Betulin Acid Ester Derivatives Inhibit Cancer Cell Growth by Inducing Apoptosis through Caspase Cascade Activation: A Comprehensive In Vitro and In Silico Study.

Betulin, or naturally occurring triterpene, possesses promising antiproliferative activity. To further explore this potential, thirty-eight betulin acid ester derivatives modified at the C-28 position were tested for antitumor activities. Four human cancer cell lines, MV4-11 (leukemia), A549 (lung), PC-3 (prostate), MCF-7 (breast) as well as the normal BALB/3T3 (mouse fibroblasts) cell line were examined using MTT and SRB assays. A few derivatives exhibited strong antiproliferative activity with IC50 values between 2 and 5 µM. Subsequent mechanistic studies revealed that some derivatives induced apoptosis by inducing caspase-3/7 activity. A strong structure-activity correlation of tested compounds has been proposed along with experimental and in silico pharmacokinetic properties.

Endogenous fluorescence can differentiate the keratoconic cornea.

The purpose of the study was to investigate the endogenous fluorescence of the keratoconic cornea in order to analyze changes in the spectra due to the keratoconic stroma abnormalities. Twenty-two corneal buttons obtained from patients with keratoconus (KC, N = 22) at the time of penetrating keratoplasty were used. As a reference, twelve normal corneas (N = 12): ten from the Eye Bank and two from enucleated eyes due to choroidal melanoma were used. The fluorescence excitation/emission matrices (EEM) in the ranges of 250-400/260-600 nm were recorded. Healthy cornea, keratoconic cornea and sclera showed three main EEM bands, which correspond to the following fluorophores: tryptophan residues in the proteoglycan fraction of corneal/scleral stromas, naturally occurring collagen cross-links and the NAD(P)H fraction present in the metabolically active cells. Relative intensity factors S1, S2 and S3 describing the contribution of each kind of fluorophore to the total fluorescence of the tissue were calculated. Normal and keratoconic corneas show qualitatively similar fluorescence matrices, but the statistically significant differences in the mean values of the S1, S2 and S3 parameters for the KC and normal corneas were observed indicating changes in contribution of different fluorophores to the whole fluorescence of the tissue. Moreover, differences between multidimensional distribution of the relative intensity factors S1, S2 and S3 between these groups were demonstrated (p < 0.001). In conclusions: Differences in the relative intensity factors calculated on a basis of the fluorescence spectra can correspond to the changes found in the KC stroma regarding natural collagen cross-links and the proteoglycan fraction. These parameters well differentiate the KC and normal corneas that could serve as an additional tool for the keratoconus characterization.

Detection of hepatocellular carcinoma by tissue resonance interaction method (TRIM).

INTRODUCTION: Diagnosis of hepatocellular carcinoma (HCC) is considerably delayed, being frequently done in the non-curative stage of disease. The reason for delayed diagnosis is indolent course in early stages and/or unspecific symptoms indistinguishable from underlying cirrhosis. Hitherto methods used for screening of HCC have important limitations. TRIMprob is a non-invasive method, which showed utility in detection of cancers located in prostate, breast, or urinary bladder. AIM: To determine the diagnostic accuracy of TRIMprob in detecting HCC in cirrhotic liver. MATERIAL AND METHODS: Forty-five patients were prospectively enrolled according to final clinical diagnosis into a group of cirrhosis and HCC or a group of cirrhosis without HCC. A control group consisted of 33 healthy subjects. Hepatocellular carcinoma was diagnosed by computed tomography (CT) or magnetic resonance (MR) and guided biopsy. The TRIMprob examination was performed in each patient. Three wave frequencies were used: 465, 930, and 1395 MHz. RESULTS: In patients with HCC the intensity of return signal using wave a frequency of 465 MHz was significantly reduced in patients with HCC in comparison to healthy subjects (p < 0.0005), but not to cirrhotic patients without HCC. Moreover, cirrhosis was associated with significantly decreased TRIMprob signal in comparison to healthy liver (p < 0.002). In ROC analysis an optimal cut-off value for detection of HCC was 106 units, which yielded 80% sensitivity. CONCLUSIONS: TRIMprob identifies HCC with good sensitivity; however, the accuracy of this method to identify HCC in screening circumstances may be hindered by attenuation of the resonance interaction signal by cirrhosis itself.

A novel approach to genome-wide association analysis identifies genetic associations with primary biliary cholangitis and primary sclerosing cholangitis in Polish patients.

BACKGROUND: Primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) are forms of hepatic autoimmunity, and risk for both diseases has a strong genetic component. This study aimed to define the genetic architecture of PBC and PSC within the Polish population. METHODS: Subjects were 443 women with PBC, 120 patients with PSC, and 934 healthy controls recruited from Gastroenterology Departments in various Polish hospitals. Allelotyping employed a pooled-DNA sample-based genome-wide association study (GWAS) approach, using Illumina Human Omni2.5-Exome BeadChips and the following novel selection criteria for risk loci: blocks of at least 10 single nucleotide polymorphisms (SNPs) in strong linkage disequilibrium, where the distance between each adjacent SNP pair in the block was less than 30 kb, and each SNP was associated with disease at a significance level of P < 0.005. A selected index SNP from each block was validated using TaqMan SNP genotyping assays. RESULTS: Nineteen and twenty-one SNPs were verified as associated with PBC and PSC, respectively, by individual genotyping; 19 (10/9, PBC/PSC) SNPs reached a stringent (corrected) significance threshold and a further 21 (9/12, PBC/PSC) reached a nominal level of significance (P < 0.05 with odds ratio (OR) > 1.2 or < 0.83), providing suggestive evidence of association. The SNPs mapped to seven (1p31.3, 3q13, 6p21, 7q32.1, 11q23.3, 17q12, 19q13.33) and one (6p21) chromosome region previously associated with PBC and PSC, respectively. The SNP, rs35730843, mapping to the POLR2G gene promoter (P = 1.2 × 10-5, OR = 0.39) demonstrated the highest effect size, and was protective for PBC, whereas for PSC respective SNPs were: rs13191240 in the intron of ADGRB3 gene (P = 0.0095, OR = 0.2) and rs3822659 (P = 0.0051, OR = 0.236) along with rs9686714 (P = 0.00077, OR = 0.2), both located in the WWC1 gene. CONCLUSIONS: Our cost-effective GWAS approach followed by individual genotyping confirmed several previously identified associations and discovered new susceptibility loci associated with PBC and/or PSC in Polish patients. However, further functional studies are warranted to understand the roles of these newly identified variants in the development of the two disorders.

Severe viral oesophagitis, pharyngitis, and stomatitis as antecedents of ileocecal Crohn's disease.

We present a 22-year-old male who developed a severe erosive oesophagitis extending to the pharynx and oral cavity without obvious risk factors. Endoscopic image suggested viral aetiology that could not be confirmed by routine serological diagnostics of infections with cytomegalovirus, Epstein-Barr virus, and Herpes simplex virus. The histopathological evaluation also gave no definite clues to the aetiology of the inflammation. Treatment with acyclovir was ineffective, but gancyclovir therapy caused spectacular clinical improvement and healing of erosions. Two months later the patient presented febrile diarrhoea that was a symptom of ileocecal Crohn's disease proven by endoscopy, enterography, and histopathology. It is the first report of severe viral oesophagitis preceding clinical manifestation of Crohn's disease. This observation warrants further study towards the viral aetiology of oral, pharyngeal, and oesophageal erosions, frequently associated with Crohn's disease.

Serum adipokines in inflammatory bowel disease.

AIM: To investigate serum adipokine levels in inflammatory bowel disease (IBD) patients before treatment and after achieving clinical remission. METHODS: Serum concentrations of six adipokines (tissue growth factor-ß1, adiponectin, leptin, chemerin, resistin, and visfatin) were studied in 40 subjects with active IBD [24 subjects with Crohn's disease (CD) and in 16 subjects with ulcerative colitis (UC)] before and after three months of therapy with corticosteroids and/or azathioprine. Clinical diagnoses were based on ileocolonoscopy, computed tomography or magnetic resonance enterography and histological examination of mucosal biopsies sampled during endoscopy. Serum levels of adipokines were assessed by an indirect enzyme-linked immunosorbent assay. The control group was comprised of 16 age- and sex-matched healthy volunteers. RESULTS: Baseline leptin concentrations were significantly decreased in both types of IBD compared to controls (8.0 ± 9.1 in CD and 8.6 ± 6.3 in UC vs 16.5 ± 10.1 ng/mL in controls; P < 0.05), and significantly increased after treatment only in subjects with CD (14.9 ± 15.1 ng/mL; P < 0.05). Baseline serum resistin concentrations were significantly higher in CD (19.3 ± 12.5 ng/mL; P < 0.05) and UC subjects (23.2 ± 11.0 ng/mL; P < 0.05) than in healthy controls (10.7 ± 1.1 ng/mL). Treatment induced a decrease in the serum resistin concentration only in UC subjects (14.5 ± 4.0 ng/mL; P < 0.05). Baseline serum concentrations of visfatin were significantly higher in subjects with CD (23.2 ± 3.2 ng/mL; P < 0.05) and UC (18.8 ± 5.3 ng/mL; P < 0.05) than in healthy controls (14.1 ± 5.3 ng/mL). Treatment induced a decrease in the serum visfatin concentrations only in CD subjects (20.4 ± 4.8 ng/mL; P < 0.05). Serum levels of adiponectin, chemerin and tissue growth factor-ß1 did not differ between CD and UC subjects compared to healthy controls and also were not altered by anti-inflammatory therapy. Clinical indices of IBD activity did not correlate with adipokine levels. CONCLUSION: IBD modulates serum adipokine levels by increasing resistin and visfatin release and suppressing leptin production.

Serum vaspin may be a good indicator of fibrosis in chronic hepatitis C and is not altered by antiviral therapy.

Vaspin was found to modulate insulin resistance (IR) and to inhibit proinflammatory and profibrogenic agents. The aim of the study was to evaluate vaspin serum concentration prior to and after antiviral treatment and to assess its relationship with morphological alterations, IR and response to antiviral therapy. The study encompassed 75 non-obese, non-diabetic chronic hepatitis C (CHC) patients, 30 of whom underwent antiviral treatment. Serum vaspin levels decreased in CHC patients and was positively associated with fibrosis stage (r = 0.44, p = 0.001). Serum vaspin was significantly higher in patients with septal fibrosis/cirrhosis or periportal fibrosis compared to those with portal fibrosis or without fibrosis (F3-4 vs. F2 vs. F1 vs. F0, p = 0.012). A marked increase in the serum vaspin level occurred in patients with periportal or more advanced fibrosis (F0-1 vs. F2-4, p < 0.001). Serum vaspin levels were also positively related to steatosis grade (r = 0.32, p = 0.03). Antiviral therapy did not change serum vaspin levels, irrespective of its efficiency. Our study showed that the serum vaspin level is decreased in CHC patients with non-advanced fibrosis, but the virus seems to have no direct effect on this finding. Progressive fibrosis is associated with rise of the vaspin level and this adipokine may serve as a predictor of advanced liver fibrosis.

[Endoscopic assessment of large bowel and safety of bowel preparation and sedoanalgesia in patients with advanced liver cirrhosis]. / Ocena endoskopowa jelita grubego oraz bezpieczenstwo przygotowania jelita i sedoanalgezji u chorych z zaawansowana marskoscia watroby.

INTRODUCTION: In patients with advanced liver cirrhosis endoscopic images of large bowel are still poorly recognized in comparison with upper digestive tract. At present, the colonoscopy is examination routinely performed during qualification to liver transplantation. The purpose of this study was to retrospectively analyze colonoscopic reports and to assess a safety of all procedures realized before and during colonoscopy. MATERIAL AND METHODS: The study included 46 patients with liver cirrhosis (males 54.4%, females 45.6%) at age of 18-66 years, hospitalized between 2007-2009 for qualification to liver transplantation. Colonoscopy was done in short general sedation, and standard bowel preparation involved 256 g of polyethylene glycol dissolved in 4 liters of fluid given to the patient one day before colonoscopy. RESULTS: In 26.1% of patients no pathology was found on colonoscopy. Anal/rectal varices were found in 41.3% of patients, lesions classified as portal colopathy in 13% of patients and sigmoid diverticula in 8.7% of patients. In 17 (37%) of patients colonoscopy disclosed 46 polyps in large bowel (38 polyps in 12 patients were retrieved for histopathological examination). In 4 (8.7%) patients polyps were hyperplastic, in 6 (about 13%) tubular adenomas of low grade dysplasia and in 2 (4.35%) tubulo-villous adenomas of low grade dysplasia. Tubulo-villous adenomas were found only in patients with alcoholic cirrhosis. Colonoscopy did not worsen the general clinical state of any patient, however, as compared with compensated cirrhotics, the patients with ascites and/or peripheral edema showed features of water retention (larger body mass changes -0.50 +1.21 kg vs 0.23 +1.38 kg; p < 0.05). After colonoscopy a significant increase of body temperature by 0.23 +0.30 degrees C; p < 0.001 was noted, while examination had no significant effect on serum creatinine level and white blood cell number. CONCLUSIONS: Liver cirrhosis may predispose to certain diseases of the large bowel, including portal colopathy and adenomatous polyps. Procedures accomplished before and during colonoscopy seem to be safe for cirrhotic patients, however, in decompensated cirrhosis exists a tendency to further water retention.

[Assessment of prevalence of intrapulmonary arterio-venous shunt in cirrhotic patients qualified to liver transplantation]. / Czestosc wystepowania plucnego przecieku krwi u chorych z marskoscia watroby kwalifikowanych do leczenia transplantacyjnego.

INTRODUCTION: Hepatopulmonary syndrome (HPS) is the clinical relationship between hepatic dysfunction and the arterial hypoxygenation caused by significant intrapulmonary arteriovenous blood leakage. HPS is responsible for increased peritransplant mortality. Data on prevalence of HPS are contradictory and its clinical risk factors are still unknown. Aim of the study was assessment of prevalence of HPS in cirrhotic patients qualified to liver transplantation and determination of clinical risk factors of this syndrome. MATERIAL AND METHODS: The study involved 30 patients with advanced liver cirrhosis of different etiology, qualified to liver transplantation. In all patients the laboratory hepatic examinations, pulmonary function tests (spirometry, gasometry) and albumin lungs-brain scintigraphy were performed. RESULTS: We did not find symptomatic HPS in the investigated group, but 2 patients (6.6%) with alcoholic cirrhosis showed arterio-venous intrapulmonary shunt. No significant differences in demographic and clinical data were found between patients with and without intrapulmonary shunt. CONCLUSIONS: Symptomatic HPS is rare complication of advanced cirrhosis. Symptom free intrapulmonary shunt occurs in less than 10% of patients, however, significance and clinical risk factors of this phenomenon remain unknown.