Simple Resistance Exercise helps Patients with Non-alcoholic Fatty Liver Disease.

To date, only limited evidence has supported the notion that resistance exercise positively impacts non-alcoholic fatty liver disease. We evaluated the effects of resistance exercise on the metabolic parameters of non-alcoholic fatty liver disease (NAFLD) in 53 patients who were assigned to either a group that performed push-ups and squats 3 times weekly for 12 weeks (exercise group; n=31) or a group that did not (control; n=22). Patients in the control group proceeded with regular physical activities under a restricted diet throughout the study. The effects of the exercise were compared between the 2 groups after 12 weeks. Fat-free mass and muscle mass significantly increased, whereas hepatic steatosis grade, mean insulin and ferritin levels, and the homeostasis model assessment-estimated insulin resistance index were significantly decreased in the exercise group. Compliance with the resistance exercise program did not significantly correlate with patient background characteristics such as age, sex, BMI and metabolic complications. These findings show that resistance exercise comprising squats and push-ups helps to improve the characteristics of metabolic syndrome in patients with non-alcoholic fatty liver disease.

Hyperhomocysteinemia enhances vascular inflammation and accelerates atherosclerosis in a murine model.

Although hyperhomocysteinemia (HHcy) is a well-known risk factor for the development of cardiovascular disease, the underlying molecular mechanisms are not fully elucidated. Here we show that induction of HHcy in apoE-null mice by a diet enriched in methionine but depleted in folate and vitamins B6 and B12 increased atherosclerotic lesion area and complexity, and enhanced expression of receptor for advanced glycation end products (RAGE), VCAM-1, tissue factor, and MMP-9 in the vasculature. These homocysteine-mediated (HC-mediated) effects were significantly suppressed, in parallel with decreased levels of plasma HC, upon dietary supplementation with folate and vitamins B6/B12. These findings implicate HHcy in atherosclerotic plaque progression and stability, and they suggest that dietary enrichment in vitamins essential for the metabolism of HC may impart protective effects in the vasculature.

Receptor for advanced glycation end products mediates inflammation and enhanced expression of tissue factor in vasculature of diabetic apolipoprotein E-null mice.

Advanced glycation end products (AGEs) and their cell surface receptor, RAGE, have been implicated in the pathogenesis of diabetic complications. Here, we studied the role of RAGE and expression of its proinflammatory ligands, EN-RAGEs (S100/calgranulins), in inflammatory events mediating cellular activation in diabetic tissue. Apolipoprotein E-null mice were rendered diabetic with streptozotocin at 6 weeks of age. Compared with nondiabetic aortas and kidneys, diabetic aortas and kidneys displayed increased expression of RAGE, EN-RAGEs, and 2 key markers of vascular inflammation, vascular cell adhesion molecule (VCAM)-1 and tissue factor. Administration of soluble RAGE, the extracellular domain of the receptor, or vehicle to diabetic mice for 6 weeks suppressed levels of VCAM-1 and tissue factor in the aorta, in parallel with decreased expression of RAGE and EN-RAGEs. Diabetic kidney demonstrated increased numbers of EN-RAGE-expressing inflammatory cells infiltrating the glomerulus and enhanced mRNA for transforming growth factor-beta, fibronectin, and alpha(1) (IV) collagen. In mice treated with soluble RAGE, the numbers of infiltrating inflammatory cells and mRNA levels for these glomerular cytokines and components of extracellular matrix were decreased. These data suggest that activation of RAGE primes cells targeted for perturbation in diabetic tissues by the induction of proinflammatory mediators.

Inhibitory effects of transforming growth factor-beta 1 on androgen-induced development of neonatal mouse seminal vesicles in vitro.

The effects of transforming growth factor-beta 1 (TGF beta 1) on the development of seminal vesicles (SVs) of neonatal mice were investigated in vitro. SVs from 0-day-old male mice were cultured for 2-8 days in serum-free, chemically defined medium with or without 5 alpha-dihydrotestosterone (DHT) at 10(-8) M in the presence or absence of TGF beta 1. Before culture, SVs from 0-day-old mice had no epithelial branches. SVs cultured in medium with DHT formed numerous epithelial branches after day 2 of culture, whereas epithelial branching did not occur in SVs cultured without DHT. The addition of TGF beta 1 (10 ng/ml) to DHT-containing medium almost completely inhibited the formation of epithelial branches. However, the removal of TGF beta 1 from DHT-containing medium on day 2 or 4 of culture initiated the formation of epithelial branches. TGF beta 1 (10 ng/ml) decreased [3H]thymidine labeling indices of both epithelium and mesenchyme of SVs cultured in medium with DHT during 8 days of culture, an effect that was reversed after the removal of TGF beta 1 from the medium on day 2 or 4 of culture. In contrast, TGF beta 1 (10 ng/ml) did not affect the labeling indices of either epithelium or mesenchyme of SVs cultured in medium without DHT. TGF beta 1 (10 ng/ml) also suppressed the normal increase in protein and DNA contents of SVs cultured in medium with DHT, whereas it did not reduce protein or DNA contents of SVs cultured in medium without DHT. The present study indicates that androgen-dependent growth of the SV can be reversibly inhibited by TGF beta 1, whereas androgen-independent growth of the SV is insensitive to TGF beta 1.

Inhibitory effects of retinoic acids on androgen-dependent development of neonatal mouse seminal vesicles in vitro.

The effects of retinoic acids (RAs) on development of seminal vesicles (SVs) of neonatal mice were investigated in vitro. SVs from 0-day-old male mice were cultured for 2-6 days in serum-free, chemically defined medium containing transferrin and BSA supplemented with 5alpha-dihydrotestosterone (DHT; 10(-8) M) and insulin (10 microg/ml), alone and in combination. Before culture, SVs from 0-day-old mice consisted of an unbranched epithelium surrounded by mesenchyme. SVs cultured in medium with DHT plus insulin or DHT alone formed numerous epithelial branches after day 2 of culture, whereas epithelial branching did not occur in SVs cultured with insulin alone. All-trans-RA or 13-cis-RA (10(-9)-10(-6) M) added to medium containing DHT plus insulin or DHT alone inhibited epithelial branching in a dose-dependent manner. This inhibitory effect was reversible after removal of the retinoids from the medium on day 4 of culture. These RAs also decreased [3H]thymidine labeling indexes of both epithelium and mesenchyme of SVs cultured in medium with DHT plus insulin or DHT alone and inhibited the increase in their protein contents. 9-Cis-RA was less inhibitory than all-trans-RA or 13-cis-RA on epithelial branching, [3H]thymidine labeling indexes of epithelium and mesenchyme, and protein content of SVs cultured in medium with DHT and insulin. In the absence of DHT (insulin alone), all-trans-RA did not affect either the [3H]thymidine labeling indexes of epithelium and mesenchyme or the protein content of cultured SVs. Reverse transcriptase-PCR demonstrated strong expression of transcripts for mouse RA receptors (RARalpha, RARgamma, and RXRalpha), with lower levels of expression of RARbeta, RXRbeta, and RXRgamma in neonatal SVs. The present results indicate that RAs reversibly inhibit androgen-dependent development of neonatal mouse SVs, most likely through RARs.

Early effects of castration on the vascular system of the rat ventral prostate gland.

Recent studies have found that blood flow to the rat ventral prostate gland is drastically reduced at an early time after castration. These observations caused us to reevaluate the effects of castration on the various cell populations of the ventral prostate, especially those in the prostatic vascular system. Sections of ventral prostate glands obtained at different times after castration were analyzed using the TUNEL (terminal deoxynucleotide transferase-mediated dUTP nick END labeling) staining method to quantify apoptosis in different cell types. The results of this analysis showed a significant increase in TUNEL staining of prostate endothelial and (nonendothelial) stromal cells as early as 12 h postcastration that continued to 24 h after castration. In contrast, TUNEL labeling of prostate epithelial cells was not significantly increased compared with control values until 72 h after castration. The use of dual immunohistochemical staining procedures (anti-CD31 for endothelial cells or antismooth muscle actin for smooth muscle cells combined with TUNEL labeling) allowed us to confirm that the TUNEL-positive vascular cells at these early times after castration were endothelial in nature, whereas smooth muscle cells surrounding the prostate glands or portions of the afferent vascular endothelium were rarely TUNEL labeled. Electron microscopic evaluation of ventral prostate tissues at 48 h after castration provided further morphological evidence for the occurrence of apoptosis in prostate endothelial cells. Finally, the Lendrum-Fraser histochemical procedure used to identify fibrin leakage in tissues with vascular damage was applied to sections of the ventral prostate gland. This stain revealed diffuse fibrin accumulation in periglandular areas outside the capillaries and blood vessels in prostates from 24-h castrated rats, but not in prostates of sham-operated rats. Our results confirm an early effect of castration on the vascular system of the rat ventral prostate identified by increased apoptosis of endothelial cells and vascular leakiness. As these changes temporally precede the loss of epithelial cells, we propose that they may be causal rather than incidental to regression of the rat ventral prostate after castration.

Idiopathic hypocomplementemic interstitial nephritis with extensive tubulointerstitial deposits.

Most forms of interstitial nephritis are cell mediated and lack tubulointerstitial immune deposits. These forms include allergic, infectious, and idiopathic interstitial nephritis. Immune complex deposits in the tubular basement membranes and interstitium most commonly are encountered in conjunction with glomerular diseases. Predominantly tubulointerstitial immune deposits without significant glomerular involvement can occur in Sjögren's syndrome and in a small subset of lupus nephritis. We report eight unusual cases of tubulointerstitial nephritis with massive tubulointerstitial immune deposits occurring in adults with hypocomplementemia and no evidence of systemic lupus erythematosus or Sjögren's disease. Most patients were older men. The renal biopsy specimens manifested a spectrum of changes ranging from tubulointerstitial nephritis to atypical lymphoid hyperplasia to changes suggestive of marginal zone B-cell lymphoma. Chronic local antigenic stimulation may predispose to lymphoma in these cases, analogous to what is postulated to occur in cases of mucosa-associated lymphoid tissue (MALT) lymphomas in extranodal sites, such as salivary gland, stomach, and thyroid. The preferential tubulointerstitial immune deposition and significant interstitial plasma cell component suggest pathomechanisms that involve local immune complex formation.

Congenital focal segmental glomerulosclerosis associated with beta4 integrin mutation and epidermolysis bullosa.

We report the occurrence of congenital nephrotic-range proteinuria secondary to focal segmental glomerulosclerosis in an infant with epidermolysis bullosa and pyloric atresia. A homozygous missense mutation, R1281W, in exon 31 of the beta4 integrin gene, ITGB4, was identified. By immunofluorescence, beta4 integrin expression was reduced in both dermal keratinocytes and glomerular podocytes. This is the first demonstration of beta4 integrin expression in human glomeruli. We postulate a role for altered beta4 integrin function in the mediation of the glomerular permeability defect.

HIV-associated nephropathy: an urban epidemic.

Human immunodeficiency virus-associated nephropathy (HIVAN) is the most common form of chronic renal disease in HIV-1-seropositive patients. Over 85% of cases of HIVAN occur in African-American patients and it is the third leading cause of ESRD in blacks age 20 to 64. Changes in incidence rates of HIVAN have coincided with changes in AIDS incidence rates. The demographics of the AIDS/HIV-1 epidemic indicate that the risk pool for HIVAN will continue to grow and that urban Nephrology centers will continue to see high rates of HIVAN. In addition, improvements in survival rates of HIV-1-seropositive patients on hemodialysis and improved treatment of HIVAN with highly active antiretroviral therapy (HAART) and angiotensin-converting enzyme (ACE)-inhibitors will result in an increased prevalence of HIVAN in the end-stage renal disease (ESRD) and pre-ESRD patient populations.

Adefovir nephrotoxicity: possible role of mitochondrial DNA depletion.

This report investigates the pathomechanism of acute renal failure caused by toxic acute tubular necrosis after treatment with the antiretroviral agent adefovir. A 38-year-old white homosexual man with human immunodeficiency virus infection and no history of opportunistic infections was maintained on highly active antiretroviral therapy (HAART), including hydroxyurea, stavudine, indinavir, ritonavir, and adefovir dipivoxil. Histologic examination of the renal biopsy showed severe acute tubular degenerative changes primarily affecting the proximal tubules. On ultrastructural examination, proximal tubular mitochondria were extremely enlarged and dysmorphic with loss and disorientation of their cristae. Functional histochemical stains for mitochondrial enzymes revealed focal tubular deficiency of cytochrome C oxidase (COX), a respiratory chain enzyme partially encoded by mitochondrial DNA (mtDNA), with preservation of succinate dehydrogenase, a respiratory chain enzyme entirely encoded by nuclear DNA (nDNA). Immunoreactivity for COX subunit I (encoded by mtDNA) was weak to undetectable in most tubular epithelial cells, although immunoreactivities for COX subunit IV and iron sulfur subunit of respiratory complex III (both encoded by nDNA) were well preserved in all renal tubular cells. Single-renal tubule polymerase chain reaction revealed marked reduction of mtDNA in COX-immunodeficient renal tubules. We conclude that adefovir-induced nephrotoxicity is mediated by depletion of mtDNA from proximal tubular cells through inhibition of mtDNA replication. This novel form of nephrotoxicity may serve as a prototype for other forms of renal toxicity caused by reverse transcriptase inhibitors.

Expression of platelet-derived endothelial cell growth factor in prostatic adenocarcinoma.

Angiogenesis is thought to play critical roles in local tumor growth and eventual metastasis. No studies have examined the expression of platelet-derived endothelial cell growth factor (PD-ECGF) in prostatic tissues. Prostatic tissues were obtained from 36 prostatic adenocarcinoma patients. We assessed the expression of PD-ECGF using ELISA and immunohistochemistry. The mean level of PD-ECGF in prostatic adenocarcinomas was higher than that in neighboring normal prostatic tissues in ELISA. Immunohistochemistry showed that the expressions of PD-ECGF, which were associated with increase of microvessel count, were found in the endothelial cells, macrophages, lymphocytes, or fibroblasts. These results suggest that PD-ECGF is involved in the development of prostatic adenocarcinoma.

Immunohistochemical study of cyclooxygenases in prostatic adenocarcinoma; relationship to apoptosis and Bcl-2 protein expression.

BACKGROUND: Cyclooxygenase (COX) is a key enzyme in the conversion of arachidonic acid to prostaglandins and other eicosanoids. A recent report has indicated a selective COX-2 inhibitor resulted in increased apoptosis and down-regulated bcl-2 expression in the androgen-sensitive human prostate cancer cell. We investigated the localization of COXs in prostatic adenocarcinoma and the possible correlation of androgen blockade with its expression. MATERIALS AND METHODS: The immunohistochemical expression of COX-1, COX-2 and bcl-2 protein was studied using paraffin-embedded archival tissues both before and after hormonal therapy. The number of apoptotic cells was also determined. RESULTS: Immunohistochemistry for COX-1, not COX-2, showed the constitutive expression in the stroma of normal prostate. The expression of COX-2 protein was detected less often than that of COX-1 protein in most cases of prostatic adenocarcinoma before hormonal therapy. Neoadjuvant hormonal therapy induced the expression of COX-2 protein in smooth muscle cells, fibroblasts and adenocarcinoma cells. The expression after hormonal therapy was possibly correlated with the bcl-2 protein expression. CONCLUSION: The present study is the first to demonstrate the immunohistochemical expression of COXs in human prostate tissue and indicated that COXs were relevant to homeostasis and tumor development of the prostate.

Immunohistochemical study of the receptors for retinoic acid in prostatic adenocarcinoma.

BACKGROUND: Retinoids play an important role in regulating cellular proliferation and differentiation. Although their effects are mediated by retinoic acid receptors (RARs) and retinoid X receptors (RXRs), limited information is available about the expression of RARs and RXRs in prostatic adenocarcinoma. We intended in this study to elucidate further the participation of those receptors in tumor progression of human prostate. MATERIALS AND METHODS: The immunohistochemical expression of RARs and RXRs proteins was studied using paraffin-embedded archival tissues obtained from patients with and without neoadjuvant hormonal therapies for prostatic adenocarcinoma. RESULTS: The immunoreactivity against RAR-alpha, RAR-gamma, RXR-alpha and RXR-gamma were detected in many, if not all, prostatic adenocarcinoma cells of the cases without and with neoadjuvant hormonal therapy, whereas less expression of both RAR-beta and RXR-beta were identified. Positively stained adenocarcinoma cells with RAR-beta and RXR-beta were found to be decreased in the cases with neoadjuvant therapy. In addition, the specimens showing positive immunoreaction of RAR-beta were limited to the cases of moderately- and poorly-differentiated but not well-differentiated, adenocarcinomas. CONCLUSION: We first demonstrated the expression of RARs and RXRs proteins in prostatic adenocarcinoma using subtype-specific antibodies. Immunohistochemistry using those antibodies might give an indication of susceptibility of the patients to retinoid therapy as a possible treatment of prostatic adenocarcinoma.

Apoptosis in prostatic adenocarcinomas; a study of relationship to Ki-67 and Bcl-2 protein expression.

BACKGROUND: Tissue samples from patients with prostatic adenocarcinoma (PCA) were examined for relationship between apoptosis and expression of Ki-67 and bcl-2 protein. MATERIALS AND METHODS: Prostatic tissue was obtained from 29 prostatic cancer patients with no prior therapy. The number of apoptotic cells (apoptotic index; AI) and the percentage of Ki-67 stained cells (Ki-67 labeling index; LI) were compared with the expression of bcl-2 which was evaluated by the proportion and intensity of tumor cell immunostaining. RESULTS: Both AI and LI of moderately and poorly differentiated PCAs were significantly higher than those of well differentiated PCAs. There was a significant correlation between AI and LI. Bcl-2 protein was expressed less in moderately and poorly differentiated PCAs than in well differentiated PCAs. Moreover, there was no correlation between the bcl-2 protein expression and the AI. CONCLUSIONS: The results suggest that, in PCAs, the degree of apoptosis is correlated with the cellular proliferation, and that expression of bcl-2 protein is not a critical factor that determines the degree of apoptosis.

The expression of basic fibroblast growth factor and vascular endothelial growth factor in prostatic adenocarcinoma: correlation with neovascularization.

BACKGROUND: Neovascularization associated with tumor invasion and metastasis may be stimulated by factors which are released from tumor cells, tumor-associated inflammatory cells or extracellular matrix. Although basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) have been characterized as promoters of angiogenesis, their precise localization in prostatic adenocarcinoma remains unclear. MATERIALS AND METHODS: In this study, the immunohistochemical expression of the growth factors and their receptors were studied using paraffin-embedded archival tissues before and after neoadjuvant hormonal therapy. The mRNA expression of the growth factors was also examined using an in situ hybridization (ISH) technique. RESULTS: The ISH study demonstrated that bFGF mRNA was present only in the stromal cells whilst that VEGF mRNA was present only in the adenocarcinoma cells. In contrast, the immunohistochemical study showed that bFGF, FGF receptor, VEGF and VEGF receptor proteins were expressed in adenocarcinoma cells and in endothelial cells. We also observed that microvessel density in prostatic adenocarcinoma was correlated with the degree of the expression of growth factors in the cases without neoadjuvant hormonal therapy. Additionally, the expression of those receptor proteins were much frequently identified in the cases with neoadjuvant hormonal therapy than those without it, while virtually no change in the expression of ligands was observed between the cases without and with neoadjuvant therapies. CONCLUSIONS: In prostatic adenocarcinoma, bFGF and VEGF may correlate with neovascularization through each characteristic pathway. In addition, we assumed that neoadjuvant hormonal therapy may have minimal inhibitory effects on bFGF, VEGF and their receptors.

[Effects of castration on apoptosis and transforming growth factor-beta 1 mRNA in the neonatal mouse seminal vesicles].

BACKGROUND: In the adult male rat prostate, castration induces apoptosis of epithelial cells concomitant with the increase in transforming growth factor-beta 1 (TGF-beta 1). In the present study, we investigated the effects of castration on apoptosis and TGF-beta 1 mRNA in neonatal mouse seminal vesicles. METHODS: 5-day-old BALB/c mice were castrated by Pfeifer's method. We estimated the weight, 3H-thymidine uptake by whole seminal vesicles, the amount of TGF-beta 1 mRNA by RT-PCR method, and the apoptotic index of both epithelium and mesenchyme. RESULTS: The castration of 5-day-old neonatal mice resulted in much less weight of seminal vesicles and DNA synthesis estimated by 3H-thymidine uptake by whole seminal vesicles compared to intact neonatal mice, indicating that the growth of neonatal mouse seminal vesicles depends on androgens secreted by the testis. The amount of TGF-beta 1 mRNA estimated by RT-PCR method increased 4 days after castration at 5 days of age. However, the castration did not induce apoptosis in the seminal vesicles. CONCLUSION: The present study indicates that castration of neonatal mice does not induce apoptosis in the seminal vesicles, although it does a transient increase in TGF-beta 1 mRNA in the seminal vesicles.

EXpression of ErbB proteins in human prostate.

ErbB proteins are widely expressed in human and animal tissues, notably in cells of epithelial or neuroendocrine origin. Protein expression and interactions of ErbBs were examined in prostate cancer specimens. Expression of ErbB1-4 proteins was determined with immunohistochemical methods using each monoclonal antibody in 20 prostatic adenocarcinomas. The 4 ErbB proteins were widely expressed in normal, hyperplastic and cancerous tissues of the prostate. ErbBs may contribute to normal development or tumor growth and progression in human prostate.