The role of a novel copper complex in overcoming doxorubicin resistance in Ehrlich ascites carcinoma cells in vivo.

One of the important pathways of resistance to anthracyclines is governed by elevated levels of glutathione (GSH) in cancer cells. Resistant cells having elevated levels of GSH show higher expression of multidrug-resistant protein (MRP); the activity of glutathione S-transferases (GSTs) group of enzymes have also been found to be higher in some drug-resistant cells. The general mechanism in this type of resistance seems to be the formation of conjugates enzymatically by GSTs, and subsequent efflux by active transport through MRP (MRP1-MRP9). MRPs act as drug efflux pump and can also co-transport drugs like doxorubicin (Dox) with GSH. Depletion of GSH in resistant neoplastic cells may possibly sensitize such cells, and thus overcome multidrug resistance (MDR). A number of resistance modifying agents (RMA) like DL-buthionine (S, R) sulfoxamine (BSO) and ethacrynic acid (EA) moderately modulate resistance by acting as a GSH-depleting agent. As most of the GSH-depleting agents have dose-related toxicity, development of non-toxic GSH-depleting agent has immense importance in overcoming MDR. The present study describes the resistance reversal potentiality of novel copper complex, viz., copper N-(2-hydroxy acetophenone) glycinate (CuNG) developed by us in Dox-resistant Ehrlich ascites carcinoma (EAC/Dox) cells. CuNG depletes GSH in resistant (EAC/Dox) cells possibly by forming conjugate with it. Depletion of GSH results in higher Dox accumulation that may lead to enhanced rate of apoptosis in EAC/Dox cells. In vivo studies with male Swiss albino mice bearing ascitic growth of EAC/Dox showed tremendous increase in life span (treated/control, T/C = 453%) for the treated group with apparent regression of tumor. Resistance to Dox in EAC/Dox cells is associated with over expression of GST-P1, GST-M1 (enzymes involved in phase II detoxification) and MRP1 (a transmembrane ATPase efflux pump for monoglutathionyl conjugates of xenobiotics). CuNG causes down regulation of all these three proteins in EAC/Dox cells. The effect of CuNG as RMA is better than BSO in many aspects.

Errors in the treatment of tuberculosis in Baltimore.

BACKGROUND: Incomplete or incorrect antibiotic therapy, especially in the initial phase of antituberculosis (anti-TB) treatment, is a major cause of acquired drug resistance and treatment failure. We determined the extent of errors in anti-TB treatment regimens by way of nonadherence to recommended treatment protocols among patients with TB in Baltimore, MD, a city with declining rates of disease. An error was defined as using too few drugs or the wrong drugs, giving inadequate doses of drugs, or prescribing an inadequate duration of treatment. METHODS: We reviewed the records of all patients with culture-positive, pulmonary TB reported in the city of Baltimore from January 1, 1994, to December 31, 1995. We determined demographic information, initial anti-TB regimen, doses and duration of therapy, history or presence of resistance to anti-TB drugs, injecting-drug or alcohol abuse, HIV status, and whether treatment was given by a private physician or by the Tuberculosis Clinic of the Baltimore City Health Department (BCHD). RESULTS: Of the 110 cases of active pulmonary TB, 17 cases (15.4%) had errors in treatment for control of their current disease. Thirteen of 34 privately treated patients (38%) had some error in their initial anti-TB regimen, compared with 4 of 76 patients (5.2%) treated by the Tuberculosis Clinic of the BCHD (p < 0.0001). Patients were otherwise similar as determined by age, sex, HIV status, drug-resistance characteristics, and injecting-drug use, regardless of whether they had erroneous anti-TB regimens. CONCLUSION: In a low-prevalence area, private physicians make frequent errors in prescribing anti-TB therapy. Additional educational resources for physicians and increased use of expert consultation may contribute to improved TB control.

Immunosuppression in hamsters with progressive visceral leishmaniasis: an evaluation of the role of nitric oxide toward impairment of the lymphoproliferative response.

The progressive visceral infection caused in golden hamsters by Leishmania donovani amastigotes led to gradual impairment of the proliferative response of their splenic (SPMC) or peripheral blood (PBMC) mononuclear cells to in vitro stimulation with leishmanial antigen, with mitogen (concanavalin A), and even with a combination of phorbol myristate acetate (PMA) and ionomycin (Io). Removal of macrophage-like adherent cells from SPMC or PBMC of infected animals, however, almost completely restored their proliferative response to PMA + Io, thus ruling out the possibility of any intrinsic defect in the signal-transduction pathways of lymphocyte activation and proliferation. Subsequent studies demonstrated that the generation of soluble mediators such as nitric oxide by these adherent cells is responsible, albeit partially, for the down-regulation of the lymphoproliferative response in hamsters with visceral leishmaniasis.

Therapeutic and prophylactic uses of protein A in the control of Leishmania donovani infection in experimental animals.

The role of the immunomodulator Protein A (PA) (from Staphylococcus aureus, Cowan I strain) in the control of leishmanial infection was studied in experimental animals. Treatment of Leishmania donovani infected hamsters with PA led to a moderate level of reduction of parasite load in their spleen (68%) and liver (46%). However, combination therapy of PA with the antileishmanial drug stibanate induced a more marked reduction of the spleen (88%) and liver (85%) parasitemia compared to that induced by PA or drug treatment alone. Similar results were also obtained with L. donovani infected BALB/c mice as the combination therapy of PA and stibanate led to a significant reduction (84%) of liver parasite load in comparison to that induced by PA (38%) or drug (61%) treatment alone. Apart from its therapeutic use, PA could also be used as a prophylactic agent in the control of leishmanial infection. Thus, treatment of hamsters with PA before leishmanial challenge significantly reduced their organ parasite load (by 59-78%) compared to that observed in infected controls without prior PA treatment. The antileishmanial effect of PA was likely to be mediated through the activation of macrophages leading to an enhancement of their phagocytic as well as leishmaniacidal activities. Subsequent studies demonstrated that PA treatment led to an increased production of nitric oxide by macrophages which could primarily be responsible for their enhanced parasite killing ability.

An insight into the structure of DNA through melting studies.

Helix-coil transition of calf thymus (CT) DNA in 0.1 M NaCl was observed under different environmental conditions for studying structural and conformational changes, if any. Pre-exposure of the DNA to different types of radiations - UV and gamma-brought different degrees of change (lowering) in the melting temperature (Tm). Interaction of drugs (daunomycin and actinomycin-D) increased the Tm inducing structural stability. There was 50% more destabilisation in case of polynucleotide-drug complex as compared to DNA-drug complex exposed to UV radiation. Thermodynamic studies on DNA in D2O (0.1 M NaCl prepared in D2O) were carried out and compared with the corresponding results in H2O. Presence of D2O increased the stability of the DNA structure. Structural and conformational aspects have been discussed in the light of the results obtained.

Spectrophotometric observations on the changes in the Y-peak of actinomycin D complexed to DNA on gamma-irradiation.

An understanding of the pi-pi * transitions of the Y-peak of the DNA bound to actinomycin D, in the presence and absence of gamma-irradiation has been attempted. It was seen that in the bound DNA the presence of the Y-peak, without irradiation was mainly due to the pi-pi * transitions associated with the DNA bases. The Y-peak was found to be more sensitive to the binding of the drug than the X-peak. After gamma-irradiation the Y-peak characteristics of the complexed DNA were different from those of the X-peak. From these studies it has been concluded that the electronic transitions (pi-pi *) of the DNA, in the far U.V., can be divided into three distinct groups: (a) Electronic transitions which are only seen at the Y-peak, (b) Transitions common both to the X and Y-peaks, and (c) Transitions which are only seen at the X-peak. This explains the difference in the behaviour of the Y- and X-peaks of the DNA due to the binding of Act.D and gamma-irradiation.

Chronic potassium depletion and sensitivity to tubocurarine.

The effect of low potassium diets with and without deoxycorticosterone (DOCA), furosemide, chlorothiazide, or ethacrynic acid on sensitivity of isolated guinea pig lumbrical nerve-muscle preparations to d-tubocurarine was examined. The ED50 of d-tubocurarine was found to fall as the potassium level was lowered by dietary restriction. Addition of DOCA or furosemide was without effect while chlorothiazide reduced and ethacrynic acid enhanced the effect of diet alone. Acute restoration of potassium levels of 5.9 mM reversed considerably, but not completely, the effect of chronic depletion. Thus, a patient suffering from chronic potassium depletion would be expected to require a decreased dose of d-tubocurarine, and although acute replacement of potassium preoperatively would tend to return the dose requirement toward normal, such reversal might not be complete.