Brief communication: body composition and hidden obesity in people living with HIV on antiretroviral therapy.

BACKGROUND: Increased incidence of lifestyle diseases as side-effects of antiretroviral therapy (ART) have been reported in people living with HIV (PLWH). Few studies have evaluated obesity and hidden obesity in Japanese PLWH and their association with ART. In order to provide more appropriate drug selection and lifestyle guidance, we investigated the relationship between the effects of HIV infection and ART on the body composition of Japanese PLWH. METHODS: PLWH who visited the outpatient clinic and had body composition measured using the body composition analyzer InBody 570 were included in this study. Medications, comorbidities, and blood test data were obtained. Body mass index (BMI), body fat percentage, and skeletal muscle mass index (SMI) were measured. RESULTS: In this study, 543 patients were included. Based on body shape, patients were classified into a thin group (13), normal weight group (14), hidden obesity group (158), apparent obesity group (14), and obesity group (218). Compared with the normal weight group, the hidden obesity group had a higher prevalence of comorbidities and a lower SMI. CONCLUSIONS: PLWH are more likely to have obesity than the general population, indicating that hidden obesity is common even among those with a normal BMI. It is important to measure body fat percentage along with body weight, as hidden obesity can be missed. Further investigation of the effects of ART on body composition is needed.

De novo malignancy after pancreas transplantation in Japan.

BACKGROUND: Long-term immunosuppression is associated with an increased risk of cancer. Especially, the immunosuppression in pancreas transplantation is more intensive than that in other organ transplantation because of its strong immunogenicity. Therefore, it suggests that the risk of post-transplant de novo malignancy might increase in pancreas transplantation. However, there have been few studies of de novo malignancy after pancreas transplantation. The aim of this study was to analyze the incidence of de novo malignancy after pancreas transplantation in Japan. METHODS: Post-transplant patients with de novo malignancy were surveyed and characterized in Japan. RESULTS: Among 107 cases receiving pancreas transplantation in Japan between 2001 and 2010, de novo malignancy developed in 9 cases (8.4%): post-transplant lymphoproliferative disorders in 6 cases, colon cancer in 1 case, renal cancer in 1 case, and brain tumor in 1 case. CONCLUSIONS: We clarified the incidence of de novo malignancy after pancreas transplantation in Japan.

MicroRNA signature of intestinal acute cellular rejection in formalin-fixed paraffin-embedded mucosal biopsies.

Despite continuous improvement of immunosuppression, small bowel transplantation (SBT) is plagued by a high incidence of acute cellular rejection (ACR) that is frequently intractable. Therefore, there is a need to uncover novel insights that will lead to strategies to achieve better control of ACR. We hypothesized that particular miRNAs provide critical regulation of the intragraft immune response. The aim of our study was to identify miRNAs involved in intestinal ACR. We examined 26 small intestinal mucosal biopsies (AR/NR group; 15/11) obtained from recipients after SBT or multivisceral transplantation. We investigated the expression of 384 mature human miRNAs and 280 mRNAs associated with immune, inflammation and apoptosis processes. We identified differentially expressed 28 miRNAs and 58 mRNAs that characterized intestinal ACR. We found a strong positive correlation between the intragraft expression levels of three miRNAs (miR-142-3p, miR-886-3p and miR-132) and 17 mRNAs including CTLA4 and GZMB. We visualized these miRNAs within cells expressing CD3 and CD14 proteins in explanted intestinal allografts with severe ACR. Our data suggested that miRNAs have a critical role in the activation of infiltrating cells during intestinal ACR. These differences in miRNA expression patterns can be used to identify novel biomarkers and therapeutic targets for immunosuppressive agents.

Gene expression profiling of MicroRNAs in small-bowel transplantation paraffin-embedded mucosal biopsy tissue.

BACKGROUND: The molecular mechanisms and regulation of immune-mediated rejection of organ allografts remains unclear. Recent studies have reported that small non-coding RNAs, microRNAs (miRNAs) play a critical role in the immune system via modulation of transcription and translation. PURPOSE: We hypothesized that particular miRNAs provide regulation of an ensuing intragraft immune effector response. The aim of our study was to detect miRNAs involved in acute cellular rejection (AR) in human small intestinal allografts. MATERIALS: We examined 12 small intestinal mucosal biopsies (AR, 7 cases, all grade 2 or 3) and non-rejecting (NR) allografts (5 cases, all grade 0) obtained from recipients after small bowel or multivisceral transplantation. RNA was isolated from the formalin-fixed paraffin-embedded (FFPE) biopsy samples and transcribed to cDNA. After preamplification we utilized a PCR based TaqMan Low Density Array (TLDA) containing 365 mature human miRNAs. Relative quantification was done based on pooled normal intestine using a comparative Ct method. RESULTS: We identified 62 miRNA upregulated genes in small bowels with ACR, and 35 were downregulated. Forty-two miRNA genes were upregulated in non-ACR small bowel biopsy samples (grade IND), and 45 were downregulated. The relative fold change ratio of ACR to non-ACR was calculated, and 50 upregulated and 8 downregulated miRNAs were detected as significant. Several interesting miRNAs will be evaluated further from this preliminary study. Our data suggests that intragraft miRNAs are potentially involved in the activation of a host alloimmune response to donor. These miRNAs may serve as targets for appropriate intervention and may be useful to monitor the allograft status.

International grading scheme for acute cellular rejection in small-bowel transplantation: single-center experience.

A standardized grading scheme for the assessment of acute cellular rejection (ACR) in small-intestine transplantation was proposed in 2003 at the Eighth International Small Intestinal Transplantation Symposium. We have implemented the current grading scheme for ACR in small-bowel transplantation since October 2003. The pathologic diagnoses of those small-intestine biopsy samples, including ACR grade and other supplementary findings were evaluated. A total of 3484 small intestine biopsy samples from 155 patients were available for evaluation in this study. Frequency of grades 0, indeterminate, 1, 2, and 3 acute cellular rejection was 33.9%, 49.1%, 12.6%, 3.7%, and 0.8%, respectively. Duration of ACR episode strongly correlated with grade of ACR episode (P < .001). Other supplementary findings included acute vascular rejection component, 2.2%; increase in lymphoplasmacytic infiltrate in lamina propria, 15.7%; mucosal fibrosis, 0.4%; and regenerative changes, 0.3%. Our data substantiate that this grading system is reliable and useful for clinical decision making in bowel transplantation. We suggest that an assessment and quantification of supplementary findings be considered a component of the International Pathology Grading Scheme.

PKC-mediated phosphorylation regulates c-FLIP ubiquitylation and stability.

Cellular FLICE-inhibitory protein (c-FLIP) proteins are crucial regulators of the death-inducing signaling complex (DISC) and caspase-8 activation. To date, three c-FLIP isoforms with distinct functions and regulation have been identified. Our previous studies have shown that the stability of c-FLIP proteins is subject to isoform-specific regulation, but the underlying molecular mechanisms have not been known. Here, we identify serine 193 as a novel in vivo phosphorylation site of all c-FLIP proteins and demonstrate that S193 phosphorylation selectively influences the stability of the short c-FLIP isoforms, as S193D mutation inhibits the ubiquitylation and selectively prolongs the half-lives of c-FLIP short (c-FLIP(S)) and c-FLIP Raji (c-FLIP(R)). S193 phosphorylation also decreases the ubiquitylation of c-FLIP long (c-FLIP(L)) but, surprisingly, does not affect its stability, indicating that S193 phosphorylation has a different function in c-FLIP(L). The phosphorylation of this residue is operated by the protein kinase C (PKC), as S193 phosphorylation is markedly increased by treatment with 12-O-tetradecanoylphorbol-13-acetate and decreased by inhibition of PKCalpha and PKCbeta. S193 mutations do not affect the ability of c-FLIP to bind to the DISC, although S193 phosphorylation is increased by death receptor stimulation. Instead, S193 phosphorylation affects the intracellular level of c-FLIP(S), which then determines the sensitivity to death-receptor-mediated apoptosis. These results reveal that the differential stability of c-FLIP proteins is regulated in an isoform-specific manner by PKC-mediated phosphorylation.

Alpha-fetoprotein mRNA detection in peripheral blood for prediction of hepatocellular carcinoma recurrence after liver transplantation.

The aim of this study was to assess the value of alphafeto protein (AFP) mRNA-expressing cells detected in peripheral blood for predicting tumor recurrence after living donor liver transplantation (LDLT) in patients with hepatocellular carcinoma (HCC). The test group consisted of 25 patients who underwent LDLT for end-stage liver disease with HCC while the control group consisted of 37 living donors. Quantitative real-time reverse-transcriptase polymerase chain reaction was used for detection of AFP mRNA-expressing cells in peripheral blood. Nine (36%) of 25 patients developed tumor recurrences (four lung; one liver; one peritoneum; two bone; one adrenal gland) during the follow-up period. Perioperatively, AFP mRNA was positive in peripheral blood of eight patients (32.0%) but only in 1 (2.7%) of the control. Preoperative AFP mRNA was positive in three cases. Univariate analyses revealed that preoperative and perioperative AFP mRNA and microscopical vascular invasion were the significant predictors for HCC recurrence (P = .007, .037, and .005, respectively). In the patients with HCC exceeding Milan criteria (n = 15), the presence of AFP mRNA-positive cells in the peripheral blood correlated significantly with HCC recurrence (P = .033). We concluded that the presence of AFP mRNA-expressing cells could be a useful predictor of HCC recurrence in liver transplant patients.

Risk factors for graft dysfunction after adult-to-adult living donor liver transplantation.

The aim of this study was to investigate the risk factors for graft dysfunction after adult-to-adult living donor liver transplantation (LDLT). Thirty-nine adults with chronic cirrhosis underwent LDLT between 1999 and 2004. Their postoperative courses were uneventful with no vascular or bile duct complications early after LDLT, except one mild hepatic artery stenosis. The preoperative MELD scores were significantly higher in the failed graft group (n=5) than the functioning graft group (n=34; P=.004), while the graft liver weight/standard liver volume ratio was similar between these groups. We concluded that a high preoperative MELD score was associated with postoperative graft failure and that graft size had little impact on graft outcome. Although large grafts would seem intuitively more suitable for sick recipients, we did not show a benefit among this cohort; the MELD score was the best predictor, a finding that is also most consistent with donor safety.

Effect of a thin HA coating on the stress/strain distribution in bone around dental implants using three-dimensional finite element analysis.

Three-dimensional finite element analysis was performed for thin hydroxyapatite (HA) coated and titanium dental implants to study the effects on stress/strain distribution in the mandible with application of axial and oblique loads. The implants were of screw and cylinder types. With an axial load, the maximum equivalent bone stresses in the titanium implants were 21.5 and 29.0 MPa for the cylinder and screw types respectively, and the stress and strain distributions differed. For the cylinder type, the highest stress was located at the implant base, and for the screw type, it was located at the top edge of the first thread within the cortical bone. For the HA-coated cylinder and screw implants, the maximum equivalent bone stresses were 7.1 and 7.2 MPa respectively. The stress and strain distributions were similar, and the highest stress was located on the upper side of the cortical bone around the implant neck for both implants. Of the implants examined, the screw type HA-coated implant had the most uniform stress distribution in bone.

Deterioration in the superelasticity of Ti sputter coated on NiTi orthodontic wire.

To evaluate the deterioration in the superelasticity of Ti-coated NiTi orthodontic wires, titanium was coated onto wires to form a 1-microm thick film using radio frequency magnetron sputtering. In superelasticity cycle tests, the sigma5.0 value (the stress at 5.0% strain) of a Ti-coated wire was lower than that of a non-coated wire by 6.2% after one cycle, and by 10.4% after 20 cycles. In a three point bend tests, the L2.0 value (the load at 2.0 mm deflection) of a Ti-coated wire was 2.55 N, and this was reduced by 39.0% of the L2.0 value of a non-coated wire (4.18 N). After the non-coated and the Ti-coated wires had been immersed in physiological saline for periods of four and eight weeks, the concentration of Ni ions released into each solution was detected using MIP-Mass, and Ni peak intensity of the wires was measured using XPS. After eight weeks, the concentration of Ni ions released from the non-coated and the Ti-coated wires was 657 and 135 ppb, respectively. In a clinical test, no inflammation was observed during the entire attached period of four weeks in both the non-coated and the Ti-coated wire. In the Ti-coated wire, little exfoliation of the Ti-coating was observed using SEM and EDS. It was concluded that Ti-coated NiTi orthodontic wire has much potential for its application, with a low probability of causing any Ni allergy.

Hand-assisted laparoscopic splenectomy for a huge splenic cyst: operative technique and case report.

We report the case of a huge splenic cyst that was successfully treated by hand-assisted laparoscopic splenectomy. A 17-year-old girl with a chief complaint of left-sided abdominal pain was admitted to our department for investigation of a splenic tumor. Ultrasonography, computed tomography, and magnetic resonance imaging revealed a huge cystic lesion in the spleen measuring approximately 10 cm in diameter. Hand-assisted laparoscopic splenectomy was safely performed to diagnose and treat the splenic tumor. The histologic diagnosis was an epithelial cyst of the spleen with no atypical cells in the cyst wall. Hand-assisted laparoscopic splenectomy may be a good method of managing a huge splenic cyst that becomes symptomatic and potentially life-threatening through enlargement, rupture, and secondary infection.

New antifungal 1,2,4-triazoles with difluoro(substituted sulfonyl)methyl moiety.

New 1,2,4-triazoles (2) having a difluoro(substituted sulfonyl)methyl moiety were designed and synthesized via alpha,alpha-difluoro-alpha-(substituted thio)acetophenones (3). Compounds (2) showed potent antifungal activities against C. albicans, C. krusei, A. flavus and A. fumigatus in vitro and against C. albicans in vivo for oral and i.v. administrations. Especially, (-)-2a, (-)-2b and (-)-2d showed potent antifungal activities.

Affinity modulation of very late antigen-5 through phosphatidylinositol 3-kinase in mast cells.

Adhesiveness of integrins is up-regulated rapidly by a number of molecules, including growth factors, cytokines, chemokines, and other cell surface receptors, through a mechanism termed inside-out signaling. The inside-out signaling pathways are thought to alter integrin affinity for ligand, or cell surface distribution of integrin by diffusion/clustering. However, it remains to be clarified whether any physiologically relevant agonists induce a rapid change in the affinity of beta1 integrins and how ligand-binding affinity is modulated upon stimulation. In this study, we reported that affinity of beta1 integrin very late Ag-5 (VLA-5) for fibronectin was rapidly increased in bone marrow-derived mast cells by Ag cross-linking of FcepsilonRI. Ligand-binding affinity of VLA-5 was also augmented by receptor tyrosine kinases when the phospholipase Cgamma-1/protein kinase C pathway was inhibited. Wortmannin suppressed induction of the high affinity state VLA-5 in either case. Conversely, introduction of a constitutively active p110 subunit of phosphatidylinositol 3-kinase (PI 3-kinase) increased the binding affinity for fibronectin. Failure of a constitutively active Akt to stimulate adhesion suggested that the affinity modulation mechanisms mediated by PI 3-kinase are distinct from the mechanisms to control growth and apoptosis by PI 3-kinase. Taken together, our findings demonstrated that the increase of affinity of VLA-5 was induced by physiologically relevant stimuli and PI 3-kinase was a critical affinity modulator of VLA-5.

[Three cases of systemic sclerosis-associated gastrointestinal dysfunction that were improved with oral bovine type II collagen].

We report three cases of enteropathy associated with systemic sclerosis that were improved with oral type II collagen. The first patient, who was diagnosed as having systemic sclerosis-polymyositis overlap syndrome at the age of 35, was hospitalized for treatment of renal crisis at the age of 36. He acquired severe enteropathy associated with systemic sclerosis. The renal crisis was improved with angiotensin-converting enzyme inhibitor, but enteropathy persisted. He was given type II collagen in order to treat ileus. Four weeks later, ileus was improved and he was able to take foods. The second patient, a 63-year-old female whose onset of systemic sclerosis was at the age of 58, complained of ileus several times and was given oral type II collagen. Six weeks later, ileus was improved. The third patient, a 19-year-old female, was diagnosed as having systemic sclerosis-polymyositis overlap syndrome. Enteropathy was so severe that she could not take foods for 1 year. She was given oral type II collagen. Six months later, ileus and skin sclerosis were markedly improved. These results suggest that oral type II collagen is effective in the treatment of gastrointestinal dysfunction associated with systemic sclerosis.

[Primary biliary cirrhosis (PBC)-CREST overlap syndrome complicated with Sjögren's syndrome].

We reported two sisters of primary biliary cirrhosis (PBC)-CREST overlap syndrome complicated with Sjögren's syndrome (SS). Both patients had Raynaud's phenomenon, sclerodactylia, telangiectasia, chronic sialoadenitis, chronic nonsuppurative destructive cholangitis by liver biopsy and were positive for anti-centromere antibodies. This is the first report of two sisters of PBC-CREST overlap syndrome complicated with SS.

[In vitro and in vivo antibacterial activities of a new quinolone derivative, FD501].

FD501 is a newly synthesized quinolone derivative with aminoazepine group at the C-7 position. The in vitro and in vivo antibacterial activities of FD501 were investigated comparing with those of norfloxacin, ofloxacin, ciprofloxacin and sparfloxacin. The in vitro antibacterial activities of FD501 against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus, were equal to or higher than those of other quinolones. But its activities against Gram-negative bacteria were equal to or lower than those of other quinolones. Minimum inhibitory concentrations of FD501 against Gram-positive bacteria were similar to that of sparfloxacin. The bactericidal activity may be due to the inhibition of the DNA super-coiling activity of DNA gyrase. The area under the serum concentration-time curve of FD501 in rats following oral administration was larger than norfloxacin and ciprofloxacin and smaller than those of ofloxacin and sparfloxacin.

In vitro antibacterial activity of FA103, a new quinolone derivative of C-7 position with 7-perhydrodiazepinone.

A number of new quinolone antibacterial agents such as ofloxacin, norfloxacin, ciprofloxacin and sparfloxacin have been developed and introduced to the market. They possess a broad spectrum of activity against Gram-positive and Gram-negative bacteria. Ciprofloxacin has the highest activity against Gram-positive is higher than other quinolones. The activity of these quinolones against Gram-negative bacteria is generally higher than against Gram-positive bacteria. The substitution group of quinolones at C-7 position is responsible to show similar antibacterial activity with broad spectrum and similar pharmacokinetic properties, and variety of the substituents have been synthesized in many laboratories. Most of the substituents are piperazinyl of six-membered ring or pyrrolidinyl of five-membered ring, being modified with an alkyl group or another group. The development of potent quinolones against bacteria involved in pneumonia was seemed to be useful, and we investigated structure-activity relationships of new quinolones with a stronger activity against Gram-positive bacteria. A quinolone derivative with a seven-membered ring, perhydrodiazepinone, at the C-7 position was found to be a candidate for further evaluation. No previous attempts have been made to synthesize this type of derivatives. The compound FA103, 5-amino-1-cyclopropyl-6,8-difluoro-7-(2,3,4,5,6,7-hexahydro-1 H-1,4-diazepin-5-on-1-yl)-1,4-dihydro-4-oxoquinoline-3-carboxylic acid, was synthesized and evaluated. Chemical structure of FA103 is shown in Fig. 1. This is a new difluoroquinolone with a broad antibacterial spectrum and improved activity against Gram-positive bacteria. In this paper, we compare the in vitro activity of FA103 with that of ofloxacin, norfloxacin, sparfloxacin against Gram-positive and Gram-negative bacteria.

[Lattice corneal dystrophy type II with familial amyloid polyneuropathy type IV].

Lattice corneal dystrophy type II associated with familial amyloid polyneuropathy type IV (Finish type, FAP-IV) has not yet been reported in Japan. We report 7 cases in a Japanese family. The proband, a 64-year-old man, suffering from itching in his limbs, impaired lip movement, and dysarthria, consulted the Department of Neurology, University of Tokyo. Neurological examination revealed bilateral facial, glossopharyngeal, vagal, and hypoglossal nerve palsy, and impaired distal vibratory perception. His vision was 1.2 and he had fine lattice corneal dystrophy in both eyes. Short glassy lines were randomly scattered in the lattice dystrophy. Corneal sensation was normal and there was no evidence of recurrent corneal erosion. Immunohistological and biochemical studies confirmed the diagnosis of FAP-IV. Six siblings were neurologically suspected to be FAP-IV patients with similar lattice corneal dystrophy. The family pedigree suggested an autosomal dominant trait of inheritance.