An analysis of histone modifications in relation to sex-specific chromatin organization in the mealybug Maconellicoccus hirsutus.

The male-specific heterochromatization of the paternal genome, observed in coccids, is an example of both genomic imprinting and differential regulation of homologous chromosomes. We observed a highly nuclease-resistant chromatin (NRC) organization of a part of the paternal genome in males of Maconellicoccus hirsutus as reported earlier in Planococcus lilacinus. The nuclease resistance of NRC is correlated with nuclear matrix association and is lost when NRC is dissociated from the matrix. We carried out a comparative analysis of epigenetic modifications of histones in matrix associated chromatin of male and female mealybugs by ELISA. We detected H3K27me3, H4K20me3, H3K9me2 and H3K4me3 in both males and females and observed significant enrichment of H3K27me3 in the nuclear matrix of males compared to that of females. To further examine the presence of NRC in the germ line, nuclei were sorted based on chromatin compaction. The analysis of sorted nuclei indicates the presence of NRC in nuclei with different DNA content including the haploid nuclei from males. We discuss these results in the light of the presence of NRC exclusively in male nuclei and the retention of the maternal genome in sperm nuclei of mealybugs.

In vitro insulin production and analysis of pancreatic transcription factors in induced human hepatic progenitor cells.

BACKGROUND: beta-Cell destruction and/or insufficient insulin production are the hallmarks of diabetes mellitus (type 1 diabetes). A hepatic progenitor from developing liver is sought to be one of the surrogate sources of insulin production as the pancreas and the liver share a common precursor and signals from the cardiac mesoderm. Production of insulin is possible by transfecting pancreatic transcription factors that play important roles in development of the pancreatic beta-cell. But, there is always the fear of using genetically manipulated cells for therapeutics. Hence, the present study was designed to analyze the feasibility of using primary human fetal hepatic progenitors as a potential source for insulin production. METHODS: Human fetal hepatic progenitors were enriched using CD-326 magnetic cell sorting. The sorted cells were cultured with different concentrations of glucose (5-30 mM) in Dulbecco's modified Eagle's medium. The amount of insulin production was estimated in the cultured cells by the chemiluminescence method. Total RNA isolated from sorted epithelial cell adhesion molecule (EpCAM)-positive cells was reverse-transcribed, and the expression of different beta-cell-producing transcriptions factors was analyzed by polymerase chain reaction (PCR). Immunocytochemical analysis was performed in cultured cells using specific insulin antibodies. RESULTS: The viability of the total liver cells isolated was found to be 95%. The average number of EpCAM-positive cells in the total liver was found to be approximately 15%. An insulin kinetics study using glucose induction with different concentrations showed increased insulin secretion in response to glucose concentrations up to 20 mM. Furthermore, results of immunocytochemical analysis demonstrated intense insulin expression in EpCAM-positive cultured cells. Expression studies of the cultured EpCAM-positive cells using reverse transcription-PCR showed positive expression of the pancreatic transcription factors essential for insulin production. CONCLUSIONS: The present study demonstrates that in vitro differentiation of induced human hepatic progenitors into insulin-producing cells without genetic manipulations may promote strategies for the treatment of type 1 diabetes.

Treatment of Crigler-Najjar Syndrome type 1 by hepatic progenitor cell transplantation: a simple procedure for management of hyperbilirubinemia.

Crigler-Najjar Syndrome (CNS) is characterized by mild, chronic unconjugated hyperbilirubinemia resulting from an autosomal-recessive inherited deficiency of hepatic uridine/diphosphoglucuronate-glucuronosyl transferase 1Al since birth. Herein we have reported a confirmed case of CNS type 1 in a 2-year-old girl with an unconjugated hyperbilirubinemia (>30 mg/dL) treated by hepatic progenitor cell infusion through the hepatic artery. No procedure-related complications were encountered. No kernicterus was observed. The total bilirubin started falling at 10 days after cell infusion. Two months after cell infusion the bilirubin fell from 29.0 to 16 mg/dL, with the conjugated bilirubin increasing approximately fivefold, the unconjugated bilirubin decreasing nearly twofold, and the SGPT also decreasing from 210 U/L to 64 U/L. This study demonstrated the efficacy of hepatic progenitor cells to manage hyperbilirubinemia in these patients. As the procedure is simple and the patient has tolerated the cell therapy, infusion can be repeated as required to manage hyperbilirubinemia, which often causes lethal kernicterus. This study was developed to assess the safety, feasibility, and efficacy of hepatic progenitor cell transplantation in a child with CNS type 1.

Management of hyperbilirubinemia in biliary atresia by hepatic progenitor cell transplantation through hepatic artery: a case report.

Cholangiodestruction of bile ducts leads to biliary atresia, a rare disease characterized by intrahepatic and extrahepatic biliary inflammation. If the intrahepatic biliary tree is unaffected, surgical reconstruction by the Kasai procedure of hepatoportoenterostomy of the extra hepatic biliary tract is possible. Untreated, this condition leads to cirrhosis and death within the first year of the life. If the atresia is complete, liver transplantation is the only option. As a result of the shortage of donor livers, hepatocytes have been infused over the past two decades, providing proof of the concept that cell therapy can be effective for the treatment of liver diseases. In the present study, we report a confirmed case of a girl of 1 year of age with increased bilirubin of 28.5 mg/dL and pediatric end-stage liver disease score 20. Biochemical liver function tests showed cholestasis (elevated cholesterol and gamma-GTs) and increased ALT, total bilirubin, conjugated bilirubin, and ALP. The patient was treated with hepatic progenitor cell infusion through the hepatic artery. The total bilirubin and conjugated bilirubin started decreasing during the first month after cell infusion. The level of total bilirubin maintained a threefold decrease after months of cell infusion. The conjugated bilirubin was 16.35 mg/dL before cell infusion, decreasing to eightfold after cell infusion. After 2 months of cell infusion, hepatobiliary scintigraphy showed increased liver cell function. This case demonstrated the efficacy and functionality of hepatic progenitor cells for the management of biliary atresia. Further, as there was a decrease in serum bilirubin, it showed that there was some percentage of the engraftment of the infused cells. As the procedure is simple and the patient has tolerated the infusion therapy, it might be repeated to manage biliary atresia.

Transcatheter arterial embolization in the management of hemobilia.

BACKGROUND: This retrospective analysis evaluated the clinical and radiologic results of transcatheter arterial embolization (TAE) in the treatment of significant hemobilia. The imaging findings, embolization technique, complications, and efficacy are described. METHODS: Thirty-two consecutive patients (21 male, 11 female, age range 8-61 years) who were referred to the radiology department for severe or recurrent hemobilia were treated by TAE. Causes of hemobilia were liver trauma (n = 19; iatrogenic in six and road traffic accident in 13), vasculitis (n = 6), vascular malformations (n = 2), and hepatobiliary tumors (n = 5). Iatrogenic liver trauma was secondary to cholecystectomy in those six patients. Four of five hepatobiliary tumors were inoperable malignant tumors and one was a giant cavernous hemangioma. Arterial embolization was done after placing appropriate catheters as close as possible to the bleeding site. Embolizing materials used were Gelfoam, polyvinyl alcohol particles or steel coils, alone or in combination. Postembolization angiography was performed in all cases to confirm adequacy of embolization. Follow-up color Doppler ultrasound and contrast-enhanced computed tomography was done in all patients. RESULTS: Ultrasonic, computed tomographic, and angiographic appearances of significant hemobilia were assessed. Angiogram showed the cause of bleeding in all cases. Three patients with liver trauma due to accidents required repeat embolization. Eight patients required surgery due to failed embolization (continuous or repeat bleeding in four patients, involvement of the large extrahepatic portion of hepatic artery in two, and coexisting solid organ injuries in two). Severity of hemobilia did not correlate with grade of liver injury. All 13 patients with blunt hepatic trauma showed the cause of hemobilia in the right lobe. No patient with traumatic hemobilia showed an identifiable cause in the left lobe. There were no clinically significant side effects or complications associated with TAE except one gallbladder infarction, which was noted at surgery, and cholecystectomy was performed with excision of the hepatic artery aneurysm. CONCLUSION: TAE is a safe and effective interventional radiologic procedure in the nonoperative management of patients who have significant hemobilia.

Gastric volvulus: acute and chronic presentation.

Diaphragmatic hernia may be congenital or traumatic in origin. Traumatic hernia may menifest immediately or several months/years after the incident. Congenital hernia usually manifests in the early years of life. Diaphragmatic hernia may be complicated by gastric volvulus. Acute gastric volvulus is surgical emergency where as chronic gastric volvulus presents with nonspecific abdominal symptoms. Diagnosis of gastric volvulus is difficult and is based on imaging studies. We describe four cases of diaphragmatic hernia complicated by gastric volvulus, diagnosed on imaging and managed surgically.

Transhiatal esophagectomy for benign and malignant conditions.

BACKGROUND: Experience with transhiatal esophagectomy (THE) for both benign and malignant diseases of the esophagus as practiced over an 18-year period is presented. METHODS: Between 1982 and 2000, 411 consecutive patients underwent THE for both benign (n = 44) and malignant (n = 367) diseases of esophagus. Surviving patients were followed up for a mean of 30.4 months. RESULTS: The overall operative mortality was 11% which had reduced to 6% for the last 111 patients. Operative mortality in the benign group was less than 5%. Respiratory complications were the most frequent cause of morbidity and mortality. Nonfatal anastomotic leaks occurred in 14%. The overall actuarial survival rates at 2, 5, and 10 years for carcinoma patients were 54%, 38%, and 18% respectively. The 2- and 5-year actuarial survival rates for postcricoid cancers were 83% and 64%, respectively. CONCLUSIONS: Transhiatal esophagectomy is safe and effective, and its results including long-term outcome are comparable with most published series.

Functional outcome and patient satisfaction after ileal pouch anal anastomosis for ulcerative colitis in a developing country.

AIM: To study the long-term outcome and patient satisfaction of patients with an ileal pouch-anal anastomosis (IPAA) for ulcerative colitis in India. PATIENTS AND METHODS: We studied 35 patients who had undergone IPAA for ulcerative colitis between 1985 and 1998 and had intestinal continuity restored for more than 6 months. These patients were asked to answer a detailed questionnaire on their bowel function, urogenital function, etc. A complete haemogram, serum iron studies, liver function tests and D-Xylose absorption test were done. In addition hepatobiliary ultrasound, stool microscopy, pouchoscopy and pouch biopsies were also performed. Patient satisfaction after the procedure was also evaluated. RESULTS: Thirty-five patients (17 men and 18 women) underwent a complete evaluation. The duration after restoration of continuity ranged from 6 months to 164 months (mean 78.6 months). The mean stool frequency was 7.2 stools per 24 hours. Five patients had urgency of stool, 9 had occasional soiling and 1 had major incontinence. Four patients had minimal restriction of social activities and 1 discontinued his employment. All patients were sexually satisfied except one man who had impotence and one woman who had dyspareunia. Fifteen patients had abnormal serum iron studies (Haemoglobin < 9 g/dl in 11). Eleven patients had D-Xylose absorption below normal values. Two patients were found to have gallstones. All pouch biopsies showed chronic inflammation and 1 patient had histological evidence of pouchitis. Eighty-five percent of patients reported that they were very satisfied with the procedure. CONCLUSION: Good functional recovery and acceptance of the procedure over the long term suggests that it is a valid procedure to be recommended for patients with ulcerative colitis in India.

Pancreatic divisum with choledochal cyst and calculus: MR appearance.

Pancreatic divisum is the most common congenital anomaly of the pancreas but its association with choledochal cyst is extremely rare. We describe here a case of pancreatic divisum with choledochal cyst with a stone which was successfully treated at surgery. The common congenital pancreaticobiliary abnormalities are briefly discussed.

Well-differentiated inflammatory liposarcoma presenting as pyrexia of unknown origin.

Liposarcomas have diverse histological appearance and clinical manifestations. Well-differentiated inflammatory liposarcoma is an uncommon sub-type, which often causes diagnostic difficulty. We report here a young female patient who presented with prolonged pyrexia and sub-diaphragmatic mass and was detected to have this uncommon soft tissue sarcoma on laparotomy.

Complications of cholecystectomy in the era of laparoscopic surgery.

Both open and laparoscopic cholecystectomy are highly safe and effective procedures for patients with symptomatic cholelithiasis. Today, adverse outcomes after open cholecystectomy are limited to the elderly patients with comorbid conditions and complicated biliary tract disease. Though underreported, major biliary tract complications still occur, more so with laparoscopic cholecystectomy and continue to be the main cause of morbidity after cholecystectomy.

Transcatheter arterial embolization in the treatment of symptomatic cavernous hemangiomas of the liver: a prospective study.

BACKGROUND: This prospective study evaluated the clinical and radiologic results of transcatheter arterial embolization (TAE) for the treatment of symptomatic cavernous hemangiomas of the liver. The technique, its complications, and effectiveness also were analyzed. METHODS: Eight patients (five male, three female; mean age +/- SD = 47.75 +/- 8.59 years) with symptomatic cavernous hemangiomas of the liver were treated by TAE with polyvinyl alcohol particles or gelfoam and steel coils (single session) followed by supportive treatment. Tumor characterization (including the extent and number of lesions) was done on triple-phase helical computed tomography or gadolinium-enhanced dynamic magnetic resonance imaging. RESULTS: The lesions were located in the right lobe in five patients, left lobe in one, and both lobes in two. The largest diameter of the lesions was 6-18 cm (9.28 +/- 5.13 cm). The treatment response was assessed on follow-up ultrasound and color Doppler and/or contrast-enhanced helical computed tomography. There were no treatment-related deaths and morbidity was minimal. Embolization was the only method of treatment in seven patients; however, one patient had surgery after TAE because the symptoms were only partly relieved. Indications for embolization were abdominal pain (eight patients), rapid tumor enlargement (four of eight), and recurrent jaundice (one of eight). Symptomatic improvement was documented in all patients after embolization. Symptoms did not worsen in any patient. The mean size of the tumor did not show any statistically significant change on follow-up radiologic examinations. However, in one patient, the tumor significantly regressed in size after embolization. CONCLUSION: TAE of hepatic cavernous hemangioma is a useful procedure in the therapy of symptomatic hemangiomas.

Current status of surgery and transplantation in the management of hepatocellular carcinoma: an overview.

Despite many therapeutic advances in the field of hepatocellular carcinoma over the past two decades, this disease continues to be a major cause of cancer-related mortality worldwide. This review focuses on the recent advances in surgical technique, perioperative management, and transplantation of cirrhotic and noncirrhotic patients with hepatocellular carcinoma. Liver resection continues to be the mainstay of curative treatment in noncirrhotic patients and selected cirrhotic patients with small tumors and preserved liver function. Transplantation should be advocated for patients with poor liver function and localized lesions or for patients with large fibrolamellar carcinomas that are otherwise unresectable. Surgery has a definite role in the management of hepatic recurrences in the absence of systemic dissemination. Newer advances in the therapeutic armamentarium, such as cryotherapy, radiofrequency ablation, microwave coagulation, and ethanol injections are discussed, and their overall efficacy assessed.

Cell cycle phase, cellular Ca2+ and development in Dictyostelium discoideum.

In Dictyostelium discoideum, the initial differentiation of cells is regulated by the phase of the cell cycle at starvation. Cells in S and early G2 (or with a low DNA content) have relatively high levels of cellular Ca2+ and display a prestalk tendency after starvation, whereas cells in mid to late G2 (or with a high DNA content) have relatively low levels of Ca2+ and display a prespore tendency. We found that there is a correlation between cytosolic Ca2+ and cell cycle phase, with high Ca2+ levels being restricted to cells in the S and early G2 phases. As expected on the basis of this correlation, cell cycle inhibitors influence the proportions of amoebae containing high or low Ca2+. However, it has been reported that in the rtoA mutant, which upon differentiation gives rise to many more stalk cells than spores (compared to the wild type), initial cell-type choice is independent of cell cycle phase at starvation. In contrast to the wild type, a disproportionately large fraction of rtoA amoebae fall into the high Ca2+ class, possibly due to an altered ability of this mutant to transport Ca2+.

Tubercular abdominal abscess following laparoscopic cholecystectomy: case report.

As laparoscopic cholecystectomy has become one of the most commonly performed operations, radiologists increasingly encounter complications resulting from these. Late abdominal abscesses developing as a result of dropped gallstones albeit unusual, have been described. Abdominal wall tuberculosis following laparoscopy has also been reported. We report a rare case of intraabdominal and abdominal wall abscesses of tubercular aetiology associated with dropped stones following laparoscopic cholecystectomy.

Unusual presentation of polycystic liver disease in a young man.

Polycystic disease of the liver is usually asymptomatic. The main symptoms of cystic disease are those of an enlarging liver or due to compression of the adjacent organs caused by a large cyst. Less than 5% of these patients present with clinically challenging and life threatening complications. We present a patient with polycystic disease of the liver who developed an aneurysm arising from the right hepatic artery presenting with surgical obstructive jaundice. In spite of intraperitoneal rupture of the aneurysm the patient was managed successfully.