Antibodies to neutrophil cytoplasma in patients with ulcerative colitis and their first-degree relatives in Thailand.

BACKGROUND: The prevalence of perinuclear antineutrophil cytoplasmic antibodies (p-ANCA) does not significantly vary in ethnically diverse populations. The prevalence of p-ANCA is high in ulcerative colitis and primary sclerosing cholangitis. While the prevalence of ulcerative colitis in Asian populations is low, it is interesting to know the prevalence of p-ANCA in such a population. METHODS: Sera from 33 cases of ulcerative colitis diagnosed during the last 10 years at the diarrhea clinic, Division of Gastroenterology, Siriraj Hospital, were prospectively compared with case controls consisting of 15 cases of diarrhea from non-inflammatory bowel diseases and 25 non-diarrheic patients. Indirect immunofluorescence assay was used to detect p-ANCA in all the sera. RESULTS: Positive p-ANCA tests were found in 13 of the 33 patients with ulcerative colitis and in one of the 40 controls. Sensitivity of the test was 39.4% and the specificity was 97.5%. The one patient with positive p-ANCA in the control group was the patient with irritable bowel syndrome. Of the 13 p-ANCA-positive ulcerative colitis patients, two cases were found to have proctosigmoiditis, seven cases had left-sided colitis, and four cases had pancolitis. Perinuclear antineutrophil cytoplasmic antibodies was one of the 22 cases of first-degree relatives of ulcerative colitis patients (22 relatives from 12 index ulcerative colitis cases). There was no correlation between the positivity of p-ANCA and disease activity, and extent of the disease. CONCLUSION: The prevalence of p-ANCA in Thai patients with ulcerative colitis (39.4%) is lower than that in the Western population. Although the prevalence of p-ANCA is low in the Thai population, it should serve as a useful tool in diagnosing ulcerative colitis in this part of the world where the disease is uncommon and difficult to diagnose. The negativity of p-ANCA in almost all first-degree relatives of Thai ulcerative colitis patients should be further elucidated.

Ulcerative colitis in Thailand: a clinical study and long term follow-up.

UNLABELLED: Ulcerative colitis (UC) is uncommon in Thailand. Few reports have been published and confirmation of the diagnosis was difficult. To make a firm diagnosis of UC in Thailand, long-term follow-up and demonstration of chronic and relapsing clinical courses should help to confirm the existence of UC in Thailand. OBJECTIVE: To review the demographic data, clinical presentation and clinical courses of the diagnosed UC cases in Thailand. MATERIAL AND METHOD: Diagnosed UC patients who were followed-up for longer than 3 months at the Diarrhea Clinic, Siriraj Hospital between 1988-2000 were included. RESULTS: Forty cases of UC were followed-up. The duration of follow-up ranged from 3-75 months (mean 27 months). Male to female ratio was 19:21 and age of onset varied from 13-77 years (mean 37.7 years). Extents of the disease was left-sided colitis in 58 per cent, left and right-sided colitis in 8 per cent, pancolitis in 21 per cent, proctosigmoiditis in 13 per cent and ileal involvement in 8 per cent. At presentation, the disease was mild in 28 per cent, moderate in 60 per cent, severe in 13 per cent and fulminant in 2. Clinical presentations were diarrhea in 97.5 per cent, lower GI bleeding in 17.5 per cent, abdominal pain in 50 per cent, fever in 27.5 per cent, weight loss in 62.5 per cent and extraintestinal manifestations in 20 per cent. Diagnoses were made in all cases by sigmoidoscopy plus barium enema or colonoscopy and biopsies were taken in all cases. Histologic findings supported the diagnoses in 69 per cent of cases. Seventy per cent responded to sulfasalazine or 5-ASA with or without corticosteroid, but in 30 per cent, azathioprine was added for a period during the treatment. Clinical courses of the diseases were chronic intermittent with remission and relapse in 44 per cent, chronic continuous activity without remission in 27 per cent, single episode without relapse in 27 per cent and acute fulminating course in 6 per cent. One patient died from fulminant pancolitis with colonic perforation. There was no colonic cancer in our cases throughout the follow-up period. CONCLUSIONS: UC is very uncommon in Thailand. Firm diagnosis of UC can be made through long-term follow-up and demonstration of chronic and relapsing clinical courses. The disease severity and extent of disease involvement was slightly different from that in Western countries. Incidence of extraintestinal manifestation was 20 per cent. Most cases responded to sulfasalazine, 5-ASA or corticosteroids.

Clofazimine-induced crystal-storing histiocytosis producing chronic abdominal pain in a leprosy patient.

Clofazimine-induced crystal-storing histiocytosis is a rare but well-recognized condition in the literature. Besides the common reddish discoloration of the skin, clofazimine produces gastrointestinal disturbances-sometimes severe abdominal pain, prompting exploratory laparotomy, because pathologic and radiologic findings can produce diagnostic difficulties if the pathologic changes caused by clofazimine are not recognized. The authors report such a case in a leprosy patient to emphasize the importance of history taking, the radiologic abnormalities of the small intestine, and the pathologic findings in small intestine and lymph node biopsies. Clofazimine crystals are red in the frozen section and exhibit bright-red birefringence. However, they are clear in routinely processed histologic sections because they dissolve in alcohol and organic solvents. They also appear as clear crystal spaces during electron microscopic study, but some osmiophilic bodies can be observed. Histiocytosis caused by clofazimine crystals produces infiltrative lesions in radiologic studies mimicking malignant lymphoma or other infiltrative disorders. Associated plasmacytosis in the histologic sections can simulate lymphoplasmacytic lymphoma or multiple myeloma with crystal-storing histiocytosis. With the knowledge of this rare condition caused by clofazimine, appropriate management to avoid an unnecessary laparotomy is possible.

Intestinal microsporidiosis in HIV infected patients with chronic diarrhea in Thailand.

Microsporidia have been recognized as emerging opportunistic agents affecting multiple organs. Intestinal microsporidiosis caused by Enterocytozoon bieneusi and Encephalitozoon intestinalis is a common disease which is associated with gastrointestinal symptoms, particularly in AIDS patients. So far, information on the frequency of this enteric disease in Thailand is not available. Therefore, the present study was undertaken to investigate the prevalence of intestinal microsporidiosis in HIV infected persons with chronic diarrhea. From 1995 to 1996, multiple diarrheal stool specimens were received and examined for the presence of the organism using Weber's modified trichrome staining method and transmission electron microscopy for confirmation. Twenty-two of 66 patients (33.3%) were positive for microsporidia which appeared as pink-red spores of 0.8-1.2 x 0.7-0.9 microm with the characteristic transverse or oblique band representing the coiled polar filament. Clinical features of these patients included chronic diarrhea (100%), weight loss (100%), abdominal pain (77%), fever (36%), vomiting (36%) and anorexia (18%). Transmission electron microscopic examination of fecal specimens from the 22 patients with positive staining results revealed E. bieneusi in 18 cases.

Causes of chronic diarrhea in patients with AIDS in Thailand: a prospective clinical and microbiological study.

A prospective study was designed to investigate the causes of chronic diarrhea in AIDS patients in Thailand. Forty-five patients from Bamrasnaradura Infectious Diseases Hospital were enrolled. Extensive investigations included multiple stool examinations for ova and parasites, using the stool formalin-ether concentration method, stool culture, stool acid-fast bacilli (AFB) stain, stool modified AFB stain, esophagogastroduoscopy with duodenal aspirate and biopsy, and colonoscopy with biopsy. Biopsied specimens were examined with H&E, Giemsa, Gram, Periodic acid Schiff, and AFB stains. Definitive causes were found in 29 patients (64.4%). Of these 29, 7 patients were found to habor more than 1 pathogen (15.5%). The most commonly found enteric pathogen was Cryptosporidium parvum (20.0%). Less frequently found pathogens were Mycobacterium tuberculosis (17.8%), Salmonella spp. (15.5%), Cytomegalovirus (11.1%), Mycobacterium avium intracellulare (6.6%), Strongyloides stercoralis (4.4%), Giardia lamblia (4.4%), Cryptococcus neoformans (2.2%), Histoplasma capsulatum (2.2%), Campylobacter jejun (2.2%), and Cyclospora cayetanensis (2.2%). Salmonella spp., Mycobacterium tuberculosis, and Mycobacterium avium intracellulare infections were shown to be more common in Thailand than in African countries.

Effects of cigarette smoking on duodenal ulcer healing and relapse rate.

104 duodenal ulcer patients were classified into non-smokers (76) and smokers (28). Their age range was between 14-72 years. They were randomly treated with cimetidine (28 non-smokers and 8 smokers), colloidal bismuth (27 non-smokers and 10 smokers) and sucralfate (21 non-smokers and 10 smokers). Follow-up endoscopic examination at 4, 6 and 8 weeks showed that overall healing rates were better in the non-smokers than in the smokers (64.5% against 46.4% at 4 weeks and 92.1% against 67.8% at 6 weeks) and almost all ulcers had healed at the end of 8 weeks (100% in non-smokers and 96.4% in smokers). Among non-smokers, there were no statistically significant differences in the healing rates by any medication at any period of time. Among smokers, colloidal bismuth had significant better healing rate at 6 weeks over cimetidine and sucralfate. (p = 0.04 and p = 0.041 respectively). Overall relapse rates were higher among smokers (32.1%) than non-smokers (10.5%). Of the 3 medications, sucralfate had the lowest relapse rate in both smokers (20%) and non-smokers (9.5%), while colloidal bismuth had the highest relapse rates (40% for smokers and 11.1% for non-smokers).

Gastric acid secretion in Thai patients with particular reference to the characteristics differentiating types of gastric and duodenal ulcer.

Acid secretion in both basal and stimulated states (using augmented histalog test) was studied in 31 normal control patients, 64 duodenal ulcer patients and 101 gastric ulcer patients. Having had the result of acid output study, the 64 DU patients could be classified according to their acid secretion results as 35 DU type I or Normosecretor (54.7%) and 29 DU type II or Hypersecretor (45.3%). For the GU patients which were classified according to the sites of lesions as GU type I (GU above angulus), GU type II (GU associated with DU) and GU type III (GU below the angulus). Their acid output study showed that the GU type I had a rather low BAO and a high MAO close to that of the Normosecretor, but the GU type II and III had their BAO and MAO significantly higher than that of the normal control and close to that of the Hypersecretor. Comparison of acid secretion in this study group to other racial groups showed that the Thai population had acid secretion patterns close to other Asian populations, except that the Chinese in Singapore had a higher proportion of Normosecretor (69.0%) than Hypersecretor (31%). The MAO of the Asian population was found to be lower than that of Europeans.

Immune activation in human rabies.

Serum cytokines were compared by enzyme-linked immunosorbent assay among (i) 28 patients with rabies, (ii) 13 patients with non-fatal encephalitis due to other viruses, (iii) 16 patients with immune-mediated neurological diseases, and (iv) 15 patients with non-viral central nervous system infections and non-HTLV-I progressive spastic paraparesis. Levels of soluble interleukin 2 receptor (S-IL2R) were comparable in groups (i)-(iii). Fewer paralytic (1/6) than encephalitic (12/22) rabies patients had elevated S-IL2R. Only one patient with rabies and one with non-fatal viral encephalitis (group ii) had elevated S-CD8. Interleukin 6 (IL-6) was elevated in 5/22 rabies patients with encephalitis and in 0/6 paralytic rabies patients. Four individuals in groups (ii) and (iii) had elevated IL-6. Patterns of cytokine responses in group (iv) were similar to those in groups (i)-(iii). The results suggest defects in immune responsiveness in paralytic rabies.

Cells with natural killer activity in human rabies.

Cytotoxic lymphocyte function in 13 patients with rabies was studied by counting the number of CD56 cells and assessing natural killer (NK) cell activity. There was no significant difference in the number of killer cells between rabies patients and 31 normal controls (P greater than 0.05). Two of six non-fatal encephalitic patients due to causes other than rabies had reduced CD56 numbers. Base-line NK cell responses versus K562 cell targets did not differ among the normal control and rabies groups (P greater than 0.05). Study of the non-rabies encephalitis group showed heterogeneous results with wide variation. Significant enhancement of NK activity was seen in four rabies patients and in 10 normal control subjects tested after interferon-alpha (IFN-alpha) and IL-2. None of the four patients with encephalitis due to causes other than rabies showed such enhancement. Our results suggest that NK cells of rabies patients are not fully stimulated and that this might contribute to the virulence of rabies. The cause of this phenomenon remains unknown.

Human immune response to rabies nucleocapsid and glycoprotein antigens.

Antibodies to two components of rabies virus, nucleocapsid (N) and glycoprotein (G), were compared in 11 rabies patients with those in nine recipients of Vero cell rabies vaccine. All rabies vaccinees had antibodies to N and G components by day 10 after the first vaccine injection. A similar but not identical response was observed in three out of 11 rabies patients. Serum antibodies appeared in rabies patients as early as 3 days after onset of the first symptoms of the disease. In these antibody-positive rabies patients, levels of both antibodies, but particularly of anti-N antibody, were lower than in the vaccinated group. Our results suggest that the process of immune recognition and of antibody development in human rabies is more likely to occur early in the pre-clinical phase, and that reactivity to N protein may be crucial for elicitation of neutralizing antibody.

Failure of interferon alfa and tribavirin in rabies encephalitis.

OBJECTIVE: To test the effect of interferon alfa and tribavirin (ribavirin) in patients with rabies encephalitis. DESIGN: An open trial of chemotherapy and intensive care in patients with early rabies. SETTING: The intensive care unit of a Bangkok hospital. PATIENTS: Four conscious men with clinical rabies encephalitis. INTERVENTIONS: Rapid virological diagnosis of rabies. Treatment with intravenous and intraventricular injections of high doses of lymphoblastoid interferon alfa in three patients and tribavirin in one patient. Intensive care was given throughout. MAIN OUTCOME MEASURES: Rabies infection confirmed by antigen detection and virus isolation. Rabies neutralising antibody and specific IgM sought in serum and cerebrospinal fluid. Interferon concentrations monitored before and during treatment in three patients. RESULTS: Interferon alfa treatment produced high concentrations in serum and cerebrospinal fluid. All four patients died after 5 1/2 to 12 1/2 days of treatment with no evidence of virostatic or clinically beneficial effects from either treatment. CONCLUSION: Interferon alfa treatment is not effective in rabies encephalitis. The use of tribavirin warrants further study, possibly combined with new therapeutic methods.

Rapid diagnosis of rabies and post-vaccinal encephalitides.

In an attempt to establish the diagnoses of rabies post-vaccinal encephalitis (PVE) and early rabies encephalitis, paired serum and CSF levels of rabies neutralizing antibody (Rab) and rabies specific-IgM (RIgM) were compared in 12 PVE, 10 rabies and five control patients with similar presenting clinical features. Rapid methods of rabies antigen detection were evaluated in 17 patients. All 12 PVE patients had Rab in their serum and in eight it was also present in the CSF. These same eight had RIgM in the serum, and in seven also in the CSF. The CSF antibodies may have originated in the plasma since six patients had a high albumin quotient indicating leakage across the blood-brain barrier. Among the rabies patients, only the two vaccinated ones had serum Rab; this was also detected in the CSF of one and RIgM was in the CSF of the other. A raised IgG Index, indicating intrathecal synthesis of IgG was seen in five of 12 PVE patients. This did not correlate with the presence of CSF rabies antibody, suggesting production of antibody to other vaccine antigens of neural origin. The diagnosis of rabies encephalitis in life was made by antigen detection in a skin biopsy. No false positive results occurred and the method was as efficient as immunofluorescence of a post-mortem brain biopsy.

Detection of Salmonella typhi protein antigen in serum and urine: a value for diagnosis of typhoid fever in an endemic area.

Using haemoculture as the gold standard, a double antibody sandwich ELISA for the detection of Salmonella typhi Barber protein antigen (BP) was compared with the Widal test. Specimens used were serum and urine obtained from normal healthy individuals and from patients with typhoid fever, paratyphoid fever, pyrexia caused by other bacteria and pyrexia with negative haemoculture. The ELISA for antigenuria gave a significantly higher sensitivity, specificity, accuracy and positive predictive value than the Widal test (p less than 0.05). The ELISA for antigenaemia gave a significantly higher sensitivity and positive predictive value only. All other values were not significantly different. The timing of specimen collection was critical for sensitivity in the ELISA for antigenaemia and antigenuria, and the best results could be obtained by carrying out both assays simultaneously. The clearance of BP from serum into urine occurred around 16 days after the onset of fever in one patient. In two patients, BP could be detected in sera up to 3 weeks after the onset of fever. In two patients, serum BP could still be detected although haemoculture was negative.

Enzyme-linked immunosorbent assay for detection of Salmonella typhi protein antigen.

A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was designed for the detection of Salmonella typhi protein antigen. The optimal concentration of antibody for coating the plate was found to be 50 micrograms/ml. The optimum conditions for antibody coating and antigen and conjugate incubation were 37 degrees C for 3 h, 37 degrees C for 2 h, and 4 degrees C overnight, respectively. The enzyme-substrate reaction was allowed to take place at 30 degrees C for 1 h. The established ELISA was found to be reproducible, with an inter-run coefficient of variation of less than 12% for the detection of an S. typhi protein antigen concentration of 0.5 to 50 micrograms/ml. The minimal detectable level of the antigen was 0.5 micrograms/ml. Cross-reactions were observed with the high level (50 micrograms/ml) of protein antigens obtained from Salmonella typhimurium, Escherichia coli, Salmonella paratyphi A, and Salmonella enteritidis. The ELISA established was used for the detection of S. typhi protein antigen in serum from 62 patients with typhoid, 30 patients with clinically diagnosed typhoid fever, 21 patients with paratyphoid, 17 patients with pyrexia caused by other bacteria, and 160 normal, healthy individuals. It was found that the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of this assay were 83.87, 89.04, 87.93, 67.53, and 95.31%, respectively.

Passive hemagglutination test for enteric fever.

A passive hemagglutination (PHA) test for serodiagnosis of enteric fever was developed by sensitizing glutaraldehyde-preserved erythrocytes with lipopolysaccharide from Salmonella serogroups A, B, C, and D singly or simultaneously. The lipopolysaccharide-sensitized erythrocytes were tested with sera from 200 blood donors, 100 patients whose hemoculture was positive for Salmonella species, and 10 patients septicemic for other members of the family Enterobacteriaceae. The PHA test was positive in 90% of 28 acute-phase serum samples from patients with enteric fever from one hospital and in 93% of 72 acute-phase serum samples from another hospital. It was also positive in 100 and 60% of early- and late-convalescent-phase sera, respectively. The PHA test was negative in all patients septicemic for other members of the Enterobacteriaceae. Absorption of sera from patients with enteric fever with lipopolysaccharide from other members of the Enterobacteriaceae did not reduce PHA titers, indicating the specificity of the PHA test. Simultaneous sensitization with lipopolysaccharide from Salmonella serogroups A, B, C, and D was useful as a screening test in a limited trial with 28 acute-phase sera, 10 early-convalescent-phase sera, and 17 late-convalescent-phase sera. The PHA test is indeed a simple, sensitive, specific, and rapid test supplementing hemoculture in laboratory diagnosis of enteric fever.

Chronic diarrhoea: a prospective study in Thai patients at Chulalongkorn University Hospital, Bangkok.

Fifty-five Thai patients with chronic diarrhoea were prospectively studied to find out the underlying causes. The aetiology was identified in 38.2%, uncertain in 29.1%, and unknown in 32.7% of the patients. In the group with a definitive aetiologic diagnosis, parasitic and infective causes were commoner than non-infective causes. Amoebiasis and giardiasis were more frequent than expected, such that empirical therapeutic trial with an antiprotozoal may be justified if initial routine investigations fail to uncover the cause of the diarrhoea. No significant clinical features were noted between the infective and the non-infective groups. Overall, repeated stool microscopy using the concentration technique was the most useful single investigation in approaching the chronic diarrhoea problem.