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CXCR5 mediates homing of both B and follicular helper T (TFH) cells into follicles of secondary lymphoid organs. We found that CXCR5+CD8+T cells are present in human tonsils and follicular lymphoma, inhibit TFH-mediated B cell differentiation, and exhibit strong cytotoxic activity. Consistent with these findings, adoptive transfer of CXCR5+CD8+T cells into an animal model of lymphoma resulted in significantly greater antitumor activity than CXCR5−CD8+T cells. Furthermore, RNA-Seq-based transcriptional profiling revealed 77 differentially expressed genes unique to CXCR5+CD8+T cells. Among these, a signature comprised of 33 upregulated genes correlated with improved survival in follicular lymphoma patients. We also showed that CXCR5+CD8+T cells could be induced and expanded ex vivo using IL-23 plus TGF-β, suggesting a possible strategy to generate these cells for clinical application. In summary, our study identified CXCR5+CD8+T cells as a distinct T cell subset with ability to suppress TFH-mediated B cell differentiation, exert strong antitumor activity, and confer favorable prognosis in follicular lymphoma patients.

Background: Tuberculosis (TB) has become a rising concern in low-income countries, particularly in those with Human Immuno Deficiency Virus (HIV) epidemics, and type 2 diabetes has emerged as a significant global chronic health problem, owing to increases in obesity, lifestyle changes, and ageing populations. Diabetes has been identified as a major risk factor for the development of TB. Despite the fact that diabetes imparts a substantially lower risk of TB (3-fold) as compared to HIV (>20-fold), in communities where the number of DM patients is high, the contribution of diabetes to TB might be bigger than HIV.Methods: This review will focus on the link between TB and diabetes, which is now one of the most important topics for physicians since diabetes impacts the clinical presentation and outcome of TB and vice versa.Results: Though TB is more common in type 1 diabetes, the extent of the problem in type 2 diabetes should be taken into account with equal care, as type 2 diabetes affects a substantially higher number of individuals.Conclusions: Diabetes patients are more vulnerable to infections because of their impaired immune systems. Increased glucose level leads to a rise in the infection status among TB patients and also leads to a rise in various complications. Extensive and increased screening for both TB and DM over years can help diagnose disease priorly and help in better management. TB, when diagnosed in its early stages, can be easily eradicated.

Steady-state visually evoked potential is one of the active explorations in the brain-computer interface research. Electroencephalogram based brain computer interface studies have been widely applied to perceive solutions for real-world problems in the healthcare domain. The classification of externally bestowed visual stimuli of different frequencies on a human was experimented to identify the need of paralytic people. Although many classifiers are at the fingertip of machine learning technology, recent research has proven that ensemble learning is more efficacious than individual classifiers. Despite its efficiency, ensemble learning technology exhibits certain drawbacks like taking more time on selecting the optimal classifier subset. This research article utilizes the Harris Hawk Optimization algorithm to select the best classifier subset from the given set of classifiers. The objective of the research is to develop an efficient multi-classifier model for electroencephalogram signal classification. The proposed model utilizes the Boruta Feature Selection algorithm to select the prominent features for classification. Thus selected prominent features are fed into the multi-classifier subset which has been generated by the Harris Hawk Optimization algorithm. The results of the multi-classifier ensemble model are aggregated using Stacking, Bagging, Boosting, and Voting. The proposed model is evaluated against the acquired dataset and produces a promising accuracy of 96.1%, 98.7%, 91.91%, and 99.01% with the ensemble techniques respectively. The proposed model is also validated with other performance metrics such as sensitivity, specificity, and F1-Score. The experimental results show that the proposed model proves its supremacy in segregating the multi-class classification problem with high accuracy.

Introduction

India has made large strides in reducing maternal mortality ratio and neonatal mortality rate, yet care-seeking behavior for appropriate care is still a challenge. We conducted a qualitative study to understand the process of recognition and care-seeking for maternal and newborn illnesses in rural India where a health intervention through women’s self-help groups (SHG) to improve maternal and newborn health behaviors is implemented by a non-governmental organization, the Rajiv Gandhi Mahila Vikas Pariyojana. The study aimed to understand the process of recognition and care-seeking for maternal and newborn illnesses from SHG and non-SHG households in the intervention area. Thirty-two illness narratives, 16 of maternal deaths and illness and 16 of newborn illnesses and deaths, were conducted. Women, their family members, and other caretakers who were present during the event of illness or death were included in the interviews. About 14 key informants, mainly frontline health workers (FLWs), were also interviewed. The interviews were conducted by two Population Council staff using a pre-tested guideline in Hindi. Our findings suggest that perceptions of causes of illness as “supernatural” or “medical” and the timing of onset of illness influence the pathway of care-seeking. Deep-rooted cultural beliefs and rituals guided care-seeking behavior and restricted new mothers and newborns’ mobility for care-seeking. Though families described experience of postpartum hemorrhage as severe, they often considered it as “normal.” When the onset of illness was during pregnancy, care was sought from health facilities. As the step of care for maternal illness, SHG households went to government facilities, and non-SHG households took home-based care. Home-based care was the first step of care for newborn illnesses for both SHG and non-SHG households; however, SHG households were prompt in seeking care outside of home, and non-SHG households delayed seeking care until symptoms were perceived to be severe. Our findings indicate that care-seeking behavior for maternal and newborn morbidities could be improved by interventions through social platforms such as SHGs.

Although maternal and newborn mortality have decreased 44 and 46% respectively between 1990 and 2015, achievement of ambitious Sustainable Development Goal targets requires accelerated progress. Mortality reduction requires a renewed focus on the continuum of maternal and newborn care from the household to the health facility. Although barriers to accessing skilled care are documented for specific contexts, there is a lack of systematic evidence on how women and families identify maternal and newborn illness and make decisions and subsequent care-seeking patterns. The focus of this multi-country study was to identify and describe illness recognition, decision-making, and care-seeking patterns across various contexts among women and newborns who survived and died to ultimately inform programmatic priorities moving forward. This study was conducted in seven countries—Ethiopia, Tanzania, Uganda, Nigeria, India, Indonesia, and Nepal. Mixed-methods were utilized including event narratives (group interviews), in-depth interviews (IDIs), focus group discussions (FDGs), rapid facility assessments, and secondary analyses of existing program data. A common protocol and tools were developed in collaboration with study teams and adapted for each site, as needed. Sample size was a minimum of five cases of each type (e.g., perceived postpartum hemorrhage, maternal death, newborn illness, and newborn death) for each study site, with a total of 84 perceived PPH, 45 maternal deaths, 83 newborn illness, 55 newborn deaths, 64 IDIs/FGDs, and 99 health facility assessments across all sites. Analysis included coding within and across cases, identifying broad themes on recognition of illness, decision-making, and patterns of care seeking, and corresponding contextual factors. Technical support was provided throughout the process for capacity building, quality assurance, and consistency across sites. This study provides rigorous evidence on how women and families recognize and respond to maternal and newborn illness. By using a common methodology and tools, findings not only were site-specific but also allow for comparison across contexts.

Objective: Peripheral artery disease (PAD) is associated with increased risk of non-healing ulcers, amputation, and mortality due to occlusive atherosclerotic plaques. Computed tomography (CT) imaging detects vascular calcification in PAD; however, quantitative vessel-by-vessel analysis of calcium burden in the feet of PAD patients has not been assessed. This study sought to perform quantitative analysis of vessel-specific calcium burden and examine the patient-level determinants of foot calcium burden in PAD patients. Approach: PAD patients (n=41) were prospectively enrolled and underwent CT imaging of the lower extremities. Manual segmentation of the medial plantar, lateral plantar, and dorsalis pedis arteries was performed. CT image Hounsfield units (HUs) were obtained for each artery to quantify vessel-by-vessel calcium mass using a cutoff value of ≥130 HU. Univariate analyses were performed to evaluate patient-level determinants of calcium burden for each foot artery. STROBE guidelines were used for reporting of data. Results: Univariate analyses revealed that body mass index, diabetes mellitus (DM), and chronic kidney disease (CKD) were significant determinants of foot calcium burden in PAD patients. Image analysis demonstrated that PAD patients with DM had significantly higher calcium mass for the medial plantar (p=0.005), lateral plantar (p=0.039), and dorsalis pedis (p=0.001) arteries compared to PAD patients without DM. Innovation: This is the first study to use CT imaging to quantify vessel-specific calcium burden in the feet of patients with PAD and evaluate the patient-level determinants of foot calcium burden in the setting of PAD. Conclusion: CT imaging quantifies vessel-specific calcification in the feet of PAD patients, which is exacerbated with concomitant DM, CKD, and/or obesity.

Introduction:Cancer treatment with chemotherapeutics can lead to off-target effects on the vasculature. Fluorine-18 (18F)-fluorodeoxyglucose (FDG) PET/CT imaging is standard of care for staging and monitoring of numerous cancers and may offer a tool for assessing vascular toxicity. We hypothesized that PET/CT imaging could quantify changes in chemotherapy-induced arterial inflammation. Thus, we sought to validate PET/CT imaging as a tool for assessing vascular toxicity in a rat model using in vivo imaging and post-mortem analyses.Methods:Male Wistar rats (n = 11 total) were divided into a control group that received twice weekly IV saline (n=5; 1 ml/kg) or a group that received twice weekly IV doxorubicin (n=6; 1 mg/kg) for 2.5 weeks (i.e., 7 mg/kg cumulative dose). Rats underwent 18F-FDG microPET/CT imaging of the abdominal aorta (AA) at 3 time points (baseline, after 4 mg/kg dosing, and after 7 mg/kg cumulative dose) to quantify serial changes in arterial inflammation, expressed as changes in the maximum standard uptake value (SUVmax). Following terminal imaging, AA tissue was harvested and cut into 3 mm sections for gamma counting of 18F-FDG uptake, hematoxylin and eosin morphological staining, and immunohistochemistry of macrophage expression.Results:In vivo image analyses demonstrated that 18F-FDG uptake remained unchanged for the control group during the treatment period, but significantly increased from baseline levels in the doxorubicin-treated group after 4 mg/kg (p=0.0007) and 7 mg/kg (p=0.001) cumulative dosing. In vivo image analyses were confirmed by post-mortem gamma counting of 18F-FDG uptake in anatomically-matched AA tissue samples. Histological analyses revealed increased levels of macrophage expression in sections with increased 18F-FDG uptake identified by in vivo and ex vivo analyses.Conclusions:18F-FDG PET/CT imaging non-invasively quantifies serial changes in chemotherapy-induced arterial toxicity that are at least partially mediated by increased macrophage activity. Routine assessment of 18F-FDG uptake on PET/CT imaging studies during cancer treatment could provide a translatable approach for characterizing the time course of vascular toxicity associated with various chemotherapeutics.

Intrinsic and extrinsic apoptotic pathways converge to activate common downstream executioner caspases (caspase-3, -6, and -7), resulting in cell death. In chronic lymphocytic leukemia (CLL), neoplastic B cells evade apoptosis owing to the overexpression of survival proteins. We hypothesized that direct activation of procaspases could bypass the apoptosis resistance induced by the upstream prosurvival proteins. The procaspase-activating compounds (PAC-1), including B-PAC-1 (L14R8), convert inactive executioner procaspases to their active cleaved forms by chelation of labile zinc ions. Both at transcript and protein levels, primary CLL cells express high levels of latent procaspases (3, -7, and -9). B-PAC-1 treatment induced CLL lymphocyte death which was higher than that in normal peripheral blood mononuclear cells or B cells, and was independent of prognostic markers and microenvironmental factors. Mechanistically, B-PAC-1 treatment activated executioner procaspases and not other Zn-dependent enzymes. Exogenous zinc completely, and pancaspase inhibitors partially, reversed B-PAC-1–induced apoptosis, elucidating the zinc-mediated mechanism of action. The cell demise relied on the presence of caspase-3/7 but not caspase-8 or Bax/Bak proteins. B-PAC-1 in combination with an inhibitor of apoptosis protein antagonist (Smac066) synergistically induced apoptosis in CLL samples. Our investigations demonstrated that direct activation of executioner procaspases via B-PAC-1 treatment bypasses apoptosis resistance and is a novel approach for CLL therapeutics.

Nano zinc ferrite powder was prepared by eco-friendly hydrothermal method. The prepared compound was well characterized by powder X-ray diffraction, scanning electron microscopy and UV-Vis spectophotometry. The compound formation was confirmed without any additional impurities. The PXRD data shows the crystallite size of the compound was about 6 nm. The SEM image showed the agglomeration of spherical nanoparticles. The UV-Vis spectrum and energy gap calculation confirmed the semiconducting behaviour of the sample. The magnetic properties of the compound revealed superparamagnetic nature at room temperature. These magnetic properties of the spherical nanoparticles are widely required in high performance devices like MRI contrast agent, cell separation and other biomedical applications.

Introduction: Although protein kinases are primary targets for inhibition in hematological malignancies, until recently their contribution to chronic lymphocytic leukemia (CLL) was poorly understood. Insights into B-cell receptor signaling and its role in regulating key cellular functions have shed light on candidate protein kinases that are aberrantly activated in CLL. In this regard, protein kinases are now considered as potential drug targets in CLL.Area covered: This review has covered signaling pathways and associated protein kinases in CLL and the kinase inhibitors currently available in preclinical and clinical investigations. Individual protein kinases that are abnormally active in CLL and the functional consequences of their inhibition are discussed.Expert opinion: A growing body of evidence suggests that protein kinases are druggable targets for patients with CLL. The emergence of novel and bio-available kinase inhibitors and their promising clinical activity in CLL underscore the oncogenic role of kinases in leukemogenesis. Further investigations directed towards their role as single agents or in combinations may provide insight into understanding the substantial role of kinase-mediated signal transduction pathways and their inhibition in B- CLL.

B-cell receptor (BCR) signaling is a critical pathway in the pathogenesis of several B-cell malignancies, including chronic lymphocytic leukemia (CLL), and can be targeted by inhibitors of BCR-associated kinases, such as Bruton tyrosine kinase (Btk). PCI-32765, a selective, irreversible Btk inhibitor, is a novel, molecularly targeted agent for patients with B-cell malignancies, and is particularly active in patients with CLL. In this study, we analyzed the mechanism of action of PCI-32765 in CLL, using in vitro and in vivo models, and performed correlative studies on specimens from patients receiving therapy with PCI-32765. PCI-32765 significantly inhibited CLL cell survival, DNA synthesis, and migration in response to tissue homing chemokines (CXCL12, CXCL13). PCI-32765 also down-regulated secretion of BCR-dependent chemokines (CCL3, CCL4) by the CLL cells, both in vitro and in vivo. In an adoptive transfer TCL1 mouse model of CLL, PCI-32765 affected disease progression. In this model, PCI-32765 caused a transient early lymphocytosis, and profoundly inhibited CLL progression, as assessed by weight, development, and extent of hepatospenomegaly, and survival. Our data demonstrate that PCI-32765 effectively inhibits CLL cell migration and survival, possibly explaining some of the characteristic clinical activity of this new targeted agent.

Forodesine, a purine nucleoside phosphorylase inhibitor, displays in vitro activity in chronic lymphocytic leukemia (CLL) cells in presence of dGuo, which is the basis for an ongoing clinical trial in patients with fludarabine-refractory CLL. Initial clinical data indicate forodesine has significant activity on circulating CLL cells, but less activity in clearing CLL cells from tissues such as marrow. In tissue microenvironments, lymphocytes interact with accessory stromal cells that provide survival and drug-resistance signals, which may account for residual disease. Therefore, we investigated the impact of marrow stromal cells (MSCs) on forodesine-induced response in CLL lymphocytes. We demonstrate that spontaneous and forodesine-induced apoptosis of CLL cells was significantly inhibited by human and murine MSCs. Forodesine-promoted dGuo triphosphate (dGTP) accumulation and GTP and ATP depletion in CLL cells was inhibited by MSCs, providing a mechanism for resistance. Also, MSCs rescued CLL cells from forodesine-induced RNA- and protein-synthesis inhibition and stabilized and increased Mcl-1 transcript and protein levels. Conversely, MSC viability was not affected by forodesine and dGuo. Collectively, MSC-induced biochemical changes antagonized forodesine-induced CLL cell apoptosis. This provides a biochemical mechanism for MSC-derived resistance to forodesine and emphasizes the need to move toward combinations with agents that interfere with the microenvironment's protective role for improving current therapeutic efforts.

Forodesine is a new and potent purine nucleoside phosphorylase (PNP) inhibitor. Patients with chronic lymphocytic leukemia (CLL) with primary resistance to fludarabine-based therapy or with progressive disease were eligible for oral forodesine (200 mg/d) for up to 24 weeks. Eight patients with median lymphocyte count of 35.9 × 109/L and median serum β2 microglobulin level of 6.45 mg/L were treated. Six had Rai stage III to IV and were previously heavily treated (median prior therapy = 5). Two had transient decrease in lymphocyte count to normal, whereas in 5, disease progressed. Adverse events were mild. Steady-state level of forodesine ranged from 200 to 1300nM and did not reach desired 2µM level. PNP inhibition ranged from 57% to 89% and steady-state 2′-deoxyguanosine (dGuo) concentration median was 1.8µM. Intracellular deoxyguanosine triphosphate (dGTP) increase was very modest, from median of 6µM to 10µM. Compared with in vivo, in vitro incubations of CLL lymphocytes with 10 or 20µM dGuo and forodesine (2µM) resulted in accumulation of higher levels of dGTP (40-250µM) which resulted in increase in apoptosis. Forodesine has biologic activity in CLL; pharmacodynamic parameters suggest that an alternate dosing schedule and/or higher doses to achieve greater intracellular dGTP may be beneficial in this patient population. This study is registered at www.clinicaltrials.govas #NCT00289549.

Marrow stromal cells (MSCs) provide important survival and drug resistance signals to chronic lymphocytic leukemia (CLL) cells, but current models to analyze CLL–MSC interactions are heterogeneous. Therefore, we tested different human and murine MSC lines and primary human MSCs for their ability to protect CLL cells from spontaneous and drug-induced apoptosis. Our results show that both human and murine MSCs are equally effective in protecting CLL cells from fludarabine-induced apoptosis. This protective effect was sustained over a wide range of CLL–MSC ratios (5:1 to 100:1), and the levels of protection were reproducible in 4 different laboratories. Human and murine MSCs also protected CLL cells from dexamethasone- and cyclophosphamide-induced apoptosis. This protection required cell–cell contact and was virtually absent when CLL cells were separated from the MSCs by micropore filters. Furthermore, MSCs maintained Mcl-1 and protected CLL cells from spontaneous and fludarabine-induced Mcl-1 and PARP cleavage. Collectively, these studies define common denominators for CLL cocultures with MSCs. They also provide a reliable, validated tool for future investigations into the mechanism of MSC–CLL cross talk and for drug testing in a more relevant fashion than the commonly used suspension cultures.

Antigenic stimulation through the B-cell antigen receptor (BCR) is considered to promote the expansion of chronic lymphocytic leukemia (CLL) B cells. The spleen tyrosine kinase (Syk), a key component of BCR signaling, can be blocked by R406, a small-molecule Syk inhibitor, that displayed activity in CLL patients in a first clinical trial. In this study, we investigated the effects of BCR stimulation and R406 on CLL cell survival and migration. The prosurvival effects promoted by anti-IgM stimulation and nurselike cells were abrogated by R406. BCR triggering up-regulated adhesion molecules, and increased CLL cell migration toward the chemokines CXCL12 and CXCL13. BCR activation also enhanced CLL cell migration beneath marrow stromal cells. These responses were blocked by R406, which furthermore abrogated BCR-dependent secretion of T-cell chemokines (CCL3 and CCL4) by CLL cells. Finally, R406 inhibited constitutive and BCR-induced activation of Syk, extracellular signal-regulated kinases, and AKT, and blocked BCR-induced calcium mobilization. These findings suggest that BCR activation favors CLL cell homing, retention, and survival in tissue microenvironments. R406 effectively blocks these BCR-dependent responses in CLL cells, providing an explanation for the activity of R406 in patients with CLL.

Resistance to apoptosis in CLL B cells is associated with overexpression of Bcl-2 family antiapoptotic proteins. Their expression is endogenous, but is also induced by signals from the microenvironment resulting in intrinsic and extrinsic drug resistance. Because AT-101 binds to the BH3 motif of all Bcl-2–family antiapoptotic proteins, we hypothesized that this molecule could overcome resistance. AT-101 treatment (20 μM for 24 hours) resulted in a median 72% apoptosis in CLL cells (patients; n = 32, P < .001). Stromal cells protected CLL B cells from spontaneous and fludarabine-induced apoptosis (P = .003) by increasing the Mcl-1 protein levels. However, AT-101 induced similar extent of down-regulation of Mcl-1 and apoptosis in CLL lymphocytes cultured in suspension or on stroma (P = .999). Stromal cells expressed undetectable levels of antiapoptotic but high levels of activated ERK and AKT proteins and had low or no apoptosis with AT-101. Collectively, these data demonstrate that AT-101 induces apoptosis in CLL B cells and overcomes microenvironment-mediated resistance while sparing normal stromal cells.

Gossypol, a cottonseed extract derivative, acts as a BH3-mimetic, binding to the BH3 pocket of antiapoptotic proteins and displacing pro-death partners to induce apoptosis. However, knowledge on the molecular underpinnings of its downstream effects is limited. Since chronic lymphocytic leukemia (CLL) cells express high levels of antiapoptotic proteins that act as a survival mechanism for these replicationally quiescent lymphocytes, we investigated whether gossypol induces apoptosis in these cells and what mechanism underlies gossypol-mediated cytotoxicity. Gossypol induced cell death in a concentration- and time-dependent manner; 24-hour incubation with 30 μM gossypol resulted in 50% cell death (median; range, 10%-80%; n = 47) that was not abrogated by pan-specific caspase inhibitor. Starting at 4 hours, the mitochondrial outer membrane was significantly permeabilized (median, 77%; range, 54%-93%; n = 15). Mitochondrial outer membrane permeabiliztaion (MOMP) was concurrent with increased production of reactive oxygen species (ROS); however, antioxidants did not abrogate gossypol-induced cell death. Mitochondrial membrane permeabilization was also associated with loss of intracellular adenosine triphosphate (ATP), activation of BAX, and release of cytochrome c and apoptosis-inducing factor (AIF), which was translocated to the nucleus. Blocking AIF translocation resulted in a decreased apoptosis, suggesting that AIF contributes to gossypol-mediated cytotoxicity in CLL lymphocytes.

Purine nucleoside phosphorylase (PNP) was recognized more than 30 years ago as a potential target for the treatment of patients with T-cell malignancies when an inherited deficiency of PNP was reported to be associated with a profound T-cell lymphopenia. The biochemical basis for this T-cell deficiency was subsequently shown to be related to the accumulation of plasma 2'-deoxyguanosine (dGuo) and intracellular dGuo triphosphate (dGTP). These observations have led to a search for PNP inhibitors that would be useful clinically in the management of T cell-derived malignancies. The most potent inhibitor of PNP described to date is forodesine, a rationally designed, transition-state analogue inhibitor. The preclinical and clinical pharmacology of forodesine showed its effectiveness in inhibiting PNP and augmenting dGuo levels in plasma. Increased dGTP concentrations in leukemia cells of different lineages provides strong support for the potential use of this agent in the treatment of patients with hematologic malignancies of both T- and B-cell origin.

Clofarabine has shown impressive response rates in patients with acute leukemias. In vitro investigations with clofarabine in combination with cyclophosphamide in primary cells have demonstrated synergistic cytotoxicity and inhibition of DNA repair. Based on these clinical and laboratory observations, we designed a mechanism-based combination protocol with clofarabine and cyclophosphamide for patients with relapsed acute leukemias. Eighteen patients were treated with cyclophosphamide (200 mg/m2) alone on day 0 and with clofarabine plus cyclophosphamide on day 1. Clinical responses, toxicity, DNA damage measured as H2AX phosphorylation, and accumulation of clofarabine triphosphate (TP) were analyzed. At dose level 1 (20 mg/m2 clofarabine + cyclophosphamide, 6 patients) and dose level 0 (10 mg/m2 clofarabine + cyclophosphamide, 12 patients) overall response rates were 50% and 30%, respectively, with responses in 4 (67%) of 6 patients with refractory acute lymphoblastic leukemia. Dose-limiting toxicity occurred at dose level 1 with prolonged marrow aplasia. Four (22%) patients died from prolonged aplasia (1), fungal pneumonia (1), or multiorgan failure (2). In 12 of 13 patient samples, increased DNA damage (γH2AX) was observed with clofarabine and cyclophosphamide compared with cyclophosphamide alone. In conclusion, pharmacodynamic end points along with clinical results suggest usefulness of this combination strategy, whereas toxicity data suggest reduction in chemotherapeutic intensity. This clinical trial is registered with the National Cancer Institute's PDQ at www.clinicaltrials.gov as no. JHOC-J0561.

Purine nucleoside phosphorylase (PNP) deficiency in humans results in T lymphocytopenia. Forodesine, a potent inhibitor of PNP, was designed based on the transition-state structure stabilized by the enzyme. Previous studies established that forodesine in the presence of deoxyguanosine (dGuo) inhibits the proliferation of T lymphocytes. A phase 1 clinical trial of forodesine in T-cell malignancies demonstrated significant antileukemic activity with an increase in intracellular dGuo triphosphate (dGTP). High accumulation of dGTP in T cells may be dependent on the levels of deoxynucleoside kinases. Because B-cell chronic lymphocytic leukemia (B-CLL) cells have high activity of deoxycytidine kinase (dCK), we hypothesized that these lymphocytes would respond to forodesine. This postulate was tested in primary lymphocytes during in vitro investigations. Lymphocytes from 12 patients with CLL were incubated with forodesine and dGuo. These CLL cells showed a wide variation in the accumulation of intracellular dGTP without any effect on other deoxynucleotides. This was associated with DNA damage-induced p53 stabilization, phosphorylation of p53 at Ser15, and activation of p21. The dGTP accumulation was related to induction of apoptosis measured by caspase activation, changes in mitochondrial membrane potential, and PARP cleavage. Based on these data, a phase 2 clinical trial of forodesine has been initiated for CLL patients.

Dilated cardiomyopathy is a disease of the heart muscle, characterized by ventricular dilation and depressed myocardial contractility in the absence of abnormal loading conditions or ischemic heart disease. Dilated cardiomyopathy may be viewed as a progressive disorder that is initiated after an ‘index event’ damages the heart muscle, thereby preventing the heart from contracting normally. This review will discuss the mechanisms of dilated cardiomyopathy, with special focus on familial dilated cardiomyopathy and virus-induced dilated cardiomyopathy.

Myeloid cell leukemia-1 (MCL-1) acts as a key survival factor for chronic lymphocytic leukemia (CLL) cells. In addition, dissipation of cellular bioenergy may impose a lethal effect on these quiescent cells. Previously, in multiple myeloma cell lines we demonstrated that halogenated adenosine (8-Cl-Ado) was phosphorylated to triphosphate (8-Cl–adenosine triphosphate [ATP]), which preferentially incorporated into mRNA and inhibited RNA synthesis by premature transcription termination. Furthermore, 8-Cl-ATP accumulation was associated with a decline in cellular bioenergy. Based on these actions, we hypothesized that 8-Cl-Ado would be ideal to target CLL lymphocytes. In the present study we demonstrate that leukemic lymphocytes incubated with 8-Cl-Ado display time- and dose-dependent increase in the accumulation of 8-Cl-ATP, with a parallel depletion of the endogenous ATP pool. Inhibition of global RNA synthesis resulted in a significant decline in the expression of transcripts with a short half-life such as MCL1. Consistent to this, protein expression of MCL-1 but not B-cell lymphoma–2 (BCL-2) was decreased. Furthermore, 8-Cl-ATP induced programmed cell death, as suggested by caspases activation, cleavage of caspase 3, and PARP (poly–adenosine diphosphate [ADP]–ribose polymerase), and increased DNA fragmentation. In conclusion, 8-Cl-Ado induces apoptosis in CLL lymphocytes by targeting cellular bioenergy as well as RNA transcription and translation of key survival genes such as MCL1.

We describe a patient with acute left ventricular wall rupture and pseudoaneurysm formation after myocardial infarction. We review the accuracy of 2-dimensional transthoracic echocardiography for detection of ventricular pseudoaneurysm and describe the use of real-time handheld 3-dimensional color Doppler echocardiography for the assessment of left ventricular rupture location, orifice geometry, and complex intracardiac flow before surgical correction.

Influence of contraceptive use on increased gap between successive births and attributed reduced risk of child deaths is well documented in developing countries. However, there is scarcity of evidence on direct contribution of contraceptive use on child survival especially in Indian context.

The B-cell receptor (BCR) and its downstream effectors have emerged as important therapeutic targets in B-cell malignancies. CC-292 is a novel, potent, covalent, and highly selective inhibitor of Btk (IC50apparentof 0.5 nM, kinact/KIratio of 7.69 × 104M-1s-1), that does not appreciably inhibit other kinases involved in BCR signaling (eg, IC50Lyn kinase, 4401 nM) (Evans et al., J Pharmacol Exp Ther. 2013). Here, we report preclinical characterization and clinical data in CLL from a single-agent phase 1 dose-escalation trial of CC-292 in B-cell malignancies, with a focus on how target engagement and downstream events correlate with clinical activity.

The functional relevance of the B cell Receptor (BCR) pathway and identification of protein kinases as therapeutic targets have recently shifted the paradigm for treatment of B cell malignancies. Inhibition of protein and lipid kinases (Bruton Tyrosine Kinase [BTK] and phosphoinositide 3-kinase [PI3K]) with ibrutinib and GS-1101 has been shown to be active in treatment of chronic lymphocytic leukemia (CLL). Importantly, differential expression and function of PI3K isoforms support isoform-selective inhibition of this kinase in CLL. Whilst PI3K-α and PI3K-β are ubiquitously expressed, PI3K-δ and PI3K-γ are primarily restricted to leukocytes. Since CLL cells generally express high levels of active PI3K-δ, great interest has been focused on inhibition of PI3K-δ. However, given the distinct and non-overlapping roles of PI3K-δ and PI3K-γ in immune cells, exploration of the therapeutic potential of combined inhibition of both PI3K-δ and PI3K-γ in CLL patients is warranted.

No relevant conflicts of interest to declare.