Modulation of cardiovascular function by adipokines.

Cardiovascular disease (CVD) and associated risk factors such as obesity remain at the forefront of health concerns. Adipose tissue has been well established as an endocrine organ that becomes dysfunctional with increased adipose tissue mass. The secretion of several adipokines is altered in subjects with abdominal adiposity and these changes to the endocrine balance may contribute to increased CVD risk. The identification and characterization of disease-specific proteins within the adipose tissue offers a novel therapeutic target for prevention or treatment of cardiovascular complications. This review will discuss the latest developments on therapeutic targets within the context of adipokines, such as adiponectin, C1q/ tumor necrosis factor (TNF) related proteins (CTRPs), visfatin, vaspin, chemerin and omentin, and their involvement in obesity-related cardiovascular complications.

Gender and locality differences in tobacco prevalence among adult Bangladeshis.

OBJECTIVES: To determine the extent of all forms of tobacco usage in adult Bangladeshis in relation to gender and locality. METHODS: Three annual urban and rural cross-sectional surveys were carried out between 2001 and 2003 involving a total of 35,446 adults, of whom 54.3% were female and 51.0% were rural dwellers. Data were collected through interview using a structured questionnaire. RESULTS: The overall prevalences of smoking, chewing tobacco and gul usage were 20.5%, 20.6% and 1.8%, respectively. Current smoking and gul usage were significantly higher in males (42.2% and 2.2%, respectively) than females (2.3% and 1.5%, respectively) while chewing tobacco was more common in females (21.6%) than males (19.4%). No significant urban-rural difference was observed in smoking rate after adjusting for sociodemographic variables, while chewing tobacco was 1.5 times more likely to occur in rural residents and gul usage was 3.6 times more likely to occur in urban residents. On average a smoker consumed 9.3 sticks a day with males and rural residents smoking more. CONCLUSIONS: Nearly a third of the population in Bangladesh use some form of tobacco. There are marked urban-rural and male-female differences. This difference is mainly accounted for by the higher prevalence of chewing tobacco in rural areas, rural female tobacco usage is close to double than the urban rate. Smoking rates were low in Bangladeshi females, more so in urban than rural areas. The tobacco awareness programme in Bangladesh might require putting emphasis on smokeless tobacco as well as smoking.

Impact of intestinal permeability, inflammation status and parasitic infections on infant growth faltering in rural Bangladesh.

A longitudinal study of 298 rural Bangladeshi infants found evidence of growth faltering starting at 3 months of age. Anthropometric status declined substantially in the first 2 years of life, with weight-for-height (WHZ) falling from - 0.49 to - 1.75, weight-for-age (WAZ) from - 1.18 to - 2.87 and height-for-age (HAZ) from - 1.00 to - 1.88. Higher concentrations of the acute-phase protein alpha-1-acid glycoprotein (AGP) and higher gut mucosal damage (as signified by raised lactulose:mannitol (L:M) ratios) were both associated with chronic malnutrition as indicated by poorer HAZ and WAZ scores (P = 0.011 and 0.005 for AGP and 0.039 and 0.019 for L:M ratio, respectively). Higher Hb levels were related to improved z-scores, while elevation of Giardia-specific IgM titre (GSIgM) was associated with poor WAZ and WHZ (P = 0.015 and 0.039, respectively). IgG did not show any significant association with z-scores and the L:M ratio did not correlate with any of the inflammation markers or Giardia infection. The prevalence of geohelminth infections was low (only 4 % in the total study period). However, the level of GSIgM indicated high endemicity of Giardia infection from early in life, although very few cysts were detected from stool samples. These findings suggest that rural Bangladeshi infants are being exposed to high levels of infection with concomitant gut damage and growth faltering.

Impact of targeted food supplementation on pregnancy weight gain and birth weight in rural Bangladesh: an assessment of the Bangladesh Integrated Nutrition Program (BINP).

OBJECTIVES: To assess whether the Bangladesh Integrated Nutrition Programme (BINP) correctly identified which pregnant women should be enrolled in the food supplementation programme, whether supplementation commenced on time and was taken on a regular basis. A second objective was to determine whether food supplementation led to enhanced pregnancy weight gain and reduction in the prevalence of low birth weight. DESIGN: A one-year community-based longitudinal study. SETTING: A rural union of Bhaluka Upazila, Mymensingh, located 110 km north-west of Dhaka City, the capital of Bangladesh.ParticipantsA total of 1104 normotensive, non-smoking pregnant women who attended Community Nutrition Centres were studied from first presentation at the centre until child delivery. RESULTS: Pregnant women who had a BMI of <18.5 kg/m(2) on first presentation should have been selected for supplementary feeding (2512 kJ (600 kcal)/d for six days per week) starting at month 4 (16 weeks) of pregnancy. However, of the 526 women who had BMI < 18.5 kg/m(2), only 335 received supplementation; so the failure rate was 36.3 %. In addition, of those receiving supplementation, only 193 women (36.7 % of 526 women) commenced supplementation at the correct time, of whom thirty-two (9.6 % of 335 women) received supplementation for the correct number of days (100 % days). There were no significant differences in mean weight gain between BMI < 18.5 kg/m(2) supplemented or non-supplemented groups or between the equivalent groups with BMI > or = 18.5 kg/m(2). Weight gain was inversely related to initial weight, so lighter women gained relatively more weight during their pregnancy than heavier women. The mean birth weight in the supplemented and non-supplemented groups was 2.63 kg and 2.72 kg, respectively. Mothers with BMI < 18.5 kg/m(2) who were or were not supplemented had almost equal percentages of low-birth-weight babies (21 % and 22 %, respectively). CONCLUSION: The study raises doubt about the efficiency of the BINP to correctly target food supplementation to pregnant women. It also shows that food supplementation does not lead to enhanced pregnancy weight gain nor does it provide any evidence of a reduction in prevalence of low birth weight.

Impact of anti-Giardia and anthelminthic treatment on infant growth and intestinal permeability in rural Bangladesh: a randomised double-blind controlled study.

In order to test the impact of Giardia and geohelminthic infection on infant growth faltering in Bangladesh, a randomised double-blind placebo controlled intervention of 36 weeks' duration was conducted in a rural community located 40 km northwest of Dhaka. Infants aged between 3 and 15 months were randomly assigned to either anti-Giardia and anthelminthic treatment, anti-Giardia treatment only, or a control. Weight and supine length were recorded every 4 weeks. Every 12 weeks intestinal permeability (lactulose/mannitol ratio), haemoglobin, plasma albumin, alpha-1-acid glycoprotein, IgG and Giardia-specific IgM (GSIgM) and eggs of the three common geohelminths and G. intestinalis cysts were determined. Data on 222 fully compliant infants were analysed. No significant differences in intestinal permeability, biochemical or anthropometric variables were found between the intervention groups, although there were associations between improvement in small intestinal mucosal function and better weight-for-age and weight-for-height (length) Z-scores. GSIgM titres indicated high endemicity with rapid re-infection of Giardia among infants; over 95% of infants were positive throughout the study, whereas the stool examination showed very few infants with either geohelminth eggs or Giardia cysts.

An easy method using glutaraldehyde-introduced fluorescence for the microscopic analysis of plant biotrophic interactions.

The method introduced in this article makes use of the glutaraldehyde-induced auto-fluorescence of proteins after cross-linking with glutaraldehyde for the analysis of cellular and sub-cellular structures. Because the interface of biotrophic interactions is rich in proteins, the method presented is particularly suitable for the analysis of such interactions; we have exemplified its usefulness by analyzing (1) the root feeding sites induced in roots from Arabidopsis thaliana by the root-knot nematode Meloidogyne incognita; (2) leaves from Cucurbita pepo infected by powdery mildew and (3) roots from Nicotiana tabacum colonized by the arbuscular mycorrhizal fungus Glomus intraradices. The use of confocal and multi-photon laser scanning microscopy allows three-dimensional reconstructions from optical sections of complex biotrophic interactions. In the case of root-knot nematode feeding sites, our method enabled us to simultaneously study the development of the plant xylem elements (using lignin auto-fluorescence), the nematode feeding site and the nematode itself.

Maternal anthropometry as a predictor of birth weight.

OBJECTIVE: To determine whether maternal anthropometry predicted birth weight, and if so, to identify which cut-offs provided the best prediction of low birth weight (LBW) in a field situation. DESIGN: Community-based longitudinal study. SETTING: A rural union of Bhaluka Upazila, Mymensingh, located 110 km north-west of Dhaka, the capital of Bangladesh. PARTICIPANTS: A total of 1104 normotensive, non-smoking pregnant women who attended community nutrition centres were studied from first presentation at the centre until delivery of their child. RESULTS: Most of the pregnant mothers were between 20 and 34 years of age. Over one-third of the women were nulliparous, while 12.8% were multiparous (parity >/= 4). Most (93%) mothers registered between the 3rd and 5th month of pregnancy. The frequency of LBW ( < 2500 g) was 17%. Polynomial regression analyses showed that the best predictors of birth weight (based on adjusted R2 values) were in general weight at registration and weight at month 9, with adjusted R2 ranging from 2.5% to nearly 20%. Sequential regression analyses with height and weight showed that there was a significant effect of height after removing the weight variables, and adjusted R2 increased in all analyses. Weight and height at registration month continued to be the best predictors of LBW. Sensitivity and specificity curves were drawn for each registration month, body mass index and different weight gain groups, and using different weight and height combinations. The results showed that, for registration month 3-5, a combination of weight (

Total energy expenditure (H218O), physical activity level and milk output of lactating rural Bangladeshi tea workers and nontea workers.

OBJECTIVE: To assess the total daily energy expenditure (TDEE), milk output and physical activity level (PAL) of chronically malnourished lactating women using the doubly labelled water method (DLW). DESIGN: Prospective, longitudinal study designed to assess the extent of malnutrition and energetics of lactating tea workers and nontea workers. SETTING: North-east Bangladesh on women working and living in the same tea estates. SUBJECTS: Of an original cohort of 150 lactating women, 30 were selected to participate in this study when they were at about 12 months postpartum. One mother subsequently dropped out. INTERVENTIONS: On day 1 each women provided a urine sample, was administered a dose of DLW and 6 h later provided another sample. Further urine samples were collected for 21 subsequent days. In addition, every 5 days the mother provided a milk sample and at the same time her baby provided a urine sample. RESULTS: Mean (s.d.) BMI was 17.4 (1.63). Mean TDEE and PAL were significantly higher in workers than nonworkers (8.42 (1.38) and 6.83 (2.09) MJ/day, P = 0.02 and 1.92 (0.34) and 1.59 (0.44), P = 0.03, respectively). Mean milk output was similar in the two groups (672 (180) ml and 749 (189) ml in workers and nonworkers, respectively). CONCLUSIONS: Based on international BMI cutoffs, 79% of mothers were suffering from some degree of chronic energy deficiency. A total of 35% of workers and 17% of nonworkers were engaged in strenuous physical activity. The mean milk output of both workers and nonworkers was not different and was high especially as most of the mothers were about 12 months postpartum. No relationship was found between menses return and any of the variables studied. SPONSORSHIP: World Health Organization, Nestle Foundation, UNICEF.

Reproducibility of prompts in computer-aided detection (CAD) of breast cancer.

AIM: We evaluated the reproducibility of prompts using the R2 ImageChecker M2000 computer-aided detection (CAD) system. MATERIALS AND METHODS: Forty selected two-view mammograms of women with breast cancer were digitized and analysed using the ImageChecker on 10 separate occasions. The mammograms were chosen to provide both straightforward and subtle signs of malignancy. Data analysed included mammographic abnormality, pathology, and whether the cancer was prompted or given an emphasized prompt. RESULTS: Correct prompts were generated in 86 out of 100 occasions for screen-detected cancers. Reproducibility was less in the other categories of more subtle cancers: 21% for cancers previously missed by CAD, a group that contained more grade 1 and small (<10 mm) tumours. Prompts for calcifications were more reproducible than those for masses (76% versus 53%) and these cancers were more likely to have an emphasized prompt. CONCLUSIONS: Probably the most important cause of variability of prompts is shifts in film position between sequential digitizations. Consequently subtle lesions that are only just above the threshold for display may not be prompted on repeat scanning. However, users of CAD should be aware that even emphasized prompts are not consistently reproducible.

The relationship between areca nut usage and heart rate in lactating Bangladeshis.

BACKGROUND: The betel-nut quid, a piece of areca nut chewed alone or mixed with tobacco and slaked lime wrapped in betel vine leaf, is widely used in Asian populations as a stimulant (due to the cholinergic agent, arecoline) or as a relaxant (due to arecaidine and guvacine). OBJECTIVES: This study, which formed part of a larger project assessing the effect of energy expenditure on the duration of post-partum amenorrhoea, provided the opportunity to assess the role of chronic areca nut usage on heart rate and oxygen consumption during resting periods and during graded stepping tests. SUBJECTS AND METHODS: The mothers (n = 47), all of whom were lactating, were aged between 19 and 39, of low nutritional status and anaemic and they all chewed betel quid daily. RESULTS: Moderate users of betel quid (defined as more than 3 times a day) were found, on average, to have a significantly lower heart rate at rest and during exercise than low betel quid users (less than 3 times a day) but there was no modification in oxygen consumption. CONCLUSION: Chronic betel quid use does not seem to affect the assessment of 24h energy expenditure provided that subjects are denied access to betel nut usage before and during calibration.

Breast cancer resistance protein expression and resistance to daunorubicin in blast cells from patients with acute myeloid leukaemia.

Breast cancer resistance protein (BCRP) is a recently described member of the ATP binding cassette transporter superfamily. It has been shown to confer resistance to mitoxantrone, topotecan, doxorubicin and daunorubicin in human tumour cell lines. We describe a study of BCRP expression in blast cells derived from 20 patients with acute myeloid leukaemia (AML). Twelve samples were from patients who had received previous cytotoxic therapy. BCRP expression was measured by immunocytochemistry using the BXP-34 monoclonal antibody. In vitro drug sensitivity was assessed using the methyl thiazol tetrazoliumbromide assay. BCRP expression varied between patients, and six out of 22 (27%) samples had > 10% cells staining positively (median 37%, range 13-95%). BCRP positivity was seen in both de novo samples and those from previously treated patients. There was a marked variation in the effect of all drugs tested between patients. Although there was no correlation between BCRP positivity and the effect of mitoxantrone, topotecan or doxorubicin, the median daunorubicin LC(50) value of BCRP(+) cells was fourfold higher than that of BCRP- cells (0.89 micromol/l compared with 0.21 micromol/l, P < 0.05). These results suggest that BCRP may be involved in resistance to the agents commonly used in AML and may explain some of the anomalous results found when studying other membrane transporters, such as P-gp or MRP.

Chemosensitivity testing predicts survival in ovarian cancer.

The aim of this study was to assess the use of the MTT assay for chemosensitity testing to identify drug resistance and predict survival in patients with advanced ovarian cancer. Samples of ascitic fluid and/or solid biopsies were taken from 120 patients with FIGO stage III or IV ovarian adenocarcinoma at presentation. Cells were exposed for 48 hours to four concentrations of clinically relevant drugs including platinums, anthracyclines and alkylating agents. Cell survival was measured using the 3-4,5-dimethyl-2, 5-diphenyl tetrazolium bromide (MTT) assay allowing patients to be grouped as "sensitive" or "resistant" in vitro. Clinical data including age, residual disease, histological grade, treatment, response after initial treatment and overall survival were collected. There was a highly significant (p<0.0001) correlation of in vitro sensitivity with in vivo response in the patients who completed their therapy, with an 83% positive predictive accuracy for resistance. This translated in the longer term to an increased survival for the patients found to be sensitive in vitro to their therapy with a 5-year survival rate of 24% compared to 12% for the resistant group (p=0.033). These results suggest that MTT chemonsensitivity testing can predict response in ovarian cancer leading to the prospect of increased survival in this devastating disease by customising therapy to individual patients.

The effect of weekly iron supplementation on anaemia and on iron deficiency among female tea pluckers in Bangladesh.

AIM: To investigate the effect of weekly iron supplementation on anaemia and iron deficiency among adult, female tea pluckers. METHOD: A randomized double-blind intervention trial was conducted in a tea estate in Bangladesh where a total of 280 women received either weekly iron supplementation (200 mg ferrous fumarate and 200 mg folic acid) for 24 weeks or a matching placebo. Capillary blood samples were drawn at baseline and post-trial to determine haemoglobin, haematocrit and ferritin concentration. Mean corpuscular haemoglobin concentration (MCHC) was calculated using the haemoglobin and haematocrit values. RESULTS: The mean haemoglobin concentration in the supplemented group increased by 5.52 g L-1 over the study period, on average, while ferritin values decreased by 0.33 microgram L-1. The control group showed a decrease in both mean haemoglobin (-0.24 g L-1) and ferritin (-5.32 micrograms L-1). Those individuals in the supplemented group with the lowest pretrial haemoglobin and ferritin values experienced the greatest improvements post-trial, whereas nonanaemic individuals showed a decrease in both haemoglobin and ferritin concentrations. A total of 62.2% of women in the supplemented group reported feeling better and more energetic compared to 51.1% in the placebo group; 14.4% of the supplemented group and 22.7% of the control group complained about side-effects. CONCLUSION: Weekly iron supplementation was logistically simpler and cheaper than daily supplementation but would have to be continued on a longer term basis in order to combat both anaemia and iron deficiency.

Dexrazoxane significantly impairs the induction of doxorubicin resistance in the human leukaemia line, K562.

Dexrazoxane combined with doxorubicin (+ 5-fluorouracil + cyclophosphamide - the FAC regime) leads to a significant decrease in doxorubicin cardiotoxicity and a significant increase in median survival time for patients with advanced breast cancer responsive to FAC. The reason for this increase in survival may be due to interference with the mechanism involved in the emergence of multidrug resistance (MDR). In order to test this hypothesis, we induced resistance to doxorubicin in the K562 cell line by growing cells in increasing concentrations of doxorubicin (10-30 nM) in the presence and absence of dexrazoxane (20 nM). The doxorubicin sensitivity of all resultant sublines was measured using the MTT assay. Flow cytometry was used to assess the MDR1 phenotype, measuring P-glycoprotein expression with MRK 16 antibody and drug accumulation in the presence and absence of PSC 833 for functional P-glycoprotein. Long-term growth in doxorubicin increased the cellular resistance (IC(50)) of K562 cells in a concentration-dependent manner (r(2 )= 0.908). Doxorubicin resistance was not induced in the presence of dexrazoxane (P< 0.0001) for several months. In parallel, the expression of functional P-glycoprotein was delayed after concomitant addition of dexrazoxane to the selecting medium (P< 0.001). Dexrazoxane did not act as a conventional modulator of P-glycoprotein. These results suggest that dexrazoxane may delay the development of MDR1, thus allowing responders to the FAC regime to continue to respond.

Circumvention of ara-C resistance by aphidicolin in blast cells from patients with AML.

Treatment failure in AML is often attributed to P-glycoprotein-associated multidrug resistance. However, the importance of increased DNA repair in resistant cells is becoming more apparent. In order to investigate the ability of the DNA repair inhibitor aphidicolin to modulate drug resistance, we continually exposed blasts cells, isolated from 22 patients with AML, to a variety of agents +/- 15 microM aphidicolin for 48 hours. Cell survival was measured using the MTT assay. Overall, there was no significant effect of aphidicolin on sensitivity to daunorubicin, doxorubicin, etoposide or fludarabine. However, there was a marked increase in sensitivity to ara-C with a median 4.75-fold increase overall (range 0.8-80-fold;P< 0.005). The effect of aphidicolin was significantly greater in blast cells found resistant in vitro to ara-C (8.9-fold compared to 2.12-fold, P< 0.01). This observation was further validated by the correlation between ara-C LC(50)and extent of modulation effect (P< 0.05). Cells isolated from 10 cord blood samples were also tested in order to establish the haematological toxicity of combining ara-C and aphidicolin. The therapeutic index (LC(50)normal cells/tumour cells) for ara-C + aphidicolin was higher than that for ara-C alone suggesting no increased myelotoxicity for the combination. Increased cytotoxicity without increased haematotoxicity makes the combination of ara-C plus aphidicolin ideal for inclusion in future clinical trials.

Dietary zinc deficiency and expression of T lymphocyte signal transduction proteins.

Impaired immune function in dietary zinc (Zn) deficiency is characterized in part by reduced lymphocyte numbers (lymphopenia) and depressed cell-mediated (T lymphocyte) immune function, however, the causative mechanisms at the molecular level have not been elucidated. This paper will focus on the role of dietary Zn in T lymphocyte signal transduction, and specifically, the early Zn-dependent steps for phosphorylation and the putative Zn-finger proteins or Zn-metalloenzymes that may be part of the molecular mechanism for explaining immune dysfunction in Zn deficiency. One of the major recent findings is that murine splenic T lymphocyte p56lck expression is elevated in dietary Zn deficiency and caloric deficiency. Based on the known functions of p56lck, it is proposed that elevated p56lck may contribute to altered thymocyte maturation, apoptosis, and lymphopenia in dietary Zn deficiency and other malnutrition syndromes.

Comparison of P-glycoprotein expression and function with in vitro sensitivity to anthracyclines in AML.

The importance of P-glycoprotein (P-gp) in AML has been well documented. Resistance to the anthracyclines can be overcome by several agents including Cyclosporin A (CsA), PSC833 and GF120918. We describe an investigation into the expression, using MRK16 and UIC2, and function of P-gp using daunorubicin with and without modulators by flow cytometric analysis on previously frozen blast cells from 27 patients with primary or secondary AML. We compared this with the in vitro chemosensitivity, using the MTT assay, of fresh blast cells from the same patients. Whilst we found a correlation between P-gp function using CsA and GF120918 and expression using MRK16 (p < 0.05) and (p < 0.02) respectively, we were unable to find any overall correlation between expression and function of P-gp with either in vitro sensitivity to the anthracyclines, previous treatment, or 1 degree or 2 degrees disease. However it was possible to identify individual patients whose cells exhibited P-gp expression and function teamed with in vitro resistance to, and modification of, the anthracyclines. Furthermore, it is possible to identify which modulator had the greatest effect. The fact that we obtained higher indications of resistance reversal using the MTT assay along with finding P-gp expression and function in patients sensitive to the anthracyclines, suggests studies of P-gp should be teemed with chemosensitivity testing to identify specific patients who will benefit.

Evidence for the involvement of the glutathione pathway in drug resistance in AML.

We have studied altered drug detoxification through the glutathione pathway as a possible mechanism of resistance in 38 patients with AML. GST alpha, mu and pi expressions were determined using immunocytochemistry, the median percentages of positive cells being 73% (range 0-98), 55% (range 0-99) and 97% (range 80-100) respectively. MRP expression was measured using MRPm6 MoAb and flow cytometry. Results were expressed as the ratio of fluorescence associated with MRP over that of an isotype matched control (median, 1.32; range 0.95-2.15). Statistical analyses showed a significant increase in GST alpha expression in blast cells showing in vitro resistance to doxorubicin, with a median value of 78% positive cells compared to 41% in the sensitive group (p < 0.02). There was a significant reduction, however, in GST mu expression from a median value of 60% in newly presenting patients to 40% in a group of patients who had received previous cytotoxic therapy (p < 0.02). Interestingly, patients with high GST mu expression appeared to co-express MRP (p < 0.05). In vitro drug modulation studies, comparing the cytotoxic effect of doxorubicin +/- ethacrynic acid at 6.5 microM resulted in only one significant increase in sensitivity (2.6-fold), out of 22 comparisons. These results support the theory that altered detoxification through the glutathione pathway contributes towards drug resistance in AML. Further studies using fresh blast cells are required to elucidate the importance of this mechanism for individual patients.

Comparison of BCL-2 and BAX protein expression with in vitro sensitivity to ARA-C and 6TG in AML.

Activity of BCL-2 protein may be antagonised by BAX protein expression, thereby affecting cellular sensitivity to chemotherapeutic drugs. We analysed the BCL-2 protein expression of blast cells from 19 patients by flow cytometry and immunocytochemistry. This was compared to in vitro sensitivity to the anthracyclines and antimetabolites using the MTT assay. We found a significant correlation between BCL-2 expression and in vitro response to two antimetabolite drugs. One of 7 patients (14%) whose cells were sensitive to ara-C expressed BCL-2 compared to 4/4 patients (100%) whose cells were resistant to ara-C in vitro (p = 0.05). Furthermore, none of the three patients whose cells were sensitive to 6-TG expressed BCL-2 compared to 6/9 patients (67%) whose cells were resistant in vitro (p = 0.045). We found no other correlation between BCL-2 expression and any other chemotherapeutic drug analysed. The ratio of BCL-2 to BAX may be more relevant clinically, therefore cells from a further 9 patients were analysed for both proteins. Whilst there was no overall relationship between BCL-2/BAX ratios and sensitivity to ara-C and 6TG, individual patients could be identified whose blast cells were resistant to ara-C and had high BCL-2/BAX ratios. Further analysis of the significance of these ratios to drug resistance may be of future prognostic value.