Comparative analysis of collagenase XI and liberase H1 for the isolation of human pancreatic islets.

BACKGROUNDS/AIMS: The aim of this study was to compare efficacy of two enzymes, Liberase HI and Collagenase XI in human adult pancreatic islet isolation. METHODOLOGY: Pancreatic tissue samples were digested either with Liberase HI or Collagenase XI, using a non-automated method. We investigated the effect of both enzymes on yield, function and percent viability of the islets. RESULTS: No significant differences were found regarding islet yield when comparing Liberase HI to Collagenase XI. Viability of Collagenase-isolated islets was initially lower, but following 3 days of culture they attained a higher viability than the Liberase treated islets. Although the stimulation index tended to be higher in the Liberase-isolated islets no significant differences were observed between the two enzymes, except on the first day of cultivation; SI values for all Liberase concentrations were significantly higher than for Collagenase (p < 0.05). CONCLUSIONS: We conclude that during the isolation procedure using Collagenase XI, the functional capacity of the isolated islets decline, but this is restored during a subsequent cultivation. On the other hand, during digestion with Liberase HI, the islets suffer less functional damage, resulting in better preservation of their functional capacity immediately after the isolation, as well as the subsequent 7 days of cultivation.

Study of molecular and granular layer of dentate gyrus of female rats neonatally treated with estrogen.

The proliferation of hippocampal dentate gyrus granule cells was investigated using (3)H-thymidine incorporation in control and estrogen-treated rats. Newborn 3-day old female Wistar rats were treated with a single dose of 1 mg of estradiol and 30 microCi (3)H-thymidine, and were sacrificed when 10 days old. The total number of neurons and the number of labeled granule cells in the granular layer and its subdivisions of both suprapyramidal and infrapyramidal limbs were analyzed using a stereological method. In both limbs, the total number of neurons as well as the total number of labeled granule cells in the granular layer were significantly increased in treated rats compared to corresponding controls. The thicknesses of the molecular and the granular layers and their subdivisions of both suprapyramidal and infrapyramidal limbs were analyzed using a stereological method. In treated female rats the molecular layer (ML) in both limbs was significantly decreased, and the granular layer (GL) was significantly increased in suprapyramidal limb. However, in the infrapyramidal limb an increased number of labeled cells in treated animals were significant in all particular zones of the granular layer. In the suprapyramidal limb's granular layer a significant increase in labeled cells was observed in subgranular zone (SGZ). Our results suggest a differential effect of estradiol on thicknesses of the ML and the GL, and dentate gyrus granule cells proliferation through the early rat life.

Proximal dystrophin gene deletions and protein alterations in becker muscular dystrophy.

Alterations in production of cytoskeletal protein dystrophin caused by in-frame gene mutations lead to the Becker muscular dystrophy. In this study we analyzed genotype-phenotype correlation in a group of Becker muscular dystrophy patients with deletions affecting the proximal part of dystrophin gene, encompassing exons 3-13. Four patients with deletions affecting N terminal dystrophin domain had early onset and faster progression of the disease, while three patients with deletions in the proximal part of dystrophin's rod domain had a more benign disease course. Our study suggests that proximal gene deletions in Becker muscular dystrophy have various phenotypic effects depending on the affected domain of protein dystrophin.

A study of pulmonary embolism after abdominal surgery in patients undergoing prophylaxis.

AIM: To determine risk factors for pulmonary embolism and estimate effects and benefits of prophylaxis. METHODS: We included 78 patients who died subsequently to a pulmonary embolism after major abdominal surgery from 1985 to 2003. A first, retrospective analysis involved 41 patients who underwent elective surgery between 1985 and 1990 without receiving any prophylaxis. In the prospectively evaluated subgroup, 37 patients undergoing major surgery between 1991 and 2003 were enrolled: all of them had received a prophylaxis consisting in low-molecular weight heparin, given subcutaneously at a dose of 2850 IU AXa/0.3 mL (body weight < 50 kg) or 5700 IU AXa/0.6 mL (body weight > or = 50 kg). RESULTS: A higher incidence of thromboembolism (43.9% and 46.34% in the two groups, respectively) was found in older patients (> 60 years). The incidence of pulmonary embolism after major abdominal surgery in patients who had received the prophylaxis was significantly lower compared to the subjects with the same condition who had not received any prophylaxis (P < 0.001, OR = 2.825; 95% CI, 1.811-4.408). Furthermore, the incidence of pulmonary embolism after colorectal cancer surgery was significantly higher compared to incidence of pulmonary embolism after other abdominal surgical procedures. Finally, the incidence of pulmonary embolism after colorectal cancer surgery among the patients who had received the prophylaxis (11/4316, 0.26%) was significantly lower compared to subjects undergoing a surgical procedure for the same indication but without prophylaxis (10/1562, 0.64%) (P < 0.05, OR = 2.522; 95% CI, 1.069-5.949). CONCLUSION: Prophylaxis with low molecular weight heparin is highly recommended during the preoperative period in patients with diagnosis of colorectal cancer due to high risk of pulmonary embolism after elective surgery.

[Mental disorders in patients with acute necrotic pancreatitis].

INTRODUCTION: The prognosis of patients with acute pancreatitis is still uncertain regardless of modern therapeutic procedures. It is even more emphasized if the acute pancreatitis is followed by psychic disorders. OBJECTIVE: The aim of the study was to provide an overview of the incidence of certain psychosomatic disorders in patients with acute pancreatitis and evaluate priority therapeutic procedures. METHOD: In this study, we analysed 16 patients with psychosomatic disorders followed by the episode of acute pancreatitis among 202 patients that were hospitalized in the period from 1993 until 2000. The diagnosis was based on anamnesis, clinical and laboratory findings and diagnostic procedures such as X-ray, US, CT and MRI. RESULTS: Among 16 patients with psychosomatic disorders followed by acute pancreatitis, 13 (81.25%) patients were operated on and 3 (18.75%) patients were medically treated. 6 patients experienced hallucinations, 5 memory deficiency, 16 disorientation and 14 confabulation. CONCLUSION: Psychosomatic disorders in patients with acute pancreatitis require complex medical treatment. Due to the already mentioned complications, the management of these conditions is very difficult and with uncertain results.

[Surgical treatment of pain in chronic pancreatitis].

INTRODUCTION: The principal indication for surgical intervention in chronic pancreatitis is intractable pain. Depending upon the presence of dilated pancreatic ductal system, pancreatic duct drainage procedures and different kinds of pancreatic resections are applied. OBJECTIVE: The objective of the study was to show the most appropriate procedure to gain the most possible benefits in dependence of type of pathohistological process in chronic pancreatitis. METHOD: Our study included 58 patients with intractable pain caused by chronic pancreatitis of alcoholic genesis. The first group consisted of 30 patients with dilated pancreatic ductal system more than 10 mm. The second group involved 28 patients without dilated pancreatic ductal system. Pain relief, weight gain and glucose tolerance were monitored. RESULTS: All patients of Group I (30) underwent latero-lateral pancreaticojejunal--Puestow operation. 80% of patients had no pain after 6 month, 13.6% had rare pain and 2 patients, i.e. 6.4%, who continued to consume alcohol, had strong pain. Group II consisting of 28 patients was without dilated pancreatic ductal system. This group was subjected to various types of pancreatic resections. Whipple procedure (W) was done in 6 patients, pylorus preserving Whipple (PPW) in 7 cases, and duodenum preserving cephalic pancreatectomy (DPCP) was performed in 15 patients. Generally, 89.2% of patients had no pain 6 month after the operation. An average weight gain was 1.9 kg in W group, 2.8 kg in PPW group and 4.1 kg in DPCP group. Insulin-dependent diabetes was recorded in 66.6% in W group, 57.1% in PPW group and 0% in DPCP group. CONCLUSION: According to our opinion, DPCP may be considered the procedure of choice for surgical treatment of pain in chronic pancreatitis in patients without dilatation of pancreas ductal system because of no serious postoperative metabolic consequences.