Increase in energy intake leads to a decrease in obestatin in restricting-type of anorexia nervosa.

Restricting-type of anorexia nervosa (AN-R) is a serious disorder affecting adolescents and young adults, and decreases quality of life over long period. Successful weight restoration is an important prognostic factor for disease outcome; however, the underlying mechanism of refeeding-resistance, a core psychopathology relevant to 'ambivalent' eating behaviors, remains unclear in this disorder. Obestatin plays an important role in the regulation of growth hormone release, appetite, and energy metabolism. However, the progress of these patients and changes in the levels of obestatin during treatment were not reported. The purpose of this study was to determine the changes in obestatin levels when energy intake increases in AN-R patients. As a result, obestatin was higher in AN-R patients than in control subjects as well as acyl ghrelin and des-acyl ghrelin. An increase in the intake calorie has decreased obestatin as well as des-acyl ghrelin. These findings indicate that the obestatin is an important factor in the diagnosis and treatment of AN-R, similarly to acyl ghrelin and des-acyl ghrelin. In the future, the research on the clinical application of the ghrelin peptide family and the receptor will be expected to progress.

Changes in acyl ghrelin, des-acyl ghrelin, and ratio of acyl ghrelin to total ghrelin with short-term refeeding in female inpatients with restricting-type anorexia nervosa.

Restricting type of anorexia nervosa (AN-R) is a serious disorder affecting adolescents and young adults and decreases quality of life over a long period. Successful weight restoration is an important prognostic factor for disease outcome; however, the underlying mechanism of refeeding resistance, a core psychopathology relevant to 'ambivalent' eating behaviors, remains unclear in this disorder. Ghrelin plays an important role in the regulation of growth hormone release, appetite, and energy metabolism. However, the early progress of these patients and changes in the levels of acyl ghrelin and des-acyl ghrelin during treatment were not reported. The purpose of this study was to determine the changes in ghrelin levels (acyl and des-acyl) during early treatment. As a result, des-acyl ghrelin in AN-R patients is higher than in control subjects before the therapy, but it decreases with treatment. The plasma des-acyl ghrelin level in AN-R patients started decreasing more rapidly and in early stage of the hospitalization than ever reported, and after 8 weeks, it is significantly lower than in control subjects. It means that des-acyl ghrelin is sensitive and changeable with their nutrition state. Furthermore, the ratio of the acyl ghrelin to total ghrelin increases with 8 weeks treatment. Eight weeks after, energy intake of the AN-R patients is recovered near the normal range with a daily energy intake of 1 700+/-93.54 kcal. These findings may be valuable for future AN-R treatments in order to increase acyl ghrelin and decrease des-acyl ghrelin, thereby influencing the refeeding outcome.

Centrally administered neuromedin S inhibits feeding behavior and gastroduodenal motility in mice.

Neuromedin S (NMS) was recently identified as an endogenous ligand for the FM-4/TGR-1 receptor in the rat hypothalamus. No previous studies have examined the effect of NMS on gut motility. We examined the effects of intracerebroventricular administration of NMS on food intake in food-deprived and free-feeding mice, and on gastroduodenal motility by using a manometric method, and gastric emptying in mice. We found that NMS decreased food intake and the gastric emptying rate. It also disrupted the motor activity in the antrum and duodenum of conscious food-deprived mice. These results suggest that NMS influences gut motility as well as feeding behavior.

Neonatal intrahepatic cholestasis caused by citrin deficiency in two Malaysian siblings: outcome at one year of life.

We report two Malaysian siblings with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). The younger sibling, a six-month-old Chinese girl, presented with prolonged neonatal jaundice, and was investigated for biliary atresia. Urine metabolic screen showed the presence of urinary-reducing sugars, and she was treated with a lactose-free formula. NICCD was suspected based on the clinical history, examination and presence of urinary citrulline. Mutation study of the SLC25A13 gene showed the compound heterozygotes, 851del4 and IVS16ins3kb, which confirmed the diagnosis of NICCD in the patient and her three-year-old female sibling, who also had unexplained neonatal cholestasis. Long-term dietary advice, medical surveillance and genetic counselling were provided to the family. The diagnosis of NICCD should be considered in infants with unexplained prolonged jaundice. DNA-based genetic testing of the SLC25A13 gene may be performed to confirm the diagnosis retrospectively. An awareness of this condition may help in early diagnosis using appropriate metabolic and biochemical investigations, thus avoiding invasive investigations in infants with neonatal cholestasis caused by NICCD.

Neonatal intrahepatic cholestasis caused by citrin deficiency: clinical and laboratory investigation of 13 subjects in mainland of China.

BACKGROUND: Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is a novel inborn error of metabolism due to dysfunction of citrin protein, and much more information about this new disease is still needed for its clinical management. AIMS: To investigate in detail the clinical and laboratory features of NICCD. PATIENTS: 13 NICCD subjects in mainland of China diagnosed in our department since 2006. METHODS: The anthropometric parameters of the patients at birth were compared with controls, representative biochemical changes and metabolome findings were investigated cross-sectionally, and mutations in the causative gene SLC25A13 were analyzed by protocols established previously. RESULTS: The patients showed reduced birth weight, length and ponderal index. Main clinical manifestations consisted of jaundice, hepato/hepatosplenomegaly and steatohepatosis on ultrasonography. Biochemical analysis revealed intrahepatic cholestasis, delayed switch of AFP to albumin, and elevated triglyceride, total cholesterol and LDL-cholesterol together with reduced HDL-cholesterol. Metabolome findings included co-existence of markers for galactosemia and tyrosinemia in urine, and elevated Cit, Met, Thr, Tyr, Lys, Arg and Orn in blood. Mutations of 851-854del, IVS6+5G>A, 1638-1660dup, A541D, IVS16ins3kb, R319X and G333D were detected in the gene SLC25A13. CONCLUSIONS: The diagnosis of NICCD cannot be established based just on the numerous but non-specific clinical manifestations and biochemical changes. The relatively specific metabolome features provide valuable tools for its screening and diagnosis, while SLC25A13 mutation analysis should be taken as one of the reliable tools for the definitive diagnosis. The body proportionality at birth, steatohepatosis on ultrasonography, delayed switch of AFP to albumin, dyslipidemia pattern, urinary metabolome features and the novel mutation G333D expanded the clinical spectrum of NICCD.

Fisetin induces transcription of NADPH:quinone oxidoreductase gene through an antioxidant responsive element-involved activation.

Induction of phase II enzymes such as NADPH:quinone oxidoreductase (QR) can reduce carcinogen-induced mutagenesis and tumor formation. In our search for novel dietary anticarcinogens, fisetin, a flavonol widely distributed in fruits and vegetables was found to induce QR activity in murine hepatoma 1c1c7 cells. The cells were treated with various concentrations of fisetin, and then were assessed for cell growth, QR activity, QR mRNA expression and transcription activation of the QR gene. The results showed that fisetin induced QR activity in time- and dose-dependent manner in the concentration range of 0.1 to 10 microM, and the activity induction was associated with QR mRNA expression as detected by reverse transcription-PCR. Furthermore, transfection studies using a human QR antioxidant/electrophile-response element (ARE/EpRE) reporter construct demonstrated that fisetin activated the ARE/EpRE. These results show that fisetin increases QR activity by transcriptional activation of the ARE/EpRE, suggesting a novel mechanism by which dietary fisetin may be implicated in cancer chemoprevention.

[A case of pediatric recurrent acute mastoiditis caused by penicillin-resistant Streptococcus pneumonia complicated by primary immunodeficiency].

Penicillin-resistant Streptococcus pneumoniae (PRSP) is a frequently detected pathogen of intractable acute otitis media and is associated with prolonged or recurrent infection. The use of antibiotics has made the incidence of secondary acute mastoiditis following acute otitis media relatively rare, but when it does occur, its severe complications may be life-threatening. We report a case of pediatric recurrent acute mastoiditis caused by PRSP in a 6-year-old boy suffering from PRSP acute mastoiditis on 4 occasions, twice undergoing simple mastoidectomy. Although we initially suspected PRSP to be the chief factor in iterative infection, immunological analysis demonstrated significantly decreased IgG and IgA antibodies in serum and the patient was diagnosed as having common variable immunodeficiency (CVID). As the first middle ear infection occurred at the age of 6 and there was no history of upper respiratory tract infection, CVID may be the main pathological factor of recurrent mastoiditis, although infection occurred, only in the ear and did not involve other organs. This suggests that recurrent mastoiditis in the present case involved the coexistence of PRSP and CVID.

Effects of clarithromycin on cultured human nasal epithelial cells and fibroblasts.

OBJECTIVE/METHODS: Long-term administration of clarithromycin has been reported to be effective in the treatment of chronic sinusitis. To investigate the mechanism underlying the anti-inflammatory activity of clarithromycin, the authors evaluated the effect of clarithromycin on the gene expression of proinflammatory cytokine and the DNA-binding activity of nuclear factor (NF)-kappa B in cultured human nasal epithelial cells and fibroblasts. Cells were incubated with endotoxin purified from nontypable Haemophilus influenzae or interleukin (IL)-1 beta in the presence of clarithromycin. RESULTS: Northern blot analysis revealed that clarithromycin suppressed IL-1 beta gene expression in human nasal epithelial cells stimulated by H. influenzae endotoxin (HIE). Intercellular adhesion molecule-1 gene expression in nasal fibroblasts stimulated by IL-1 beta was also suppressed by clarithromycin. Furthermore, electrophoretic mobility shift assay demonstrated that clarithromycin reduced DNA-binding activity of NF-kappa B in both human nasal epithelial cells and fibroblasts stimulated by HIE or IL-1 beta, respectively. CONCLUSION: The present results suggest that clarithromycin may reduce gene expression of proinflammatory cytokines and adhesion molecules from nasal mucosa at the transcriptional factor level and exert an anti-inflammatory effect on nasal mucosa in chronic sinusitis.

Identification of two novel mutations in the SLC25A13 gene and detection of seven mutations in 102 patients with adult-onset type II citrullinemia.

Adult-onset type II citrullinemia (CTLN2) is characterized by a liver-specific deficiency of argininosuccinate synthetase (ASS) protein. We have recently identified the gene responsible for CTLN2, viz., SLC25A13, which encodes a calcium-binding mitochondrial carrier protein, designated citrin, and found five mutations of the SLC25A13 gene in CTLN2 patients. In the present study, we have identified two novel mutations, 1800ins1 and R605X, in SLC25A13 mRNA and the SLC25A13 gene. Diagnostic analysis for the seven mutations in 103 CTLN2 patients diagnosed by biochemical and enzymatic studies has revealed that 102 patients had one or two of the seven mutations and 93 patients were homozygotes or compound heterozygotes. These results indicate that CTLN2 is caused by an abnormality in the SLC25A13 gene, and that our criteria for CTLN2 before DNA diagnosis are correct. Five of 22 patients from consanguineous unions have been shown to be compound heterozygotes, suggesting a high frequency of the mutated genes. The frequency of homozygotes is calculated to be more than 1 in 20,000 from carrier detection (6 in 400 individuals tested) in the Japanese population. We have detected no cross-reactive immune materials in the liver of CTLN2 patients with any of the seven mutations by Western blot analysis with anti-human citrin antibody. From these findings, we hypothesize that CTLN2 is caused by a complete deletion of citrin, although the mechanism of ASS deficiency is still unknown.

Demonstration of antibodies against human papillomavirus type-11 E6 and L2 proteins in patients with recurrent respiratory papillomatosis.

Recurrent respiratory papillomatosis (RRP) is highly prevalent in Thailand, with the human papillomavirus (HPV) type-11 being the most widespread. In this study, we isolated the HPV type-11 (HPV-11) genome from subjects with RRP and subcloned the E6 and L2 open reading frames (ORFs) with the expression vectors pEX1 and pEX3. The recombinant E6/beta-gal and L2/beta-gal fusion proteins were expressed in E. coli. Using the recombinant proteins, we demonstrated the presence of antibodies against HPV-11 E6 and L2 in RRP patients by Western blot analysis. The prevalence of seropositivity for HPV-11 E6 and L2 were 5% (1/20) and 10% (2/20), respectively. Although RRP is caused by infection on the mucosal surface, it appears that an immune response occurs against viral proteins expressed in the epithelial lesions.

[Reconstructive surgery for head and neck cancers under multiple facility cooperation system].

The problems with the system for reconstructive surgery for head and neck cancers in our department where plastic surgeons are regularly invited from distant facilities were discussed. Especially, the importance of preoperative discussion between the plastic surgeons and ENT doctors was stressed. The computer system with Internet will play an important role in exchanging information such as CT scans and MRI scans between the plastic surgeons and ENT doctors. Moreover, the establishment of a system to prevent postoperative troubles such as closure of anastomosed vessels was stressed. Twenty-seven re-vascularized free flaps had been transferred with a 96.3% success rate (26 of 27 cases). We concluded that the system for reconstructive surgery under the multiple facility cooperation system worked well.

[A case report of congenital stapes fixation accompanied by symphalangism and hypermetropia].

A 32-year-old female with bilateral congenital stapes fixation accompanied by bilateral proximal symphalangism and bilateral hypermetropia is reported. This is the 19th case of congenital stapes fixation and symphalangism in Japan. Hypermetropia was speculated to be one of the cardinal symptoms of the disease based on the present case and cases previously reported. In this case hypermetropia was due to pure microphthalmos and this was the first report that revealed the cause of hypermetropia in the syndrome. Bilateral stapedotomy were carried out and her hearing loss was improved to the satisfactory level.

[Preoperative autologous blood donation in patients with malignant tumors of the head and neck region].

Twelve instances of preoperative autologous blood donation were assessed in 10 patients with malignant tumors of the head and neck region. All patients received preoperative radiation therapy and chemotherapy and most of the cases were also given recombinant human erythropoietin (EPO) and iron sulfate. Hemoglobin concentrations immediately before starting the 800ml blood donations were higher than 13g/dl in 5 cases and less than 13g/dl in 6 cases. The mean hemoglobin concentration just prior to donation and one week after donation in these two groups changed from 14.4g/dl to 12.5g/dl and 11.7g/dl to 11.7g/dl, respectively. A 1000ml blood donation was carried out in only one patient, and the hemoglobin concentrations just before donation and one week after donation were 11.4g/dl and 10.5g/dl, respectively. Only half of the blood volume scheduled, 400ml, was achieved in 3 of the 12 cases attempted because of fever, diarrhea and a change of the day of surgery, respectively. The mean estimated blood loss in the 10 patients that underwent surgery as planned was 898ml, and allogenic blood transfusion was avoided in all cases. We concluded that 800-1000ml preoperative autologous blood donation can be performed safely in patients with advanced malignant tumors of the head and neck region who have undergone preoperative radiation therapy and chemotherapy by giving EPO and iron sulfate.

trans activation by the full-length E2 proteins of human papillomavirus type 16 and bovine papillomavirus type 1 in vitro and in vivo: cooperation with activation domains of cellular transcription factors.

Papillomaviral E2 genes encode proteins that regulate viral transcription. While the full-length bovine papillomavirus type 1 (BPV-1) E2 peptide is a strong trans activator, the homologous full-length E2 product of human papillomavirus type 16 (HPV-16) appeared to vary in function in previous studies. Here we show that when expressed from comparable constructs, the full-length E2 products of HPV-16 and BPV-1 trans activate a simple E2- and Sp1-dependent promoter up to approximately 100-fold in human keratinocytes and other epithelial cells as well as human and animal fibroblasts. Vaccinia virus-expressed, purified full-length HPV-16 and BPV-1 E2 proteins bound a consensus E2 site with high specific affinities (Kd = approximately 10(-9) M) and stimulated in vitro transcription up to six- to eightfold. In vivo and in vitro trans activation by either E2 protein required cooperation with another activator, such as Sp1, or other factors that interact with papillomavirus promoters, such as AP-1, Oct-1, nuclear factor 1/CTF, transcriptional enhancer factor 1, or USF. The glutamine-rich domain B of Sp1 or the mutually unrelated activation domains of other transcription factors were necessary and sufficient for cooperation with either E2 factor. We conclude that like BPV-1 E2, the HPV-16 E2 protein has the potential to function as a strong activator of viral gene expression in cooperation with cellular transcription factors.

Localization of IL-1 beta mRNA and cell adhesion molecules in the maxillary sinus mucosa of patients with chronic sinusitis.

Interleukin-1 beta (IL-1 beta) is a predominant cytokine in retained paranasal sinus fluid of chronic sinusitis where infiltration by polymorphonuclear neutrophils (PMNs) of nasal and paranasal mucosa is characteristic. The authors investigated the localization of IL-1 beta messenger RNA (mRNA) in the maxillary sinus mucosa of patients with chronic sinusitis, using digoxigenin-labeled oligonucleotide probes. IL-1 beta mRNA was detected in some extravascular PMNs and small numbers of mononuclear leukocytes but was not detected in other tissue cells or intravascular leukocytes. The expression and distribution of the cell adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and endothelial-leukocyte adhesion molecule-1 (ELAM-1), were also studied in cultured human mucosal microvascular endothelial cells and in the maxillary sinus mucosa in chronic sinusitis by immunohistochemistry using monoclonal antibodies against these cell adhesion molecules. Only ICAM-1 was expressed on cultured human mucosal microvascular endothelial cells without IL-1 beta stimulation. With IL-1 beta activation of these cells, ELAM-1 was expressed strongly and the expression of ICAM-1 was enhanced. In the maxillary sinus mucosa, ICAM-1 was strongly and universally expressed on endothelial cells of all small vessels, whereas ELAM-1 was expressed only in the subepithelial region. These findings suggest that IL-1 beta, one of mediators in chronic sinusitis, is produced by PMNs, induces the expression of ICAM-1 and ELAM-1 on endothelial cells, and, thereby, stimulates PMN infiltration in chronic sinusitis.

Detection and cloning of human papillomavirus DNA associated with recurrent respiratory papillomatosis in Thailand.

Recurrent respiratory papillomatosis (RRP) is highly prevalent in Thailand. In this study, we examined the presence of human papillomavirus (HPV) DNA in 25 RRP patients in Thailand by means of dot blot analysis and/or polymerase chain reaction. Eighty-four percent (21/25) of cases and 4% (1/25) of cases were positive for HPV-11 DNA and HPV-6 DNA, respectively. Three cases (3/25) were negative for all of the examined HPV types. No cases were positive for HPV-16 or 18. Furthermore, we isolated the recombinant HPV-11 DNA clone from a genomic library constructed with the DNA of RRP tissue. The restriction map of the cloned HPV DNA was identical with the map of known HPV-11 DNA. These results suggest at least that no specific HPV type or subtype is likely to be associated with RRP in Thailand.

Oculomotor parasympathetic pathway to the accessory ciliary ganglion bypassing the main ciliary ganglion by way of the trigeminal nerve.

When an HRP or WGA-HRP solution was injected into the rostral midbrain including the oculomotor visceral nuclei, densely distributed HRP/WGA-HRP-positive granules were observed around the ganglion neurons in the accessory ciliary ganglion (ACG) and ectopic neurons in the communicating branch from the long ciliary nerve to the ACG. The same injections labeled fibers within the communicating branch as well as the fibers between the ACG and the main ciliary ganglion (CG). These findings indicate that some oculomotor parasympathetic preganglionic fibers reach the ACG bypassing the CG by way of the trigeminal nerve.

[A clinical and immunological study on non-Hodgkin's malignant lymphoma in nasosinus region].

Twenty-eight cases of non-Hodgkin's lymphoma in nasosinus region were studied for tumor-surface marker phenotype, anti-HTLV-I antibody titer and histopathologic correlation with clinical features. The incidence of these cases was 25% of all lymphomas in head and neck region from 1978 to 1987. In 16 cases, 12 (75%) had T-cell markers and 4 (25%) had B-cell markers. Of 11 cases, 6 (54.5%) were anti-HTLV-I antibody carriers. Five year survival ratio of all patients was 4.5%. The survival curve for T-cell lymphoma patients was characterized by a rapid initial decline. The survivals of the group of anti-HTLV-I antibody carriers was estimated inferior to those of non-carriers. Correlation of HTLV-I to prognosis of these patients was discussed.

A study on the relationship between adenoid vegetation and rhinosinusitis.

The relationship between adenoidal-nasopharyngeal ratios (AN ratios) and sinusitis were evaluated in 404 children. The AN ratios of children with snoring, mouth breathing, or nasal obstruction (major symptoms of adenoidal enlargement) were significantly higher than those of children without these symptoms. However, the AN ratios of children with sinusitis were almost equal to those of normal children.