Urinary concentration of human chorionic gonadotrophin and its fragments as a prognostic marker in bladder cancer.

OBJECTIVE: To examine the prognostic significance of elevated urinary beta human chorionic gonadotrophin (beta-hCG) in patients with bladder cancer. PATIENTS AND METHODS: Total beta-hCG was measured in the urine of 142 patients referred for cystoscopic examination. Patients were followed for a minimum of 17 months and grouped according to stage of disease. Because the water output by individual patients varied, urinary creatinine levels were measured as an indicator of the concentration of the urine sample. Patient outcomes were correlated with urinary total beta-hCG levels both corrected and uncorrected for creatinine concentration. After correcting for urinary creatinine levels, 40 patients were excluded because the sample was too dilute (undetectable beta-hCG and a creatinine level of < 4 mmol/L). A further four patients were excluded as they had concurrent malignancies not in the bladder and one patient was lost to follow up. RESULTS: None of the 52 patients with benign conditions, nine of the 27 with Ta-T1, and nine of the 25 with T2-T4 bladder disease had urinary total beta-hCG levels > 3.74 IU/mmol/L creatinine. There was no significant association between urinary total beta-hCG concentrations and the rates of recurrence or progression for Ta-T1 disease at 17 months of follow-up. For patients with T2-T4 disease there was a significant association with widespread metastasis (P < 0.01) and mortality (P < 0.01) at 17 months of follow-up. These associations persisted when urinary total beta-hCG levels were not corrected for urinary creatinine concentration (metastasis, P < 0.01; mortality, P = 0.07; Kaplan-Meier survival time analysis, uncorrected for creatinine P = 0.027, corrected for creatinine P < 0.001). This association could not be accounted for by differences in age, histopathology or treatment. CONCLUSION: Although sample concentration was a serious confounding factor, after correcting for dilution using the creatinine content, the elevated urinary levels of total beta-hCG indicated those T2-T4 lesions which were likely to metastasize and those patients likely to die early. If this test is to be used clinically, concentrated samples, i.e. early-morning urine, and a more sensitive beta-hCG assay are required. Nevertheless, for T2-T4 bladder tumours, an elevated pre-treatment level of urinary beta-hCG is a marker of poor prognosis and may prove useful in deciding appropriate therapy.

Plasmid profile and phage type of Salmonella typhimurium strains encountered in different regions of India.

A total of 190 Salmonella typhimurium strains encountered in different parts of India were characterized on the basis of plasmid profile, phage type and antimicrobial resistance pattern. Recent trends in the epidemiology of R-plasmids were also studied. The majority of S. typhimurium strains (90.5%) were untypable by phage typing. Only 18 strains (9.5%) were phage typable. The phage untypable strains isolated from northern (57) central (65), and southern (50) regions of India could be subgrouped into 24, 12 and 16 different plasmid profiles respectively. Heterogeneity was the prominent feature although most of the plasmid profiles were related among strains isolated from particular place. A great diversity among small plasmids (2.7-8.3 kb) made subgrouping of majority strains (71%) with R-pattern ApCmKmSmSuTcTp possible. Conjugation studies and plasmid profile analysis of transconjugants revealed all the strains to harbour non conjugative non-auto transmissible plasmids with the exception of 7.2 and 2.7 kb plasmids which were not mobilizable.

Enterotoxin production and plasmid profiles in Salmonella typhimurium isolated from man.

One hundred twenty-six isolates of Salmonella typhimurium from various clinical sources were tested for enterotoxin production and characterization of plasmid profile. Cell-free culture supernates and polymyxin B extracts of all the strains were assayed by rabbit ileal loop and skin permeability tests. Enterotoxic activity was detected in culture supernates of 32 strains. Twenty-one strains by both rabbit ileal loop and skin permeability tests, nine strains by skin permeability test, and two strains by rabbit ileal loop test were positive. Live culture of three enterotoxic strains, positive in culture supernates produced ileal secretion. Polymyxin B extracts from 6 hours and 18 hours broth cultures of all the strains were devoid of enterotoxicity. Ileal mucosa exposed to culture supernate of enterotoxigenic strains showed swollen and blunted villi with submucosal oedema while those exposed to polymyxin B extracts showed shortening of villi and sloughing of epithelial lining. Plasmid profiles of enterotoxigenic strains were heterogenous and grouped into 20 different profiles. No correlation could be established between plasmid profile, R-pattern, and enterotoxin production.

Pregnancy-associated plasma protein A levels in maternal serum, extraembryonic coelomic and amniotic fluids in the first trimester.

First trimester maternal serum levels of pregnancy-associated plasma protein A(PAPP-P) are reduced in women with a Down's syndrome pregnancy. We have examined the concentration of this molecule in the amniotic (AF) and extra-embryonic coelomic (EECF) fluids surrounding the developing fetus. Maternal serum levels of PAPP-A were elevated in all samples and steadily rose from a median of 480 mIU/1 at 8 weeks to a median of 6375 mIU/1 at 14 weeks gestation. Levels of PAPP-A were low in EECF and undetectable in the AF until 14 weeks gestation. This pattern of distribution is in contrast to that of most other trophoblast-associated antigens. This may reflect PAPP-A physiology and its specific production by the syncytiotrophoblast.

Plasmid profiles, adhesive & invasive properties of Salmonella ser. Typhimurium strains: phage types 99 & 36.

Adhesive and invasive properties were compared with plasmid profile in S. Typhimurium strains of phage type 99(10) and 36(10). All strains of phage type 99 were multiple drug resistant (ApCmKmSmSuTcTp) and harboured small plasmids of 2.4-5.2 MDal. Six strains of the phage type 36 had ApCmTc R-pattern and two with only ampicillin resistance, carried plasmids of molecular size 2.6-5.2 MDal; two strains were sensitive to all antibiotics and devoid of plasmids. None of the strains were found to harbour high molecular weight plasmids. All plasmid positive strains of phage types 99 and 36 could be divided into two groups of three plasmid patterns each, which were phage type specific. All plasmid positive and negative strains adhered and invaded HeLa cells to different degrees. No correlation could be established between plasmid profile and adhesion invasion characteristics. High molecular weight plasmids therefore are unlikely to be essential for adhesion and invasion.

Plasmid profile analysis of Salmonella ser. Typhimurium phage untypable strains encountered in diverse geographic regions of India.

Plasmid profile analysis and antibiotic resistance pattern determination were carried out for 117 phage untypable S. Typhimurium strains. Majority of the strains (82%) were resistant to all the seven antibiotics tested, R-pattern being ApCmKmSmSuTcTp, rest (12%) showed heterogenous R-patterns. Plasmid DNA analysis revealed phage untypable strains to harbour large (58.8-114.3 MDal), intermediate size (36 MDal, 42 MDal) and small (1.8-5.2 MDal) plasmids with varying molecular weights. All the phage untypable strains could be subgrouped by plasmid profile analysis into 23 plasmid patterns. Plasmid profile analysis could discriminate large number of phage untypable strains on the basis of their plasmid pattern.

Genetic and molecular characterization of an epidemic plasmid coding for multidrug resistance in Salmonella typhimurium of human origin.

All 201 multidrug resistant Salmonella typhimurium strains isolated from epidemics in India contained nonconjugative (157 strains) or conjugative (44 strains) Inc F1me multiresistance plasmids. Two small R-plasmids of 7 MDa which coded for resistance to either ampicillin or streptomycin and sulfamethoxazole were also detected along with other plasmids. The small plasmids were members of group 1 and group 2 incompatibility groups. Restriction endonuclease analysis of conjugative (96 MDa) and nonconjugative (88 MDa) Inc F1me plasmids showed considerable similarity except for the presence of unique fragments among both the groups and the loss of fragments corresponding to the smaller size of the nonconjugative plasmid. A single Inc F1me plasmid appears responsible for various outbreaks of multiresistant S. typhimurium in different parts of India.

Characterization of self-transmissible plasmids determining lactose fermentation and multiple antibiotic resistance in clinical strains of Klebsiella pneumoniae.

The lactose fermentation (Lac+) and antibiotic resistance (R+) phenotypes were conjugally transferred from Klebsiella pneumoniae strains (K166, K182, K186, K218, and K220) to Salmonella typhi, S. typhimurium, Shigella flexneri, and Vibrio cholerae. The genes for lactose fermentation and antibiotic resistance were located on the plasmids. Further analysis of plasmid DNA from these isolates indicated the presence of multiple plasmids (Mr ranged less than 2.7 to 70 X 10(6)). The Lac+R+ plasmids p166 and p182 were members of the FII incompatibility group. The fertility inhibition property of plasmids, p182, p218, and p220 was fi+ type. Furthermore, phage typing experiments showed that plasmids p166 and p218 (Lac+R+) conferred the ability to inhibit the multiplication of bacteriophages 12 and 13 in S. typhimurium. However, the plasmids p182, p186, and p220 (Lac+R+) could inhibit the visible lysis of all the 30 phages in S. typhimurium. This study describes the characterization of Lac+R+ plasmids and the medical significance of an intergeneric transfer of lactose fermentation to non-lactose-fermenting pathogens.

A comparison of lectin binding in rat and human peripheral nerve.

Eleven different fluorescein- or peroxidase-conjugated lectins with different sugar-binding affinities were employed to analyze and compare glycoconjugates of rat and human peripheral nerves at the light microscopic level. A majority of lectins showed a distinct binding pattern in different structures of the nerve. Lectin binding was similar but not identical in rat and human nerves. Limulus polyhemus agglutinin did not stain any structures in rat or human nerves. In both species, all other lectins bound to the perineurium. Perineurial staining was intense with Canavalia ensiformis (Con A), Triticum vulgaris (WGA), Maclura pomifera (MPA); moderate with Glycine max (SBA), Griffonia simplicifolia-I (GS-I) and GS-II; weak with Ulex europaeus (UEA), Dolichos biflorus (DBA), and Ricinus communis (RCA). In the endoneurium of both species, ConA staining was intense, MPA and WGA moderate, SBA, GS-II, PNA, and RCA weak, and UEA and DBA absent. Interestingly, GS-I stained rat but not human endoneurium. Most lectins bound to blood vessels. GS-I bound to rat but not human, whereas UEA bound to human but not rat vessels. The results show that lectins can be used to reveal heterogeneity in sugar residues of glycoconjugates within neural and vascular components of nerves. They may therefore be potentially useful in detecting changes in glycoconjugates during nerve degeneration and subsequent regeneration after trauma or in pathological states.

Studies of streptococcal membrane antigen--binding cells in acute rheumatic fever.

Blood specimens from patients with rheumatic heart disease in both India and New Mexico were typed for the presence of B cell alloantigen 883 by use of a mouse monoclonal antibody with identical specificity to the original 883 human alloantiserum. Strong relative segregation was recorded for 883 positive B cell typing in patients with rheumatic heart disease in both geographic locations as compared with that in normal unaffected controls. In patients with acute rheumatic fever, studies of actual B-lymphocyte membrane binding by anti-883 monoclonal antibody and sonicated group A streptococcal membrane antigens showed separate but contiguous localization on isolated cell surfaces. Although physically distinct, 883 B cell alloantigen and sonicated group A streptococcal membrane antigens moved together in cell capping studies after incubation at 37 degrees C. These findings reaffirm the apparent close association between 883 B cell alloantigen and rheumatic heart disease. They also demonstrate that the B cell alloantigen 883 itself is physically distinct from but very close to sites on antigen-reactive B cells actually binding to group A streptococcal membrane antigens.