A novel gene delivery method transduces porcine pancreatic duct epithelial cells.

Gene therapy offers the possibility to treat pancreatic disease in cystic fibrosis (CF), caused by mutations in the CF transmembrane conductance regulator (CFTR) gene; however, gene transfer to the pancreas is untested in humans. The pancreatic disease phenotype is very similar between humans and pigs with CF; thus, CF pigs create an excellent opportunity to study gene transfer to the pancreas. There are no studies showing efficient transduction of pig pancreas with gene-transfer vectors. Our objective is to develop a safe and efficient method to transduce wild-type (WT) porcine pancreatic ducts that express CFTR. We catheterized the umbilical artery of WT newborn pigs and delivered an adeno-associated virus serotype 9 vector expressing green-fluorescent protein (AAV9CMV.sceGFP) or vehicle to the celiac artery, the vessel that supplies major branches to the pancreas. This technique resulted in stable and dose-dependent transduction of pancreatic duct epithelial cells that expressed CFTR. Intravenous (IV) injection of AAV9CMV.sceGFP did not transduce the pancreas. Our technique offers an opportunity to deliver the CFTR gene to the pancreas of CF pigs. The celiac artery can be accessed via the umbilical artery in newborns and via the femoral artery at older ages--delivery approaches that can be translated to humans.

Management of infants with idiopathic dilatation of the right atrium and atrial tachycardia.

Idiopathic dilatation of the right atrium (IDRA) is a rare anomaly defined as isolated enlargement of the right atrium in the absence of other cardiac lesions known to cause right atrial dilatation. IDRA is a congenital anomaly with unknown pathogenesis and highly variable clinical presentation. Optimal management of severe IDRA is controversial and individualized. Literature reports of long-term follow-up have been limited. We describe a child with IDRA with rapid atrial tachycardia (AT) refractory to both medical and surgical management, and we provide long-term follow-up on our two previously reported cases, both of whom had documented AT. For infants with AT, the clinical course is unpredictable, and medical therapy is the first line of treatment. The decision to proceed with surgical resection of a giant right atrium should be made on an individual basis. Atrial resection along with a modified right atrial MAZE procedure could be considered in infants with life-threatening atrial tachyarrhythmia refractory to medical treatment. Surgical scarring of the right atrium may produce substrate for atrial arrhythmia, which may also be refractory to medical therapy. Histological examination of excised atrial tissue remains inconsistent and not contributory to the determination of the etiology of IDRA. Our three infants with IDRA illustrate unique features of their variable clinical courses, as well as continued difficulties with establishing clear guidelines with regard to surgical management of this unusual disorder.

Risk factors for laparoscopically confirmed pelvic inflammatory disease: findings from Mumbai (Bombay), India.

OBJECTIVES: Sexually transmitted diseases (STDs) are an important cause of pelvic inflammatory disease (PID) but have often not been detected in microbiological studies of Indian women admitted to hospital gynaecology wards or private clinics. In this cross sectional study, women living in the inner city of Mumbai (Bombay) were investigated for socioeconomic, clinical, and microbiological risk factors for PID. METHODS: Microbiological tests and laparoscopic examination were carried out on 2736 women aged < or = 35 years who came to a health facility with suspected acute salpingitis or infertility or for laparoscopic sterilisation. 86 women with a clinical diagnosis of PID were not referred for laparoscopy although their characteristics are described. Associations between various risk factors and PID status were investigated and logistic regression performed on all factors that remained significant. RESULTS: Of women with a laparoscopically confirmed evaluation, 26 women had acute and 48 chronic pelvic infection. Independent risk factors for PID were later age at menarche (> or = 14 years), a history of stillbirth and no previous pregnancy, history of tuberculosis, STD, dilatation and curettage or previous laparoscopy, and presence of Gardnerella vaginalis. CONCLUSIONS: It is concluded that STD related risk factors applied to only a small proportion of PID cases and that other determinants of PID are important, including obstetric complications, invasive surgical procedures such as laparoscopy, and tuberculosis.

Seroepidemiological investigations on human immunodeficiency virus infections in some parts of India.

PIP: 2597 serum samples from individuals belonging to various groups were screened for antibodies to human immunodeficiency virus (HIV). The majority of the sera screened were from residents of India; 16 were from foreigners. Screening was done using ELISA kits from 4 different commercial sources. Samples which were reactive initially were retested using the same kit. 4 samples were reactive repeatedly in all the kits used. 2 of these were from patients with Acquired Immune Deficiency Syndrome (AIDS), 1 from a patient with AIDS-related complex, and 1 from an apparently healthy female prostitute living in Bombay. These 4 samples were confirmed to be positive by Western Blot, immunofluorescence, and the Karpas AIDS test. Among the sexually promiscuous persons screened for antibodies to HIV in India, female prostitutes appear to be the only risk group in whom antibodies to HIV virus have been detected. This also has been reported from Tamil Nadu. Positive reactors among blood donors screened even in areas of high incidence of AIDS has been very low. There were no positive reactors among the tribals, naval personnel, and individuals from jails. Overall, the data and an earlier report from Delhi suggest that the activity of AIDS retrovirus remains low in India, but the possible threat of spread of this disease should be considered. As prostitutes have been the only risk group with positive serological evidence of HIV infection, surveillance of this group is indicated.^ieng

Potentiation by guanine nucleosides of the growth-inhibitory effects of adenosine analogs on L1210 and sarcoma 180 cells in culture.

The growth-inhibitory effect of 6-methylmercaptopurine riboside (MMPR) against leukemia L1210 cells in culture was dramatically potentiated by the addition of guanine nucleosides to the medium. In the presence of either deoxyguanosine or guanosine, the concentration of MMPR that caused 50% inhibition of growth was 35 times lower than in the absence of these nucleosides. Similar potentiation was also observed against Sarcoma 180 cells in culture by guanosine. The metabolic basis of this synergism was approached in a study of the incorporation of [14C]glycine into 5'-phosphoribosyl-N-formylglycinamide in Sarcoma 180 cells. The results show that the site of inhibition resulting in synergism is an early step in purine biosynthesis, probably phosphoribosyl pyrophosphate amidotransferase (EC 2.4.2.14). In the L1210 cell system, the addition of hypoxanthine to the medium prevented the potentiation of MMPR by guanine nucleosides supporting the conclusion that the site of the synergistic interaction involves purine biosynthesis de novo. While hypoxanthine partially reversed the growth-inhibitory effects of MMPR, an even higher degree of protection was observed in the presence of both uridine and hypoxanthine, suggesting that MMPR may have additional sites of action concerned with pyrimidine metabolism.

Adenosine phosphyorylase activity as distinct from inosine-guanosine phosphorylase activity in Sarcoma 180 cells and rat liver.

Adenosine phosphorylase (EC 2.4.2.-) activity present in Sarcoma 180 cells grown in culture and in rat liver, is shown to be distinct from inosine-guanosine phosphorylase by several criteria: (a) treatment of Sarcoma 180 cell extract with p-chloromercuribenzoate inhibited the two activities to a different extent, (b) adenine selectively protected the adenosine phosphorylase activity of Sarcoma 180 and rat liver extract against heat inactivation, while hypoxanthine selectively protected inosine-guanosine phosphorylase activity, (c) at nearly saturating substrate concentrations and using Sarcoma 180 extract, the rates of ribosylation of a mixture of adenine + hypoxanthine or adenine + guanine, but not of hypoxanthine + guanine, were found to be almost equal to the sum of their individual rates as measured separately, (d) inosine selectively inhibited the ribosylation of hypoxanthine and guanine catalysed by Sarcoma 180 and rat liver extract while 2-chloroadenosine selectively inhibited the ribosylation of adenine and N6-furfuryladenine, (e) pH vs. activity curves were similar with hypoxanthine or guanine as the substrate but they were markedly different from the curve with adenine as the substrate. The potential role of adenosine phosphorylase activity in vivo is discussed.