genital ambiguity. We studied 64 Japanese boys with isolated micropenis (age, 0-14 yr; median, 7 yr), whose stretched penile lengths were between -2.5 and -2.0 SD (borderline micropenis) in 31 patients (age, 0-13 yr; median, 8 yr) and below -2.5 SD (definite micropenis) in 33 patients (age, 0-14 yr; median, 6 yr). Mutation analysis of the AR gene was performed for exons 1-8 and their flanking introns, except for the CAG and GGC repeat regions at exon 1, by denaturing HPLC and direct sequencing, identifying a substitution of cytosine to thymine at a position -3 in the 3' splice site of intron 1 in a patient with definite micropenis. CAG repeat length at exon 1 was determined by electrophoresis with internal size markers and direct sequencing, revealing no statistically significant difference in the distribution of CAG repeat lengths [median (range) and mean +/- SE: total patients with isolated micropenis, 24 (14-34) and 23.5 +/- 0.38; patients with borderline micropenis, 24 (15-29) and 23.5 +/- 0.53; patients with definite micropenis, 23 (14-34) and 23.5 +/- 0.56; and 100 control males, 23 (16-32) and 23.5 +/- 0.29] or in the frequency of long CAG repeats (percentage of CAG repeats > or =26 and > or =28: total patients with isolated micropenis, 17.2 and 4.7%; patients with borderline micropenis, 19.4 and 6.5%; patients with definite micropenis, 15.2 and 3.0%; and 100 control males, 21.0 and 10.0%). These results suggest that an AR gene mutation is rare and that CAG repeat length is not expanded in children with isolated micropenis.

Nucleotide sequence and genomic structure of duck acyl-CoA binding protein/diazepam-binding inhibitor: co-localization with S-acyl fatty acid synthase thioesterase.

A computer-aided homology search of the GenBank nucleotide database using the amino acid sequence of human acyl CoA-binding protein (ACBP)/diazepam-binding inhibitor (DBI)-endozepine as a probe revealed that a genomic fragment containing the gene encoding the mallard duck (Anas platyrhynchos) S-acyl fatty acid synthase thioesterase also contains sequences which encode the duck homolog of ACBP/DBI. The duck ACBP/DBI gene is positioned downstream of the thioesterase gene in a tail-to-tail orientation separated from the 3' end of the thioesterase gene by only several hundred nucleotides. Three exons were identified that have strong homology to the published cDNA sequences of human and bovine ACBP/DBI. These exons define all of the coding region except for the amino-terminal domain, which was subsequently cloned by polymerase chain reaction (PCR) amplification. The encoded amino acid sequence of the duck ACBP/DBI is 62-68% homologous to mammalian ACBP/DBI sequences. While the mammalian ACBP/DBI is expressed mainly in the liver, with smaller amounts in the brain and heart, mRNA transcripts of duck ACBP/DBI were detected only in the brain with no evidence for expression in the liver or heart. The close proximity of the genes for ACBP/DBI and S-acyl fatty acid synthase thioesterase raises the possibility of co-regulation of expression.

Effect of cardioplegic arrest and reperfusion on coronary reserve and autoregulation.

BACKGROUND: The effects of cardioplegic arrest and reperfusion on the coronary vasculature remain to be characterized. This study was designed to investigate changes in coronary reserve and autoregulation after hypothermic cardioplegic arrest and reperfusion. METHODS: Isolated rabbit hearts were perfused in a retrograde manner with Krebs-Henseleit bicarbonate buffer solution at a pressure of 80 cm H2O. Baseline measurements were performed for (1) coronary flow; (2) vasodilatory response to 5-hydroxytryptamine (10(-7) mol/L) and nitroglycerin (10(-4) mol/L); (3) autoregulatory capacity, quantified as closed-loop gains; and (4) isovolemic left ventricular function. Hearts were then subjected to cardioplegic arrest for 90 minutes. Twenty minutes after reperfusion, measurements were repeated. RESULTS: Coronary flow decreased significantly after reperfusion (6.2 +/- 1.1 versus 5.3 +/- 1.1 mL.min-1.g-1; p < 0.01). The response to 5-hydroxytryptamine as percentage increase of flow decreased significantly after reperfusion (134.0% +/- 12.0% versus 109.1% +/- 6.8%; p < 0.01). However, there was no significant change in the response to nitroglycerin after reperfusion (121.3% +/- 17.6% versus 136.6% +/- 13.3%). The closed-loop gain demonstrated negative values before arrest but became positive after reperfusion, indicating loss of autoregulation after reperfusion. There was no significant change in left ventricular function. CONCLUSIONS: The coronary flow reserve in response to 5-hydroxytryptamine and autoregulation were impaired after cardioplegic arrest and reperfusion, whereas nitroglycerin-induced vasodilatory response and left ventricular function were preserved.

Evaluation of autonomic nervous system function with spectral analysis of heart rate variability in a case of tetanus.

The autonomic nervous system is affected in a wide variety of neurological disorders. Its dysfunction may play an important role in the clinical course and may result in serious complications, such as cardiac arrest. We report a case of tetanus who presented with severe autonomic nervous system dysfunction which was detected by spectral analysis of heart rate variability monitored over 24 h. This is a semi-quantitative method for evaluation of the status of the autonomic nervous system. In the present case, the analysis revealed profoundly decreased activity of both sympathetic and parasympathetic nervous system modulation of cardiac rhythm. The parasympathetic nervous system activity was more severely impaired than that of the sympathetic nervous system. The relative predominance of the sympathetic nervous system in the present case may have resulted in unopposed sympathetic nervous system hyperactivity manifested in this patient by tachycardia and excessive sweating. We further infer that the documented diminished buffering capacity of the autonomic nervous system may have lead to a sudden cardiac arrest in our case. Thus, spectral analysis of heart rate variability is a non-invasive and sensitive method for evaluating the status of the autonomic nervous system of critically ill patients in the hospital setting.

Intraoperative sustained limited expansion (ISLE) as an immediate reconstructive technique.

Intraoperative sustained limited expansion (ISLE) has provided ideal tissue replacement for coverage of small defects with an acceptably low complication rate. The ISLE technique may have more general applicability to the reconstructive surgeon than does chronic expansion because of the frequent need for additional local tissue at the time of surgery.

Tissue expansion in reconstruction of acquired auricular defects.

Over the past 10 years, tissue expansion has gradually become accepted as a useful adjunct to ear reconstruction with an autologous framework. The advantage of expanded tissue in these cases has been the addition of more tissue with similar color, texture, and sensation. In traumatic cases, the creation of well-vascularized tissue allows an improved anterior draping of skin over the cartilaginous framework and production of a well-defined posterior sulcus. Encroachment of hair onto the reconstructed ear is minimized because of the extra tissue produced by expansion. Under favorable conditions, tissue expansion is an acceptable and reliable method for partial or total ear reconstruction.

Noninvasive monitoring of tissue viability.

Noninvasive monitoring techniques are procedures of little or no morbidity that may be repeated frequently to assess tissue viability. The ideal noninvasive technique would be safe, sensitive, reliable, reproducible, simple to use, and inexpensive. Although no such ideal technique yet exists, several currently available methods have clinical usefulness. Visual fluorescein, fluorescein photography, and surface fluorometry all work by the same principle. We prefer the surface fluorometer because of its greater sensitivity, the more objective nature of the data it provides, and the greater frequency with which it can be repeated. Ultrasound Doppler techniques are useful preoperatively in locating vessels and assessing their patency, and we have found the bidirectional Doppler quite helpful in the postoperative monitoring of replanted or revascularized digits. The laser Doppler, photoplethysmograph, and surface thermometer have not been as helpful in our experience. Transcutaneous oxygen measurement shows great promise for the postoperative monitoring of replants and free tissue transfers.

Pharmacologic manipulation of the microcirculation in cutaneous and myocutaneous flaps in pigs.

The vascular effects of isoxsuprine, diazoxide, and isoproterenol were studied in arterial buttock flaps and latissimus dorsi myocutaneous flaps in pigs. Capillary blood flow to the skin and muscle of these flaps was measured by the radioactive microsphere (15-mu diameter) technique 6 hours postoperatively under pentobarbital anesthesia. It was observed that isoproterenol, a beta-adrenergic receptor agonist, was not effective in augmentation of skin blood flow in the arterial buttock flaps. However, isoproterenol significantly increased capillary blood flow to the arterialized portion of latissimus dorsi myocutaneous flaps compared with controls. Isoxsuprine and diazoxide (vascular smooth muscle relaxants) significantly (p less than 0.05) increased total capillary blood flow to the skin of arterial buttock flaps and to the skin and muscle of the latissimus dorsi myocutaneous flaps. However, the increase in capillary blood flow occurred mainly in the arterialized portion of these flaps. The capillary blood flow, which was supplied by the small arteries in the distal portion of the arterial buttock and latissimus dorsi flaps, was not increased by treatment with isoxsuprine or diazoxide. Therefore, there was also no increase in the maximum distance of capillary blood flow from the pedicle to the distal end of the flaps. These observations led us to hypothesize that different sizes (diameters) of arteries in the skin and muscle have different reactivity (or sensitivity) to vasodilatory drugs. In the present experiment, the large dominant artery of the arterial buttock and latissimus dorsi flaps responded to isoxsuprine or diazoxide (vascular smooth-muscle relaxants), resulting in an increase in blood supply to the capillaries in the proximal portion of the flaps. On the other hand, the small arteries in the distal portion (random portion) of these flaps were not sensitive to isoxsuprine or diazoxide. Therefore vasodilation did not occur, and there was no increase in blood supply to the capillaries in the distal portion of arterial buttock and latissimus dorsi flaps. This explanation also lent support to our previous report that treatment with isoxsuprine did not augment skin viability in the distal portion of arterial buttock and latissimus dorsi flaps. It is suggested that research in pharmacologic manipulation of skin blood flow and viability in skin flap surgery should emphasize the sensitivity of small arteries to various drug actions.(ABSTRACT TRUNCATED AT 400 WORDS)

Pathophysiology of skin flaps raised on expanded pig skin.

The pathophysiology of skin flaps raised on nonexpanded and expanded skin was studied in pigs. Skin expansion was achieved by gradual inflation of a subcutaneously implanted silicone tissue expander with sterilized isotonic saline over a period of 4 to 5 weeks. Comparisons were made of the skin flap viability and capillary blood flow among random acute and delayed skin flaps and skin flaps constructed on skin pockets implanted with noninflated or inflated tissue expanders. The extents of skin flap viability and capillary blood flow were similar among delayed skin flaps and skin flaps constructed on skin pockets but were significantly higher than those of acute random skin flaps raised on normal skin. It was concluded that skin viability and capillary blood flow in skin flaps raised on expanded skin were not compromised and the regulation of blood flow in skin flaps raised on nonexpanded or expanded skin pockets was similar to that seen in delayed skin flaps. The mechanisms of the delay phenomenon was discussed.

Hemodynamics and viability of acute neurovascular island skin flaps in rats.

Hemodynamics and skin survival were studied in acute 8 X 8 cm2 neurovascular island skin flaps in rats. Maximum blood flow and extent of fluorescein and 15 mu microsphere penetration in the skin flap occurred at about 24 hours after flap construction. However, maximum A-V shunt flow occurred at about 12 hours. It is concluded that ischemia in the acute skin flap, probably due to vasoconstriction, did occur in the early stage of the postoperative period. The opening of A-V shunts and vasodilation of the capillary bed in the later stage may be controlled by the same mechanism(s). It is also suggested that treatment which results in vasodilation in the skin flap in the early stages of the postoperative period may augment the extent of skin viability.

Experimental evidence for involvement of prostaglandins in viability and acute skin flaps: effects on viability and mode of action.

Treatment with prostaglandin synthesis inhibitors alone or combined with exogenous PGE2 significantly (p less than 0.05) increased skin viability in acute island skin flaps. Increased cutaneous blood flow was observed 15 minutes after skin flap construction in rats injected with ibuprofen (5 mg/kg) intraarterially before surgery compared with that in control rats. It is hypothesized, therefore, that prostaglandins may influence skin viability and the microcirculatory level.

Pathogenesis and treatment of infant skin strawberry hemangiomas: clinical and in vitro studies of hormonal effects.

Corticosteroid treatment of infant strawberry hemangiomas produced premature regression of growing lesions in patients less than a year of age, but not in patients with cavernous or port-wine hemangiomas. Abnormally elevated serum estradiol-17 beta levels were found in strawberry hemangiomas, fourfold higher than in control, cavernous, and port-wine hemangiomas which were adjusted for age and sex of the patients. Specific estradiol-17 beta receptor binding activity was studied in biopsy tissues obtained from normal prepubertal skin and skin from various kinds of hemangiomas. Minimal specific estradiol-17 beta binding activity was detected in tissues of normal skin and involuting strawberry, cavernous, or port-wine hemangiomas. Abnormally high levels of specific estradiol-17 beta binding sites were demonstrated by receptor assays and by in vitro tissue culture technique in nine tissue samples obtained from strawberry hemangiomas, seven of which responded definitely or probably to corticosteroid therapy. The in vitro estradiol-17 beta binding activity in these tissue explants was inhibited by low (5 micrograms/ml) and high (100 micrograms/ml) doses of cortisone. The present data seem to suggest that there may be a causal relationship between the presence of elevated serum E2 and specific estradiol-17 beta receptors in the pathogenesis of strawberry hemangiomas and in response to corticosteroid treatment of the hormone-sensitive hemangiomas.

Photoradiation therapy of cutaneous angiosarcomas in mice.

Cutaneous angiosarcoma is a relatively rare but devastating malignant vascular tumor. It has a high incidence of recurrence following conventional therapeutic modalities applied either singly or in combination. The increased vascularity of cutaneous angiosarcomas, facilitating selective uptake and retention of a photosensitizing agent, such as hematoporphyrin derivative (HPD), suggests that these tumors would respond well to photoradiation therapy. To study the feasibility of this treatment modality, transplantable hemangiosarcomas were implanted in B6C3F1 female mice. Within 2.5 to 3.5 hours after intraperitoneal administration of HPD, fluorescence was recorded in the tumor as compared with surrounding normal skin. When these photosensitized tumors were exposed to 70 J/cm2 of laser energy from an argon-pumped dye laser at 630 nm, the tumors showed marked necrosis within 24 hours. In another series, the tumors were initially photosensitized with HPD for 3 hours and then treated with laser energy ranging from 0 to 96 J/cm2. A dual labeling procedure demonstrated a dose-related decrease in DNA synthesis rate in tumors that were exposed to 0 to 30 J/cm2 at 24 hours after treatment. Furthermore, tumor tissue exposed to laser energy in excess of 30 J/cm2 showed no significant cellular DNA synthesis. These data, supported by histologic evidence of tissue destruction, suggest that photoradiation therapy has a great potential as a therapeutic modality for cutaneous angiosarcomas.

Quantitative evaluation of mast cells in cellularly dynamic and adynamic vascular malformations.

Mast cells were counted in 78 histologic specimens from 70 patients with various vascular malformations showing cellularly dynamic and cellularly adynamic lesions. In growing stages of strawberry hemangiomas, there was an increased number of mast cells (mean 11.0 cells per high-power field in stage III and 23.7 in stage IV), as well as a high number of mast cells in the initial involution of strawberry hemangiomas (stage V, mean 21.0 cells per high-power field). In later involuting stages (stages VI and VII), the number of mast cells decreased (mean 9.3 in stage VI; mean 4.7 in stage VII). In cellularly adynamic lesions, i.e., port wine stains, the mean number of mast cells was 4.8, and in congenital arteriovenous malformations, it was 3.6. In normal skin, the mean number of mast cells was 3.2. In cellular hemangiomas that showed active growth (stages III to IV), the number of mast cells was strikingly low (mean 1.3). It seems that the mast cells are not responsible for the proliferation of the endothelium or for growth of the hemangioma. The markedly increased number of mast cells in the growing stages and initial involuting stage of strawberry hemangiomas parallels the gradual growth of fibrous connective tissue inside the tumor. Mast cells may thus be a precursor of the beginning of the involution of a strawberry hemangioma.

Hemodynamics and vascular sensitivity to circulating norepinephrine in normal skin and delayed and acute random skin flaps in the pig.

Cutaneous circulation in 4 X 10 cm skin samples and delayed and acute random skin flaps constructed on the flanks of castrated Yorkshire pigs (13.3 +/- 0.7 kg; n = 12) were studied during intravenous infusion (0.5 ml per minute) of 5% dextrose solution (vehicle) and 5% dextrose containing norepinephrine (1 microgram/kg per minute). Total and capillary blood flow and A-V shunt flow were measured by the radioactive microsphere technique 6 hours after the raising of 4 X 10 cm single-pedicle acute and delayed random skin flaps using the technique and calculations published previously. Fluorescein dye test was also performed to assess vascular perfusion. It was observed that the capillary blood flow in the single-pedicle delayed skin flaps was similar to that in the normal skin, and the maintenance of this normal skin blood flow was not due to the closing of A-V shunt flow in the delayed skin flaps. Similarly, the significant (p less than 0.01) decrease in capillary blood flow and distal perfusion in the acute skin flaps compared with the delayed skin flaps was not due to the opening of A-V shunts in the acute skin flaps. There was no evidence to indicate that A-V shunt flow per se was the primary factor for the regulation of capillary blood flow in the acute and delayed skin flaps in the pig. Our data seemed to indicate that tissue ischemia in the distal portion of acute skin flaps was likely the result of vasoconstriction of the small random arteries which supplied blood to arterioles and A-V shunts, and locally released neurohumoral substances may play an important role in the pathogenesis of vascular resistance and ischemia in the acute skin flaps.