IDegLira fixed-ratio combination in the real world: a retrospective observational single-center Italian experience.

OBJECTIVE: An improvement of type 2 diabetes treatment is represented by the recent availability of a fixed-ratio combination of slow insulin degludec and GLP-1 RA liraglutide (IDegLira), which shows encouraging clinical trial results. This work represents a real-world evidence study to evaluate if the obtained clinical results are also confirmed in the clinical practice, in an Italian type 2 diabetes patients' population. PATIENTS AND METHODS: This retrospective observational study was conducted in the Diabetology Service of the Umbria local sanitary agency (USL Umbria 1) in Perugia. The study investigated all diabetic patients >18 years, who underwent anti-diabetic treatment with basal insulin with or without the concomitant consumption of one or more oral anti-diabetic agent (BOT group) or GLP-1 RA or rapid-acting insulin bolus (BB group), with unsatisfactory glycemic control for either hypoglycemic episodes or weight gain. The observation period was February 2018 to April 2019. RESULTS: IDegLira results to be effective in reducing HbA1c and fasting plasma glucose, especially among GLP-1 RA and BOT subgroup. In BOT group, a statistical difference was noted from the first month of treatment, also for post-prandial glycemia. Obtained results were achieved at a moderate dose of IDegLira reported during the study, which also represents a significant reduction of the amount of basal insulin in BB patients. CONCLUSIONS: Obtained results suggest that in a real-world setting, the switch to IDegLira treatment is a valid option for patients with unsatisfactory glycemic control, or who experienced side effects such as weight gain and hypoglycemia of other insulin therapies.

Thymidine phosphorylase expression and benefit from capecitabine in patients with advanced breast cancer.

BACKGROUND AND AIM: Capecitabine is an orally bioavailable prodrug that is converted to 5-fluorouracil through several enzymatic steps, the last of which is mediated by thymidine phosphorylase (TP). TP has been reported to be expressed at higher levels in cancer tissue compared with normal counterpart. The present study aimed at evaluating the potential relationship between TP expression and benefit from capecitabine in patients with metastatic breast cancer (BC). METHODS: Immunohistochemistry for TP and other biological markers was carried out on paraffin-embedded cancer tissues of 61 patients with BC treated with at least three cycles of capecitabine as single agent for metastatic disease. All patients had received capecitabine 1000 mg/m(2) b.i.d. days 1-14 every 21 days. The following variables were analyzed as potential determinants of benefit from capecitabine: TP expression, estrogen receptor (ER) and progesterone receptor status, human epidermal growth factor receptor-2 (HER-2) status, MIB-1 expression, performance status at the beginning of capecitabine treatment, stage at diagnosis, grade, presence of visceral metastases at the beginning of capecitabine treatment, and previous chemotherapy. RESULTS: Overall, median time to progression (TTP) was 6.5 months (range 1.4-33). On multivariate analysis, ER status [hazard ratio (HR) for progression = 0.31; 95% confidence interval (CI) = 0.15-0.64; P = 0.002], presence of visceral metastases at the beginning of capecitabine treatment (HR = 2.30; 95% CI = 1.21-4.39; P = 0.01), and capecitabine as first- or second-line treatment (HR = 2.28; 95% CI = 1.21-4.32; P = 0.01) independently predicted TTP. TP was highly expressed in 34 of 61 cases (55.7%). In the subgroup of patients with TP-expressing tumor, TTP was significantly longer in patients who received anthracyclines and taxanes before capecitabine (median TTP 7.5 versus 3.3 months, P = 0.01, log-rank test). Similarly, patients with a TP-positive tumor showed a longer TTP if they received taxanes before capecitabine than patients with TP-positive tumor who did not receive this treatment (7.3 versus 3.4 months, P = 0.03). CONCLUSIONS: These data provide further evidence that TP expression in BC could represent a biomarker of sensitivity to capecitabine treatment. Prospective studies with translational approach are desirable to confirm the predictive and prognostic role of TP.

Thymidine phosphorylase expression is associated with time to progression in patients receiving low-dose, docetaxel-modulated capecitabine for metastatic breast cancer.

BACKGROUND: Preclinical data have indicated a synergistic interaction between docetaxel and capecitabine by means of taxane-induced up-regulation of thymidine phosphorylase (TP). On the basis of such premises, we conducted a phase II trial to determine the activity and tolerability of weekly docetaxel plus capecitabine in patients with metastatic breast cancer (MBC). Furthermore, we explored the relationship between TP tumor expression and benefit from this regimen. PATIENTS AND METHODS: Patients received docetaxel 36 mg/m(2) i.v. on days 1, 8, and 15 and capecitabine orally 625 mg/m(2) b.i.d. from days 8 to 21. Cycles were repeated every 4 weeks. In the correlative study, we evaluated the TP expression by immunohistochemistry and the TP messenger RNA expression by real-time RT-PCR in the primary tumor. RESULTS: Forty-seven women were enrolled. In the intention-to-treat analysis, objective responses were achieved in 24 patients (51%). Fourteen additional patients (30%) had stable disease. The median time to progression (TTP) was 6 months (range 1-44 months). Median survival was 17 months (range 1-48 months). Overall, the treatment was well tolerated. The most common clinical adverse events (all grades) were alopecia (55%), nail changes (53%), fatigue/asthenia (51%), nausea/vomiting (51%), neutropenia (49%), and neuropathy (49%). A significantly higher TTP was observed in patients with TP-positive tumors (log-rank test, P = 0.009). Interestingly, a subgroup analysis confirmed this TTP benefit in patients with TP-positive tumors obtaining a tumor response (log-rank test, P = 0.03), whereas the statistical significance was lost in nonresponders (log-rank test, P = 0.3). CONCLUSIONS: This study indicates that a regimen with low doses of capecitabine plus weekly docetaxel is active against MBC. The correlative analysis provides preliminary evidence that TP expression may be a predictive marker for therapeutic benefit.

Myocyte death in the failing human heart is gender dependent.

Cardiovascular disease is delayed and less common in women than in men. Myocyte death occurs in heart failure, but only apoptosis has been documented; the role of myocyte necrosis is unknown. Therefore, we tested whether necrosis is as important as apoptosis and whether myocyte death is lower in women than in men with heart failure. Molecular probes were used to measure the magnitude of myocyte necrosis and apoptosis in 7 women and 12 men undergoing transplantation for cardiac failure. Myocyte necrosis was evaluated by detection of DNA damage with blunt end fragments, whereas apoptosis was assessed by the identification of double-strand DNA cleavage with single base or longer 3' overhangs. An identical analysis of these forms of cell death was performed in control myocardium. Heart failure showed levels of myocyte necrosis 7-fold greater than apoptosis in patients of both sexes. However, cell death was 2-fold higher in men than in women. Heart failure resulted in a 13-fold and 27-fold increase in necrosis in women and men, respectively. Apoptosis increased 35-fold in women and 85-fold in men. The differences in cell death between women and men were confirmed by the electrophoretic pattern of DNA diffusion and laddering of isolated myocytes. The lower degree of cell death in women was associated with a longer duration of the myopathy, a later onset of cardiac decompensation, and a longer interval between heart failure and transplantation. In conclusion, myocyte necrosis and apoptosis affect the decompensated human heart; each contributes to the evolution of cardiac failure. However, the female heart is protected, at least in part, from necrotic and apoptotic death signals.

Expression of thyroid-specific transcription factors TTF-1 and PAX-8 in human thyroid neoplasms.

TTF-1 and PAX-8 are tissue-specific transcription factors expressed in the thyroid follicular cells, contributing to the maintenance of the differentiated phenotype. In fact, it has been demonstrated that TTF-1 and PAX-8 are able to activate transcription from thyroglobulin and thyroperoxidase (TPO) promoters, the transcriptional activity of which is in vivo restricted only to the thyroid follicular cell. In order to gain insight into how these transcription factors control in vivo the differentiation of the thyroid cell and to have a better molecular characterization of human thyroid tumors, TTF-1, PAX-8, thyroglobulin, and TPO mRNA levels were measured in nonmalignant and malignant human thyroid tissues. Results indicate that the expression of TTF-1 and PAX-8 is not sufficient per se for the expression of the thyroid-differentiated phenotype. Furthermore, in follicular adenomas, PAX-8 mRNA levels are strictly related to TPO mRNA levels, suggesting that the amount of PAX-8 could play a role in the modulation of TPO gene expression. TTF-1 mRNA is always well detectable in papillary carcinomas and, in contrast, always absent in anaplastic carcinomas. Identical results were obtained when the expression of TTF-1 protein was investigated using immunohistochemistry. Thus, TTF-1 gene expression could be a molecular marker in order to distinguish these two types of thyroid neoplasms.

A polymorphism in endostatin, an angiogenesis inhibitor, predisposes for the development of prostatic adenocarcinoma.

We have performed association studies between a novel coding single nucleotide polymorphism (D104N) in endostatin, one of the most potent inhibitors of angiogenesis, and prostate cancer. We observed that heterozygous N104 individuals have a 2.5 times increased chance of developing prostate cancer as compared with homozygous D104 subjects (odds ratio, 2.4; 95% confidence interval, 1.4-4.16). Modeling of the endostatin mutant showed that the N104 protein is stable. These results together with the observation that residue 104 is evolutionary conserved lead us to propose that: (a) the DNA segment containing this residue might contain a novel interaction site to a yet unknown receptor; and (b) the presence of N104 impairs the function of endostatin.

Nuclear alterations of cells and atypical metaplastic cells in cervical smears are predictive criteria of high-grade cervical intraepithelial neoplasia.

Diagnoses based on the screening of cervical smears show low interobserver reproducibility and are frequently discordant with the final histological diagnosis. The aim of this study was to identify which of the cytomorphologic criteria used in the screening of cervical smears were most predictive of the histopathological grades of cervical intraepithelial neoplasia. The abnormal cervical smears of 206 women were reviewed blindly according to 22 pre-established cytomorphological criteria. Colposcopic evaluation was carried out in all cases. The marked presence of several nuclear criteria frequently found together in the same smear was associated with high grade intraepithelial neoplasia regardless of the presence of any other criteria. On the other hand, when the nuclear criteria were less evident, the cluster of criteria related to metaplastic cells was predictive of a diagnosis of high-grade intraepithelial neoplasia. Focusing on selected cytological criteria can aggregate predictive value to cervical smear diagnoses.

TTF-1 gene expression in human lung tumours.

Tissue-specific transcription factors control cell determination and differentiation. TTF-1 is a tissue-specific transcription factor expressed in the thyroid and lung. We investigated the expression of TTF-1 in normal human lung, and in various histopathological types of lung cancers by immunohistochemistry. In normal lung, TTF-1 expression was restricted to bronchial and alveolar epithelial cells. TTF-1 expression was found in 7 of the 29 cases of non-small cell lung carcinomas. In these tumours, the expression of TTF-1 did not correlate with the histological degree of differentiation. Results obtained using RNase protection assay confirmed that TTF-1 was expressed only in a subset of non-small cell carcinomas. TTF-1, as expected, was not expressed in neoplasms having a neuroendocrine cell origin, such as carcinoids. Interestingly, TTF-1 was always expressed in small cell lung carcinomas. These findings indicate that: (i) small cell lung carcinomas could originate from the endothermal cell lineage and (ii) dedifferentiation processes that operate in these neoplasms do not affect molecular mechanisms necessary for TTF-1 gene expression.

Large core biopsy for diagnostic and prognostic evaluation of invasive breast carcinomas.

Large core biopsy is a recently introduced method for pre-operative evaluation of breast lumps. The aim of this study was to evaluate the usefulness of this technique in providing pre-operative diagnostic and prognostic information that can lead to a correct line of treatment. We compared 41 cases of breast carcinomas diagnosed both by core biopsies and surgically removed samples. A high (93%) diagnostic agreement was obtained. Moreover, we found a significant correlation for mitotic count (r = 0.76), oestrogen receptor (r = 0.78), progesterone receptor (r = 0.80), p53 (r = 0.86) and c-erbB-2 (r = 0.90) analysis between core biopsy and definitive surgical pathology. An agreement for histological grading evaluation between the two techniques was obtained in 32 out of 40 cases (k = 0.65) whereas in the other cases, a lower grade was assigned by evaluating core biopsies. These findings suggest that percutaneous core breast biopsy is a valid tool for pre-operative management of breast lesions, but this should be confirmed in larger, prospective studies.

TSH controls Ref-1 nuclear translocation in thyroid cells.

Ref-1 (called also APE) is a bifunctional protein playing a role in a large variety of cell functions. It is a major member of the DNA base excision repair system. Moreover, through reduction of cysteine residues, Ref-1 controls the activity of several transcription factors. It has been previously demonstrated that TSH up-regulates Ref-1 gene expression in thyroid cells. By using the rat FRTL-5 cell line, we demonstrate that TSH controls Ref-1 intracellular localization. Western blot experiments indicate that addition of TSH to the culture medium increases the Ref-1 cytoplasm-to-nucleus translocation. This phenomenon occurs at early times of TSH stimulation and is not dependent on protein neosynthesis. The Ref-1 cellular compartmentalization was also investigated in human thyroid tumors. A Ref-1 nuclear/cytoplasmic ratio difference between normal and cancerous thyroid tissues was observed. These results suggest that Ref-1 localization may have a critical role in the control of thyroid cell functions.

TTF-1 protein expression in pleural malignant mesotheliomas and adenocarcinomas of the lung.

TTF-1 is a tissue-specific transcription factor expressed in the epithelial cells of thyroid and lung. This study investigates the immunohistochemical expression of TTF-1 in pleural malignant mesotheliomas (MM) and adenocarcinomas (AC) of the lung, respectively. For this purpose, 33 biopsy specimens of pulmonary AC and 24 specimens of MM were studied. TTF-1 immunoreactivity was identified in 19 of 33 cases of AC (57.5%) and in none of the 24 cases of MM. Positivity for TTF-1 was 100% specific and 57.5% sensitive for lung AC. Alternatively, negativity for TTF-1 was 57.5% specific and 100% sensitive for MM. These results suggest that TTF-1 can be favourably added to the immunohistochemical diagnostic panel for distinction between AC of the lung involving the pleura and pleural MM.

Image-guided core breast biopsy: a suitable method for preoperative biological characterization of small (pT1) breast carcinomas.

Multiple prognostic indicators, namely histological grade and immunostaining for estrogen (ER) and progesterone receptors (PgR), MIB 1, bc1-2, and p53, were retrospectively determined on preoperative core biopsies from 75 patients with pT 1 breast carcinoma. The association of the preoperatively evaluated factors with those on the corresponding resected tumors (i.e. nodal status, histological grade, presence or absence of vascular invasion and necrosis) was assessed. In univariate analysis, histological grade on resected tumors was significantly associated with histological grade on core biopsy, p53 expression, MIB1 immunostaining. An inverse association was found between postoperative histologic grade and ER, PgR, and bc1-2. Necrosis was significantly associated with grade, p53, MIB1, and inversely with ER, PgR, and bc1-2. Nodal involvement and vascular invasion were significantly associated with MIB1. In multivariate analysis, histological grade and ER were the only independent core biopsy variables associated with postoperative histological grade and necrosis, respectively. This study showed that image-guided core biopsy is a suitable method that can be used to reveal some characteristics of the tumor biology in a preoperative stage.

Combined analysis of MIB-1 and thyroid transcription factor-1 predicts survival in non-small cell lung carcinomas.

The prognostic value of combined immunohistochemical analysis for the thyroid transcription factor-1 (TTF-1) and the proliferation marker MIB-1 was assessed in a consecutive series of non-small cell lung carcinomas (NSCLC). Tumor immunoreactivity for TTF-1 and MIB-1 was classified in three groups (-,+,++) and in two groups (-,+), respectively. Comparison across groups for TTF-1 reactivity showed significantly different survival curves (P=0.04). In particular, the best prognosis was associated with a TTF-1 negative pattern, whereas the TTF-1 '++' cases showed the worst prognosis. A trend towards better prognosis was observed for MIB-1 negative cases (P=0.09). Multivariate analysis confirmed independent prognostic significance of TTF-1 (P=0.002), MIB-1 (P=0.01) and pStage (P=0.04). Accordingly, analysing TTF-1 and MIB-1 together, a better prediction of survival was obtained (P=0.02), with the poorest prognosis for the 'TTF-1++/MIB-1+' cases.

Diagnostic accuracy of perforated compression grid approach for mammographically guided core needle biopsy of breast lesions.

The aim of this study was to evaluate the diagnostic accuracy of large core breast biopsy with the use of the perforated compression grid (PCG) in a series of 106 consecutive mammographically detected breast lesions. The PCG consists in a fenestrated paddle that replaces the usual mammographic compressor. Each hole in the grid is marked by letters and numbers in order to obtain the coordinates of the area to biopsy. By analysing the two orthogonal projections, the depth of the lesion in the breast is calculated. With the breast positioned in the PCG, a skin incision is made. After calculations about the depth of needle insertion, the tip of the needle is inserted proximally to the target area. In such a way, taking into consideration the total running of the needle (23 mm), a correct sampling is assured, and multiple needle passes are performed. Sensitivity was of 90.62 and 90.90%, specificity was of 100 and 95.45% and positive predictive value was of 100 and 90.90% by excluding and including suspicious diagnoses, respectively, for each computation. In conclusion, the study indicated that image-guided core biopsy performed by the approach of PCG is a cost-effective, simple and accurate technique for the diagnosis of breast lesions, and it could be proposed as a low-cost alternative where the stereotactic equipment is not available.

Human mammary gland and breast carcinoma contain immunoreactive inhibin/activin subunits: evidence for a secretion into cystic fluid.

OBJECTIVE: Inhibins and activins are members of the transforming growth factor beta superfamily and are known to modulate the growth and differentiation of several cell types. The present study investigated the localization of inhibin and activin subunits in human normal and pathological breast tissues. DESIGN: A cross-sectional study comparing the expression of inhibin/activin subunits alpha, betaA and betaB in surgical specimens from women undergoing reductive mammoplasty (classified, according to the phase of the menstrual cycle, as follicular, luteal, or postmenopausal), and patients submitted to lumpectomy for fibrocystic disease, benign (intraductal papilloma, adenomyoepithelioma, and hamartoma) or malignant breast neoplams (intraductal, intralobular, and invasive carcinoma). METHODS: Immunohistochemistry was used to localize inhibin alpha and activin betaA and betaB subunits in the cytoplasm of epithelial cells of mammary glands. Dimeric activin A, inhibin A and inhibin B were measured by specific two-site enzyme immunoassay in the cystic fluid collected from patients with fibrocystic disease. RESULTS: An intense staining for the alpha inhibin subunit and a mild staining for betaA and betaB subunits were present in samples obtained from normal breast tissue regardless of menstrual cycle phase, and in fibrocystic disease and benign neoplasms. Carcinoma cells stained weakly to moderately for alpha subunit and were negative for betaA and betaB subunits. Fibrocystic disease was associated with absence of betaA subunit expression in normal epithelial cells and intense staining for all subunits in the apocrine cells. Immunoreactive inhibin A, inhibin B, and activin A were also present in cystic fluid, suggesting a local secretion of these proteins. CONCLUSION: These data suggest a local expression and secretion of inhibin and activin in human normal, fibrocystic disease and neoplastic breast tissues. The low expression of these proteins may facilitate abnormal cell proliferation in breast carcinoma.

Correlation between visual clues, objective architectural features, and interobserver agreement in prostate cancer.

Three pathologists evaluated a number of designated architectural features to assign grades to 41 cases of well- to moderately differentiated adenocarcinoma, and their opinions were compared. The consensus opinion was obtained and evaluated against objective measurements of glandular architecture that were obtained by morphometric techniques. The observers agreed on gland size, gland uniformity, and the number of glands per field in only 49%, 31%, and 39% of cases, respectively. There were significant differences in the Gleason grades assigned by observers. Paired matching of individual Gleason grades showed agreement among observers in 44% (18 of 41), 56% (23 of 41), and 75% (31 of 41) of cases, respectively. This level of interobserver disagreement occurred even though cases with predominant patterns were selected carefully and those with variable patterns were excluded. A direct relationship appears to exist between increasing Gleason grade and increasing glandular variability, and there is an inverse relationship between Gleason grade, gland lumen area, and the number of glandular nuclei, as assessed by a group of pathologists.

B cell lymphoma of the thymus and salivary gland.

A case of primary low grade B cell lymphoma of the salivary gland associated with a low grade B cell lymphoma of the thymus and involvement of the skin is reported. The lesions in the salivary gland and in the thymus showed the typical features of a lymphoma arising from the mucosa associated lymphoid tissue (MALT) and comprised lymphatic follicles, centrocyte-like (CCL) cells and lymphoepithelial lesions. Immunohistochemistry and Southern blot analysis supported the hypothesis that these lesions can originate from the same cellular clone. These findings confirm the occurrence of low grade B cell MALT lymphoma in the thymus and the possibility of spread of MALT lymphoma to other mucosal sites.

Immunocytochemical expression of tissue specific transcription factor-1 in lung carcinoma.

AIMS: To investigate the immunocytochemical expression of the tissue specific transcription factor-1 (TTF-1) on cytological specimens of small cell lung carcinoma (SCLC) and to establish its value in the cytological diagnosis of lung cancer. METHODS: For each case, the diagnosis was made on cytological specimens and confirmed on subsequent bronchial biopsy specimens. TTF-1 was detected immunocytochemically using the avidinbiotin complex technique with a rabbit antiserum. Expression of TTF-1 was evaluated in 41 cases of SCLC and 17 cases of non-small cell carcinoma (NSCC). The latter were subdivided into eight cases of adenocarcinomas and nine cases of squamous cell carcinomas (SCC). RESULTS: Positive nuclear immunoreactivity to TTF-1 was identified in 38 (92.7%) of the 41 cases of SCLC, in five (62.5%) of eight cases of adenocarcinoma, and one (11%) of nine cases of SCC. A significant difference was observed between the two main groups, SCLC and NSCC. A comparison between SCLC and adenocarcinoma and SCC showed that TTF-1 expression was significantly different. TTF-1 immunoreactivity was not detected in the inflammatory cells of the same cases. CONCLUSIONS: TTF-1 is strictly associated with SCLC; it was weakly expressed in the various subtypes of NSCC. Although TTF-1 is not specific for SCLC, it can be used to highlight neoplastic cells to good effect when a large inflammatory component is present, and to differentiate SCLC from lymphoid infiltrates.

Expression of p53 and bcl-2 and response to preoperative chemotherapy and radiotherapy for locally advanced squamous cell carcinoma of the oesophagus.

AIMS: To investigate the immunohistochemical expression of p53 and bcl-2 proteins in squamous cell carcinoma (SCC) of the oesophagus and to assess whether expression of these oncoproteins can be used to stratify patients into groups with a favourable or unfavourable response to preoperative chemo/radiotherapy. METHODS: The initial diagnostic biopsy and the corresponding resected samples were obtained from 22 consecutive patients with SCC. All patients underwent preoperative chemo/radiotherapy. Tumour sections were incubated with a monoclonal antibody directed against p53 (DO-7). Twenty four non-neoplastic oesophageal biopsy specimens immunostained for p53 served as controls. Twelve randomly chosen sections from the 22 SCC samples were immunostained to test for bcl-2 protein expression. RESULTS: After chemo/radiotherapy, 12 (55%) of the 22 patients had no evidence of tumour in the resected oesophagus. Before chemoradiotherapy, however, 17 (77%) patients were p53 positive. After treatment, residual carcinoma was detected in seven (41%) of the 17 p53 positive patients. All non-responsive cases had the same p53 immunopattern as before treatment. Bcl-2 immunoexpression was detected in six (50%) of 12 patients. Residual tumour was detected in the residual oesophagus in two (33%) of the six bcl-2 positive patients. After treatment, bcl-2 expression was no longer detected in the residual neoplastic cells of a previously bcl-2 positive tumour. Using Fisher's exact test no significant association was found between oncoprotein expression and response to preoperative treatment. CONCLUSION: This study confirms the observation that p53 protein is frequently expressed in SCCs of the oesophagus, probably as a result of a mutation of the TP53 gene. However, no significant association was found between oncoprotein expression and response to chemo/radiotherapy. Anticancer agents do not seem to modify the expression of p53 and bcl-2 proteins.

BCL-2 immunohistochemical evaluation in B-cell chronic lymphocytic leukemia and hairy cell leukemia before treatment with fludarabine and 2-chloro-deoxy-adenosine.

Bcl-2 overexpression has been shown to be associated with several malignancies, including B-cell chronic lymphocytic leukemia (CLL) and non-Hodgkin's lymphomas (NHL), mainly low-grade and follicular in type. It has as yet not been described in hairy cell leukemia (HCL). In 30 patients with CLL and 14 with HCL who were consecutively selected for treatment with purine analogues (Fludarabine in CLL and 2-chloro-deoxy-adenosine in HCL), we evaluated bcl-2 oncoprotein expression in leukemic cells on marrow sections that were taken before treatment and stained immunohistochemically with a monoclonal antibody (Dakopatts 124 clone), by the avidin-biotin-peroxidase method. All samples were found to be bcl-2 positive, with a staining intensity that was moderate to strong in CLL and weak to moderate in HCL. 83% of CLL and 100% of HCL patients were responsive to purine analogues. These findings show that bcl-2 is overexpressed in almost all cases CLL and HCL and that bcl-2 overexpression does not predict a poor response to purine analogues, which are believed to induce apoptosis.