TCFL5 knockdown sensitizes DLBCL to doxorubicin treatment via regulation of GPX4.

BACKGROUND: Resistance to chemo-drug is a major cause of bad outcome in diffuse large B-cell lymphoma (DLBCL). It was reported that TCFL5 may be related to chemoresistance in childhood acute lymphoblastic leukemia. However, it is still unclear whether TCFL5 is involved in DLBCL drug-resistance. METHODS: To explore the underlying mechanism of doxorubicin resistance, recombinant lentivirus was applied to control expression of TCFL5 in DLBCL cells. CCK-8 assay was perfomed to investigate the influence of doxorubicin on proliferation of TCFL5-overexpressed or sh-TCFL5 DLBCL cells. Correlation between TCFL5 and GPX4 was analyzed with bioinformatic methods, which was further confirmed by qPCR and western blot. TCFL5 overexpression conferred doxorubicin resistance via regulating GPX4 and was verified by TUNEL assay and western blot in vitro and mice model in vivo. RESULTS: TCFL5 was enriched in DLBCL cells and conferred doxorubicin resistance through binding to GPX4. Inhibition of TCFL5 enhanced the sensitivity of DLBCL cells to doxorubicin. GPX4 knockdown reversed doxorubicin resistance in TCFL5-overexpressed DLBCL cells. CONCLUSION: DLBCL cells overexpress TCFL5 that promotes chemoresistance by regulating GPX4. Targeting TCFL5 may provide a prospective therapeutic strategy for doxorubicin-resistant DLBCL.

Advanced Strategies of Drug Delivery via Oral, Topical, and Parenteral Administration Routes: Where Do Equine Medications Stand?

While the global market for veterinary products has been expanding rapidly, there is still a lack of specialist knowledge of equine pharmaceutics. In many cases, the basic structure of the gastrointestinal tract (GIT) and integumentary system of the horse shares similarities with those of humans. Generally, the dosage form developed for humans can be repurposed to deliver equine medications; however, due to physiological variation, the therapeutic outcomes can be unpredictable. This is an area that requires more research, as there is a clear deficiency in literature precedence on drug delivery specifically for horses. Through a careful evaluation of equine anatomy and physiology, novel drug delivery systems (NDDSs) can be developed to adequately address many of the medical ailments of the horse. In addition to this, there are key considerations when delivering oral, topical, and parenteral drugs to horses, deriving from age and species variation. More importantly, NDDSs can enhance the duration of action of active drugs in animals, significantly improving owner compliance; and ultimately, enhancing the convenience of product administration. To address the knowledge gap in equine pharmaceutical formulations, this paper begins with a summary of the anatomy and physiology of the equine gastrointestinal, integumentary, and circulatory systems. A detailed discussion of potential dosage-form related issues affecting horses, and how they can be overcome by employing NDDSs is presented.

Epidemiologic Characteristics, Prognostic Factors, and Treatment Outcomes in Primary Central Nervous System Lymphoma: A SEER-Based Study.

OBJECTIVE: This study was conducted in order to study the clinical characteristics, prognostic factors, and treatment outcomes in patients with primary central nervous system lymphoma (PCNSL). MATERIALS AND METHODS: The data of a total of 5,166 PCNSL patients diagnosed between 2000 and 2018 from the Surveillance, Epidemiology, and End Results (SEER) database were obtained. RESULTS: The mean age was 63.1 ± 14.9 years, with a male to female ratio of 1.1:1.0. The most common histologic subtype was diffuse large B-cell lymphoma (DLBCL) (84.6%). The 1-, 3-, and 5-year overall survival (OS) rates were 50.1%, 36.0%, and 27.2%, respectively, and the corresponding disease-specific survival (DSS) rates were 54.4%, 41.3%, and 33.5%, respectively. Multivariate analysis with Cox regression showed that race, sex, age, marital status, surgical resection, and chemotherapy were independent prognostic factors for OS and DSS, but radiotherapy was only for OS. Nomograms specially for DLBCL were established to predict the possibility of OS and DSS. The concordance index (C-index) values of OS and DSS were 0.704 (95% CI 0.687-0.721) and 0.698 (95% CI 0.679-0.717), suggesting the high discrimination ability of the nomograms. CONCLUSION: Surgical resection and/or chemotherapy was favorably associated with better OS and DSS. However, radiotherapy was not beneficial for OS and DSS in the long term. A new predictive nomogram and a web-based survival rate calculator we developed showed favorable applicability and accuracy to predict the long-term OS for DLBCL patients specifically.

Poststroke depression and risk of recurrent stroke: A meta-analysis of prospective studies.

BACKGROUND: Conflicting results have been reported on the association of poststroke depression with recurrent stroke events. This meta-analysis of prospective studies aims to evaluate whether poststroke depression is an independent predictor of stroke recurrence among stroke patients. METHODS: A systematic search of articles in PubMed and Embase databases from their inception to October 2018 was conducted. Prospective studies reporting risk estimates of stroke recurrence by depression status in stroke patients were included and pooled risk ratio (RR) with 95% confidence intervals (CIs) of stroke recurrence was calculated for patients with or without poststroke depression. RESULTS: Six studies with 4648 stroke patients were finally included, and the prevalence of poststroke depression was found to from 15.9% to 40.5%. The pooled adjusted RR for stroke recurrence in patients suffering from poststroke depression was 1.48 (1.22-1.79) in a fixed-effect model. Subgroup analyses indicated that poststroke depression significantly increased stroke recurrence (RR 1.64; 95% CI, 1.28-2.10) among ischemic stroke patients but not in total stroke patients (RR 1.28; 95% CI, 0.96-1.73). CONCLUSIONS: This meta-analysis suggests that poststroke depression may be an independent predictor of stroke recurrence among ischemic stroke patients. Further studies are required to investigate whether treatment of poststroke depression can reduce the risk of stroke recurrence.

Long Non-Coding RNA CASC2 Improves Diabetic Nephropathy by Inhibiting JNK Pathway.

It's known that long non-coding RNA CASC2 overexpression inhibit the JNK pathway in some disease models, while JNK pathway activation exacerbates diabetic nephropathy. Therefore we speculate that long non-coding RNA CASC2 can improve diabetic nephropathy by inhibiting JNK pathway. Thus, our study was carried out to investigate the involvement of CASC2 in diabetic nephropathy. We found that serum level of CASC2 was significantly lower in diabetic nephropathy patients than in normal people, and serum level of CASC2 showed no significant correlations with age, gender, alcohol consumption and smoking habits, but was correlated with course of disease. ROC curve analysis showed that serum level of CASC2 could be used to accurately predict diabetic nephropathy. Diabetes mellitus has many complications. This study also included a series of complications of diabetes, such as diabetic retinopathy, diabetic ketoacidosis, diabetic foot infections and diabetic cardiopathy, while serum level of CASC2 was specifically reduced in diabetic nephropathy. CASC2 expression level decreased, while JNK1 phosphorylation level increased in mouse podocyte cells treated with high glucose. CASC2 overexpression inhibited apoptosis of podocyte cells and reduced phosphorylation level of JNK1. We conclude that long non-coding RNA CASC2 may improve diabetic nephropathy by inhibiting JNK pathway.

Treating diabetic patients complicated with early cognitive impairment by using the repetitive transcranial magnetic stimulation combined with cognitive rehabilitation / 中华物理医学与康复杂志

Objective To explore the clinical effect of cognitive rehabilitation therapy combined with repetitive transcranial magnetic stimulation (rTMS) on diabetes mellitus patients complicated with early cognitive impairment.Methods Eighty patients with type 2 diabetes mellitus complicated with early cognitive impairment admitted to our hospital between February 2016 and February 2018 were randomly divided into a control group and an observation group.Patients in both groups were treated with conventional medications and comprehensive rehabilitation training,while the observation group was additionally treated with rTMS for 4 weeks.The fasting blood glucose (FBG) was detected before and after the treatment.The cognitive status,as well as the latency and amplitude of P300 were evaluated using Montreal cognitive assessment (MoCA),NTS-2000 Instrument for electromyography and evoked potentials before and after the treatment.Results After the treatment,significant decrease was observed within both groups in terms of FBG compared to before the treatment,but no significant differences between the two groups were revealed (P＞0.05).After the treatment,the sub-scores and total scores of the MoCA scale of the two groups were significantly higher than those before treatment,with those of the observation group significantly higher than the control group.After the treatment,the latency of P300 decreased significantly,while the amplitude of P300 increased significantly in both group,with significantly higher improvement in the observation group than the control group (P＜0.05).Conclusion rTMS,in addition to cognitive function rehabilitation,significantly improved cognitive function status in patients with type 2 diabetes mellitus complicated with early cognitive impairment.Such combination is worthy of clinical promotion with its safety,reliability and patient compliance.

MicroRNA-335 is downregulated in papillary thyroid cancer and suppresses cancer cell growth, migration and invasion by directly targeting ZEB2.

MicroRNAs (miRs) are a group of short, endogenous, non-protein-coding and single-stranded RNAs that regulate gene expression by binding to the 3'-untranslated region (3'UTR) of mRNAs, which results in their degradation or translational repression. The aim of the present study was to investigate the expression and function of miR-335 in human papillary thyroid cancer (PTC). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to quantify the relative miR-335 expression levels in PTC tissues and cell lines. The effect of miR-335 on the proliferation, migration and invasion of PTC cells was assessed by an MTT assay, and transwell migration and invasion assays, respectively. Dual-luciferase reporter assays were employed to explore whether miR-335 directly targeted the 3'UTR of the potential target gene zinc finger E-box binding homeobox 2 (ZEB2). RT-qPCR and western blotting were adopted to assess the effect of miR-335 on the mRNA and protein expression of ZEB2. RT-qPCR revealed that miR-335 was downregulated in PTC tissues and cell lines. The MTT assay and transwell migration and invasion assays demonstrated that the overexpression of miR-335 significantly inhibited the proliferation, migration and invasion of PTC cells. ZEB2 was identified as a direct target of miR-335 with computational analysis, which was confirmed with a dual-luciferase reporter assay, RT-qPCR and western blotting. The knockdown of ZEB2 significantly inhibited the proliferation, migration and invasion of PTC cells, indicating that ZEB2 may be a functional target of miR-335. Taken together, these findings suggested that miR-335 functioned as a tumor suppressor and suppressed the growth and metastatic behavior of PTC cells by targeting ZEB2.

Different intracellular signalling pathways triggered by an anti-prolactin receptor (PRLR) antibody: Implication for a signal-specific PRLR agonist.

In this work, we prepared a panel of monoclonal antibodies directed against prolactin receptor (PRLR) using the hybridoma technique. Of these monoclonal antibodies (Mabs), the Mab designated B6 was chosen for further characterization based on its biological activity. We first demonstrated that B6 can specifically bind to the prolactin receptor (PRLR) expressed on target cells by immunoprecipitation and Western blotting analysis. Subsequently, epitope mapping studies using a competitive receptor-binding assay indicated that B6 epitopes partially overlapped with those of prolactin (PRL). We then examined the resulting signal transduction pathways activated by this antibody in T-47D and CHO-PRLR cells and found that B6 induced different intracellular signalling compared with prolactin, which activates serine-threonine kinase (AKT), extracellular signal-regulated kinase 1/2 (ERK1/2), signal transducer and activator of transcription1 (STAT1) and STAT3 but not STAT5. The present study suggests that: (a) B6 may be a signal-specific prolactin receptor (PRLR) agonist; (b) B6 may be a biological reagent that can be used to explore the mechanism of PRLR-mediated intracellular signalling. In addition, this work also implies a strategy for preparing signal-specific cytokine agonists.

The clinical analysis of glycemic control and changes of pulmonary diffusion capacity in elderly type 2 diabetes / 中华老年医学杂志

Objective To investigate the effect of blood glucose level on pulmonary diffusion capacity in elderly patients with type 2 diabetes and its clinical significance.Methods Totally 132 older adults with type 2 diabetes were enrolled in this study.According to blood glucose level,the patients were divided into well-controlled group (n =57) and poor controlled group (n =75).Additionally,52 age-matched non-diabetic healthy subjects were selected as control group.Levels of fasting blood glucose (FBG),glycosylated hemoglobin (HbA1c),fasting insulin (FINS) and 2-hour postprandial glucose (2h PG) in the diabetic patients were detected,and homeostasis model assessment of insulin resistance (HOMA IR) was calculated.The patients with type 2 diabetes underwent urinary albumin excretion rate (UAER) detection,fundus examination and nerve conduction velocity test.pulmonary ventilation,diffusion of the lungs for carbon monoxide (DLCO) and DLCO corrected by alveolar volume (DLCO/VA) were examined in all subjects.Results Levels of FBG,2h PG,HbA1c and HOMA-IR were higher in poor-controlled group than in good glycemicwell-controlled group (all P＜0.05).Compared with the control group,body mass index (BMI) was increased in diabetic groups (both P＜0.05).The pulmonary ventilation function in the three groups had no significant differences (P＞0.05).DLCO and DLCO/VA were lower in diabetic groups than in the control group(all P＜0.05).DLCO and DLCO/VA in poor-controlled group were lower than those in well controlled group (both P＜0.05).DLCO and DLCO/VA were lower in patients with microangiopathy score ≥ 2 than those without microangiopathy (both P ＜ 0.05).Multiple liner regression analysis showed that DCLO and DLCO/VA were negatively correlated with HbA1c,HOMA-IR,duration of diabetes and microangiopathy (r=-2.51,-2.35,-2.42,-2.37,-2.41,-2.52,-2.47,-2.36,all P＜0.05).Conclusions The pulmonary diffusion capacity is significantly impaired in elderly patients with type 2 diabetes.Pulmonary diffusion capacity is negatively correlated with the blood glucose level.The lung may be one of the target organs of type 2 diabetes mellitus.

[Efficient reprogramming of human cord blood CD34(+) cells for formation of induced pluripotent stem cells with non-integrating plasmid system].

This study was to establish the episomal vector reprogramming method to reprogram iPSC from human cord blood (CB) CD34(+) cells. The non-integrating plasmids of pEB-C5 and pEB-Tg were transfected into short-term cultured CB CD34(+) cells by using the nucleofector, so as to demonstrate efficient reprogramming of CB CD34(+) cells. Within 14 days of one-time transfection by two plasmids together, up to 200 iPSC-like colonies per 2 million transfected CB CD34(+) cells were generated. The results showed that the pluripotency of iPSC-derived CB CD34(+) cells was similar to that of hESC and the karyotypes of iPSC were normal. In addition, no vector integration was found in iPSC of 9th and 10th passages. Furthermore, hiPSC formed teratoma with three embryonic germ layers. It is concluded that the integration-free method to generate human iPSC from CB CD34(+) cells is reliable and can provide new ways for both research and future clinical applications.

Efficient reprogramming of human cord blood CD34(+) cells for formation of induced pluripotent stem cells with non-integrating plasmid system / 中国实验血液学杂志

This study was to establish the episomal vector reprogramming method to reprogram iPSC from human cord blood (CB) CD34(+) cells. The non-integrating plasmids of pEB-C5 and pEB-Tg were transfected into short-term cultured CB CD34(+) cells by using the nucleofector, so as to demonstrate efficient reprogramming of CB CD34(+) cells. Within 14 days of one-time transfection by two plasmids together, up to 200 iPSC-like colonies per 2 million transfected CB CD34(+) cells were generated. The results showed that the pluripotency of iPSC-derived CB CD34(+) cells was similar to that of hESC and the karyotypes of iPSC were normal. In addition, no vector integration was found in iPSC of 9th and 10th passages. Furthermore, hiPSC formed teratoma with three embryonic germ layers. It is concluded that the integration-free method to generate human iPSC from CB CD34(+) cells is reliable and can provide new ways for both research and future clinical applications.

WE-G-BRCD-02: Speed Up Standard Optimization of IMPT Treatment Planning Through the Application of GPUs.

PURPOSE: While intensity-modulated proton therapy (IMPT) has great potential to deliver highly conformal tumoricidal dose to targets whilst minimizing dose to nearby organs-at-risk, IMPT optimization is very time consuming and memory extensive due to finer dose grids and a large number of energy layers used compared to intensity-modulated radiation therapy (IMRT). In this presentation, for the first time, a new approach is introduced to speed up the IMPT treatment planning through application of parallel computing with Graphic Processor Units (GPUs). METHODS: Parallel computation with GPUs, which are affordable and can be plugged in a workstation easily, is potentially a good way to improve the computation efficiency. In our approach, we used the standard quadratic objective function to optimize the intensity map of beamlets. The objective function and gradient equations, which are the most time consuming parts of the optimization, were calculated with GPUs. We compared the computation time of optimization done by an Intel ® Core™ i7 CPU and that by the same CPU accelerated by GPUs (TESLA C1060). The influence matrix was pre- calculated before optimization with an in-house proton pencil beam dose calculation engine. Two clinical cases were studied: one base-of-skull (BOS) case (clivus chordoma) and one prostate case (adenocarcinoma). The dose volume histogram (DVH) data for the tumor and critical organs were derived for comparison of optimization results generated by CPU and GPUs. RESULTS: For the BOS case, application of GPUs for the optimization and overall gained 54 and 36.5 times speedup. For the prostate case, application of GPUs for the optimization and overall gained 69 and 28.5 times speedup. CONCLUSIONS: The application of GPUs for the parallel computing of IMPT treatment plan optimization can dramatically improve the computation efficiency. The optimization time can be reduced from typically half to one hour to only several minutes.

WE-G-BRCD-03: Improvement in Robustness and Delivery Efficiency of Intensity-Modulated Proton Therapy Using Single-Field Optimization with Energy Absorber.

PURPOSE: Intensity-modulated proton therapy (IMPT) using multi-field optimization (MFO) could generate highly conformal dose distributions but it is more sensitive to setup and proton range uncertainties than IMPT using single-field optimization (SFO). This work evaluates the effectiveness of SFO treatment plans with the use of energy absorbers (EAs) to improve the robustness and delivery efficiency of IMPT for head and neck cancers. METHODS: IMPT treatment plans were generated using 2-field SFO with an EA in each field (EA-SFO) for four patients with head and neck cancers. We compared the plan quality, robustness, and delivery efficiency of the EA- SFO plan with a 3-field MFO plan that was used to treat the patient. Robustness analysis of each plan was performed to generate two dose distributions, consisting of the highest and the lowest possible doses from spatial and range perturbations at every voxel. Dosimetric indices and the numbers of energy layers required in the EA-SFO and MFO plans were compared. RESULTS: All the nominal EA-SFO plans are clinically acceptable. They achieved similar levels of target coverage compared to the MFO plans; the differences in D95 of the GTV and CTVs between the two plans were within 3.5%. Although some of the OARs received higher dose in the EA- SFO plan, they were all within tolerance. The EA-SFO plans yielded an average of 38.5% reduction of plan sensitivity to uncertainties in the targets and 18.5% overall. The EA-SFO plans used an average of 79 (46%) fewer energy layers than the MFO plans, which corresponds to nearly 3 minutes shorter delivery time. CONCLUSIONS: The use of energy absorber greatly facilitated the design of clinically acceptable SFO treatment plans. Compared to MFO, EA-SFO not only improved the robustness to setup and range uncertainties, but also reduced the time required for delivery and patient QA.

A faster ordered-subset convex algorithm for iterative reconstruction in a rotation-free micro-CT system.

We present a faster iterative reconstruction algorithm based on the ordered-subset convex (OSC) algorithm for transmission CT. The OSC algorithm was modified such that it calculates the normalization term before the iterative process in order to save computational cost. The modified version requires only one backprojection per iteration as compared to two required for the original OSC. We applied the modified OSC (MOSC) algorithm to a rotation-free micro-CT system that we proposed previously, observed its performance, and compared with the OSC algorithm for 3D cone-beam reconstruction. Measurements on the reconstructed images as well as the point spread functions show that MOSC is quite similar to OSC; in noise-resolution trade-off, MOSC is comparable with OSC in a regular-noise situation and it is slightly worse than OSC in an extremely high-noise situation. The timing record shows that MOSC saves 25-30% CPU time, depending on the number of iterations used. We conclude that the MOSC algorithm is more efficient than OSC and provides comparable images.

Fluorescence Property of Reaction Productsof Sparfloxacin with Halide and Its Application / 分析化学

The fluorescence characteristics of reaction products of sparfloxacin with halide (F, Cl, Br, I) have been studied. It was found that sparfloxacin was oxidized by nitrous acid then reacted with halide further to formstrong fluorescence substances in acid media. Their fluorescence intensitites enhanced with aggrandizing of atom radii of halides, which could emit the strong fluorescence 58～151 fold more than that of sparfloxacin itself. Thus a new sensitive method for the direct determination of sparfloxacin in human urine by derivative-synchronous fluorescence was presented. A plausible mchanism was proposed to explin this behavior.

Comparison of biomechanical and biochemical properties of cartilage from human knee and ankle pairs.

Cartilage was obtained from eight matched knee (tibiofemoral and femoropatellar) and ankle (talocrural) joints of five different donors (both left and right from donors 14, 22, and 38 years of age, and left only from donors 31 and 45 years of age) within 24 hours of death. All cartilage was graded as normal by the macroscopic visual Collins' scale and the histological Mankin scale. Cylindrical disks of cartilage were harvested from 10 sites within the tibiofemoral and femoropatellar joint surfaces and four sites within the talocrural joint, and uniaxial confined compression measurements were performed to quantify a spectrum of physical properties including the equilibrium modulus, hydraulic permeability, dynamic stiffness, streaming potential, electrokinetic coupling coefficient, and electrical conductivity. Matched specimens from the same 14 sites were used for complementary measurements of biochemical composition and molecular interaction, including water content, hypotonic swelling behavior, and sulfated glycosaminoglycan and collagen contents. In comparison of the top 1-mm slices of talar cartilage with the top 1-mm of tibiofemoral cartilage, the talar cartilage appeared denser with a higher sulfated glycosaminoglycan content, lower water content, higher equilibrium modulus and dynamic stiffness, and lower hydraulic permeability. The equilibrium modulus increased with increasing sulfated glycosaminoglycans per wet weight and decreased with increasing water content for all joint surfaces. Multiple linear regression showed that greater than 80% of the variation in the equilibrium modulus could be accounted for by variations in the biochemical parameters (water content, sulfated glycosaminoglycans/wet weight, and hydroxyproline content/wet weight) for each joint surface. Nonhomogeneous depth-dependent changes in the physical properties and biochemical composition of full-thickness distal femoral cartilage were consistent with previous reports. Since the compressive deformation of cartilage during cyclic loading is confined to the more superficial regions, the differences in properties of the upper regions of the talar compared with tibiofemoral or femoropatellar cartilage may be important in the etiology of osteoarthritis.

Intravenous aspirin at reperfusion does not reduce infarct size in the dog with a residual critical stenosis.

Platelet-related events being associated with the increment of infarct size at reperfusion in the presence of a residual stenosis, we tested in dogs whether intravenous aspirin (ASA) could limit infarct size. The left anterior descending coronary artery was occluded for 90 min and reperfused for 6 h in the presence of a residual critical stenosis. Controls received saline, and treated groups were given 2, 6, or 12 mg/kg ASA, i.v., 5 min before reperfusion. Infarct size did not differ significantly between groups (control, 43.80+/-6.28%; ASA, 2 mg/kg: 41.07+/-7.78%; ASA, 6 mg/kg: 37.55+/-3.44%; ASA, 12 mg/kg: 29.40+/-5.41%), as well as transmural collateral blood flow and [111In]-platelet accumulation in the infarcted myocardium (2.5-3.6 x 10(5) platelets/g). However, myocardial neutrophil accumulation was significantly reduced (p < 0.05) in groups given 6 (15.0+/-2.6 x 10(6)/g tissue) and 12 mg/kg (18.4 +/-3.8) ASA, but not in the 2-mg/kg group (21.0+/-5.2), as compared with control group (32.0+/-7.2). Ex vivo platelet aggregation to collagen was abolished during reperfusion in all treated groups (p < 0.05). Transcardiac arteriovenous differences in 6-keto-PGF1alpha were reduced significantly 1 h after reperfusion in groups given 6 or 12 mg/kg ASA (94.7+/-13.1 and 71.7+/-19.2 pg/ml, respectively) but not in the 2-mg/kg group (178.3+/-78.2 pg/ml), as compared with control (405.4+/-171.6 pg/ml). ASA-insensitive platelet activation at the site of stenosis or inhibition by ASA of prostacyclin production by jeopardized myocardium may explain the observed lack of benefit of ASA.

The low molecular weight heparin, enoxaparin, limits infarct size at reperfusion in the dog.

OBJECTIVE: Heparin (HEP) is used in the post-thrombolytic state to prevent vessel reocclusion, thereby aiding myocardial salvage. Side effects limit its benefits, but besides anticoagulant activity HEP has diffuse actions that may be potentially beneficial to jeopardized reperfused myocardium. This study compares the effect of therapeutic doses of HEP and enoxaparin (ENOX), a low molecular weight heparin, and to streptokinase (SK), on infarct size. METHODS: The left anterior descending coronary artery was occluded in dogs for 90 min, followed by 6 h of reperfusion with a residual critical stenosis in place. Five min before reperfusion, HEP (2800 IU) was injected i.v., and perfused at 500 IU/h until sacrifice in group 2, while groups 3 and 4 received ENOX (2128 anti-Xa IU i.v.) followed by 380 anti-Xa IU/h. Group 4 was also given 500,000 IU SK over 30 min before reperfusion beginning at 55 min of occlusion (ENOX + SK), while group 5 received only SK. Controls (CON, group 1) received saline. P-selectin mediated platelet-neutrophil rosettes formation was also tested in vitro in the presence of HEP and ENOX. RESULTS: The area at risk delimited by dye perfusion was statistically similar among groups. Covariance analysis between infarct size (% of area at risk) delimited with triphenyltetrazolium and collateral flow measured with radioactive microspheres confirmed that groups given ENOX (21.6 +/- 5.5%) and ENOX + SK (24.9 +/- 3.9%) developed smaller infarcts (P < 0.05) than CON (48.1 +/- 4.5%), as opposed to HEP (32.2 +/- 3.6%) and SK (46.8 +/- 3.4%) groups. 111In-platelet counts in the infarct were reduced significantly by 64% in the ENOX group as compared to CON, and to a lesser extent (42%, n.s.) in the ENOX + SK group, but were not reduced by HEP and SK treatments. Neutrophil accumulation in the infarcts was decreased significantly and by more than 75% in the ENOX and ENOX + SK groups versus CON, but not in the HEP and SK groups. Also, only ENOX (10-100 micrograms/ml) significantly inhibited platelet-neutrophil rosettes formation in a plasmatic milieu. CONCLUSIONS: The ENOX treatment, as opposed to that of HEP, reduces myocardial platelet and neutrophil accumulations, and limits infarct size when given just before and during reperfusion. The benefits of ENOX on infarct size were not modified by SK, and may be related, at least in part, to an interaction with P-selectin-mediated cell adhesion.

Postnatal development of the vasopressinergic system in golden hamsters.

Adult golden hamsters, as compared to rats, lack several parvicellular vasopressinergic cell groups. We looked at the development of the vasopressinergic system in hamsters to draw comparisons with maturing rats. Arginine-vasopressin-immunoreactive (AVP-ir) neurons, their fibers and associated AVP binding sites were observed at several intervals after birth. Different rates of maturation were observed between different populations of vasopressinergic neurons. Within the suprachiasmatic nucleus (SCN), small AVP-ir neurons, their fibers and related binding sites maturated gradually during the first month after birth. In comparison, large AVP-ir neurons were apparent in newborn animals. Similarly, AVP-ir fibers and AVP binding sites were also present in the brain of newborns within areas not related to small vasopressinergic neurons from the SCN, such as the central amygdala (CeA) or the cerebral cortex. During the following weeks, a heterogenous pattern of development was observed within such areas. As the neurosecretory vasopressinergic system appeared to develop gradually, projections to the brain and their associated binding sites developed rapidly during the first week of life. Transient patterns of maturation were observed within certain sites. Indeed, some of the labelling observed in newborns regressed later. As similar reports were made in rats, our observations draw analogies between the vasopressinergic systems of these two species, beside their apparent dissimilarities in adult animals. Furthermore, our data also reinforce the concept that large vasopressinergic neurons do not constitute a homogenous population.

Cell-type-specific transactivation of the parathyroid hormone-related protein gene promoter by the human T-cell leukemia virus type I (HTLV-I) tax and HTLV-II tax proteins.

The human T-cell leukemia virus type I (HTLV-I) and HTLV-II Tax proteins are potent transactivators of viral and cellular gene expression. Using deletion mutants, the downstream parathyroid hormone-related protein (PTHrP) promoter is shown to be responsive to both HTLV-I and HTLV-II Tax as well as the AP1/c-jun proto-oncogene. Transactivation of PTHrP by Tax was seen in T cells but not in B-cell lines or fibroblasts. A carboxy terminal Tax deletion mutant was deficient in transactivation of both the PTHrP and IL2R alpha promoters but not the HTLV-I long terminal repeat (LTR). Exogenous provision of NFkB rescued IL2R alpha expression but not the PTHrP promoter. Thus, HTLV-I Tax, HTLV-II Tax, and c-jun transactivate PTHrP and may contribute to the pathogenesis of hypercalcemia in adult T-cell leukemia.