Larger amounts of nitrite and nitrate-reducing bacteria in megaesophagus of Chagas' disease than in controls.

In the megaesophagus of Chagas' disease, chronic esophagitis is caused by stasis of swallowed food and saliva. In this environment, the overgrowth of aerobic and anaerobic bacteria, including nitrate-reducing bacteria, is observed. The reduction of nitrate into nitrite by the action of these bacteria has been associated with the formation of volatile nitrosamines in different situations of gastric bacterial overgrowth. We have hypothesized that this phenomenon could occur in the esophageal lumen of patients with megaesophagus. To evaluate the concentration of nitrite, the presence of volatile nitrosamines and the concentration of nitrate-reducing bacteria in the esophageal lumen of patients with non-advanced megaesophagus of Chagas' disease and in a group of patients without esophageal disease. Fifteen patients with non-advanced megaesophagus [megaesophagus group (MG)] and 15 patients without any esophageal disease [control group (CG)] were studied. Saliva samples were taken for nitrate and nitrite quantitative determination and esophageal stasis liquid samples were taken for nitrate and nitrite quantitative determination, volatile nitrosamines qualitative determination and reductive bacteria quantitative determination. MG and CG were equivalent in nitrate and nitrite saliva concentration and in nitrate esophageal concentration. Significant difference was found in nitrite (p = 0.003) and reductive bacteria concentration (p < 0.0001), both higher in MG. Volatile nitrosamines were identified in three MG patients and in none of the CG patients, but this was not significant (p = 0.113). There is a higher concentration of reductive bacteria in MG, responsible for the rise in nitrite concentration at the esophageal lumen and, eventually, for the formation of volatile nitrosamines.

An evaluation of gastric scintigraphy pre- and postpyloroduodenal peptic stenosis dilation.

BACKGROUND: The usual treatment of pyloroduodenal peptic stenosis has been mainly surgical, through pyloroplasty or gastric resection, with or without vagotomy. Since the first description of treatment for this peptic complication by endoscopic balloon dilation perfomed by Benjamin in 1982 [2], this procedure has become a therapeutic option in association with the medical treatment of peptic disease. The aim of this study is to evaluate the results involving clinical, endoscopic, and gastric emptying scintigraphy parameters. METHODS: Between August 1998 and February 2000, 20 patients with pyloroduodenal stenosis refractory to conservative treatment were treated at the Gastrointestinal Endoscopy Unit of the University of São Paulo Medical School. All patients who presented clinical manifestations of pyloroduodenal stenosis underwent upper gastrointestinal endoscopy to confirm peptic stenosis. Biopsy of the narrowing for the confirmation of a benign disease and gastric biopsy for Helicobacter pylori detection were performed. The treatment consisted of dilation of the stenosis with type TTS (Through The Scope) hydrostatic balloon under endoscopic control, treatment of Helicobacter pylori infection, and gastric acid suppression with oral administration of proton pump inhibitor. All patients, except one who was excluded from this study, were submitted to a clinical endoscopic assessment and gastric emptying evaluation by ingestion of (99m)Tc before and after the treatment. Endoscopic evaluation considered the diameter of the stenotic area before and after treatment. A scintigraphic study compared the time of gastric emptying before and after balloon dilation. RESULTS: Nineteen patients completed treatment by hydrostatic balloon dilation. Clinical symptoms such as bloating (p < 0.0001), epigastric pain (p = 0.0159), gastric stasis (p < 0.0001), and weight gain (p = 0.036) showed significant improvement. The diameter of the stenotic area increased significantly (p < 0.01) after the dilation treatment as well as a better gastric emptying of (99m)Tc (p < 0.0001). CONCLUSION: The dilation of the peptic pyloroduodenal stenosis using a hydrostatic balloon is a safe and effective procedure. The evaluation with gastric scintigraphy by ingestion of (99m)Tc is an effective method of assessment for the improvement of gastric function, because its results corresponded to the clinical improvement after endoscopic treatment.

Lipid peroxidation in bariatric candidates with nonalcoholic fatty liver disease (NAFLD) -- preliminary findings.

BACKGROUND: Pathogenesis of nonalcoholic fatty liver disease (NAFLD) remains incompletely known, and oxidative stress is one of the mechanisms incriminated. The aim of this study was to evaluate the role of liver oxidative stress in NAFLD affecting morbidly obese patients. METHODS: 39 consecutive patients with BMI >40 kg/m2 submitted to Roux-en-Y gastric bypass were enrolled, and wedge liver biopsy was obtained during operation. Oxidative stress was measured by concentration of hydroperoxides (CEOOH) in liver tissue. RESULTS: Female gender was dominant (89.7%) and median age was 43.6 +/- 11.1 years. Histology showed fatty liver in 92.3%, including 43.6% with nonalcoholic steatohepatitis (NASH), 48.7% with isolated steatosis and just 7.7% with normal liver. Liver cirrhosis was present in 11.7% of those with nonalcoholic steatohepatitis. Concentration of CEOOH was increased in the liver of patients with NASH when compared to isolated steatosis and normal liver (0.26+/- 0.17, 0.20+/- 0.01 and 0.14+/- 0.00 nmol/mg protein, respectively) (P < 0.01). Liver biochemical variables were normal in 92.3% of all cases, and no difference between NASH and isolated steatosis could be demonstrated. CONCLUSIONS: 1) Nonalcoholic steatosis, steatohepatitis and cirrhosis were identified in substantial numbers of morbidly obese patients; 2) Concentration of hydroperoxides was increased in steatohepatitis, consistent with a pathogenetic role for oxidative stress in this condition.

Severe protein-calorie malnutrition after bariatric procedures.

BACKGROUND: Serious nutritional complications after Roux-en-Y gastric bypass (RYGBP) are infrequent. In a retrospective study of patients operated during a 68-month period, malnutrition was investigated to analyze circumstances associated with nutritional failure. METHODS: In 236 consecutive RYGBPs, 11 patients with severe malnutrition were identified (4.7%) with age 45.1 +/- 10.6 years (10 females/1 male) and initial BMI 54.6 +/- 8.4 kg/m(2). RESULTS: In these 11 patients, the derangement was diagnosed 17.9 +/- 15.8 months after RYGBP, following defined events in 63.6% (gastric stenosis, associated diseases ) or mostly exaggeration of expected symptoms in 36.4% (vomiting without endoscopic abnormalities). BMI then was 31.4 +/- 8.6 kg/m(2) (42.5 +/- 9.9% total reduction, or 2.4 +/- 2.1% decrease/month), and serum albumin and hemoglobin were 24.0 +/- 8.2 g/L and 97.0 +/- 23.0 g/L respectively. Edema was present in 45.4% (5/11), hospitalization was required in 54.5% (6/11), and 18.2% (2/11) eventually died. CONCLUSIONS: Serious malnutrition was unusual but not exceedingly rare in this series. Exogenous precipitating factors were clearly identified in 63.6% of the patients. Careful clinical and nutritional follow-up is recommended to prevent these uncommon but potentially dangerous complications.

Prognostic significance of intraperitoneal free cancer cells obtained by laparoscopic peritoneal lavage in patients with gastric cancer.

Laparoscopy is a safe and useful method for examining the local extent and regional spread of disease in patients with gastric cancer. Peritoneal dissemination remains a frequent type of recurrence after surgical treatment. The aim of this study was to determine the prognostic value of intraperitoneal free cancer cells (IFCCs) detected by laparoscopic peritoneal lavage. Forty-nine patients with advanced gastric cancer underwent laparoscopy with cytologic examination for staging. Peritoneal lavage was performed when ascites was not present. Aspirated fluid from the peritoneal cavity was centrifuged and subjected to cytologic examination using Giemsa and Papanicolaou staining methods. Patients were surgically treated and followed for a minimum of 5 years. IFCCs were detected in 41% of the patients. In eight cases (16.3%) laparoscopy revealed carcinomatosis and/or multiple liver metastases, so laparotomy was not performed. Patterns of recurrence after curative resection included the following: peritoneal (n = 3), local (n = 4), liver (n = 1), and other (n = 1). All patients who tested positive for IFCCs had peritoneal recurrence. The absence of IFCCs was associated with improved overall survival (21 months for a 95% confidence interval of 7.4 to 34.6 vs. 4 months for a 95% confidence interval of 2.4 to 5.6). Overall survival adjusted for type of resection also demonstrated a favorable outcome for patients who were negative for IFCCs. The following conclusions were drawn: (1) laparoscopic peritoneal lavage cytology may be useful in identifying patients at high risk for peritoneal relapses and may alter treatment, and (2) lFCCs provide additional prognostic information in patients with gastric cancer.

Preoperative assessment in cases of adult megacolon suffering from moderate malnutrition.

Acquired megacolon is a chronic disease associated with constipation and malnutrition. Surgical treatment may be required for the alleviation of the intestinal symptoms, emphasizing the interest of nutritional assessment in this population. In a prospective study of 33 patients suffering from acquired megacolon and requiring either anterior resection or pull-through operation, standard anthropometric and biochemical measurements as well as the PNI of Buzby and Mullen were preoperatively assessed. Mean age of the population was 49 +/- 13 y, with 17 males and 16 females. The combined nutritional score indicated 63.6% of the population were malnourished (21/33) of the patients, whereas the Prognostic nutritional index (PNI) revealed 3 high-risk cases (9.1%), 9 with moderate risk (27.3%), and reduced risk for all others. Indeed, 39.4% (13/33) of the patients displayed surgical or septic problems. Chi-square analysis confirmed that both studied criteria were significantly associated with complications (p < 0.05), as morbidity was restricted predominantly to cases with unfavorable Prognostic nutritional index (PNI) results or clear signs of nutritional deficit. Both PNI and conventinoal nutritional assessment are valuable tools for the screening of these surgical candidates. Artificial alimentation was not used in this experience, but deserves consideration in selected patients. Nutritional status improved in the late postoperative period, with normalization of bowel function.

Refeeding procedures after 43 days of total fasting.

Refeeding syndrome encompasses fluid and electrolyte imbalances and metabolic, intestinal, and cardiorespiratory derangements associated with appreciable morbidity and mortality. Although refeeding syndrome has been well documented in concentration-camp subjects, and more recently during parenteral therapy of critically ill patients, little is known about the importance of refeeding syndrome during recovery from a hunger strike. Thus, we studied the response to a four-step dietary replenishment routine in eight hunger strikers who refused food for 43 d. In this retrospective, observational study, we assessed the safety and efficacy of the refeeding procedure and analyzed the clinical and nutritional course of the cohort during both starvation and refeeding, mainly on the basis of clinical as well as a few biochemical determinations. During starvation, average weight loss was about 18% and, with the exception of occasional oral vitamins and electrolytes, the subjects consumed only water. Available body-composition and biochemical profiles showed no clinically significant changes during starvation, but one-half of the group displayed spontaneous diarrhea at some time before refeeding. Stepwise nutritional replenishment lasted for 9 d, after which all patients tolerated a full, unrestricted diet. Only one episode of diarrhea occurred during this phase, and both clinical and biochemical indexes confirmed a favorable clinical course, without any manifestation of refeeding syndrome. In conclusion, we observed the following: 1) Hypophosphatemia and other micronutrient imbalances did not occur, nor was macronutrient intolerance detected. 2) Despite some episodes of diarrhea, nutritional replenishment was not associated with significant enteral dysfunction. 3) There was some fluid retention, but this was mild. 4) Acute-phase markers were abnormally elevated during the refeeding phase, without associated sepsis or inflammation.

Cell lines used for the selection of recombinant baculovirus.

Four insect cell lines were used to isolate two recombinant baculoviruses which had the beta-galactosidase (beta-gal) gene for colorimetric assay purposes. Plaque assays were performed using two Trichoplusia ni cell lines: BTI-TN-5B1-4 and TN-368, and two Spodptera frugiperda cell lines: IPLB-SF-21AE and SF9. The number of plaques (occlusion positive and blue beta-gal+ recombinants) formed in the Trichoplusia cells was higher than in the Spodoptera cells. The appearance of Autographa californica NPV polyhedra was also faster in the T. ni cell lines. The effect of cell passage on the plaque formation proved to be critical when two different passages of the SF9 cells were tested. The higher passage produced a lower viral titration. The size and time of appearance of the plaques was also different.

An evaluation of cold chain in Maharashtra & Karnataka states by potency testing of field samples of oral poliovirus vaccine.

The cold chain for oral poliovirus vaccine was monitored in Maharashtra and Karnataka by potency testing of vaccine vials collected from various stages of the delivery system. Results showed that cold chain maintenance improved in the state of Maharashtra within a period of three years as the monitoring began in 1987. Of the 6289 samples of trivalent OPV collected from all 30 districts of the state during 1990 to 1992, 5834 (92.8%) had retained virus titre of at least 10(5.81) TCID50/dose. In comparison, 72 per cent of the 1660 samples collected from the state of Karnataka during the same period were found to contain this minimum required virus titre. Defects in cold chain maintenance in Karnataka could be demonstrated by plotting virus titre of samples of individual batches collected from different outlets. It was concluded that potency retesting of OPV samples for cold chain monitoring will ensure proper storage, transport and use of potent vaccine in the field.

Prevalence of antibodies to polioviruses & enteroviruses excreted by healthy children in Bombay.

A total of 132 healthy children between the ages one month and 12 yr were surveyed to determine the prevalence of antibodies to the three poliovirus serotypes. Among infants up to six months of age, 73.2, 85.4 and 56.1 per cent had antibodies to poliovirus types 1, 2 and 3, respectively. In children of age groups 7 months to 3 yr and above 3 yr, antibody prevalence to the three poliovirus serotypes was 90.2, 86.9 and 57.4, and 83.3, 96.7 and 76.7 per cent, respectively. Immunization coverage with three doses of OPV exceeded 85 per cent in children above 7 months of age. Low seroprevalence to type 3 poliovirus in the children was conspicuous. Of the 80 faecal samples studied from these children, 24 (30%) were positive for virus. Among these isolates, 16 were poliovirus type 1 and three type 2. Intratypic differentiation revealed that 15 of the 16 poliovirus type 1 isolates were of wild origin. Two out of the three poliovirus type 2 isolates were of oral poliovaccine origin. Our data indicate that in spite of good vaccination coverage wild poliovirus type 1 circulation was endemic in Bombay and; that a large number of children were susceptible to poliovirus type 3 infections.

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

Increasing trend of HIV seroprevalence among pulmonary tuberculosis patients in Pune, India.

A total of 4618 tuberculosis patients attending the TB clinic at the Sassoon General Hospitals, Pune between 1991 and 1996 were screened for anti-HIV antibodies. Of these 694 were found reactive in enzyme immuno assay (EIA) and 624 were further confirmed by a second test, either rapid EIA or Western blot. HIV-1 reactivity was predominant among tuberculosis patients with HIV-2 reactivity appearing only in 1995. HIV-2 seroreactivity accounted for 0.54 and 1.02 per cent of all HIV reactive samples in 1995 and 1996. HIV-1 and HIV-2 dual reactivity accounted for 1.63 and 2.04 per cent of all infections in 1995 and 1996. The overall seroprevalence of HIV among newly diagnosed tuberculosis patients rose from 3.2 per cent in 1991 to 20.1 per cent in 1996.

PIP: A total of 4618 tuberculosis (TB) patients attending the TB clinic at the Sassoon General Hospitals, Pune, India, between 1991 and 1996 were screened for anti-HIV antibodies. Of these, 694 were found reactive in enzyme immunoassay (EIA) and 624 were further confirmed by a second test, either rapid EIA or Western blot. HIV-1 reactivity was predominant among tuberculosis patients, with HIV-2 reactivity appearing only in 1995. HIV-2 seroreactivity accounted for 0.54% and 1.02% of all HIV reactive samples in 1995 and 1996. HIV-1 and HIV-2 dual reactivity accounted for 1.63% and 2.04% of all infections in 1995 and 1996. The overall seroprevalence of HIV among newly diagnosed tuberculosis patients rose from 3.2% in 1991 to 20.1% in 1996.

Absence of wild poliovirus circulation among healthy children in a rural area with high oral poliovirus vaccination coverage.

Poliovirus circulation in a rural community was studied by a stool sample survey. No acute paralytic poliomyelitis case had been reported from the study area during the previous 5 months. Immunization coverage in age groups 7 to 12 months and 12 to 60 months was 95.8 and 94 per cent, respectively. Of the 257 children from whom stool samples were collected (about 6% of the child population), 161 (62.6%) were positive for virus isolation. Poliovirus was isolated from 60 (23.3%) children. All three poliovirus types were detected (41 type 1, 16 type 2 and 3 type 3). Intratypic differentiation tests classified these isolates as vaccine-like. Among the children excreting poliovirus, the proportion of those who did not receive polio vaccine within 30 days prior to the sample collection was 46.3, and 68.7 per cent for poliovirus type 1 and 2, respectively. It was concluded that these poliovirus excreting children were infected by the vaccine strains circulating in the environment. The survey showed that wild poliovirus was not detectable within five months after the last case of acute poliomyelitis. Displacement of the wild virus from the environment and circulation of vaccine virus was achieved by high vaccination coverage in this area.

Evidence for high prevalence & rapid transmission of HIV among individuals attending STD clinics in Pune, India.

Prevalence and incidence of HIV-1 infection among persons attending two STD clinics in Pune between May 1993 and October 1995 are reported. On screening 5321 persons, the overall prevalence of HIV-1 infection was found to be 21.2 per cent, being higher in females (32.3%) than in males (19.3%). Analysis of behavioural and biological factors showed that old age, sex work, lifetime number of sexual partners, receptive anal sex, lack of circumcision, genital diseases and lack of formal education were related to a higher HIV-1 seroprevalence. The observed incidence rate of 10.2 per cent per year was very high, much higher in women than in men (14.2% and 9.5% per year respectively) and over three times higher among the sex workers. Females in sex work, males having recent contacts with female sex workers (FSWs) and living away from the family and persons with previous or present genital diseases had a higher risk of seroconversion. Condom usage was shown to have a protective effect in seroprevalence and seroincidence analysis. With limited available resources and lack of a suitable vaccine or a drug, long-term prevention policy of creating awareness in the community must be supplemented by strengthening STD control measures and promotion of condom use and safe sex. Factors related to availability and utilization of condoms must be carefully investigated.

Molecular cloning and characterization of the ribosomal RNA genes of the cestode Echinococcus granulosus.

1. The analysis of total protoscolex DNA and some rDNA recombinants of Echinococcus granulosus by restriction endonuclease mapping and hybridization to rDNA probes indicated the complex organization of the ribosomal RNA genes and that some repeat units are larger than 15 kb. The non-transcribed spacer can be up to 13 kb in length in some repeat units. 2. Restriction site polymorphism was detected mainly in the nontranscribed spacer regions although some polymorphism was also observed in the 28S rRNA coding region. 3. On the basis of Southern blot hybridization using EcoRI-digested genomic DNA, we conclude that the repeat units containing an extra EcoRI site are present almost in the same proportion as the repeat units without the extra EcoRI site in the 28S rRNA coding region.

Serological surveillance for HIV-l infection in Bombay.

BACKGROUND: Screening of populations for the antibody to the human immunodeficiency virus has been recommended to be a practical method for assessing the prevalence of the infection and its potential for spread. METHODS: Between 1987 and 1989 we carried out a serological survey for the detection of antibody to the human immunodeficiency virus type 1, using the enzyme linked immunosorbent assay, to screen 2123 persons belonging to different high-risk groups from various parts of Bombay. RESULTS: Eighty-four repeatedly positive samples were subjected to the Western blot test for confirmation and 58 were found to be positive-4 (0.6%) out of 670 in 1987, 9(1.2%) out of 740 in 1988 and 45 (6.3%) out of 713 in 1989. CONCLUSION: These data indicate that the human immunodeficiency virus type 1 infection has already spread in the cityof Bombay among the various high-risk groups and that its prevalence rose between 1987 and 1989.

Endoscopic treatment of benign postanastomotic annular stricture of the cervical esophagus.

Although most patients presenting with a fibrotic stricture at an esophagocolic or esophagogastric anastomosis respond well to the classical dilation therapy, a number of them do not and are therefore considered to have a condition refractory to the classical method. In these cases, we performed fiberesophagoscopic section and segmentation of the stenosing ring itself, which did not exceed one cm in length, using appropriate equipment such as an electric endoscopic scalpel. Widening of the strictured segment was completed with successive dilations using a Maloney type bougie. Dysphagia disappeared after the endoscopic procedure in 21 (84%) of the 25 patients with stricture of the cervical anastomosis, after esophagocoloplasty or esophagogastroplasty, without any serious complications.

Two-way cross-protection between West Nile and Japanese encephalitis viruses in bonnet macaques.

Cross-protection between Japanese encephalitis (JE) and West Nile (WN) viruses was tested in bonnet macaques (Macaca radiata) immunized either with JE virus (JEV) or WN virus (WNV). JEV immunized monkeys were challenged by intranasal (i.n.) route with WNV and vice versa. Four control unimmunized monkeys were similarly infected either with WNV or JEV. Two of three control monkeys infected with WNV, developed paralysis followed by death. Virus was recovered from the central nervous system (CNS) of the both dead control monkeys and the histopathological examination of CNS revealed changes suggestive of viral encephalitis. The control monkey infected with JEV developed encephalitis and the virus was recovered from the blood and CNS. All the 3 JEV-immunized monkeys withstood WNV challenge, whereas only 2 of the 5 WNV immunized monkeys withstood the challenge with JEV. Out of 3 WNV-immunized monkeys surviving challenge with JEV, 2 revealed symptoms suggestive of mild encephalitis followed by complete recovery. The third monkey died on the 60th day post-infection (p.i.) without any symptoms and virus was recovered only from the olfactory lobe. These studies indicate that the immunization with JEV protects the bonnet macaques against WNV, whereas the WNV immunization only reduces the severity of the disease due to JEV.

Susceptibility of BL6 nude (congenitally athymic) mice to Japanese encephalitis virus by the peripheral route.

Studies on the susceptibility of adult BL6 nude (congenitally athymic, nu/nu) mice, their euthymic littermates (+/nu) and Swiss mice to Japanese encephalitis (JE) virus inoculated subcutaneously were carried out. The mice were observed over a period of 60 days p.i. for sickness and/or death, which was noticed only in nu/nu mice. However, the onset and the duration of sickness varied and no definite pattern was observed. Thirty four of 53 nu/nu, 28 of 42 +/nu and 30 of 52 Swiss mice bled during the first 5 days p.i. exhibited viraemia. Interestingly, only nu/nu mice had secondary viraemia during the period of sickness. The cause of sickness and death in nu/nu mice was confirmed by recovering the virus from the blood, the brain and other organs. Antibodies were detected in the sera of +/nu and Swiss mice from the 10th day p.i. onwards but not in nu/nu mice. These findings indicate the important role of functional T cells both in induction of active immunity and protection against JE virus infection in mice.

Localization of arboviral antigen in brain tissues from patients with encephalitis.

Brain tissues from 38 patients with a clinical suspicion of encephalitis or encephalopathy were examined by two immunoenzymatic techniques for the detection of arboviral antigen. Group B arboviral antigen was identified in 23 of these tissues. This simple method could be used for the diagnosis of the causal agent of encephalitis.