The cystic fibrosis transmembrane conductance regulator as a marker of human pancreatic duct development

BACKGROUND & AIMS: The cystic fibrosis transmembrane conductance regulator (CFTR) protein is a small conductance adenosine 3',5'-cyclic monophosphate (cAMP)-activated chloride ion channel found in the apical membranes of epithelia within the pancreas, airway, intestine, bile duct, sweat gland, and male genital ducts. Pancreatic insufficiency is a feature of about 85% of patients with cystic fibrosis and is believed to be caused by pancreatic autolysis after pancreatic duct obstruction. The aim of this study was to investigate the expression of CFTR in the pancreas from early development to postnatal life to establish whether the CFTR plays a key role in development of the pancreatic duct epithelium. METHODS: Expression of CFTR from the start of the mid-trimester of human development through term to adult life by messenger RNA (mRNA) in situ hybridization was examined. RESULTS: CFTR mRNA is detected throughout the pancreatic duct epithelium and its pattern of expression follows the differentiation of the duct system. CONCLUSIONS: CFTR is a valuable marker of human pancreatic duct cell development and differentiation.     

Cryopreservation of all prezygotes in patients at risk of severe hyperstimulation does not eliminate the syndrome, but the chances of pregnancy are excellent with subsequent frozen-thaw transfers

In-vitro fertilization patients (n = 15) at risk of ovarian hyperstimulation syndrome (OHSS) (oestradiol > or =4500 pg/ml on the day of human chorionic gonadotrophin administration and 25 or more follicles of intermediate or large size) underwent aspiration of all follicles and cryopreservation of all fertilized oocytes at the pronuclear stage. Patients were monitored for up to 2 weeks post-retrieval. Subsequent transfer of cryopreserved-thawed embryos was performed in programmed cycles using exogenous oestrogen and progesterone for endometrial preparation. Two patients (13%) developed OHSS necessitating hospitalization and vaginal aspiration of ascitic fluid. Two other patients (13%) developed moderate OHSS requiring ascitic fluid vaginal aspiration in the office setting, with dramatic improvement of the condition. Subsequent transfer of cryopreserved-thawed embryos yielded a clinical pregnancy rate of 58% per transfer and ongoing or delivery rates of 42 and 67% per transfer and per patient respectively. By eliminating pregnancy potential with cryopreservation of all prezygotes and examining the pregnancy potential with subsequent cryopreserved-thawed transfers, it is concluded that OHSS is reduced, but not eliminated for patients at risk. Subsequent transfer of cryopreserved-thawed prezygotes in a programmed cycle with exogenous steroids yields an excellent pregnancy rate.     

Dissecting the order of bacteriophage T4 DNA polymerase holoenzyme assembly

Most biological organisms rely upon a DNA polymerase holoenzyme for processive DNA replication. The bacteriophage T4 DNA polymerase holoenzyme is composed of the polymerase enzyme and a clamp protein (the 45 protein), which functions as a processivity factor by strengthening the interaction between DNA and the holoenzyme. The 45 protein must be loaded onto DNA by a clamp loader ATPase complex (the 44/62 complex). In this paper, the order of events leading to holoenzyme formation is investigated using a combination of rapid-quench and stopped-flow fluorescence spectroscopy kinetic methods. A rapid-quench strand displacement assay in which the order of holoenzyme component addition is varied provided data indicating that the rate-limiting step in holoenzyme assembly is associated with the clamp loading process. Pre-steady-state analysis of the clamp loader ATPase activity demonstrated that the four bound ATP molecules are hydrolyzed stepwise during the clamp loading process in groups of two. Clamp loading was examined with stopped-flow fluorescence spectroscopy from the perspective of the clamp itself, using a site-specific, fluorescently labeled 45 protein. A mechanism for T4 DNA polymerase holoenzyme assembly is proposed in which the 45 protein interacts with the 44/62 complex leading to the hydrolysis of 2 equiv of ATP, and upon contacting DNA, the remaining two ATP molecules bound to the 44/62 complex are hydrolyzed. Once all four ATP molecules are hydrolyzed, the 45 protein is poised on DNA for association with the polymerase to form the holoenzyme.     

Clonal expansion of T-cells in measles

Videolaparoscopic cholecystectomy is considered the treatment of choice for simple cholelithiasis. Now many surgeons consider the laparscopic procedure usable also in the complicated biliary lithiasis like acute cholecystitis and choledocholithiasis. The authors report their recent experience of the laparoscopic treatment of biliary lithiasis, regarding 221 non-selected patients (69% symptomatic cholelithiasis, 20% chronic cholecystitis, 4.5% acute cholecystitis, 4.5% coledocolithiasis, 2% hydrops). The diagnostic-therapeutic protocol and the results are described and compared with the beginning of their experience, when they treated only symptomatic gallbladder stone disease, and with the reports of the literature. The authors concluded that the laparoscopic procedure is a good chance for the surgeon in the treatment of all cases of benign biliary disease. But, in particular for patients with choledocholithiasis, he has be able to know all the diagnostic and therapeutic possibilities, to choose the best in every single case.     

Functional defects of Cx26 resulting from a heterozygous missense mutation in a family with dominant deaf-mutism and palmoplantar keratoderma

BACKGROUND: There is controversy about the impact on morbidity from delayed diagnoses of blunt hollow viscus injuries. A recent study suggested that the increased morbidity was primarily from delayed diagnosis of blunt duodenal injury (BDI). STUDY DESIGN: We studied the medical records from a 10-year period from June 1987 to June 1997 examining the data on 22,163 cases of blunt trauma. We assessed the incidence and consequences of delayed diagnoses of BDI, and identified preoperative factors associated with these delayed diagnoses. RESULTS: Thirty-five patients (0.2%) were identified in the retrospective study of the records from 22,163 blunt trauma patients to have sustained BDI. Of these, 25 patients (71%) were male. Ages ranged from 1 to 58 years (mean 18.8 years), and the predominant mechanism was motor vehicle accident in 18 patients (51%). Seven patients (20%) (group I) had a diagnostic delay of > 6 hours; 28 patients (80%) (group II) were diagnosed in < 6 hours. Six of the seven group I patients (86%) were evaluated initially with CT scans, and five (83%) showed findings suggestive of BDI. Among the 28 group II patients, 14 (50%) underwent initial diagnostic peritoneal lavage (DPL), and 14 (50%) had a CT scan. In seven of the group II patients (50%) who were initially evaluated by CT scan, there were findings suggestive of BDI. Diagnostic peritoneal lavage was initially equivocal (red blood cell count=5,000 to 100,000) in the remaining one group I patient compared with three of the group II patients who had DPL. Deterioration found on physical examinations prompted followup CT scans in 6 group I patients (86%), and the scans were diagnostic for BDI in all cases. CONCLUSIONS: Blunt duodenal injury is an uncommon entity. Despite the presence of suggestive CT and DPL findings, the diagnosis was delayed in 20% of the 35 patients whose records were examined in the study; this delayed diagnosis was associated with increased abdominal complications. Patients with persistent abdominal complaints and equivocal CT or DPL findings should undergo laparotomy or repeat CT scan evaluations.     

Further analysis of the reliability of the posterior tangent lateral lumbar radiographic mensuration procedure: concurrent validity of computer-aided X-ray digitization

OBJECTIVE: To investigate the reliability of a specific method of radiographic analysis of the geometric configuration of the lumbopelvic spine in the sagittal plane, and to investigate the concurrent validity of a computer-aided digitization procedure designed to replace the more tedious and time-consuming manual measurement process. DESIGN: A blind, repeated-measures design was used. The results of radiographic measures derived through the traditional manual marking method were compared with measures derived by computer-aided digitization of lateral lumbopelvic radiographs. SETTING: Private chiropractic clinic. MAIN OUTCOME MEASURES: Pearson's product-moment correlation coefficients, paired sample t tests and intraclass correlation co-efficients (ICC) were used to examine intraexaminer reliability, and repeated measures of analysis of variance were used to examine interexaminer reliability for relative rotation angles for T12-L1, L1-L2, L2-L3, L3-L4, L4-L5, L5-S1, overall lordosis measurement [absolute rotation angle (ARA)] from L1-L5 and Cobb angle of overall lordosis measured from the inferior surface of T12 to the superior surface of S1, Ferguson's sacral base angle to horizontal, angle of pelvic tilt (arcuate angle) to horizontal and anteroposterior thoracic translation (Sz) in millimeters. RESULTS: ICC estimates for intraexaminer reliability were in the range of 0.96-0.98 for the L1-L5 ARA, a range of 0.87-0.99 for the arcuate angle measurement, 0.83-0.94 for the Ferguson's angle measurement, 0.88-0.95 for the Cobb angle measurement from the inferior surface of T12 compared with the superior surface of S1 and 0.98-1.00 for the translation measurement of the lower thoracic spine to S1 (Sz). The intersegmental measurement's (T12-L1, L1-L2, L2-L3, L3-L4, L4-L5, L5-S1) correlations ranged from a low of 0.55 to a high of 0.97. Examination of these findings suggests that the reliability for the three doctors is acceptable with only the T12-L1 intersegmental measure falling below 0.70 for the least experienced examiner. Average ICC of interexaminer reliability for manual and computer-aided digitizing examiners were the following: 0.96 for the L1-L5 ARA; 0.84 for the arcuate angle measurement; 0.82 for the Ferguson's angle measurement; 0.88 for the Cobb angle measurement; 1.00 for the Sz translation measurement; and values of 0.65, 0.73, 0.74, 0.75, 0.89 and 0.81 for relative rotation angle measurements T12-L1, L1-L2, L2-L3, L3-L4, L4-L5 and L5-S1, respectively. CONCLUSION: The data tend to support the reliability of this method of radiographic analysis of the geometric configuration of the lumbopelvic spine as viewed on lateral lumbopelvic radiographs. The additional data presented here tend to support the concurrent validity of the computer-aided digitization method of analysis inasmuch as the measures determined by the digitizing examiners are essentially identical to those determined by the manual method plus or minus the average standard error of measure of each value.     

The rapid inhibitory effect of glucocorticoid on cytosolic free Ca2+ increment induced by high extracellular K+ and its underlying mechanism in PC12 cells

The effect of glucocorticoid(GC) on peak cytosolic free calcium net increment (delta[Ca2+]i) induced by high-K+ was detected with MiraCal Image System. The main results were as follows: (1) Corticosterone(B) could inhibit delta[Ca2+]i in a time-dependent and concentration-dependent manner. (2) The inhibitory effect of B could be mimicked by bovine-serum albumin conjugated corticosterone (B-BSA) also in a dose-dependent manner. (3) G-protein inhibitor, either PTX or GDP beta S significantly reduced the inhibitory effect of B and B-BSA on delta[Ca2+]i (4) PMA, a stimulator for protein kinase C(PKC), could inhibit delta[Ca2+]i. (5) Although the inhibitors of PKC, chelerythrine chloride and bisindolylamide I per se had no influence on delta[Ca2+]i, but they significantly antagonized the inhibitory effect of B and B-BSA on delta[Ca2+]i. It is postulated that GC inhibit delta[Ca2+]i induced by high-K+ through a membrane mechanism and by a pathway involving G-protein and PKC.     

Stress and coping behaviors of substance-abusing mothers

BACKGROUND: The guided regeneration of periodontal tissues demonstrated to represent a therapeutical technique with predictable results. It has been observed that different materials, used as regenerative membranes, offer very similar results. Unconventional materials too, like the rubber dam, seem to be useful in the guided tissues regeneration technique. The object of the present study has been to comparatively evaluate the effectiveness of Gore-Tex and rubber dam-made membranes in the therapy of intra-osseous periodontal defects. MATERIALS AND METHODS: Six patients with two similar intra-osseous defects, participated in the study; one defect has been treated using, during the surgical intervention, a Gore-Tex membrane, while the other has received, a fragment of sterile rubber dam membranes. The principal clinical parameters of the periodontal health (probing depth -PD- and attachment loss -AL-) has been evaluated in both the defects before and 6 months after the periodontal surgery. RESULTS AND CONCLUSIONS: The results have showed that there are not statistically significant differences (p > 0.05) in the healing of the intra-osseous defects treated by rubber dam or Gore-Tex. The conclusion is drawn that the rubber dam can represent a valid and cheap alternative to the materials traditionally used in the regenerative surgery of the periodontal tissues.     

Comparison of mitochondrial respiratory chain enzyme activities in rodent astrocytes and neurones and a human astrocytoma cell line

This study has found that mitochondrial NADH-CoQ1 reductase (complex I) activity is significantly lower in C57 mice astrocytes compared with Wistar and Sprague-Dawley rat astrocytes, and a human astrocytoma cell line. In addition, complex I activity is 4-fold greater in Sprague-Dawley neurones when compared to Wistar or C57 neurones. These findings have important implications for mitochondrial studies involving rodent or human cell line systems, and in particular, indicate the importance of choosing an appropriate model when investigating the mitochondrial respiratory chain.