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.

Neurogenic stunned myocardium and transient severe tricuspid regurgitation in a child following nonaccidental head trauma.

Neurogenic stunned myocardium is widely reported in the adult literature and is typically associated with subarachnoid hemorrhage. We present a case of neurogenic stunned myocardium and transient severe tricuspid regurgitation in a 7-month-old infant following nonaccidental head trauma.

High rates of syphilis among STI patients are contributing to the spread of HIV-1 in India.

BACKGROUND: Recent syphilis outbreaks have raised concern regarding the potential enhancement of HIV transmission. The incidence of syphilis and its association with HIV-1 infection rates among a cohort of sexually transmitted infection (STI) clinic attendees was investigated. METHODS: 2732 HIV-1 seronegative patients attending three STI and one gynaecology clinic, were enrolled from 1993-2000 in an ongoing prospective cohort study of acute HIV-1 infection in Pune, India. At screening and quarterly follow up visits, participants underwent HIV-1 risk reduction counselling, risk behaviour assessment and HIV/STI screening that included testing for serological evidence of syphilis by RPR with TPHA confirmation. Patients with genital ulcers were screened with dark field microscopy. RESULTS: Among 2324 participants who were HIV-1 and RPR seronegative at baseline, 172 participants were found to have clinical or laboratory evidence of syphilis during follow up (5.4 per 100 person years, 95% CI 4.8 to 6.5 per 100 person years). Independent predictors of syphilis acquisition based on a Cox proportional hazards model included age less than 20 years, lack of formal education, earlier calendar year of follow up, and recent HIV-1 infection. Based on a median follow up time of 11 months, the incidence of HIV-1 was 5.8 per 100 person years (95% CI 5.0 to 6.6 per 100 person years). Using a Cox proportional hazards model to adjust for known HIV risk factors, the adjusted hazard ratio of HIV-1 infection associated with incident syphilis was 4.44 (95% CI 2.96 to 6.65; p<0.001). CONCLUSIONS: A high incidence rate of syphilis was observed among STI clinic attendees. The elevated risk of HIV-1 infection that was observed among participants with incident syphilis supports the hypothesis that syphilis enhances the sexual transmission of HIV-1 and highlights the importance of early diagnosis and treatment of syphilis.

Rate of hospitalization and inpatient care costs for HIV-1-infected patients in Pune, India.

BACKGROUND: The transition of human immunodeficiency virus (HIV) infection to acquired immune deficiency syndrome (AIDS) has begun in India, and an increase in AIDS-related hospitalizations and deaths is an anticipated challenge. We estimated the rates of hospitalization and inpatient care costs for HIV-1-infected patients. METHODS: Data were analysed on 381 HIV-1-infected persons enrolled in a HIV-1 discordant couples' cohort between September 2002 and March 2004. Inpatient care costs were extracted from select hospitals where the study patients were hospitalized and the average cost per hospitalization was calculated. RESULTS: A majority of the patients were in an advanced state of HIV-1 disease with the median CD4 counts being 207 cells/cmm (range: 4-1131 cells/cmm). In all, 63 participants who did not receive antiretroviral therapy required hospitalization, 53 due to HIV-1-related illnesses and the remaining 10 due to worsening of pre-existing conditions. The overall HIV-1-related hospitalization rate was 34.2 per 100 person-years (95% CI: 26.94-42.93). The median duration of HIV-1-related hospitalization was 10 days (range 2-48 days) and the median cost was Rs 17,464 (range: Rs 400-63,891). CONCLUSION: It is necessary to strengthen the inpatient care infrastructure and supporting diagnostic set-up, and work out economically optimized treatment algorithms for HIV-1-infected patients. Although this analysis does not cover all costs and may not be generalizable, these baseline data might be a useful reference while planning related studies accompanying the government-sponsored programme to roll out antiretroviral therapy to AIDS patients.

Low HIV-1 incidence among married serodiscordant couples in Pune, India.

Unlike commercial sex workers and patients attending sexually transmitted infection (STI) clinics, married couples are not typically targeted for HIV risk reduction programs in India. Thus, married partners of HIV-infected persons are at particularly high risk for HIV infection. Between September 2002 and November 2004, 457 HIV-1 sero-discordant, married couples were enrolled in a one-year prospective study of HIV transmission in Pune, India. The HIV incidence among uninfected partners was 1.22 per 100 person-years (95% CI 0.45-2.66), which is much lower than what has been previously reported among discordant couples in Africa. This may be due to higher rates of condom use, lower rates of STIs and higher CD4 T lymphocyte counts, among the Indian HIV sero-discordant couples.

High-risk behaviour in young men attending sexually transmitted disease clinics in Pune, India.

The present study reports sexual risk factors associated with HIV infection among men attending two sexually transmitted disease (STD) clinics in Pune, India and compares these behaviours between young and older men. Between April 1998 and May 2000, 1872 STD patients were screened for HIV infection. Data on demographics, medical history and sexual behaviour were collected at baseline. The overall HIV prevalence was 22.2%. HIV risk was associated with being divorced or widowed, less educated, living away from the family, having multiple sexual partners and initiation of sex at an early age. The risk behaviours in younger men were different to older men. Younger men were more likely to report early age of initiation of sex, having friends, acquaintances or commercial sex workers as their regular partners, having premarital sex and bisexual orientation. Young men were more educated and reported condom use more frequently compared with the older men. Similar high HIV prevalence among younger and older men highlights the need for focused targeted interventions aimed at adolescents and young men and also appropriate interventions for older men to reduce the risk of HIV and STD acquisition.

Pulmonary atresia with intact ventricular septum and major aortopulmonary collaterals: association with deletion 22q11.2.

We describe the first association of pulmonary atresia, intact ventricular septum, and absent central pulmonary arteries with deletion 22q11.2. The pulmonary blood flow was derived from major aortopulmonary collaterals. The role of the deletion in pulmonary arborization is discussed.

Prevalence and incidence of hepatitis B virus infection in STD clinic attendees in Pune, India.

OBJECTIVES: To estimate the prevalence and incidence of hepatitis B virus (HBV) infection among patients attending three STD clinics in Pune, India, and to identify associated risk factors. METHODS: Of the 2098 patients screened at STD clinics in Pune during 1996, 497, who returned for at least one follow up visit, were screened for various markers of HBV infection (HBsAg, anti-HBs, anti-HBc), HIV antibody, and VDRL. RESULTS: Of the 497 participants 3.6%, 26.5%, and 43.2% were positive for HBsAg, anti-HBs, and anti-HBc respectively. Tattooing (AOR 1.64, 95% CI 1.03 to 2.64) was found to be independently associated with presence of core antibody. Additionally, history of being in commercial sex work and history of a genital ulcer were independently associated with a positive anti-HBc antibody test (AOR 12.45, 95% CI 5.58 to 27.82 and AOR 1.70, 95% CI 1.09 to 2.66, respectively). 72 out of 497 (14.5%) participants were HIV positive at baseline. HIV-1 antibody positive patients were more likely to have a positive anti-HBc test (69.4% v 39.0%, p<0.001). 30 out of 282 participants, negative for anti-HBc antibody at enrolment, seroconverted subsequently, resulting in an incidence of 10.86 per 100 person years (95% CI 7.2%, 14.5%) (mean and accumulated follow up of 11.7 months and 276.17 person years, respectively). CONCLUSIONS: A high prevalence and incidence of HBV infection, seen in STD clinic attendees underscore the need to provide HBV vaccine to commercial sex workers and their clients in India.

Changing trends in clinical presentations in referred human immunodeficiency virus infected persons in Pune, India.

AIMS: To study profile and trends of clinical presentations among human immunodeficiency virus (HIV) infected individuals seen in a HIV Reference Clinic in Pune. METHODOLOGY: In a cross-sectional study, 3574 subjects were seen at a HIV Clinic in Pune from January 1997 to December 1999. Data on clinical presentation of 2801 (78.4%) HIV seropositive subjects were evaluated. RESULTS: Clinical conditions like oral thrush, tuberculosis, skin rash and sexually transmitted diseases showed decreasing trends during the three years study period (p=0.03, 0.02, < 0.01 and < 0.01, respectively). Conversely a significant increase in the number of asymptomatic HIV positive persons at the time of detection was observed over the same period (p < 0.01). CONCLUSION: Temporal change in the clinical presentations in the HIV positive persons referred to our clinic probably reflects increased awareness and a high index of suspicion among clinicians. Early diagnosis of HIV infection in asymptomatic phase might help the clinicians to make timely decisions on prescribing chemoprophylaxis for prevention of opportunistic infections and to take appropriate measures for prevention of secondary HIV transmission to the uninfected sex partners/spouses.

A serological survey of arboviral diseases among the human population of the Andaman and Nicobar Islands, India.

In an attempt to determine the prevalence of certain arthropod-borne viruses of public health importance amongst the human population of the Andaman and Nicobar Islands of India, 2,401 sera were collected from six major localities. The sera were analysed by the hemagglutination inhibition (HI) and neutralization (N) tests, using Chikungunya (CHIK), Japanese encephalitis (JE), West Nile (WN), dengue (DEN-2), Langat (TP-21) and Kyasanur Forest disease (KFD) viral antigens. The highest prevalence of HI antibodies was detected against KFD virus (22.4%), followed by Langat (20.2%), JE (5.9%), DEN-2 (3.1%), CHIK (2.9%) and WN (0.8%) viruses. Cross-reactions to the viral antigens were also noted. The results of N tests indicated a high prevalence of DEN-2 (25.4%) virus, followed by Langat (17.5%), CHIK (15.3%), KFD (12%), JE (2.19%) and WN (1.8%). These results are discussed in relation to important epidemiological parameters like age, sex and geographical location. To our knowledge, this is the first report of an extensive serosurvey of arthropod-borne viruses on these islands.

Low carotenoid concentration and the risk of HIV seroconversion in Pune, India.

Low vitamin A and carotenoid levels could increase the risk of sexual HIV acquisition by altering the integrity of the genital epithelium or by immunologic dysfunction. We addressed this issue by measuring serum vitamin A and carotenoid levels in patients who were at risk of subsequent HIV infection. In a nested case-control study in individuals attending two sexually transmitted disease (STD) clinics in Pune, India, serum micronutrient levels were measured in 44 cases with documented HIV seroconversion (11 women and 33 men) and in STD patients matched for gender and length of follow-up with no subsequent HIV seroconversion (controls). STD patients in Pune had low vitamin A and carotenoid levels, and low serum beta-carotene levels were independently associated with an increased risk of subsequent HIV seroconversion. STD patients with beta-carotene levels less than 0.075 micromol/L were 21 times more likely to acquire HIV infection than those with higher levels (adjusted odds ratio = 21.1; p =.01). No such association was observed in case of other non-provitamin A carotenoids. This study reports the first evidence of an association between low serum provitamin A carotenoid levels and an increased risk for heterosexual HIV acquisition in STD patients in Pune, India.

Abnormal right coronary artery flow and multiple right ventricular myocardial infarctions associated with severe right ventricular systolic hypertension.

The impact of left ventricular hypertension on coronary flow patterns in adult patients has been well described. In contrast, few reports exist regarding the association of right coronary flow abnormalities with right ventricular hypertension. The observation of myocardial infarcts, right ventricular hypertension, and abnormal coronary flow pattern-as well as its normalization with relief of right ventricular hypertension-lends support to a causal relation between ventricular hypertension and coronary flow abnormalities.

Relationship between clinical conditions and CD4 counts in HIV-infected persons in Pune, Maharashtra, India.

BACKGROUND: A decade after the detection of human immunodeficiency virus (HIV) infection in India, a steady increase in the number of patients with acquired immunodeficiency syndrome (AIDS) has been observed. The therapeutic options for patients with AIDS in developing countries include chemoprophylaxis and identifying and treating opportunistic infections. CD4 counts help in clinical monitoring and making decisions about initiating antiretroviral therapy or chemoprophylaxis. Flowcytometry is expensive and available only at specialized laboratories. Therefore, the possibility of using clinical indicators to predict low CD4 counts and disease progression needs to be explored. METHODS: This cross-sectional study was conducted among 137 HIV-infected persons investigated at an HIV reference centre in Pune. The study methods comprised pre-test counselling, informed consent, blood withdrawal and clinical evaluation. Serum samples were tested for HIV and CD4 counts were estimated on FACSort. RESULTS: Study participants commonly reported with oral candidiasis, herpes zoster, pulmonary tuberculosis, lymphadenopathy, weight loss, rash, diarrhoea and fever. CD4 counts were significantly lower among men, symptomatic patients and those with oral candidiasis, weight loss and multiple clinical conditions. The sensitivity of most of the clinical conditions was low, the specificity was high and the positive predictive value of oral candidiasis and weight loss for low CD4 counts was > 75%. CONCLUSION: The presence of oral candidiasis and weight loss were highly predictive of low CD4 counts and these can be considered as markers of HIV disease progression. Absence of clinical conditions was found to be a good predictor of high CD4 counts. Larger systematic natural history studies may help in identifying clinical conditions that could have a prognostic significance among HIV-infected people.

Disease prevalence in women attending the STD clinic in Mumbai (formerly Bombay), India.

Our objectives were to determine the prevalence of Neisseria gonorrhoeae and its association with other STD causing organisms. Three hundred and thirty-six consecutive women (female sex workers (FSWs) and married contacts), attending a sexually transmitted disease (STD) clinic in Mumbai, were screened for N. gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis. Per speculum examination was performed and clinical signs were recorded. Symptoms perceived by the women were also recorded. The mean age for married contacts, FSWs and gonorrhoea-positive women was 27.9, 29.7 and 27.5, respectively. 9.7% of the women were positive for N. gonorrhoeae, 23.2% were chlamydia-positive and 5.9% had trichomoniasis. N. gonorrhoeae was isolated more frequently from FSWs as compared to the married contacts. The prevalence of HIV was significantly higher among women with multiple sex partners (FSWs) (P<0.001). Gonococcal infection is significantly associated with the presence of HIV. A significant association between sexual habits and prevalence of gonorrhoea, trichomoniasis and HIV was observed. The prevalence of gonorrhoea over 1988 to 1996 remained approximately the same.

PIP: This clinical trial was conducted to determine the prevalence of Neisseria gonorrhea and its correlation with other sexually transmitted disease (STD) causing organisms. A total of 336 consecutive women (female sex workers (FSWs) and married contacts) attending an STD clinic in Mumbai, India, were screened for N. gonorrhea, Chlamydia trachomatis, and Trichomonas vaginalis. Speculum examinations were performed, the vulva and external genitalia were assessed for ulcers and warts, and clinical signs were recorded. The symptoms reported by the women were also documented. The mean age of married contacts was 27.9 years, while that of FSWs was 29.7 years. The mean age of gonorrhea-, chlamydia-, and trichomonas-positive women was 27.3, 28.0, and 28.8 years, respectively. This study showed that 9.7% of the women were gonorrhea-positive, 23.3% were chlamydia-positive, and 5.9% were trichomonas-positive. N. gonorrhea was more prevalent among FSWs than married contacts. Moreover, HIV was more prevalent among FSWs (P 0.001). Gonococcal infection is significantly associated with HIV. An important correlation between sex behavior and the prevalence of gonorrhea, trichomoniasis, and HIV was found. Between 1988 and 1996 the prevalence of gonorrhea remained approximately the same.

Pediatric echocardiography: applications and limitations.

Echocardiography is an extraordinarily useful imaging technique in fetuses, infants, children, and adolescents. Recent technologic innovations have expanded its versatility in the pediatric population. However, limited societal resources, limitations inherent to ultrasound imaging, and numerous imaging options even within the field of pediatric echocardiography necessitate the discriminate and thoughtful use of echocardiography in children. The clinical assessment remains a critical prelude to echocardiographic examination of the pediatric cardiovascular system.

Association between auxotypes, serogroups, and antibiotic susceptibilities of Neisseria gonorrhoeae isolated from women in Mumbai (formerly Bombay), India.

OBJECTIVES: Gonococcal isolates were differentiated based on susceptibility pattern, penicillinase production (PPNG or non-PPNG), serogroup, auxotype, protein, and plasmid profile. The association between serogroup and auxotype and PPNG was determined. STUDY DESIGN: Women attending tertiary level health centers and the sexually transmitted disease (STD) clinic in Mumbai, India, were screened for Neisseria gonorrhoeae. Minimal inhibitory concentration testing was performed according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines. Auxotypes, serogroups, protein profile, and plasmid content were also studied. RESULTS: Of the 33 isolates, 16 (48.5%) were resistant to penicillin, and 28 (84.8%) showed a chromosomally mediated resistance to tetracycline. Five (15.2%) isolates showed resistance to ciprofloxacin, whereas 12 (36.4%) showed a reduced susceptibility. Twenty-seven (81.8%) isolates belonged to the WI serogroup, and 15 (46.7%) were penicillinase producers (PPNG). Seventeen (51.5%) isolates were of the nonrequiring auxotype, whereas seven (21.2%) were proline requiring. Fifteen (55.6%) of the isolates belonged to the nonrequiring-WI auxotype/serogroup (A/S) class. Ten of the PPNG isolates possessed the 4.4 MDa plasmid, whereas four had the 3.2 MDa plasmid. Increases in the molecular weight of the major outer membrane protein were observed. CONCLUSION: A high prevalence of chromosomal resistance to penicillin and tetracycline was observed. The 4.4 MDa plasmid was the most prevalent among the PPNG isolates. We observed ciprofloxacin resistance, which has not been reported in previous studies in India. The nonrequiring auxotype was the most prevalent, followed by the proline requiring auxotype. WI serogroup was the most commonly observed among the isolates studied. The nonrequiring/WI A/S class was the most prevalent among the PPNG.

PIP: This study aims to determine the prevalence of penicillinase-producing Neisseria gonorrhea (PPNG), as well as to differentiate gonococcal isolates based on susceptibility pattern, penicillinase production, serogroup, auxotype, protein and plasmid profile among women attending tertiary level health centers and sexually transmitted disease (STD) clinics in Mumbai (formerly Bombay), India. An association between auxotypes, serogroups and antibiotic susceptibilities of Neisseria gonorrhea was determined. Subjects were screened for Neisseria gonorrhea; the minimal inhibitory concentration testing was performed according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines. The antibiotics tested were penicillin, tetracycline, ciprofloxacin, and spectinomycin. The ability of various typing methods to distinguish different isolates was calculated using the discrimination index. Results showed that in the susceptibility testing group 16 (48.5%) were resistant to penicillin and 28 (84.8%) showed chromosomally mediated resistance to tetracycline. 5 isolates (15.2%) showed resistance to ciprofloxacin, while 12 (36.4%) showed reduced susceptibility. 27 (81.8%) isolates belonged to the WI serogroup, and 15 (46.7%) were penicillinase producers. In auxotyping, 17 (51.5%) isolates were of the nonrequiring auxotype, while 7 (21.2%) were proline requiring. 15 (55.6%) of the isolates belonged to the nonrequiring-WI auxotype/serogroup (A/S) class. In plasmid pattern, 10 of the PPNG isolates possessed the 4.4 MDa plasmid, while 4 had the 3.2 MDa plasmid. Increases in the molecular weight of the major outer membrane protein were observed. In conclusion, the combined use of auxotyping and serogrouping offers a good method for discriminating gonococcal isolates. A high prevalence of chromosomal resistance to penicillin and tetracycline was observed. The 4.4 MDa plasmid was the most prevalent among the PPNG isolates. The nonrequiring auxotype was the most prevalent, followed by the proline requiring auxotype. WI serogroup was commonly observed among the isolates, while the nonrequiring/WI A/S class was the most prevalent among PPNG isolates.