Comparison of the effectiveness of WHO short-course chemotherapy and standard Russian antituberculous regimens in Tomsk, western Siberia.

BACKGROUND: There has been a resurgence of tuberculosis in Russia in the past decade. Traditional Russian services for treatment of tuberculosis are very different from those in the west. We aimed to compare the effects of WHO short-course chemotherapy with standard Russian antituberculous regimens. METHODS: New tuberculosis patients aged 18 years or older were included in a trial and systematically allocated to traditional Russian tuberculosis treatments or WHO short-course chemotherapy in the two largest tuberculosis diagnostic and treatment centres of Tomsk Oblast, western Siberia. Standard WHO tuberculosis outcomes and rates of sputum conversion were used as primary outcomes. Analyses were by intention-to-treat. FINDINGS: 646 new cases were enrolled into the trial, of which 356 patients were given Russian tuberculosis treatment (155 smear positive) and 290 were given WHO short-course chemotherapy (155 smear positive). There was no statistical difference between the proportion cured or completing treatment (63% for both groups [difference in proportion=0%, 95% CI -11 to 11%]); or dying (short-course chemotherapy, 8% vs Russian, 11% [difference in proportion=-3%, 95% CI -9 to 4%]). There was no statistical difference with respect to sputum conversion rate at 6 months (91% vs 85% [difference in proportion=6%, 95% CI -2 to 13%]). Overall, outcomes were worse among patients with multidrug resistant isolates than non-resistant isolates. INTERPRETATIONS: WHO short-course chemotherapy treatment for tuberculosis can work well in Russia.

Sulindac sulfone inhibits K-ras-dependent cyclooxygenase-2 expression in human colon cancer cells.

Both the sulfide and sulfone metabolites of sulindac, a nonsteroidal anti-inflammatory drug, display anticarcinogenic effects in experimental models. Sulindac sulfide inhibits cyclooxygenase (COX) enzyme activities and has been reported to suppress ras-dependent signaling. However, the mechanisms by which sulindac sulfone suppresses cancer growth are not as defined. We studied the effects of these sulindac metabolites in human colon cancer-derived Caco-2 cells that have been transfected with an activated K-ras oncogene. Stable transfected clones expressed high levels of COX-2 mRNA and protein, compared with parental cells. K-ras-transfected cells formed tumors more quickly when injected into severe combined immunodeficiency disease mice than parental cells, and this tumorigenesis was suppressed by treatment with sulindac. Sulindac sulfone inhibited COX-2 protein expression, which resulted in a decrease in prostaglandin synthase E2 production. Sulindac sulfide had little effect on COX-2 in this model, but did suppress prostaglandin synthase E2 production, presumably by inhibiting COX enzyme activity. These data indicate that the sulfide and sulfone derivatives of sulindac exert COX-dependent effects by distinct mechanisms.

Influence of K-ras activation on the survival responses of Caco-2 cells to the chemopreventive agents sulindac and difluoromethylornithine.

The nonsteroidal anti-inflammatory drug sulindac and the ornithine decarboxylase inhibitor difluoromethylornithine (DFMO) are both potent inhibitors of colon carcinogenesis in experimental models of this disease. The combination of these two agents is undergoing evaluation as a strategy for colon cancer chemoprevention in humans with resected colon polyps. We evaluated the effects of the major sulfide and sulfone metabolites of sulindac and DFMO alone, or in combinations, on the growth and survival of Caco-2 colon cancer-derived cells and in clones of these cells transfected with an activated K-ras oncogene. Both the sulfide and sulfone metabolites of sulindac reduced cell viability, measured by colony-forming assays, primarily by inducing apoptosis. Expression of an activated K-ras oncogene caused cells treated with either sulindac sulfide or sulfone to undergo apoptosis earlier than nontransfected controls. However, clonogenic survival, measured 2 weeks after drug treatment, was the same in both Caco-2 and ras-transfected Caco-2 cells treated with sulindac metabolites. A 24-h treatment with DFMO caused a dose-dependent decrease in the colony-forming ability of cells expressing an activated K-ras but had no effect on the viability of the parental Caco-2 cells. The DFMO-dependent decrease in colony formation in K-ras-activated cells occurred in the absence of apoptosis. Assessment of cell survival by colony-forming assays indicated that these two agents acted in an additive manner when combined. These data indicate that K-ras can influence the kinetics of apoptosis induction by sulindac metabolites and cell survival in response to DFMO. However, cytotoxicity induced by these agents occurs via unique mechanisms. These studies suggest that the combination of DFMO and sulindac may be useful in human cancer prevention strategies.

APC-dependent changes in expression of genes influencing polyamine metabolism, and consequences for gastrointestinal carcinogenesis, in the Min mouse.

The colorectal mucosa of pre-symptomatic individuals with familial adenomatous polyposis (FAP) contains elevated levels of the proliferation-associated polyamines. The Min mouse, like humans with FAP, expresses an abnormal genotype for the APC tumor suppressor gene. In order to determine how APC mutation influences intestinal tissue polyamine content, we measured steady-state RNA levels of ornithine decarboxylase (ODC), the first enzyme in polyamine synthesis, antizyme (AZ), a protein which negatively regulates ODC, and the spermidine/spermine N(1)-acetyltransferase (SSAT), the first enzyme in polyamine catabolism. RNA content was increased 6- to 8-fold in both the small intestine and colon for ODC, decreased significantly in the small intestine but not the colon for AZ and was not statistically different in either intestinal tissue for SSAT in Min mice compared with normal littermates. Consistent with the changes in ODC and AZ gene expression, small intestinal, but not colonic, polyamine content was elevated in Min mice compared with normal littermates. Treatment of Min mice with the specific ODC inhibitor difluoromethylornithine (DFMO) suppressed small intestinal, but not colonic, polyamine content and tumor number. These data indicate that small intestinal tissue polyamine content is elevated in Min mice by a mechanism involving APC-dependent changes in ODC and AZ RNA. Further, ODC enzyme activity, which is influenced by both ODC and AZ RNA levels and inhibited by DFMO, is consequential for small intestinal tumorigenesis in this model. In the FAP population, DFMO may be of value in the chemoprevention of small intestinal adenocarcinoma that remains a risk following colectomy.

Expression of the human spermidine/spermine N1-acetyltransferase in spermidine acetylation-deficient Escherichia coli.

A cDNA encoding the human spermidine/spermine N1-acetyltransferase (N1SSAT) was conditionally expressed in a strain of Escherichia coli deficient in spermidine-acetylating activity. Conditional expression of this cDNA was performed under the control of the lac promoter, by addition of the non-hydrolysable lactose analogue isopropyl beta-D-thiogalactoside. Expression of the N1SSAT cDNA oriented in the sense direction resulted in the acetylation of spermidine at the N1 but not the N8 position and a decrease in endogenous spermidine contents and growth rates in these bacteria. When this cDNA was expressed in the antisense orientation, spermidine acetylation was not detected and endogenous spermidine contents and growth rates were unaffected. Increasing the endogenous N1-acetylspermidine concentration by addition of this amine to the culture medium did not suppress growth, and increasing endogenous spermidine pools by exogenous addition was not sufficient to restore optimal growth in cells expressing the human N1SSAT. Exogenous spermidine, but neither N1- nor N8-acetylspermidine, stimulated cell growth in strains unable to synthesize spermidine. These results suggest that one physiological consequence of spermidine acetylation in E. coli is growth inhibition. The mechanism of this inhibition seems to involve the formation of acetylspermidine, and is not simply due to a decrease in the intracellular concentration of non-acetylated spermidine.

Tuberculosis in Siberia: 2. Diagnosis, chemoprophylaxis and treatment.

OBJECTIVE: To assess tuberculosis diagnosis, chemoprophylaxis and therapy in Siberia as a paradigm for the Russian Federation. DESIGN: Data was obtained from official sources and through visits to dispensaries and hospitals in 1994. RESULTS: Tuberculosis disease and cure is classified according to a Dispensary Group Register based principally on clinical and radiological criteria. Isoniazid is widely used for chemoprophylaxis and post-therapy and may be linked to high levels of isoniazid resistance. Combination drug therapy is individualized, frequently changed, and given orally, parenterally or intra-bronchially. Galvanization, autotransfusion of ultra-violet irradiated blood, antioxidants and steroids are used as adjunct treatment. Ambulatory treatment is uncommon. Surgical treatments including lobectomy and pneumonectomy are used in 5-10% of patients. CONCLUSION: Tuberculosis is increasing in Siberia. An improved drug supply using short course standardized regimens is required supported by high quality co-ordinated bacteriological services. Surgery retains a useful role, but many adjunct therapies should be abandoned.

Tuberculosis in Siberia: 1. An epidemiological and microbiological assessment.

SETTING: Siberia, Russian Federation. OBJECTIVE: To assess the situation regarding tuberculosis as a paradigm for the Russian Federation. DESIGN: Data was obtained from official sources and through visits to dispensaries and hospitals in 1994. RESULTS: The downward trend in notifications of tuberculosis throughout Russia reversed in 1990/91, the rate increasing from 34/100,000 to 42.9/100,000 in 1993. Incidence rates are higher in Siberia, varying from approximately 43 to 108/100,000; prevalence is 250-300/100,000. The tuberculosis service is centralized and based on specialized polyclinics and dispensaries. An extensive surveillance system employs regular fluorography and tuberculin testing: half of the cases diagnosed are detected by fluorography, against 1% through contact tracing. Patients are classified principally on clinical and radiological grounds. Bacille Calmette-Guérin immunisation is performed at birth and at age 7, and again at 13, 21, and 28 years if Mantoux test is negative. Microscopy and culture services are organisationally separate, and direct comparison of smear and culture data is not possible. Drug resistance to isoniazid and streptomycin is probably high and resistance to rifampicin low, but data on susceptibility of isolates from new cases are not available. CONCLUSION: Tuberculosis is increasing in Siberia. Homelessness, unemployment and alcoholism are important factors, but concurrent human immunodeficiency virus (HIV) infection appears to be uncommon. Prisons probably form a significant reservoir of infectious cases.

Growth arrest- and polyamine-dependent expression of spermidine/spermine N1-acetyltransferase in human tumor cells.

Polyamines are essential for optimal cell growth. The regulation of polyamine biosynthetic, but not catabolic, enzymes has been studied in detail. Because intracellular polyamine contents depend on both synthesis and catabolism, we studied the regulation of spermidine/spermine N1-acetyltransferase (N1SSAT), the first enzyme in polyamine catabolism. Steady-state RNA levels of N1SSAT increased 3-5 fold as human colon tumor-derived HCT116 cells traversed the log phase and entered the plateau phase. Depletion of cellular polyamines, using alpha-difluoromethylornithine, caused a decrease in the steady-state levels of both the 1.3-kb N1SSAT transcript and its 3.5-kb precursor, without affecting the stability of either RNA. N1SSAT enzyme activity was low in cells with normal polyamine contents but could be induced by heat shock. The level of induction of N1SSAT enzyme activity by heat shock on different days of growth correlated with N1SSAT RNA levels prior to heat shock and occurred without changes in levels of message after heat shock. Although non-heat-shocked cells containing normal polyamine contents expressed N1SSAT RNA but not enzyme activity, exogenous spermidine restored both RNA levels and enzyme activity in polyamine-depleted cells. This result suggests that the expression of N1SSAT enzyme activity, but not RNA, requires a change in the intracellular compartmentalization of spermidine. These data demonstrate that N1SSAT is regulated at both the transcriptional and posttranscriptional levels by conditions that arrest growth in HCT116 cells, and that both of these mechanisms are affected by endogenous polyamine contents.

[International concept of antituberculous care organization for the Tomsk region's population]. / Mezhdunarodnaia kontseptsiia organizatsii protivotuberkuleznoi pomoshchi naseleniiu Tomskoi oblasti.

The priorities of the international concept of organization of antituberculous care to the population of the Tomsk Region are as follows: detection of bacillar patients and their short-term intensive chemotherapy by the WHO categories; formation of a 6-month reserve of tuberculostatics and complete control over patients' drug use; diagnosis of diseases by the general medical network for patients' referrals by introducing indications for clinical examinations for tuberculosis and sputum tests for Mycobacterium tuberculosis, fluorographic studies of only risk group populations; tuberculin diagnosis among children aged 1-14 years from risk groups; BCG vaccination of the newborn and revaccination at the age of 6-7 years. Resource mobilization is essential. This includes: strengthening of basic service institutions and centralization of its structures; fund saving by reasonably reducing the beds in sanatoria and hospitals; extension of therapeutical and diagnostic services at the outpatient stage of a follow-up; decrease in the length of patients' medical examinations and in the number of groups registered at a tuberculous dispensary; intensification the work of a phtx11p4trist and a nurse with their salary increases.

[Genetic polymorphism of the erythrocytic enzymes in the Buryat populations]. / Geneticheskii polimorfizm éritrotsitarnykh fermentov v populiatsiiakh Buriat.

Polymorphism of seven erythrocytic enzymes PGM1, ESD, CLO1, PGD and PGP were studied in five samples of Buryats. The main investment into differentiation between populations has been made by the following systems: CLO1, PGD and PGM1. Analysis of genetic distances between populations demonstrated that there was some parallelism among the genetic and anthropological differentiation in the Buryat populations. The groups of the Agingsky county (the area to the east from the Baikal Lake) have probably the largest proportion of the Caucasian genes as compared to other populations studied. One of the characteristics of the Buryats, especially for the population to the east from the Baikal Lake, is high frequency of the PGD allele. The rate of the genetic variability on the intra-population level is higher than the difference between populations. This means that the divergence between the Buryats populations is not very strong. Consideration of the genetic variability on the intra-population level seems to be more perspective for ecogenetic estimation of the adaptive genetic processes than analysis of the differences between populations studied.

[Genetic studies of phosphoglucomutase-1 (PGM1) subtypes: population aspects]. / Geneticheskie issledovaniia subtipov fosfogliukomutazy-1 (PGM1): populiatsionnye aspekty.

Distribution of the subtypes and gene frequencies of phosphoglucomutase-1 among some populations of Buryats, Kirghizes of the Pamir and Russians of Moscow district was analysed. The frequencies of PGM1 genes vary in Buryats being PGM1+(1) 0.647-0.743, PGM1-(1)-0.100-0.132, PGM2+(1)-0.122-0.199 and PGM2-(1)-0.007-0.037. Following frequencies of PGM1 genes were established for Kirghizes: PGM1+(1) = 0.614, PGM1-(1) = 0.114, PGM2+(1) = 0.217 and PGM2-(1) = 0.054; in Russian populations the frequencies were: PGM1+(1) = 0.578, PGM1-(1) = 0.110, PGM2+(1) = 0.253 and PGM2-(1) = 0.059. Peculiarities of PGM1 polymorphism in the USSR and all over the world were analysed. Parallel biodemographic investigations in Buryat population demonstrated differences in intensities of selection, related to concrete PGM genotypes.

Ornithine decarboxylase activity and polyamine content in adenocarcinomas of human stomach and large intestine.

Ornithine decarboxylase (ODC) activity and differential polyamine composition have been studied in macroscopically normal mucosa, adjacent mucosa, and neoplastic tissue of 95 patients with adenocarcinomas of stomach and large intestine. In tumors, we found increased ODC activity and polyamine content as compared with surrounding mucosa. ODC activity in macroscopically normal tissue of patients with tumors of stomach and large intestine increased as the disease progressed. An inverse relationship was observed between ODC activity in adenocarcinomas and differentiation.

[Ornithine decarboxylase activity and polyamine contents in 1,2-dimethylhydrazine-induced carcinogenesis of the intestines in rats]. / Aktivnost' ornitindekarboksilazy i soderzhanie poliaminov pri indutsirovannom 1,2-dimetilgidrazinom kantserogeneze kishechnika u krys.

The ornithine decarboxylase (ODC) activity and concentration of polyamines have been studied in small and large intestine during 1,2-dimethylhydrazine-induced carcinogenesis in rats. Changes in the polyamine biosynthesis consisting both in enhanced ODC activity and an increase of the intracellular content of putrescine, spermidine and spermine. Process of the polyamine synthesis activation proceeds in two phases: the most expressed and similar changes in polyamine metabolism factors have been observed at early (the 1st month) and late (the 5th-6th months) stages of carcinogenesis. It is supposed that intensification of the polyamine synthesis is a typical feature of malignization in the gastrointestinal tract.

[Ornithine decarboxylase activity and polyamine content in human adenocarcinomas of the stomach and large intestine]. / Aktivnost' ornitindekarboksilazy i soderzhanie poliaminov v adenokartsinomakh zheludka i tolstogo kishechnika cheloveka.

Ornithine decarboxylase (ODC) activity and polyamine content were studied in the intact mucosa and tumours of human stomach and large intestine adenocarcinomas. ODC activity has been found to be much higher in the neoplastic tissue than in the corresponding intact mucosa. Concentrations of polyamines in neoplasms were also higher than in mucosa. Interrelation was found between ODC activity polyamine content and degree of differentiation of the tumour cells in rectal adenocarcinomas.

[Increase in ornithine decarboxylase activity and polyamine levels of the rat liver during an early period of hepatocarcinogenesis]. / Uvelichenie aktivnosti ornitindekarboksilazy i soderzhaniia poliaminov v pecheni krys v rannii period gepatokantserogeneza.

The ornithine decarboxylase activity and the polyamine content in the fraction of plasma membranes and cytosol of the rat liver are studied in the early period (1-4 weeks) of nitrosodiethyl amine-induced hepatocarcinogenesis. The enzyme activity and polyamine levels in the cytosol of the rat liver cells are found to increase sharply during the first month of the disease, the maximum being observed the first-second week. By the end of the fourth week these indices become lower but they remain significantly higher than the normal levels. The polyamine level increases considerably in the fraction of plasma membranes with the maximum observed the second week.