A role of a lymphocyte tryptase, granzyme A, in experimental ulcerative colitis

In the large-intestinal mucosae of rats orally administered dextran sulfate sodium, which induces an enteritis resembling ulcerative colitis (UC), the activity for granzyme A, a lymphocyte tryptase, increased at an earlier stage than that at which UC markers (growth-regulated gene product/cytokine-induced neutrophil chemoattractant-1 and caspase-3) increased. This suggests involvement of the enzyme in the exacerbation and perpetuation of enteritis.     

M1 Macrophage exosomes MiR-21a-5p aggravates inflammatory bowel disease through decreasing E-cadherin and subsequent ILC2 activation

Abnormal immune regulation is a key feature of the complex pathogenic mechanism of ulcerative colitis (UC). In particular, macrophages and group 2 innate lymphoid cells (ILC2s) are important components of natural immunity that have been shown to play important roles in the pathogenesis of UC, as well as decreased E-cadherin expression on the colonic mucosa. However, it remains unclear how these components interact with each other. In this study, we investigated the molecular mechanisms of UC mediated by macrophage-derived exosomes. We showed for the first time that miR-21a-5p expression is increased in the peritoneal exosomes of mice with dextran sulphate sodium induced enteritis and that miR-21a-5p expression correlates negatively with E-cadherin expression in enterocytes. Moreover, we confirmed that miR-21a-5p was mainly derived from M1 macrophages and demonstrated that KLRG1, a surface inhibitory receptor on ILC2s, participated in excessive ILC2 activation in UC by promoting GATA-3. In conclusion, our results suggest molecular targets and provide a theoretical basis for elucidating the pathogenesis of UC and improving its treatment.     

Influence of unilateral castration and increased plane of nutrition on sexual development of Holstein bulls. II. Histologic development of the testes

Sixty-five Holstein bull calves were assigned in an experiment to determine the effects of unilateral castration (UC) at 1 week of age and of two levels of nutrition after 6 months on reproductive development to 16 months. Five animals were killed at 1 wk and half the remainder UC at that age. Groups of 5 in all factorial groups were killed at 2, 4, 8 and 16 months. Tubular diameter increased with age (P<.01) and at 8 and 16 months with UC (P<.01). Epithelial area at 8 and 16 months increased with age and UC (P<.01). The percents of tubular and intertubular tissue varied with age (P<.01) with the tubular tissue having its highest value at 8 months. Indexes of both total tubular and intertubular tissue were increased with age and UC (P<.01). The number of type A spermatogonia per cross section of stage 1 tubules of 16-month bulls was increased by UC (P<.05).     

Association of DNA Methyltransferases 3A and 3B Polymorphisms,and Plasma Folate Levels with the Risk of Urothelial Carcinoma

Background

Interindividual genetic variations of human DNA methyltransferases (DNMTs), which involve the methyl donor from the folate-related one-carbon metabolism pathway, are hypothesized as a risk factor for urothelial carcinoma (UC). Therefore, we evaluated the role of gene-environment interaction in UC carcinogenesis.

Methods

A hospital-based case-control study was conducted by recruiting 192 patients with UC and 381 controls. Their plasma folate levels were measured using a competitive immunoassay kit. In addition, DNMT3A −448A>G and DNMT3B −579G>T genotyping was evaluated using a polymerase chain reaction-restriction fragment length polymorphism technique. Multivariate logistic regression and 95% confidence intervals (CIs) were applied to estimate the UC risk.

Results

We observed that patients with UC exhibited a higher prevalence rate of folate insufficiency (folate levels ≤6 ng/mL) compared with the controls (35.94% and 18.37%, respectively). Furthermore, folate levels were higher in the prevalent UC patients than in the incident UC patients. However, folate insufficiency was similarly associated with a nearly two-fold increase in the risk of UC regardless of the UC patient group. In addition, the frequencies of the variant alleles for DNMT3A and DNMT3B were 0.80 and 0.92, respectively, and no association was observed with UC risk. However, participants with a variant homozygous genotype of DNMT3B −579G>T and folate insufficiency or with high cumulative cigarette smoking exhibited an increased risk of UC.

Conclusion

Overall, environmental factors may contribute more significantly to UC carcinogenesis compared with genetic susceptibility. Future studies should investigate other polymorphisms of DNMT3A and DNMT3B to determine genetic susceptibility.     

A novel model of ischemic enteritis induced in rats by stenosis of the superior mesenteric artery

AimsWe established a new model of ischemic enteritis in rats and evaluated its usefulness for screening prophylactic drugs.Main methodsMale SD rats were used after 18 h of fasting. Under ether anesthesia, the superior mesenteric artery (SMA) was exposed, and a calibrated stenosis was produced by placing a needle on a blood vessel, ligating both the vessel and needle, and then removing the needle from the ligature.Key findingsThe stenosis caused severe damage on the anti-mesenteric side of the small intestine within 3 days; the severity of the damage increased with the gauge of a needle. No damage occurred in the small intestine following the stenosis with a needle of less than 21 gauge. Multiple hemorrhagic lesions occurred at an incidence of 100% when a 23-gauge needle was used. The development of enteritis was accompanied by enterobacterial invasion in the mucosa, with an up-regulation of inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production. The ischemia-induced enteritis was significantly prevented by repeated treatment with aminoguanidine (a selective iNOS inhibitor), L-NAME (a nonselective NOS inhibitor), ampicillin, and aztreonam (a gram-negative bacterium antibiotic), but not vancomycin (a gram-positive bacterium antibiotic).SignificanceThese results showed that a novel model of ischemic enteritis is induced in rats by stenosis of the SMA, this model may be useful for screening drugs against ischemic enteritis, and gram-negative bacteria as well as iNOS/NO are involved in the pathogenesis of enteritis in this model.     

Effects of unilateral castration on morphologic characteristics of the testis in one-, two-, and three-year-old stallions

The effects of unilateral castration on testicular compensatory hypertrophy were measured in 12 Morgan stallions, four each at one, two, and three years of age. They were randomized within age to intact (IN) or unilaterally castrated (UC) groups. Allotment and surgery were in January 1983 and total castration was in June 1983, 150 d after unilateral castration. Testis weight increased linearly with age (P < 0.01) and was increased by unilateral castration (P < 0.07). Epididymal weight also increased linearly with age (P < 0.05) and was heavier in UC animals (P = 0.15). Tubule diameter (P < 0.10) and epithelial height (P < 0.03) were greater in UC than in IN stallions. In conclusion, testes of stallions underwent compensatory hypertrophy after unilateral castration.     

Altered angiogenic balance in ulcerative colitis: a key to impaired healing?

Angiogenesis is an essential component of ulcer healing since it assures delivery of oxygen and nutrients to the healing site. Previous studies demonstrated increased serum and tissue levels of vascular endothelial growth factor (VEGF, the most potent angiogenic growth factor) in patients with active ulcerative colitis (UC) and animal models of UC. However, there is no explanation why the healing of UC-related mucosal injury is impaired despite increased expression of VEGF. Expression of angiogenesis inhibitors, angiostatin and/or endostatin, in UC has not been determined before. We examined expression of VEGF, angiostatin, and endostatin in two models of experimental UC. The results revealed that in addition to increased VEGF, both endostatin and angiostatin levels were markedly (2-3-folds) increased in colonic mucosa at early stage of experimental UC. This is the first demonstration that colitis triggers increase in angiostatin and endostatin levels. The results may explain why mucosal lesions heal slowly despite increased VEGF levels, and may provide a novel and mechanistic insight into UC.     

A variable undecad repeat domain in cavin1 regulates caveola formation and stability

Caveolae are plasma membrane invaginations involved in transport, signalling and mechanical membrane sensing in metazoans. Their formation depends upon multiple interactions between membrane‐embedded caveolins, lipids and cytosolic cavin proteins. Of the four cavin family members, only cavin1 is strictly required for caveola formation. Here, we demonstrate that an eleven residue (undecad) repeat sequence (UC1) exclusive to cavin1 is essential for caveolar localization and promotes membrane remodelling through binding to phosphatidylserine. In the notochord of mechanically stimulated zebrafish embryos, the UC1 domain is required for caveolar stability and resistance to membrane stress. The number of undecad repeats in the cavin1 UC1 domain varies throughout evolution, and we find that an increased number also correlates with increased caveolar stability. Lastly, we show that the cavin1 UC1 domain induces dramatic remodelling of the plasma membrane when grafted into cavin2 suggesting an important role in membrane sculpting. Overall, our work defines a novel conserved cavin1 modular domain that controls caveolar assembly and stability.     

Serum follicle stimulating hormone, luteinizing hormone, and testosterone in sexually stimulated intact and unilaterally castrated rams

Ten two-year-old intact (IN) and unilaterally castrated (UC) Targhee rams were exposed to an estrogenized ewe each week from June to October. Each week the rams were subjectively evaluated for libido (10 for high interest and 1 for no interest). Semen was collected from all cooperating rams and evaluated for volume, concentration, and motility. Every 2 wk, blood samples were obtained at -30 and 0 min before and 30 and 60 min after ewe access. Serum was harvested; follicle stimulating hormone (FSH), luteinizing hormone (LH), and testosterone concentrations were quantified by radioimmunoassay (RIA). Week 5 of ewe access was assigned as Week 1. Libido scores rose from a low on Week 1, with eight rams ejaculating, to a high on Week 12, with all rams ejaculating (Week 1, 5.0 +/- 1.0; Week 12, 10.0 +/- 0.0). The product of testis length and width was significantly greater in UC compared with IN rams (88.4 +/- 1.4 versus 73.2 +/- 1.0 cm(2), respectively). Serum FSH concentrations (ng/ml) were greater (P < 0.05) in UC than IN rams and dropped over the experimental period. Serum LH concentrations (ng/ml) were significantly greater in UC compared with IN rams. This difference was more pronounced in Weeks 1 and 3 compared with Weeks 11 and 13. Serum testosterone concentrations (ng/ml) were similar in UC and IN rams throughout the experiment. In conclusion, serum testosterone was not altered in UC rams; however, serum FSH and LH concentrations were increased in UC rams. Unilateral castration did not enhance the normal changes in semen quantity and quality in the rams from July to October.     

Necrotizing lesions in the intestine, gizzard, and liver in captive capercaillies (Tetrao urogallus) associated with Clostridium perfringens.

During the period from 1982 to 1991, 863 captive and 32 wild capercaillies (Tetrao urogallus) were necropsied. The most common cause of death in captive capercaillies was necrotizing enteritis, diagnosed in 110 (13%) birds. Of these, 31 (28%) birds also had necrotizing lesions in the liver. Necrotizing gastritis occurred in 29 birds, two of which had concurrent necrotizing enteritis. In the capercaillies with necrotizing enteritis, Clostridium perfringens type A was isolated more frequently and in larger numbers than in birds which died from other causes. Thus, Clostridium perfringens type A may be of etiological importance in necrotizing enteritis. Necrotizing enteritis was not diagnosed in wild capercaillies.     

Soybean meal induces intestinal inflammation in common carp (Cyprinus carpio L.)

The development of soybean meal (SBM) induced enteritis in the hindgut of the omnivorous common carp (Cyprinus carpio L.). The developed condition was assessed when carp, continuously fed on animal protein, were transferred to a diet in which 20% of the protein was replaced by SBM. After week 1, most of the inflammation parameters were already present, but at week 3, a strong aggravation of the condition was observed which included a shortening of the mucosal folds, the disappearance of the supranuclear vacuoles, an increased number of goblet cells, a thickened lamina propria and sub-epithelial mucosa with increased numbers of basophilic granulocytes as well as a decreased uptake capacity of enterocytes (impaired endocytosis and microvilli). Contrary to previous observations made with respect to Atlantic salmon, common carp start to recover from the fourth to the fifth week after switching to SBM feeding. At this stage, the supranuclear vacuoles refill and most of the parameters revert to basal levels. During the enteritis process, a real-time quantitative PCR analysis was conducted to measure the expression of inflammatory and anti-inflammatory cytokine genes in the isolated intraepithelial lymphocytes (IEL). The pro-inflammatory interleukin 1 beta (IL-1 beta) and tumour necrosis factor alpha1 (TNF-alpha1) genes were up-regulated during the inflammation process while the anti-inflammatory interleukin 10 (IL-10) was down-regulated after an initial up-regulation at week 1. Transforming growth factor beta (TGF-beta) expression showed an up-regulation from week 3 onwards despite the high Ct value and the low primer efficiency shown. This study confirms the contribution of IEL (mainly T-like cells) and basophils in the enteritis process. In addition, the results show a clear involvement of up- and down-regulated cytokine genes in both the onset and recovery of the SBM-induced enteritis in the hindgut of carp.     

Effect of combining an ACE inhibitor and a VDR activator on glomerulosclerosis, proteinuria, and renal oxidative stress in uremic rats

Angiotensin-converting enzyme (ACE) inhibitors ameliorate the progression of renal disease. In combination with vitamin D receptor activators, they provide additional benefits. In the present study, uremic (U) rats were treated as follows: U+vehicle (UC), U+enalapril (UE; 25 mg/l in drinking water), U+paricalcitol (UP; 0.8 μg/kg ip, 3 × wk), or U+enalapril+paricalcitol (UEP). Despite hypertension in UP rats, proteinuria decreased by 32% vs. UC rats. Enalapril alone, or in combination with paricalcitol, further decreased proteinuria (≈70%). Glomerulosclerosis and interstitial infiltration increased in UC rats. Paricalcitol and enalapril inhibited this. The increase in cardiac atrial natriuretic peptide (ANP) seen in UC rats was significantly decreased by paricalcitol. Enalapril produced a more dramatic reduction in ANP. Renal oxidative stress plays a critical role in inflammation and progression of sclerosis. The marked increase in p22(phox), a subunit of NADPH oxidase, and decrease in endothelial nitric oxide synthase were inhibited in all treated groups. Cotreatment with both compounds inhibited the uremia-induced increase in proinflammatory inducible nitric oxide synthase (iNOS) and glutathione peroxidase activity better than either compound alone. Glutathione reductase was also increased in UE and UP rats vs. UC. Kidney 4-hydroxynonenal was significantly increased in the UC group compared with the normal group. Combined treatment with both compounds significantly blunted this increase, P < 0.05, while either compound alone had no effect. Additionally, the expression of Mn-SOD was increased and CuZn-SOD decreased by uremia. This was ameliorated in all treatment groups. Cotreatment with enalapril and paricalcitol had an additive effect in increasing CuZn-SOD expression. In conclusion, like enalapril, paricalcitol alone can improve proteinuria, glomerulosclerosis, and interstitial infiltration and reduce renal oxidative stress. The effects of paricalcitol may be amplified when an ACE inhibitor is added since cotreatment with both compounds seems to have an additive effect on ameliorating uremia-induced changes in iNOS and CuZn-SOD expression, peroxidase activity, and renal histomorphometry.     

Effects of management methods,social organization,and physical space on primate behavior and health

The incidence of clinical treatment in a large colony of pigtailed macaques (Macaca, nemestrina) was studied retrospectively to assess the effectiveness of three sequential housing and management protocols. In the initial standard protocol, groups were housed in single rooms, with each room occupied by a different group after daily cleaning. In the experimental protocol, animals were housed in two-room suites dedicated exclusively to the resident group. This protocol was meant to reduce enteritis due to communication of Shigella flexneri across groups. Isolation of groups was coupled with the doubling of available space in an attempt to reduce stress. Under a final compromise protocol, a two-room suite was dedicated to each group, but the group occupied only one room at a time. The compromise protocol maximized isolation and reduced the available space to the level under the standard protocol. Thus, the study involved two main factors, the degree of group isolation and the amount and configuration of physical space available to each group. Under the experimental conditions enteritis increased, other communicable diseases did not decrease, and trauma from fighting increased by over 45% even though more space was available. Under the compromise conditions, trauma, enteritis, respiratory disorders, and inflammation all decreased markedly. Behavioral studies identified beneficial social behavior patterns associated with spatial reduction as the basis of the decrease in aggression, trauma, and disease under the compromise conditions.     

Genes involved in differentiation, stem cell renewal, and tumorigenesis are modulated in telomerase-immortalized human urothelial cells

The expression of hTERT, the catalytic subunit of telomerase, immortalizes normal human urothelial cells (NHUC). Expression of a modified hTERT, without the ability to act in telomere maintenance, did not immortalize NHUC, confirming that effects at telomeres are required for urothelial immortalization. Previous studies indicate that inhibition of telomerase has an immediate effect on urothelial carcinoma (UC) cell line viability, before sufficient divisions to account for telomere attrition, implicating non-telomere effects of telomerase in UC. We analyzed the effects of telomerase on gene expression in isogenic mortal and hTERT-transduced NHUC. hTERT expression led to consistent alterations in the expression of genes predicted to be of phenotypic significance in tumorigenesis. A subset of expression changes were detected soon after transduction with hTERT and persisted with continued culture. These genes (NME5, PSCA, TSPYL5, LY75, IGFBP2, IGF2, CEACAM6, XG, NOX5, KAL1, and HPGD) include eight previously identified as polycomb group targets. TERT-NHUC showed overexpression of the polycomb repressor complex (PRC1 and PRC4) components, BMI1 and SIRT1, and down-regulation of multiple PRC targets and genes associated with differentiation. TERT-NHUC at 100 population doublings, but not soon after transduction, showed increased saturation density and an attenuated differentiation response, indicating that these are not acute effects of telomerase expression. Some of the changes in gene expression identified may contribute to tumorigenesis. Expression of NME5 and NDN was down-regulated in UC cell lines and tumors. Our data supports the concept of both telomere-based and non-telomere effects of telomerase and provides further rationale for the use of telomerase inhibitors in UC.     

Effects of unilateral castration on serum luteinizing hormone, follicle stimulating hormone, and testosterone concentrations in one-, two-, and three-year-old stallions

The endocrine control of compensatory hypertrophy was investigated in 12 Morgan stallions, four each at one, two and three years of age. Half were assigned to be unilaterally castrated (UC) in January and half to remain intact (IN). Nine blood samples were taken from each stallion at half-hour intervals 30, 90, and 150 d after unilateral castration for radioimmunoassay of serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), and testosterone. Mean serum LH concentration was greater (P<0.06) in UC than IN stallions; however, the difference was greatest at 30 d and least at 150 d. Serum LH was greater (P<0.01) in two- and three-year-olds than in one-year-olds. The mean log(10) for serum FSH concentration was greater (P<0.06) in UC than IN stallions. Mean serum testosterone concentrations were similar in UC and IN stallions for all sample days, suggesting that the single testes of the UC stallions produced as much testosterone as the two testes of the IN stallions. Two- and three-year-old stallions had greater (P<0.01) serum testosterone than one-year-old stallions. Unilateral castration of stallions was associated with a significant increase in serum LH and FSH concentrations and, perhaps, higher intratesticular testosterone, which may explain, in part, the compensatory hypertrophy noted in the remaining testis.     

Mc3 和Mc5 在溃疡性结肠炎癌变过程中的意义*

目的:探讨Mc3和Mc5在溃疡性结肠炎-上皮内瘤变-癌变过程中的表达和临床意义。方法:利用免疫组化染色检测168例溃疡性结肠炎(UC)、溃疡性结肠炎伴上皮内瘤变(UC+IN)、结直肠癌(CRC)中Mc3和Mc5的表达情况,分析Mc3和Mc5的表达水平在溃疡性结肠炎-上皮内瘤变-癌变过程中变化趋势,探讨潜在的临床意义。结果:Mc3和Mc5在UC的表达阳性率分别为30%和29%,在UC+IN的阳性率分别是48%和46%,在CRC组的阳性率分别是91%和89%。统计分析发现,Mc3和Mc5在溃疡性结肠炎、上皮内瘤变和结直肠癌的表达逐渐增加,差异具有统计学意义(P<0.05)。结论:Mc3和Mc5在溃疡性结肠炎-上皮内瘤变-癌变过程中表达逐渐增加,检测Mc3及Mc5的表达有助于判断癌变风险,且Mc3和Mc5具有较好的一致性。     

Pre-Illness Isoflavone Consumption and Disease Risk of Ulcerative Colitis: A Multicenter Case-Control Study in Japan

Introduction

Previous studies have suggested that estrogens play a role in the development of ulcerative colitis (UC). Because isoflavones have a similar structure to 17β-estradiol, dietary consumption of isoflavones may have similar influences on the development of UC. We examined the association between pre-illness isoflavone consumption and the risk of UC.

Materials and Methods

We conducted a hospital-based case control study, and compared the dietary habits of 126 newly diagnosed UC cases with those of 170 age- and gender-matched hospital controls. Information on dietary factors was collected using a self-administered diet history questionnaire. To consider potential changes in dietary habits due to disease symptoms, the habits were assessed separately during the previous 1 month and at 1 year before the recruitment.

Results

In the assessment of dietary habits during the previous 1 month, the highest tertile of isoflavone consumption revealed an increased odds ratio (OR) for UC (OR = 2.79; 95% confidence interval (CI), 1.39–5.59; Trend P = 0.004). A significant association was also observed for the dietary assessment at 1 year before, when most UC cases had not yet experienced their first disease symptoms (OR = 2.06; 95% CI, 1.05–4.04; Trend P = 0.04). Associations were more pronounced in females (OR in highest tertile of isoflavone consumption at 1 year before = 4.76; 95% CI, 1.30–17.5; Trend P = 0.02) but were obscured in males (corresponding OR = 1.21; 95% CI, 0.49–3.01; Trend P = 0.63).

Conclusions

Dietary isoflavone consumption may be associated with an increased risk of UC, particularly in females. Prospective cohort studies are warranted to confirm these findings.     

Influence of unilateral castration and increased plane of nutrition on sexual development of Holstein bulls. I. Growth and sperm production

Sixty-five Holstein bull calves were used to study the effects of unilateral castration (UC) and increased plane of nutrition on the growth and development of the reproductive system. Bulls were slaughtered at 1 wk., 2, 4, 8 and 16 months. Half of each slaughter group above one week was unilaterally castrated at 7 days of age. Half of the bulls remaining at 6 months of age received 90% of their recommended daily TDN allowance while the remainder received 120%. Compensatory hypertrophy was evident as early as 2 months and the degree of compensation increased for the duration of the experiment (Age x UC, P<.01). By 16 months of age the remaining testis of UC animals was 73% heavier than the average testis weight of intact bulls. While epididymal weight was significantly increased by UC, seminal vesicle weight was not. UC bulls produced significantly more sperm per testis than intact bulls both from the onset of puberty to slaughter and for the 16 week period prior to slaughter. Testis sperm concentration was similar in UC and intact bulls. UC at one weel of age caused greater testis growth and greater sperm production per testis, but did not promote earlier puberty.     

Hsp10, Hsp70, and Hsp90 immunohistochemical levels change in ulcerative colitis after therapy

Ulcerative colitis (UC) is a form of inflammatory bowel disease (IBD) characterized by damage of large bowel mucosa and frequent extra-intestinal autoimmune comorbidities. The role played in IBD pathogenesis by molecular chaperones known to interact with components of the immune system involved in inflammation is unclear. We previously demonstrated that mucosal Hsp60 decreases in UC patients treated with conventional therapies (mesalazine, probiotics), suggesting that this chaperonin could be a reliable biomarker useful for monitoring response to treatment, and that it might play a role in pathogenesis. In the present work we investigated three other heat shock protein/molecular chaperones: Hsp10, Hsp70, and Hsp90. We found that the levels of these proteins are increased in UC patients at the time of diagnosis and decrease after therapy, supporting the notion that these proteins deserve attention in the study of the mechanisms that promote the development and maintenance of IBD, and as biomarkers of this disease (e.g., to monitor response to treatment at the histological level).