Leptin reverses nonalcoholic steatohepatitis in patients with severe lipodystrophy

Severe lipodystrophy is characterized by diminished adipose tissue and hypoleptinemia, leading to ectopic triglyceride accumulation. In the liver, this is associated with steatosis, potentially leading to nonalcoholic steatohepatitis (NASH). We investigated the prevalence of NASH and the effect of leptin replacement in these patients. Ten patients with either generalized lipodystrophy (8 patients) or Dunnigan's partial lipodystrophy (2 patients) were included in this analysis. Paired liver biopsy specimens were obtained at baseline and after treatment with recombinant methionyl human leptin (r-metHuLeptin), mean duration 6.6 months. The extents of portal and parenchymal inflammation, steatosis, ballooning, presence of Mallory bodies, and fibrosis in liver biopsy specimens were scored using a previously validated system developed to assess NASH activity. Histological disease activity was defined as the sum of ballooning, steatosis, and parenchymal inflammation scores. We concurrently tested serum triglycerides and aminotransferases and estimations of liver volume and fat content by magnetic resonance imaging. Eight of 10 patients met histological criteria for NASH at baseline. After treatment with r-metHuLeptin, repeat histological examinations showed significant improvements in steatosis (P = .006) and ballooning injury (P = .005), with a reduction of mean NASH activity by 60% (P = .002). Fibrosis was unchanged. Significant reductions were seen in mean serum triglycerides (1206-->226 mg/dL, P = .002), glucose (220-->144 mg/dL, P = .02), insulin (46.4-->24.8 muIU/mL, P = .004), ALT (54-->24 U/L, P = .02), AST (47-->22 U/L, P = .046), liver volume (3209-->2391 cm(3), P = .007), and liver fat content (31-->11%, P = .006). In conclusion, r-metHuLeptin therapy significantly reduced triglycerides, transaminases, hepatomegaly, and liver fat content. These reductions were associated with significant reductions in steatosis and the hepatocellular ballooning injury seen in NASH.     

Ultrasonic strain/strain rate imaging--a new clinical tool to evaluate the transplanted heart.

OBJECTIVE: The aim of this study was to investigate the clinical applicability of strain and strain rate imaging (epsilon/SRI) in heart transplantation (Htx) patients and to establish "normal" post-Htx regional systolic deformation values. BACKGROUND: Epsilon/SR indices have been shown to be a more sensitive measure of regional systolic function than standard echo measurements. Thus, they might provide a new tool to better define both normal cardiac graft function and detect changes due to post-Htx complications. However, prior to investigating the role of epsilon/SRI in detecting abnormalities, "normal" post-Htx regional deformation values must be established as graft regional function can be altered by a number of factors such as ischemic time, surgical technique or accelerated graft ageing. METHODS: A total of 57 Htx patients (age 36+/-12 years; post-Htx 5.5+/-3 years) without any documented complication were studied. Epsilon/SRI data were acquired from the septum, left ventricular (LV) free walls and right ventricular free wall (RVFW). A total of 29 age-matched healthy subjects served as controls. RESULTS: Htx longitudinal peak systolic velocities (Vsys) were lower in inferior, septal and RVFW segments compared to controls. Peak systolic epsilon/SR (epsilonsys/SRsys)) did not differ from controls except in septum and RVFW in which the values were significantly reduced. Radial Vsys) in the Htx group were higher than controls while epsilonsys/SRsys were reduced. There was a significant decrease in SR(sys) in apical LV segments with increasing time post-Htx, whereas those measured in RVFW showed an increase by that time. CONCLUSION: Epsilon/SRI demonstrated that "healthy" Htx hearts have normal global systolic function but altered regional systolic deformation indices compared to normal hearts. Post-Htx time has a diminishing effect on the regional systolic deformation indices in LV segments but an improving effect in RVFW. These "normal" Htx values should provide the basis for subsequent studies into the role of epsilon/SRI in the non-invasive detection of post-Htx complications.     

Histopathology and immunohistochemistry of gills of Van fish (Alburnus tarichi Güldenstädt, 1814) infected with myxosporean parasites

ABSTRACT

The Van fish are a cyprinid species endemic to Turkey’s largest soda lake, Lake Van, and have great economic value because they are a food source. Once a year, the fish take part in reproductive migration to the fresh waters flowing into the lake. The fish migrate from an extreme environment with high salinity (2.2%) and high pH (9.8). These fish are unable to reproduce in this alkaline environment and must migrate to fresh water during their breeding season. The aim of the present study is to report the presence of the myxosporean parasites on the gills and the pathological changes. Changes in gill histopathology, mucocytes, mitochondria-rich cells, expression of Heat Shock Protein 70 (Hsp70), and ATPase (NKA) were observed in the gill tissue. As a result of the histopathological changes in gills, infected fish had abundant plasmodia with different sizes. Plasmodia were found on gill filaments inside white ovoid-shaped structures. It was observed that plasmodia were contained on the primary filament which changed the histological structure of the gill tissue to a large extent. It was determined that the density and size of mucocytes in the infected areas of the gill tissue increased, whereas the number of mitochondria-rich cells decreased. Hsp70, an indicator of stress, was not different between normal and infected fish.     

Induced sputum-retrieved matrix metalloproteinase 9 and tissue metalloproteinase inhibitor 1 in granulomatous diseases

Matrix metalloproteinases (MMPs) capable of degrading various components of connective tissue matrices, and tissue inhibitor metalloproteinases (TIMPs) are considered important in lung parenchymal remodeling and repair processes in pulmonary diseases. Induced sputum (IS) is a reliable noninvasive method to investigate pathogenesis, pathophysiology and treatment of lung disease. This study was designed to determine whether IS-MMP9/TIMP1 levels demonstrate lung parenchymal remodeling in sarcoidosis (SA) and Crohn's disease (CRD) patients. Sputum was induced and processed conventionally in 13 SA patients, 18 CRD patients and 9 controls. Two-hundred cells were counted on Giemsa-stained cytopreps, and T lymphocytes subsets (CD4 = T helper and CD8 = T suppressor cytotoxic cells) were analysed by FACS using monoclonal antibodies.MMP-9 and TIMP-1 were measured using commercial ELISA kits. MMP-9 concentrations, but not those of TIMP-1, were significantly greater in the sputum supernatant in SA and CRD patients compared to controls (P = 0.018 and P = 0.0019, respectively). The molar ratio, MMP-9/TIMP-1, was significantly higher in SA and CRD patients compared to controls (P = 0.008 and P = 0.024, respectively). Gelatinase species having a molecular weight similar to that of MMP-9 were demonstrated by zymographic analysis. MMP-9 levels were highly correlated with the CD4/CD8 ratio and DLCO capacity in SA but less in CRD patients. MMP-9 levels in IS provide a sensitive marker for pulmonary damage.     

The effect of specific immunotherapy on T-cell receptor repertoire in patients with allergy to house-dust mite

BACKGROUND: The precise mechanism of specific immunotherapy (SIT), long used for treating allergic diseases, remains undefined. SIT was shown to act by modifying the immune response of T lymphocytes to antigens. We examined the effect of SIT on the expression and use V-alpha, -beta, -gamma and -delta chains of T-cell receptors (TCR) in patients allergic to house-dust mite. METHODS: Peripheral venous blood was taken for lymphocyte TCR analysis from 10 house-dust mite (HDM) allergic adults before initiating SIT and 6 months after initiating the treatment. Twelve similarly allergic patients without SIT served as controls. TCR chains were identified by fluorescence-activated cell sorter (FACS) using the following monoclonal antibodies: CD3, CD14, CD8, pan alpha-beta, pan gamma-delta, V-alpha2, V-alpha12.1, V-beta5a, V-beta5b, V-beta5c, V-beta8a, V-beta8b, V-beta3.1, V-beta13, V-beta12, V-beta6.7, V-delta1, V-delta2, V-gamma9, and V-gamma4. RESULTS: Analyzed before and 6 months after SIT initiation, lymphocyte TCR showed significantly increased V-beta5b, V-beta12 and V-alpha12.1 values compared to controls (without significant changes in other markers). CONCLUSIONS: SIT caused selective expansion of certain V-beta- and V-alpha-expressing T cells in patients allergic to HDM. Our results support the notion that the effect of SIT in patients with allergic rhinitis may be achieved by modifying the T lymphocyte response through the modulation of TCR usage.     

Investigation of genotype–phenotype correlation in patients with AZF microdeletion in a single-reference centre

In this study, we aimed to elucidate the relationship between AZF deletion type and clinical information of azoospermic patients with AZF microdeletion in the Turkish population. Azoospermic patients with normal karyotype and AZF microdeletion were analysed retrospectively by collecting clinical data including hormone profile, demographic characteristics and micro-TESE results. As a result of the AZF microdeletion tests of 42 cases with 46 XY karyotype, AZFa deletion was detected in 3 cases, AZFb deletion in 2 cases, AZFc deletion in 31 cases, AZFb + AZFc deletion in 4 cases and AZFa + AZFb + AZFc deletion in 2 cases respectively. Spermatozoon was obtained in 16 cases with AZFc microdeletion with micro-TESE. Pregnancy was achieved in 2 cases. There was no statistically significant difference between the type of deletion and age, height, weight, body mass index, hormone profile and testicular volume. When AZF is evaluated according to the type of microdeletion, it will be appropriate to plan the medical and surgical options more carefully in a multidisciplinary manner in cases with deletions including AZFa, AZFb or their combinations. Also, genotype–phenotype correlation was found to be consistent with the literature; particularly patients having AZFc deletions were found to have a chance for pregnancy.     

Inhibition of autophagy enhances DENSpm-induced apoptosis in human colon cancer cells in a p53 independent manner

Purpose

One of the recently developed polyamine (PA) analogues, N ¹ ,N¹¹-diethylnorspermine (DENSpm), has been found to act as an apoptotic inducer in melanoma, breast, prostate and colon cancer cells. Also, its potential to induce autophagy has been established. Unfolded protein responses and starvation of amino acids are known to trigger autophagy. As yet, however, the molecular mechanism underlying PA deficiency-induced autophagy is not fully clarified. Here, we aimed to determine the apoptotic effect of DENSpm after autophagy inhibition by 3-methyladenine (3-MA) or siRNA-mediated Beclin-1 silencing in colon cancer cells.

Methods

The apoptotic effects of DENSpm after 3-MA treatment or Beclin-1 silencing were determined by PI and AnnexinV/PI staining in conjunction with flow cytometry. Intracellular PA levels were measured by HPLC, whereas autophagy and the expression profiles of PA key players were determined in HCT116, SW480 and HT29 colon cancer cells by Western blotting.

Results

We found that DENSpm-induced autophagy was inhibited by 3-MA treatment and Beclin-1 silencing, and that apoptotic cell death was increased by PA depletion and spermidine/spermine N¹-acetyltransferase (SSAT) upregulation. We also found that autophagy inhibition led to DENSpm-induced apoptosis through Atg5 down-regulation, p62 degradation and LC3 lipidation in both HCT116 and SW480 cells. p53 deficiency did not alter the response of the colon cancer cells to DENSpm-induced apoptotic cell death under autophagy suppression conditions.

Conclusions

From our results we conclude that DENSpm-induced apoptotic cell death is increased when autophagy is inhibited by 3-MA or Beclin-1 siRNA through PA depletion and PA catabolic activation in colon cancer cells, regardless p53 mutation status.