μL In Group Research Articles

The profile of T-cells (CD4 +, CD8 +) and natural killer cells (CD16 / 56+) in STEMI patients with poor prognosis

Abstract Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): Institute of Cardiology, Kyiv, Ukraine The role of lymphocytes in the progression of atherosclerosis is still unclear. T-cells helper (CD4+) and suppressor (CD8+) subtypes are activated in acute cardiac injury and myocardial infarction (AMI),including cytotoxicity mediated by CD8+ T-cells and immunoregulatory activity mediated by CD4+ T-cells (which can also enhance the CD8+ T-cell cytotoxicity). The natural killer (NK) cells are important in the infarcted myocardium, where it affects heart remodeling. The aim of the study was to identify the difference in the dynamic profile of T-cells (CD4 +, CD8 +) and NK cells (CD16 / 56+) in STEMI patients with and without complications during 1-year follow-up. Methods The composition of individual subtypes of T-cells (CD4 +, CD8 +) and NK cells (CD16 / 56+) in the venous blood was evaluated retrospectively in 60 pts with STEMI, treated with primary PCI, at admission and on day 7. The following cardiovascular events were considered as the end point in remote (1 year) observation: death, AMI, stroke, revascularization, development/decompensation of heart failure and cardiac ischemia. All patients were divided into 2 groups depending on the presence (1st group - 35 pts) and absence of endpoint (2nd group - 25 pts). Both groups were comparable by the clinical profile. Results: There were lower number of T-suppressor on day 7 in patients with cardiovascular events during long-term follow-up (357.4 ± 36.2 /μl in group 1 vs 779.7 ± 222.8 /μl in group 2, p <0,01) with the trend in their percentage (23.6 ± 1.5% vs. 27.6 ± 2.3% in group 2, p <0.1). In pts with poor prognosis NK cells number and percentage has tendency to increase at the admission– 45,6% vs 37,3% in the group 2. Tab 1 Conclusion We can assume that the initial tendency to increase in the number and percentage of NK cells (CD16 / 56 +) followed by a decrease in the number and percentage of T cells suppressor (CD8+) on STEMI day 7 are associates with an undesirable 1-year prognosis. Dynamics of T-cells subtypes in STEMI pGroup 1 (n = 35)Group 2 (n = 25)Р group 1 vs group 2Day1Day 7Day 1Day7T-cells suppressor (CD8+) %20,9 ± 1,623,6 ± 1,524,2 ± 2,127,6 ± 2,3<0.1 on day 7T-cells suppressor (CD8+) /μl341,4 ± 40,1357,4 ± 36,2338,4 ± 46,8779,7 ± 222,8<0.02 on day 7T-cells helper (CD4+) %42,0 ± 1,948,0 ± 1,345,3 ± 2,248,0 ± 2,1>0.1T-cells helper (CD8+) /μl671,9 ± 72,9771,7 ± 68,0666,6 ± 94,2900,9 ± 115,8>0.1Natural killer (CD16 / 56+) %18,4 ± 1,912,5 ± 1,413,4 ± 2,09,73 ± 0,96<0.1 on day 1Natural killer (CD16 / 56+) /μl263,7 ± 31,9219,7 ± 33,8181,1 ± 26,7170,6 ± 25,9<0.1 on day 1

Role of caspase-1 in spinal cord in a rat model of incisional pain

Objective To evaluate the role of caspase-1 in the spinal cord in a rat model of incisional pain. Methods Eighteen adult male Sprague-Dawley rats, weighing 250-280 g, in which intrathecal catheters were successfully implanted, were divided into 3 groups(n=6 each)using a random number table: incision pain group(group I), incision pain plus dimethyl sulfoxide group(group ID)and incision pain plus caspase-1 inhibitor(Ac-YVAD-CMK)group(group IA). At 10 min before establishment of the model, 0.9% normal saline 20 μl was intrathecally injected in group I, dimethyl sulfoxide 20 μl was intrathecally injected and then the catheter was washed with 0.9% normal saline 10 μl in group ID, and Ac-YVAD-CMK 1 nmol/μl(dissolved in 20 μl dimethyl sulfoxide)and then the catheter was washed with 0.9% normal saline 10 μl in group IA.The mechanical paw withdrawal threshold(MWT)of the ipsilateral hind paw was measured at 2 h before intrathecal catheterization(T0), 3 days after intrathecal catheterization(T1)and 2, 6, 24 and 48 h after establishment of model(T2-5). The rats were sacrificed after the last measurement of pain threshold at T5, and lumbar enlargement segments of the spinal cord were removed for detection of caspase-1(p20)expression and interleukin-1beta(IL-1β)content by Western blot and enzyme-linked immunosorbent assay, respectively. Results Compared with the baseline at T0, the MWT was significantly decreased at T2-5 in I and ID groups(P 0.05). Compared with I and ID groups, the MWT at T2-5 was significantly increased at T2-5, and the caspase-1(p20)protein expression and IL-1β content were decreased in group IA(P 0.05). Conclusion The activation of caspase-1 in the spinal cord can promote the release of IL-1β and thus is involved in the incision pain in rats. Key words: Pain, postoperative; Caspase1; Interleukin-1beta; Spinal cord

Hypokalemia during acute metabolic acidosis on hemodilution with succinylated gelatin in rats

BackgroundExtracellular metabolic acidosis of mineral origin is commonly associated with plasma hyperkalemia. Nevertheless, in previous experiments, animals subjected to acute metabolic acidosis induced by normovolemic hemodilution using a colloidal volume replacement solution containing succinylated gelatin (gelafundin), developed a hypokalemic state with concomitant marked increases in diuresis and renal potassium excretion. In the present study, the succinylated gelatin's impact on diuresis and consequently potassium excretion was studied. Material and methodsAnesthetized Wistar rats were subjected to acute metabolic acidosis either due to normovolemic hemodilution with gelafundin (group I) or HCl application (groups II and III). Animals of group III received mannitol in addition. Blood gas analyses were performed regularly. Urine was continuously collected, and the excreted volume as well as potassium concentration was measured. ResultsIn all groups, mean base excess value was about −3.0 mEq/L. Plasma potassium concentration decreased from 5.0 mM to 4.5 mM in group I, whereas it was almost constant in groups II and III. The urine volume amounted to 2300 μL in groups I and III and 1000 μL in group II. Excreted total amount of potassium in urine was 340 μmol (group I), 125 μmol (group II), and 230 μmol (group III), respectively. ConclusionsThe employed volume replacement solution leads to increased diuresis induced by excretion of succinylated gelatin, which also sufficiently accounts for enhanced potassium loss into urine and decreased plasma potassium concentration. Therefore, generalization of the connection between acute metabolic acidosis and plasma hyperkalemia, as often stated in literature, is not justified.

Role of the vascular endothelial growth factor in the inverse relationship between increased nuchal translucency thickness and fetomaternal transfusion

To elucidate the possible etiological role of the vascular endothelial growth factor (VEGF) in the inverse correlation between nuchal translucency (NT) thickness and fetomaternal transfusion (FMT). The level of FMT was determined prospectively in 80 viable, singleton pregnancies in which 10-14-week ultrasonographic scanning, NT thickness measurement; chorionic villus sampling (CVS) for fetal karyotyping and VEGF concentration determination were performed. The grouping procedures were based either on NT thickness (<2 MoM in Group I, and ≥2 MoM in Group II), or on karyotype (euploid in Group A, and aneuploid in Group B). The level of FMT was determined via maternal serum α-fetoprotein levels before and after CVS. The FMT and the VEGF concentration of the chorionic tissue were analysed in comparisons between Groups I and II, and between Groups "A" and "B". The mean level of FMT after CVS was 72.5±21.3 μL and 19.28±5.4 μL in Groups I (n=44) and II (n=36), respectively (P<0.02). The VEGF concentration of the chorionic tissue in Groups I and II was 40.6±16.7 pg/mg protein and 21.1±6.3 pg/mg protein, respectively (P=0.28). The mean level of FMT was 57.9±15.0 μL and 8.1±3.9 μL in Groups A and B, respectively (P<0.003). The VEGF concentration of the chorionic tissue in Groups A and B was 25.9±10.7 pg/mg protein and 21.3±11.3 pg/mg protein, respectively (P=0.77). No difference exists in the VEGF concentration in the aspirated chorionic tissue between Groups I and II and between Groups A and B. A higher level of FMT was observed among the aneuploid pregnancies after CVS than among the euploid cases. Chorionic VEGF does not influence the inverse relationship between the pre-CVS NT thickness and FMT.

A Pilot Study of Denileukin Diftitox (DD) in Combination With High-dose Interleukin-2 (IL-2) for Patients With Metastatic Renal Cell Carcinoma (RCC)

High-dose (HD) IL-2 is approved to treat renal cell carcinoma (RCC) with modest response rates and significant toxicity. Enhancement of cytotoxic T-cell activity by IL-2 is 1 mechanism of action. IL-2 also stimulates regulatory T lymphocytes (Tregs), which are associated with poor prognosis. Favorable outcomes are associated with greater rebound absolute lymphocyte count (Fumagalli 2003). DD depletes IL-2 receptor (CD25 component) expressing cells. We hypothesized that sequential therapy could complement each other; DD would deplete Tregs so IL-2 could more effectively stimulate proliferation and activity of cytotoxic T lymphocytes. Patients (n=18) received standard HD IL-2 and 1 dose of DD daily for 3 days; periodic flow cytometry and complete blood counts were performed. Group A included 3 patients to assess safety only with DD 6 μg/kg between the IL-2 courses. Group B included 9 patients at 9 μg/kg DD before the IL-2 courses. Group C included 6 patients at 9 μg/kg DD between the IL-2 courses. Efficacy using the RECIST criteria was assessed after the treatment. Fifteen patients from a study of IL-2 without DD served as controls for toxicity comparison and 13 of these for flow cytometry comparisons. No unusual toxicity was noted. For group B/C patients receiving DD, the median decline in Tregs was 56.3% from pre-DD to post-DD (P=0.013). Peak absolute lymphocyte count change from baseline was +9980/μL for group B, +4470/μL for group C, and +4720/μL for the controls (P=0.005 B vs. C). The overall response rate was 5 of 15 (33%); 3 of 9 (33%) and 2 of 6 (33%) for groups B and C, respectively, including 2 patients with sarcomatoid RCC and 1 with earlier sunitinib therapy.

Oral Eltrombopag for the Long-Term Treatment of Patients with Chronic Idiopathic Thrombocytopenic Purpura: Results of a Phase III, Double- Blind, Placebo-Controlled Study (RAISE)

INTRODUCTION: Eltrombopag (PROMACTA®/REVOLADE®; GlaxoSmithKline, Collegeville, PA, USA) is a first-in-class, oral, small molecule, non-peptide, thrombopoietin receptor agonist being studied for the treatment of thrombocytopenia related to a variety of conditions.METHODS: RAISE was a 6-month, randomized, double-blind, placebo-controlled, phase III study that evaluated the efficacy and safety of eltrombopag in previously treated adults with chronic idiopathic thrombocytopenic purpura (ITP) with platelet counts <30,000/μL. It was estimated that approximately 189 patients randomized 2:1 (eltrombopag:placebo) would provide sufficient statistical power. Patients were stratified by splenectomy status, use of baseline ITP medication, and platelets □15,000/μL. Patients initiated treatment with eltrombopag 50 mg (or matching placebo) once daily, and the dose was individualized based upon each patient's platelet response, from a maximum of 75 mg once daily to 25 mg once daily or less frequently. Patients could reduce concomitant medications and receive rescue therapy as dictated by local standard of care. The primary endpoint was the odds of responding (platelets 50,000 to 400,000/μL) during the treatment period for patients receiving eltrombopag relative to placebo. Bleeding symptoms were prospectively evaluated using the WHO Bleeding Scale: Grade 0 = no bleeding, Grade 1 = mild bleeding, Grade 2 = moderate bleeding, Grade 3 = gross bleeding, and Grade 4 = debilitating blood loss.RESULTS: One hundred ninety-seven patients (eltrombopag, 135; placebo, 62) were enrolled in RAISE, and baseline characteristics were balanced: in both arms ~50% of patients had platelet counts □15,000/μL, ~50% were receiving concomitant ITP therapies, ~35% were splenectomized, and >15% had received at least 3 prior ITP medications. Patients who received eltrombopag were 8 times more likely to achieve platelet counts 50,000 to 400,000/μL during the 6-month treatment period compared with patients on placebo (OR [95% CI] = 8.2 [4.32, 15.38]; P <0.001). Baseline median platelet counts were 16,000/μL in both groups and never exceeded 30,000/μL in the placebo group. In contrast, platelets rose to 36,000/μL after 1 week in the eltrombopag group (Figure 1) and subsequently ranged from 52,000 to 91,000/μL for the remainder of the study. Median platelet counts returned to near baseline 2 weeks after stopping eltrombopag. Patients responded to eltrombopag regardless of splenectomy status, use of baseline ITP medications, or baseline platelet counts. Significantly fewer patients treated with eltrombopag had any bleeding (WHO Grades 1–4; P <0.001) or clinically significant bleeding (WHO Grades 2–4; P <0.001) throughout the trial compared with patients treated with placebo. More patients in the eltrombopag group (59%) stopped or dose-reduced their concomitant ITP medications than in the placebo group (32%; P = 0.016). Patients in the eltrombopag group (19%) required less rescue therapy compared with the placebo group (40%; P = 0.001) during the treatment phase of the study. The overall incidence of adverse events (AEs) was similar between the eltrombopag (87%) and placebo groups (92%), and the AEs were mostly mild to moderate. Headache was the most common AE in both groups (30%). Of note, 2 steroid-associated AEs (dyspepsia and peripheral edema) were significantly less likely to occur in the eltrombopag group compared with the placebo group. A higher incidence of hepatobiliary laboratory abnormalities were reported in the eltrombopag group (13%) compared with the placebo group (7%). There were no clinical or laboratory symptoms suggestive of bone marrow fibrosis. One death due to brain stem hemorrhage was reported in the placebo group.DISCUSSION: Long-term eltrombopag therapy significantly increased platelet counts, decreased bleeding symptoms, allowed for a reduction or discontinuation of baseline ITP therapies, and reduced the use of rescue ITP medications compared with placebo. Eltrombopag was well-tolerated, with a similar safety profile to placebo, and is an important new treatment option for patients with chronic ITP. [Display omitted]

Effect of collection frequency on quantitative and qualitative characteristics of pigeon (Columba livia) semen

1. The aim was to estimate the optimal frequency of semen collection from pigeons in relation to ejaculate volume, sperm concentration, total spermatozoa in ejaculate and percentage of live morphologically normal cells. 2. The study was carried out on 455 ejaculates collected from two groups of pigeons, each of 10 males (group I: meat-type breed; group II: fancy pigeon). The birds were selected and kept individually in cages under a natural photoperiod. 3. A two-person technique was used for semen collection (lumbo-sacral and cloacal region massage). Semen was collected once, twice or three times per week. 4. Colour, consistency and volume of ejaculates were evaluated macroscopically immediately after collection. Sperm concentration and total number of cells in the ejaculate were estimated after dilution with Ringer's solution. A live–dead stain technique (nigrosin–eosin) was used to determine the percentage of live and normal spermatozoa. 5. Semen collected 3×/week was of high quality. The average volume of a single ejaculate was small (21 µl in group I and 19 µl in group II), but sperm concentration was high—1·58 × 109/ml and 1·96 × 109/ml, respectively. The mean number of spermatozoa per ejaculate was 30·48 × 106 in group I and 39·49 × 106 in group II. An increased percentage of live and normal spermatozoa in semen collected more frequently was also observed. 6. Collecting pigeon semen 3×/week provides spermatozoa in larger amounts and of better quality than less frequent collections (1×/week or 2×/week) and is recommended for obtaining more insemination doses.

Number and Volume of Islets Transplanted in Immunobarrier Devices

Immunobarrier devices may prevent immune destruction of transplanted islets, but there are concerns about survival within such devices. Islets were transplanted in diffusion chambers that employed two laminated polytetrafluoroethylene membranes held together with titanium rings. Five hundred syngeneic mouse islets placed in devices were transplanted into the epididymal fat pads of streptozotocin (STZ) diabetic mice (B6AF1). After 2 wk the devices were removed. Sections were made parallel to the membrane surface. Eight to 13 systematically selected sections of each device were analyzed by planimetry to determine the area of the device space and of the islets within that space. From these data we estimated total volume of the device, volume of islets, and number of islets in a device. The data were segregated into two groups: group I (blood glucose less than 100 mg/dL 2 wk after implantation), and group II (over 150 mg/dL). The volume (mean ± SE) of devices implanted for 2 wk was 2.1 ± 0.4 μL in group I and 2.2 ± 0.2 μL in group II. The islet volume and number within devices were 0.30 ± 0.06 and 0.17 ± 0.01 μL, or 340 ± 50 and 230 ± 20 islets in group I and group II, respectively. The volume of fibrous tissue in devices was about 0.50 μL. About 10% of the islet tissue had central necrosis. The beta cell volume in a membrane device needed for cure is comparable to that required with islets under the kidney capsule (0.25–0.80 μL). The mass of islets contained within membrane devices needed to cure diabetes is equivalent to that of a graft in an optimal transplant site such as under the kidney capsule.

Number and volume of islets transplanted in immunobarrier devices.

Immunobarrier devices may prevent immune destruction of transplanted islets, but there are concerns about survival within such devices. Islets were transplanted in diffusion chambers that employed two laminated polytetrafluoroethylene membranes held together with titanium rings. Five hundred syngeneic mouse islets placed in devices were transplanted into the epididymal fat pads of streptozotocin (STZ) diabetic mice (B6AF1). After 2 wk the devices were removed. Sections were made parallel to the membrane surface. Eight to 13 systematically selected sections of each device were analyzed by planimetry to determine the area of the device space and of the islets within that space. From these data we estimated total volume of the device, volume of islets, and number of islets in a device. The data were segregated into two groups: group I (blood glucose less than 100 mg/dL 2 wk after implantation), and group II (over 150 mg/dL). The volume (mean +/- SE) of devices implanted for 2 wk was 2.1 +/- 0.4 microL in group I and 2.2 +/- 0.2 microL in group II. The islet volume and number within devices were 0.30 +/- 0.06 and 0.17 +/- 0.01 microL, or 340 +/- 50 and 230 +/- 20 islets in group I and group II, respectively. The volume of fibrous tissue in devices was about 0.50 microL. About 10% of the islet tissue had central necrosis. The beta cell volume in a membrane device needed for cure is comparable to that required with islets under the kidney capsule (0.25-0.80 microL). The mass of islets contained within membrane devices needed to cure diabetes is equivalent to that of a graft in an optimal transplant site such as under the kidney capsule.

Anticoagulation in Patients Treated by Continuous Venovenous Hemofiltration: A Retrospective Study

The most adequate anticoagulation regimen during extracorporeal renal replacement therapy can be difficult to define. Two hundred fifty-five critically ill patients with a mean age (+/- SD) of 58.2 +/- 16.3 years were treated by continuous venovenous hemofiltration (CVVH) between 1986 and 1992 in our intensive care units. Blood was circulated through hemofilters, either polyacrylonitrile (AN 69; Hospal, Lyon, France) or polyamide (FH 66; Gambro, Lund, Sweden), using a roller pump and an air safety system. The patients were classified into three subgroups according to the amount of heparin needed to achieve an adequate anticoagulation (ie, prevention of extracorporeal circuit clotting without inducing a patient's bleeding tendency): group 1, 37 patients who received no heparin (14.5%); group 2, 189 patients who received 100 to 700 IU/hr of heparin (74.1%); and group 3: 29 patients who received more than 700 IU/hr of heparin (11.4%). We analyzed the filter survival, the routine coagulation parameters, and the evolution of the patients for each group. Median duration of treatment was 144 hours (range, 4 to 1,152 hours). There were no differences in requirement of heparin among the two types of membrane: AN 69 (mean +/- SD), 393 +/- 106 IU/hr v FH 66, 374 +/- 35.3 IU/hr (range, 0 to 2,000 IU/hr). There were no relationships between the amount of heparin the patients received and the mean survival of the filters (group 1, 22.1 +/- 14.8 hr; group 2, 24.7 +/- 13.2 hr; group 3, 23 +/- 9.6 hr).(ABSTRACT TRUNCATED AT 250 WORDS)