Gangliosides enhance migration of mouse B16-melanoma cells through artificial basement membrane alone or in presence of laminin or fibronectin.

The migration of B16LuF1 cells, B16-melanoma cells of lower metastatic potential to lung was enhanced through artificial basement membrane in presence of gangliosides of B16LuF1 cells as well as gangliosides of B16-melanoma cells of higher metastatic potential to lung, namely, B16LuF5 and B16LuF10 cells. The same concentration (50 microM) of gangliosides of B16LuF1, B16LuF5 and B16LuF10 cells gradually increased the migration of B16LuF1 cells through basement membrane. Moreover, B16LuF10 cell gangliosides modified the migratory effect of laminin and fibronectin on B16LuF1 cells. Laminin alone increased migration of B16LuF1 cells whereas fibronectin alone decreased migration of the same cells. When B16LuF10 cell gangliosides were used in combination with fibronectin, gangliosides removed the migration inhibitory effect of fibronectin resulting in net enhancing effect. Gangliosides in association with laminin also increased the enhancing effect of laminin on migration of B16LuF1 cells. Thus, gangliosides showed additive enhancing effect when used in combination with laminin. However, effect of individual gangliosides were different. Out of six gangliosides isolated from B16LuF10 cells only two gangliosides corresponding to standard gangliosides GM2 and GM3 enhanced migration of B16LuF1 cells. The migration of B16LuF1 cells in presence of each of the remaining four gangliosides corresponding to GT1b, GD1b, GD1a and GM1 was not altered and was comparable to that of untreated control. Thus, gangliosides of B16 melanoma cells alone or in combination with laminin or fibronectin enhanced migration of B16 melanoma cells through artificial basement membrane, suggesting possible role of tumor gangliosides during invasion of metastatic tumor cells through basement membrane of the surrounding tissues in vivo.

Secretion of metastasis related gangliosides by mouse B16-melanoma in circulation in vivo and in culture media in vitro.

Mouse B16LuF1 melanoma cells of lower metastatic potential to lung were treated in vitro with same concentration (50 microM) of gangliosides prepared from plasma of mice bearing lung metastasis of B16LuF5, B16LuF9 or B16LuF10 melanoma cell lines of increasing metastatic potential to lung (LuF1 < LuF5 < LuF9 < LuF10) and injected to normal mice through tail vein. The number of metastatic tumor nodules formed in lung increased gradually in mice receiving B16LuF5, B16LuF9 and B16LuF10-ganglioside-treated B16LuF1 cells compared to mice receiving B16LuF1 cells without any ganglioside treatment. Similarly, mouse B16LuF1 melanoma cells treated in vitro with 50 microM concentration of gangliosides prepared from spent culture media of B16LuF5, B16LuF9 or B16LuF10 cells cultured in vitro were injected to normal mice through tail vein. The number of metastatic tumor nodules formed in lung increased gradually in mice receiving B16LuF5, B16LuF9 and B16LuF10-ganglioside-treated B16LuF1 cells compared to mice receiving B16LuF1 cells without any ganglioside treatment. The results indicated that metastasis-associated gangliosides present in plasma and culture media of B16-melanoma of increasing metastatic potential to lung enhanced metastatic potential of B16LuF1 cells. The increasing concentration of metastasis-associated gangliosides present in plasma and in culture media of B16-melanoma of increasing metastatic potential possibly determined increase in metastatic potential of B16LuF1-melanoma cells.

Changing trends in management of HIV and TB.

Since the isolation of HIV in 1983, the TB graph which was on the decline saw a spurt since 1990s. The fall in immunity caused by HIV leads to reactivation as well as new infections. The developing and developed countries showed two specific patterns with TB first and HIV later and vice versa respectively. Clinical presentation of TB in HIV showed a distinctive pattern with fulminant disease in extrapulmonary forms. The correlation between HIV and TB is important. Treatment of TB in HIV and non-HIV patients remains the same. DOTS therapy has shown significant success in HIV-TB co-infection.

Correlation of gangliosides GM2 and GM3 with metastatic potential to lungs of mouse B16 melanoma.

Ten B16 mouse melanoma cell lines with increasing metastatic potential to lungs (B16LuF1 to B16LuF10) were generated by in-vitro & in-vivo selection technique starting with B16F1 melanoma cell line. The number of metastatic tumor nodules in lungs rose with increasing metastatic potential. Tumor cell gangliosides of B16LuF1 to B16LuF10 cell lines, analysed and compared with TLC, showed eight major ganglioside bands. Band1 to band6 corresponded with standard gangliosides GT1b, GD1b, GD1a, GM1, GM2 and GM3 respectively. Band7 and Band8 could not be identified. The concentration of total as well as individual ganglioside bands of B16LuF1 to B16LuF10 cells appeared to rise with increasing metastatic potential. Gangliosides from the plasma of these cell lines (B16LuF1 to B16LuF10) maintained in-vivo in C57BL/6 mice on TLC analysis gave eight major ganglioside bands, similar to those of cells. Plasma gangliosides appeared to rise with increasing metastatic potential. However, it was interesting to see that only band5 and band6 gangliosides in plasma increased almost linearly with increasing metastatic potential. The remaining six ganglioside bands in the plasma did not show such correlation. Band5 and Band6 gangliosides corresponded with standard gangliosides GM2 and GM3 respectively. Gangliosides of the spent culture media, secreted by these cell lines in-vitro in tissue culture also gave eight major ganglioside bands, similar to that of cells. Spent culture media gangliosides appeared to increase with increasing metastatic potential. However, concentration of only band5 and band6 gangliosides of spent culture media increased almost linearly with increasing metastatic potential, thus further confirming the role of band5(GM2) and Band6(GM3) gangliosides in regulating metastatic potential of B16-melanoma cells to lung.

Enhancement of metastatic potential of mouse B16-melanoma cells to lung after treatment with gangliosides of B-16-melanoma cells of higher metastatic potential to lung.

Mouse B16LuF1 melanoma cells of lower metastatic potential to lung were treated in vitro with same concentration (50 microM) of gangliosides isolated from B16LuF5, B16LuF9 or B16LuF10 cells with higher metastatic potential to lung (LuF1< LuF5< LuF9< LuF10) and injected to groups of normal mice through tail vein. The number of metastatic tumor nodules formed in lung increased in mice receiving B16LuF5, B16LuF9 and B16LuF10-ganglioside-treated B16LuF1 cells compared to mice receiving B16LuF1 cells without any ganglioside treatment. Metastatic potential of B16LuF1 cells gradually increased after treatment with gangliosides of B 16-melanoma cells of increasing metastatic potential to lung. The six major gangliosides isolated from B16LuF10 cells corresponded with standard gangliosides GT1b, GD1b, GD1a, GM1, GM2 and GM3 respectively on TLC-analysis. When B16LuF1 cells were treated in vitro with each of these six individual gangliosides and injected to groups of normal mice through tail vein the number of tumor nodules formed in lung varied. The four groups of mice receiving B16LuF1 cells treated with each of four gangliosides corresponding to GT1b, GD1b, GD1a or GM1 produced lung metastasis comparable to that of untreated control group. Only remaining two gangliosides which corresponded with standard gangliosides GM2 and GM3 increased metastatic potential of B16LuF1 cells. Thus, these results indicated that gangliosides GM2 and GM3 of B16-melanoma cells are definitely associated with metastatic potential of these tumor cells.

Immunohistochemistry and tinctorial affinity of adenohypophysial cells of the rat snake Ptyas mucosus (Colubridae).

Immunocharacteristics of the adenohypophysial cells of the rat snake Ptyas mucosus (Colubridae) were studied with an unlabeled antibody enzyme technique (PAP method) using rabbit antisera against mammalian/synthetic hypophysial hormones. Adenohypophysial cells were identified on the basis of their specific immunoreactivity with various heterologous antisera. As in other reptiles, there was regional localization of pars distalis (PD) cell types in this snake. The gonadotropic (GTH) cells were identified by their specific immunoreactivity with anti-porcine (p)LH beta serum and were distributed uniformly throughout the PD. The thyrotropic (TSH) cells, recognized by their specific immunoreactivity with anti-human (h)TSH beta serum, were found in the medial PD. The prolactin (PRL) and growth hormone (GH) cells were revealed by their specific immunoreactivity with anti-ovine (o)PRL and anti-hGH sera, respectively; the former were confined to the anterior two-thirds of the PD, and the latter were restricted to the posterior third of the PD. The corticotropic (ACTH) cells, identified by their specific immunoreactivity with the anti-ACTH1-24 serum, were localized in the anterior two-thirds of the PD. Though both ACTH and PRL cells were confined to the same area of the PD, they could be distinguished by their distinctive morphology and distribution pattern. Cells of the pars intermedia were revealed by their immunoreactivity to anti-alpha MSH (melanophore-stimulating hormone) and anti-ACTH1-24 sera. Among each adenohypophysial cell type, there was variation in the intensity of immunoreactivity and morphological features, which may be due to their heterogeneity, reflecting various stages of cellular activity. Unlike most other snake species, the occurrence of a rudimentary pars tuberalis in P. mucosus containing a few immunoreactive GTH and TSH cells appeared to be a novel finding. The occurrence of the PRL-like and TSH-like immunoreactivity seen in certain neuronal perikarya and fibers of the hypothalamus and median eminence conforms with earlier observations in other tetrapods. To resolve certain discrepancies in the literature, the tinctorial affinities of immunohistochemically identified adenohypophysial cell types of P. mucosus were studied using various conventional staining methods and were compared with those of other reptilian species studied earlier, including snakes of the family Colubridae.

The cost effectiveness of treatment of genital warts with podophyllotoxin.

The cost effectiveness of 25% podophyllin resin and 0.5% podophyllotoxin solution in the treatment of genital warts in Genitourinary Medicine Clinic attenders was studied. Although the average treatment cost for a course of podophyllotoxin was more than that for podophyllin resin (20.75 pounds v. 14.95 pounds respectively) the overall cure rate with podophyllotoxin was 66% as opposed to 34.6% with podophyllin. When the costing of secondary treatment options was considered the cost per patient cured of warts with podophyllin resin was 27.15 pounds compared with 25.73 pounds for podophyllotoxin solution (not significant).

Controlled trial of ciprofloxacin in short-term chemotherapy for pulmonary tuberculosis.

The study was undertaken in patients with newly diagnosed bacteriologically positive pulmonary tuberculosis. The patients were randomly allocated to receive one of two regimens. In one of the regimens, patients received 2 months of daily treatment with streptomycin, isoniazid, rifampin, and pyrazinamide (SHRZ) followed by isoniazid plus rifampin for 4 months (4 HR). In the other regimen, patients received 2 months of daily streptomycin, isoniazid, pyrazinamide, and ciprofloxacin (SHZ Cipro) followed by isoniazid plus ciprofloxacin for 4 months. All patients in the SHRZ/HR regimens and all but one in the SHZ Cipro/H Cipro regimens had a favorable bacteriologic response during chemotherapy. In the patients in the SHZ Cipro/H Cipro regimen, ciprofloxacin was well tolerated during the period of treatment. After chemotherapy, one (5.9 percent) of 17 patients in the SHRZ/HR group and three (16.6 percent) of 18 patients in the SHZ Cipro/H Cipro group relapsed bacteriologically.

Cold coagulation therapy in the treatment of histologically diagnosed subclinical human papilloma virus (HPV) infection of the cervix.

The efficacy of cold coagulation therapy in the treatment of histologically diagnosed subclinical human papilloma virus (HPV) infection of the cervix, was studied among 65 patients who also had genital warts. This therapy was found to be successful as the failure rate was only 8%. There were no serious side-effects. The treatment can be administered easily in an out-patient setting after histological confirmation. This study also showed that if, on the other hand, a subclinical HPV infection is not treated, it may develop into a clinical disease (genital warts) within two years. Therefore long-term follow-up of this condition may be necessary in untreated cases.

Cryotherapy in the management of histologically diagnosed subclinical human papilloma virus (HPV) infection of the cervix.

The aim of the study was to establish the efficacy of cryotherapy in the management of histologically diagnosed subclinical human papilloma virus (HPV) infection of the cervix. Women with genital warts who had subclinical HPV infection of the cervix and no other sexually transmitted diseases were followed up at six weeks, 12 weeks, six months, 12 months and 18 months after treatment with cryotherapy. They came to the out-patient department of genito-urinary medicine (GUM) at St Luke's Hospital, an open clinic without any appointment system. Two hundred and fifty-five patients with genital warts took part in the study. They underwent clinical genital examination, along with cytology, colposcopy and colposcopically directed cervical punch biopsies. Sixty-three of them had subclinical HPV infection of the cervix only, and were considered suitable for cryosurgery. Twelve weeks after treatment only one patient developed clinical warts on the cervix, which increased to three patients (4.8%) after 12 months. No further patients developed any clinical cervical warts. In the untreated group development of clinical warts was noticed in five patients (20%) after 18 months. The difference was significant. The failure rate for the histologically diagnosed subclinical HPV infection was 38% in the treated group, whereas it persisted in 76% of the untreated group. Cryotherapy was found to be successful in the treatment of subclinical HPV infection and prevented the development of clinical disease. There were no serious side-effects and the treatment can be administered easily in an out-patient setting after histological confirmation.

Efficacy of ceftizoxime in the treatment of uncomplicated gonorrhoea: comparison with amoxycillin.

Four centres contributed a total of 466 patients to a study comparing the efficacy of oral amoxycillin with that of probenecid and intramuscular ceftizoxime. Both treatments were equally efficacious, except in the small number of penicillinase-producing Neisseria gonorrhoeae patients, all of whom responded to ceftizoxime and none to amoxycyllin. Both treatments were well tolerated, the only two adverse effects occurring in the ceftizoxime group.

Sexually transmitted diseases among patients seeking HIV antibody test for AIDS.

A total of 489 patients who had blood tests for antibodies to the human immunodeficiency virus (HIV) were given medical advice for sexually transmitted disease (STD) screening. Of 378 patients who had STD screening, STDs were found to be present in 47%. Among various STDs, non-specific urethritis (NSU) in men and chlamydial genital infection in women were found to be common. When the prevalence of these two diseases were compared with those of a control group the difference was significant.