Decrease In WBC Count Research Articles

Is therapeutic hypothermia immunosuppressive?

Is therapeutic hypothermia immunosuppressive?

Phase I results from a phase I/II study of orteronel, an oral, investigational, nonsteroidal 17,20-lyase inhibitor, with docetaxel and prednisone (DP) in metastatic castration-resistant prostate cancer (mCRPC).

4656 Background: The investigational agent orteronel (ortl, TAK-700) is a selective 17,20-lyase inhibitor that blocks androgen production. Since DP is standard chemotherapy in mCRPC, this phase 1/2 study examined ortl + DP in men with mCRPC. Methods: The primary phase I objective was to determine the maximum dose of ortl 200–400 mg BID that can be administered safely with DP in castrate men (testosterone [T] <50 ng/dL) with mCRPC, ≤1 prior chemotherapy and no ketoconazole/abiraterone ≤30 d prior. A 3+3 dose escalation was used with ortl 200 then 400 mg BID. Ortl dosed daily and D (75 mg/m2 q3w) + P (5 mg BID) was started on d8 of cycle 1 (28 d); cycles ≥2 = 21 d. Results: 14 men, median age 68 yrs (range 53–81), ECOG PS 0/1 (71%/29%), median PSA 59.4 ng/mL (5.2–1052), T 8.1 ng/dL (1.2–16.7), 9 with measurable disease, were treated. Median ortl exposure was 33.3 wks (6.3–59); 6 and 8 men received ortl 200 and 400 mg BID + DP, in cohorts 1 and 2, respectively. No dose-limiting toxicities (DLTs) occurred in cohort 1 (2 x 3 pts); in cohort 2 (2 x 4 pts), 1 pt in group 1 had DLT (Gr3 related febrile neutropenia); no DLTs occurred in group 2. 3 pts discontinued due to AEs (2 D infusion reactions, 1 at each dose; 1 unrelated cardiorespiratory death at 200 mg). All men had at least 1 AE Gr ≥3 which were treatment-related in 11 men. The most common Gr ≥3 AEs were seen at both doses and included neutropenia 50% (n = 7), hyperglycemia 21% (n = 3), febrile neutropenia, infusion reaction, hyponatremia, decreased neutrophil count (2 each); at 400 mg only: fatigue, hypophosphatemia, and decreased WBC count (2 each). 7 men had serious AEs (SAEs), which were drug-related in 5. The most common SAE was febrile neutropenia in 2 men. At 3 mo, PSA50 and PSA90 rates were 86% and 36%, respectively; 12/14 men had a median PSA decline ≥70%; and median T declined to ≤0.2 ng/dL (0–10.7). The Cmax of D + ortl was similar to D alone. Conclusions: Ortl 200 mg and 400 mg BID + DP appears safe and tolerable with androgen-lowering activity at the maximum planned dose in mCRPC. There is no evidence of AE potentiation when ortl is administered with DP. The phase II portion of the study will use ortl 400 mg BID + DP.

Evaluation of prognostic value of selected biochemical markers in surgically treated patients with acute mediastinitis

SummaryBackgroundMonitoring of biochemical markers of inflammation in acute mediastinitis (AM) can be useful in the modification of treatment. This study was a retrospective evaluation of selected biochemical parameters with negative impact on the prognosis in surgically treated patients.Material/MethodsThere were 44 consecutive patients treated surgically due to AM of differentiated etiology. Selected biochemical markers (WBC, RBC, HGB, HCT, PLT, CRP, PCT, ionogram, protein and albumins) were assessed before surgery and on the 3rd day after surgery. ANOVA was applied to find factors influencing observations. Numerical data [laboratory parameters] were compared by means of medians.ResultsThe overall hospital mortality rate was 31.82%. In the group of dead patients, there were observed statistically significant lower mean preoperative values of RBC [p=0.0090], HGB [p=0.0286], HCT [p=0.0354], protein [p= 0.0037], albumins [p=0.0003] and sodium [p<0.0001] and elevated values of CRP [P=0.0107] and PCT p<0.0001]. High level of inflammatory markers on day 3 after surgery was found to increase the risk of death – for WBC (by 67%), for CRP (by 88%) and for PCT (by 100%).ConclusionsPoor prognosis was more frequent in patients with preoperative high levels of CRP, PCT, anemia, hypoproteinemia and hyponatremia. The risk of death increases significantly if in the immediate postoperative period no distinct decrease in WBC count and of the CRP and PCT level is observed. In such a situation the patients should be qualified earlier for broadened diagnostic workup and for reoperation.

A Retrospective Analysis of 111 Cases of Pneumococcal Pneumonia: Clinical Features and Prognostic Factors

Streptococcus pneumoniae is an important and common pathogen of acute pneumonia. The urinary pneumococcal antigen test has been increasingly used for the diagnosis of pneumococcal pneumonia, but there have been few studies on pneumococcal pneumonia after this test became prevalent. The present study was conducted to characterize the clinical features of pneumococcal pneumonia after the introduction of the urinary antigen test. We retrospectively analyzed 111 cases from 105 patients with pneumococcal pneumonia in our hospital between 2007 and 2010, and collected data regarding background characteristics, laboratory data, isolated bacteria, and clinical courses. The cases analysed included 77 of community-acquired pneumonia (CAP), 21 of healthcare-associated pneumonia (HCAP), and 13 of hospital-acquired pneumonia (HAP). Penicillin-resistant Streptococcus pneumoniae was isolated in 12.5 and 55.6% of non-HAP and HAP cases, respectively. When the cases were divided into 5 groups according to A-DROP scores (0, 1, 2, 3 and 4 points), critical events occurred at 0, 0, 8.7, 15.4, and 75.0% in groups of cases of 0, 1, 2, 3, and 4 points, respectively. Similarly, critical events occurred at 66.7, 4.7, 3.8, and 9.1% in groups of cases according to WBC counts of <4,000, 4,000-9,999, 10,000-19,999, and ≥20,000 cells/µL, respectively. Most of the pneumococcal pneumonia cases occurred as CAP and were treated successfully. HAP cases were frequently penicillin resistant. Elevated A-DROP scores and decreased WBC counts were found to be predictive of critical events.

Chromium (III) induced haematological alterations in Indian common carp, Labeo rohita (Ham.)

The principle source of chromium in water bodies is discharged from industries which use large amount of chromates or dichromates. The freshwater fish, Labeo rohita was used as a model in the present studies to investigate the responses to chromium metal salt contamination in water bodies as it is the most common fish consumed widely in India. Haematological tests were carried out as these are the important diagnostic tools and are equally valuable as indicators of disease or stress due to pollutants and environmental fluctuations. The fishLabeo rohita were exposed to sublethal concentration of chromium chloride for 10, 20 and 30 days to study alterations in certain haematological parameters. The parameters under study were total erythrocyte count (TEC), haemoglobin (Hb gm%), total leucocyte count (TLC), differential leucocyte count (DLC) and pack cell volume (PCV). The studies revealed that chromium chloride-treated Labeo rohita exhibited decreased levels of total erythrocyte count (TEC), haemoglobinn (Hb gm%) and PCV whereas, total leucocyte count (TLC) was increased under thestress of chromium metal The decreased WBC count found after 30 days along with depleted Hb content and RBC count indicate dysfunctioning of haemopoietic systems along with dysleucopoiesis. Leucocytosis observed after 10 and 20 days exposure has been considered to be an adaptation to meet stressful conditions by animals. Increase in macrophages and basophils appears to be protective response during chromium exposure. An increasing trend in the mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were evident in fish exposed to sublethal doses of chromium chloride when compared with the control.

Correlation between maternal milk and infant serum levels of chlorinated pesticides (CP) and the impact of elevated CP on bleeding tendency and immune status in some infants in Egypt

Chlorinated pesticides (CP) are environmentally persistent pollutants that (prenatally through the placenta and post-natally via breastfeeding) are transferred from mother to child. Considering the significant bleeding tendency noted in infants of CP-intoxicated mothers in Egypt, this study aimed to investigate any correlation between levels of these xenobiotics in mothers’ milk and bleeding tendencies of their infants, as well as a possible role of any related immunosuppression in this phenomenon. This study examined 180 newborns presenting with altered bleeding tendencies and their mothers, and 180 normal newborns and their mothers (serving as a controls), selected from the Breastfeeding Unit, Center for Social and Preventive Medicine at the Cairo University Pediatric Hospital. Chlorinated pesticides (e.g., hexachlorocyclohexane, DDT, hepta-chloroepoxide, α- and β-endosulfan, aldrin, endrin, dieldrin) levels and their derivatives were measured in mothers’ milk as well as in serum of neonates using gas chromatography/high resolution mass spectrometry. To link bleeding tendency with lactational intoxication of neonates by CP, newborns’ blood was assessed for: platelet count, bleeding and prothrombin time, liver enzymes, Vitamin K, TNFα, and IL-10. Breast milk CP levels were associated with a higher incidence of bleeding in infants. Interference with the coagulation cascade was supported by changes in prothrombin time (prolonged), platelet counts (decreased), liver enzymes (increased), and serum Vitamin K concentrations (decreased). Moreover, the significant decrease in WBC count and lymphocytes added to depressed cytokine secretion, i.e., TNFα and IL-10, suggested an organochlorine-induced immunotoxicity in infants developmentally exposed to the agents. We conclude that maternal transfer of CP, via breastfeeding or across the placenta, was sufficient to achieve similar CP levels in the serum of their infants; this correlated with a manifesting of altered bleeding tendencies and perturbed cytokine biology in these infants.

Footprints of Response to Hydroxyurea Are in the Hemogram.

Abstract 4619 IntroductionThe salutary effects of hydroxyurea (HU) in sickle dell anemia (SS) include decreased frequency of painful crises, decreased incidence of acute chest syndrome, decreased blood transfusion, and decreased mortality & morbidity. Markers of the response to HU include increased Hb F level, macrocytosis, improved RBC survival & deformability, and decreased WBC count. In this study we report markers of early response to HU by using mixed (random effects) models to determine the relationship between HU usage and the longitudinal trajectories of RBC parameters and the corresponding longitudinal trajectories of reticulocytes parameters. Materials and MethodsRBC and reticulocyte parameters were determined on peripheral blood from selected patients (10 HU subjects and 11 placebo subjects) enrolled in the placebo controlled double blind multicenter study of hydroxyuea (MSH) in SS. Multiple blood samples were obtained during enrollment in the study at a rate of about once every two weeks. The mean weeks of follow-up from enrollment date was 86.6 weeks (range 2-172 weeks). RBC and Reticulocyte indices were measured on a Siemens H*3 RTX (H*3) hematology analyzer. Laser light scattering was used to quantify cell volume, Hb concentration, and the light absorbance of cells stained with oxazine 750 to detect reticulocytes and distinguish them from mature cells. A total of 20,000 cells were counted for each sample. After the volume (V) and Hb concentration (HC) of individual mature RBC and reticulocytes were measured, the Hb content (CH) of individual cells was calculated. Mixed (random effects) models were used to determine the relationship between the longitudinal trajectories of RBC and reticulocyte indices and HU usage. ResultsGoodness-of-fit criteria for the mixed (random effects) models [Akaike Information Criterion (AIC), Akaike Information Criterion, Small Sample Size Corrected Version (AICC) and the Bayesian Information Criterion (BIC)] indicated that the RBC indices modeled more precisely than those of reticulocytes. However, both the RBC indices and reticulocytes provided useful information regarding the relationship between their longitudinal trajectories and HU usage. There was a significant relationship between HU usage and the following longitudinal trajectories: mean cell hemoglobin (CH) (p<.0001), reticulocyte mean cell hemoglobin (CHr) (p<.0001); Cell hemoglobin concentration (CHCM) (p=.0394); Reticulocyte mean cell volume (MCVr) (p=.0019); hemoglobin distribution width (HDW) (p=.0003); and reticulocyte hemoglobin distribution width (HDWr) (p=.0008). HU usage produced higher value trajectories than those generated by placebo usage for the following: CH, CHr; and MCVr. These changes were first detected 10 weeks after starting HU and before the increase in Hb F levels. ConclusionTogether the data suggest that subtle and early markers of response to HU reside in the hemogram. This is a fast, simple and inexpensive methodology provided the lab in question has the capability to determine the indices of both reticulocytes and RBC. Disclosures:No relevant conflicts of interest to declare.

DRDE-07 and its analogues as promising cytoprotectants to nitrogen mustard (HN-2)—An alkylating anticancer and chemical warfare agent

Nitrogen mustard (HN-2), also known as mechlorethamine, is an alkylating anticancer agent as well as blister inducing chemical warfare agent. We evaluated the cytoprotective efficacy of amifostine, DRDE-07 and their analogues, and other antidotes of mustard agents against HN-2. Administration of 1 LD(50) of HN-2 (20mg/kg) percutaneously, decreased WBC count from 24h onwards. Liver glutathione (GSH) level decreased prominently and the maximum depletion was observed on 7th day post-HN-2 administration. Oxidised glutathione (GSSG) level increased significantly at 24h post-administration and subsequently showed a progressive decrease. Hepatic malondialdehyde (MDA) level and percent DNA damage increased progressively following HN-2 administration. The spleen weight decreased progressively and reached a minimum on 3-4 days with subsequent increase. The antidotes were administered repeatedly for 4 and 8 days after percutaneous administration of single sublethal dose (0.5 and 0.25 LD(50)) of HN-2. Treatment with DRDE-07, DRDE-30 and DRDE-35 significantly protected the changes in spleen weight, WBC count, GSH, GSSG, MDA and DNA damage following HN-2 administration (0.5 and 0.25 LD(50)). There was no alteration in the transaminases (AST and ALT), and alkaline phosphatase (ALP) activities, neither with HN-2 nor with antidotes. The present study shows that HN-2 is highly toxic by percutaneous route and DRDE-07, DRDE-30 and DRDE-35 can partially protect it.

A Phase I Dose Escalation Study of KW-2449, An Oral Multi-Kinase Inhibitor against FLT3, Abl, FGFR1 and Aurora in Patients with Relapsed/Refractory AML, ALL and MDS or Resistant/Intolerant CML

Background: Activating mutations of FMS-like tyrosine kinase 3 (FLT3) occur in 30% of patients with de novo AML and confer a poor prognosis. KW-2449 is an oral multi-kinase inhibitor which is highly potent against mutant FLT3 (IC50=0 1–7 nmol/L) and other tyrosine kinases including FGFR1, TrkA, Abl (including T315I) and Aurora A serine threonine kinase. Based on the activity of KW-2449 and its metabolite (M1) in both in vitro and in vivo preclinical leukemia models, KW-2449 was evaluated in patients with leukemia and MDS in this first-in-man study. M1 is formed via monoamine oxidase-B and aldehyde oxidase mediated oxidation of KW-2449.Methods: The objectives were to assess the safety, tolerability, pharmacokinetic (PK) and pharmacodynamic effects of KW-2449 in patients with refractory/relapsed AML, ALL and MDS, or resistant/intolerant CML. A range of daily doses of KW-2449 (12.5–250 mg twice daily, i.e. 25–500 mg/day) on 2 treatment schedules (14 or 28 days) with a recovery period of 7–28 days between cycles were evaluated. Dose limiting toxicity (DLT) and maximum tolerated dose (MTD) were assessed for the 1st cycle. The 28-day schedule was later eliminated. The plasma concentration of KW-2449 and M1 were analyzed by LC-MS/MS method. A plasma inhibitory activity (PIA) assay [Blood 108(10) 3477–83] for P-FLT3 and P-STAT5 was used to measure FLT3 inhibition.Results: 37 patients aged 26–88 years (16 male) were treated at 7 dose levels: 25, 50, 100, 200, 300, 400, and 500 mg daily. Thirty-one patients had AML, 5 CML and 1 ALL. The mean duration of therapy was 2 cycles in AML patients and 4 cycles in CML patients. KW-2449 was rapidly absorbed and metabolized to M1. The half-lives for KW-2449 and M1 were not dose-related and ranged from 2.4 to 4.9 hours and 2.6 to 6.6 hours, respectively. Administration of KW-2449 in a BID regimen led to minor accumulation and was consistent with the short half-life. The PIA assay demonstrated the near complete down-regulation of P-FLT3 and P-STAT5 2 hours post-dose at a dosing level of 400 mg daily. The extent of inhibition was lower at 8 hours and generally absent at 12 hours post-dose. The most frequently reported adverse events (AEs; any grade, regardless of causality) were nausea (70.3%), vomiting (48.6%), fatigue (45.9%), diarrhea (32.4%), dyspnea (29.7%), febrile neutropenia (29.7%), pain in extremity (29.7%), and arthralgia (27.0%). Febrile neutropenia (24.3%), pneumonia (10.8%), and thrombocytopenia (10.8%) were the most frequently reported Grade 3/4 AEs. DLTs occurred in 2 patients: Grade 3 atrial fibrillation (100 mg daily) and Grade 3 nausea and vomiting (500 mg daily). A total of 70 SAEs were reported in 27 patients including 11 on-study deaths; only atrial fibrillation and pleural effusion were considered possibly related to KW-2449. Eight of 31 patients with AML (26%) (FLT3 mutation: 5 positive and 3 negative) and 1 of 5 patients with CML (20%) exhibited a ³ 50% reduction in peripheral blasts and/or bone marrow blasts from baseline to the end of Cycle 1. One patient (500 mg daily) with AML exhibited a > 50% decrease in peripheral blasts, increased platelets, and ANC, and decreased WBC count. A patient with CML (Bcr-Abl T315I +) lost the mutant clone while on KW-2449 treatment.Conclusions: KW-2449 was safe and well tolerated at the dose levels evaluated. There were no complete or partial responses, but transient decreases in peripheral blood and bone marrow blasts were observed, justifying continued investigation of this agent. Sustained inhibition of P-STAT5 and P-FLT3 was not achieved at trough at the highest BID dose evaluated. TID and QID dosing schedules should be evaluated to accommodate the short t1/2 and to achieve sufficient target inhibition.

Rebamipide enema therapy for left-sided ischemic colitis patients accompanied by ulcers: Open label study

To attempt rectal administration of rebamipide in the treatment of ischemic colitis patients with ulcers, and evaluate its effects. We compared 9 ischemic colitis patients (2 men, 7 women) with ulcers treated by bowel rest only from 2000 to 2005 (conventional therapy group), with 6 patients (2 men, 4 women) treated by rebamipide enema therapy in 2006 (rebamipide enema therapy group) and analyzed the mean duration of fasting and hospitalization, degree of ulcer healing, and decrease in WBC count for the two groups. The mean duration of fasting and hospitalization were 2.7+/-1.8 d and 9.2+/-1.5 d in the rebamipide group and 7.9+/-4.1 d and 17.9+/-6.8 d in the control group, respectively, and significantly reduced in the rebamipide group (t= -2.915; P=0.0121 and t= -3.054; P=0.0092). As for the degree of ulcer healing at 7 d after admission, the ulcer score was reduced by 3.5+/-0.5 (points) in the rebamipide group and 2.8+/-0.5 (points) in the control group (t=1.975; P=0.0797), while the decrease in WBC count was 120.0+/-55.8 (x 10(2)/microL) in the rebamipide group and 85.9+/-56.8 (x 10(2)/microL) in the control group (t=1.006; P=0.3360). In left-sided ischemic colitis patients with ulcers, rebamipide enema therapy significantly reduced the duration of fasting and hospitalization, recommending its use as a new and effective therapeutic alternative.

Emergency therapeutic leukapheresis in a case of acute myeloid leukemia M5

Cell separators in India are routinely used for plateletpheresis, peripheral blood stem cell collections and therapeutic plasma exchange. Therapeutic leukapheresis, particularly as an emergency procedure, has been uncommonly performed and reported. Here, a case of a 53-year-old male, diagnosed with acute myeloid leukemia subtype M5 (AML M5) with hyperleukocytosis, who underwent emergency leukaphereis, is reported. After two procedures, there was a decrease of WBC count by 85%, which enabled cytotoxic therapy to be initiated.

The Effects of Guided Imagery on the Immune System: A Critical Review

The research on the effect of guided imagery (GI) on immune system is reviewed and, accordingly, a direction for future research is proposed. Studies suggest that: GI can reduce stress and elevate the immune system; cell-specific imagery affects corresponding WBCs, neutrophils, or lymphocytes; decreases in WBC count occur in the initial stages of GI and relaxation due to fluctuations in WBC production or margination; and changes in WBC count or adherence occur earlier in medical patients. Directions for further investigations: Definition of the ideal WBC count; the effects of long-term practice of GI; and influence of cell-specific imagery on WBCs.

The Mouse Mutation “Thrombocytopenia and Cardiomyopathy” (trac) Is within the ABCG5 Gene: A Single Gene Model for Human Mediterranean Macrothrombocytopenia.

Homozygosity for a new spontaneous mouse mutation named “thrombocytopenia and cardiomyopathy” (trac) results in thrombocytopenia, dilated cardiomyopathy, and infertility. A/J-trac/trac mice show a precipitous drop in platelet numbers and increases in platelet volume by 4 weeks of age. By 2–3 months of age, trac/trac mice have a 20-fold decrease in platelet number, a 3-fold increase in platelet volume, and a greatly increased bleeding time. Blood smears showed abnormally large platelets and megakaryocytoid cells. Homozygotes (trac/trac) also developed mild microcytic anemia accompanied by the presence of stomatocytes, a doubling of reticulocyte numbers, and a two-fold decrease in WBC counts. Increased numbers of megakaryocytes were present in bone marrow, spleen, and lungs. Ultrastructural studies of trac/trac megakaryocytes showed a poorly developed demarcation system and a failure to form platelet territories. The trac/trac platelets were enlarged, spherical, and contained numerous small alpha granules. The thrombocytopenia was not associated with defects intrinsic to bone marrow progenitor cells. Although thrombopoietin (TPO) levels were decreased, TPO treatment failed to reverse the thrombocytopenia. To identify the responsible gene, we produced a fine structure genetic map of a 5 megabase interval containing the trac locus on mouse chromosome 17. The human syntenic region is Chr 2p21–p22. Analyses of 1100 F2 progeny from intercross matings of (A/J x C57BL/6) F1 +/trac mice narrowed the interval to 0.3 Mb containing 4 genes. Sequencing of these genes revealed a G to A mutation at base 1435 (refseq nm031884) of Abcg5 (ATP-binding cassette sub-family G, member 5). This G>A base change results in a tryptophan codon (UGG) at amino acid position 463(uniprot) being changed to a premature stop codon (UAG). The transmembrane helices prediction program, TMHMM, predicts that the premature stop codon would truncate the last four of the six transmembrane domains of the ABCG5 protein. No DNA alterations were found in any of the other candidate genes. Genetic crosses of +/trac mice with mice doubly transgenic for the closely linked human ABCG5 and ABCG8 genes (stock B6SJL-Tg(ABCG5/ABCG8)14-2Hobb/J) showed that the transgenes normalized platelet counts and volumes in trac/trac mice. ABCG5 (sterolin-1) functions as part of a heterodimer, with ABCG8, that regulates plant sterol uptake. The trac/trac mutant mice have greatly elevated plasma levels of plant sterols. When placed on a phytosterol-free diet, the thrombocytopenia is reversed. Recent studies have shown that Mediterranean Macrothrombocytopenia is caused by mutations in ABCG5 or ABCG8. Identification of the molecular basis of the mouse Abcg5trac mutation provides a new model for studying the role of phytosterols in pathogenic changes in the hematopoietic, cardiovascular, and reproductive systems.

Immunological Responses in CLL Patients Vaccinated with Autologous Dendritic Cells Loaded with Apoptotic Bodies (Apo-DC).

We have previously established the suitability of dendritic cells that have endocytosed apoptotic B-CLL cells (Apo-DC) as an antigen presentation platform to stimulate autologous antileukemic immune responses. The safety as well as immunological and clinical efficacy of Apo-DC vaccination was tested in a phase I/II clinical trial. CLL patients (Rai stage 0–2) who did not require concurrent therapy were leukapheresed and CD14+ monocytes as well as CD19+ B-CLL cells were purified immunomagnetically. Dendritic cells were generated ex vivo with GM-CSF and IL-4. Apoptotic bodies were obtained by irradiating B-CLL cells and overnight culture. DC were allowed to endocytose the apoptotic bodies and further matured with TNFα. The first cohort of 5 patients received 4 doses of the vaccine 2 weeks apart and a 5th dose at week 14. Each vaccine dose comprised of 107 autologous Apo-DC. The second cohort of 5 patients received the vaccine in a similar schedule with the addition of 75 μg of GM-CSF (Leukine™, Berlex Laboratories, Richmond VA) with every vaccine administration. Patients are scheduled to be monitored for a minimum of 52 weeks which has been completed for all of the patients in cohort 1 and is ongoing for cohort 2 The vaccine was well tolerated and occasional, low-grade toxicity was associated with the administration of GM-CSF. 4/5 patients in cohort 1 and 3/5 patients in cohort 2 responded immunologically in terms of increase in T cell proliferation or γ-IFN ELISPOT against autologous leukemic targets compared to prevaccination levels. Analysis of T cell responses at week 8 and week 16 following vaccination did not demonstrate an obvious augmentation of immunity by GM-CSF. One patient in cohort 1 demonstrated a transient decrease in WBC counts but no significant clinical responses have been noted thus far. Our results demonstrate that it is feasible to enrich monocyte precursors and generate Apo-DC for vaccinating patients with high count B-CLL. No significant toxicity was associated with this approach to vaccination therapy. The clinical efficacy of Apo-DC vaccination in CLL remains to be determined at the termination of the clinical trial.

Association of the glycophorin A gene mutation in peripheral erythrocytes and chronic benzene poisoning

To explore the association of the glycophorin A(GPA) gene mutation in peripheral erythrocytes and chronic benzene poisoning. Sixty-three patients with chronic benzene poisonings and 45 benzene-exposed workers who were engaged in the same job title were investigated. Fluorescence immunolabeling technique and flow cytometry were used to detect GPA mutation frequency in peripheral read blood cell. A significant decrease in WBC count and neutrophil count was found in patients with chronic benzene poisoning compared with control individuals (P<0.01). The WBC count and neutrophil count both decreased along with the GPA-NN frequency, and the trends were significant(P<0.05).Both WBC counts and neutrophil counts decreased as the frequency, and trends were significant(P<0.05). GPA-NN frequency increased along with the accumulative exposure score, and the trend was significant (P = 0.0026). There was no significant trend between the GPA-Nphi frequency and the accumulative exposure score (P = 0.2037). A decrease in WBC count and neutrophil count is found in patients with chronic benzene poisoning, which can arise from genetic damage in bone marrow stem cells, namely gene-duplicating mutations (NN) at the GPA locus in bone marrow cells of MN-heterozygous subjects, GPA-NN mutagens contributed to the pathogenesis of chronic benzene poisoning.

In Vivo Phycocyanin Treatment of S180 Bearing-Mice: Increase in Immunic Function and Inhibition of Tumor Growth.

Objectives: To explore the effect of Phycocyanin (PC), with or without Cyclophosphamide(CY), on the growth inhibition of S180 cells and on the immunity improvement of S180-burdened mice. And to explore the anti-tumor mechanism of Phycocyanin.Methods: Each 0.2ml of S180 cells, a sarcoma cell line cells, at the concentration of 2×107 cells/ml, were inoculated into the right axilla of each test mouse. These mice were randomizely divided into 4 groups: PC-treated group (including 50−, 75− and 100mg/kg·d- subgroups); CY-treated group (20mg/kg·d); PC plus CY-treated group and normal saline(NS)-treated group which served as control group. 24 hours after the inoculation, PC or CY or PC plus CY or NS, at a final volume of 0.2ml, were injected to the abdominal cavity of each mouse in different group, and consecutively for 7 days. These mice were killed after the seventh treatment, then the peripheral blood, the whole tumor and the spleen were collected from each mouse. Each mouse's tumor weight was measured and blood routine exam was done. The serum level of IL-2, IFN-γ and TNF-α in each mouse was measured with ELISA method and the result was shown as pg/ml. Spleen cells from each mouse were collected and cultured respectively with ConA for 36 hours, and splenic T lymphocyte proliferation was measured.Results: PC inhibited the growth of S180 cells at all three tested dosages, 50mg/kg·d, 75mg/kg·d and 100mg/kg·d, and the inhibition rates were 41.41%, 37.37% and 36.36%, respectively. PC-treatment increased WBC counts in PB of tumor-bearing mice. The results also showed that PC-treatment and tumor-bearing situation increased the spleen weight and enhanced the splenic T lymphocyte proliferation induced by ConA. PC-treatment did not reduce the body weight of tumor-bearing mice. Among these three dosage, 50mg/kg·d was shown to have the best effect in the above mentioned parameters, as well as in the increase of IL-2, IFN-γ and TNF-α secretion. PC at dosages of 75mg/kg·d and 100mg/kg·d just increased the serum level of IFN-γ and TNF-α, without the increase of IL-2. Compared with PC-treatment, CY-treatment diminished serum level of IL-2, IFN-γ and TNF-α. PC (75mg/kg·d) combined with CY-treatment increased the tumor inhibition rate to 92.93%.Conclusions: 1, PC-treatment inhibited the growth of S180 cells. PC plus CY treatment showed more potential effect of growth inhibition. 2, PC enhanced markedly the immune function of S180-bearing mice in terms of increasing the spleen weight and the serum level of IFN-γ and TNF-α, and enhancing T lymphocyte proliferation induced by ConA. Those might account for the anti-tumor mechanism of PC. 3, PC-treatment antagonized the decrease in WBC count induced by CY-treatment. No obvious side effect of PC was found in the present study. Therefore, PC alone or combine with other chemotherapy drugs may offer a new approach in the treatment of hematological malignancies.

A Ph-Negative Myeloproliferative Disorder with JAK2 Mutation Disclosed by Imatinib Therapy in Two Patients with Ph+ CML.

Protein tyrosine kinases play a crucial role in normal and neoplastic cell development. Recently, a unique clonal mutation in the JAK2 gene that results in a valine to phenylalanine substitution at position 617 (V617F), leading to constitutive JAK2 activation and abnormal signaling, was found in the majority of BCR-ABL negative chronic myeloproliferative disorders (CMPD) patients. Although JAK2 activation by tyrosine phosphorilation is involved in the BCR-ABL transformation of CML cells, V617F mutation is not usually observed in Ph-positive leukemias.We describe two patients with BCR/ABL+ (p190) CML which followed a Ph- CMPD characterized by JAK2 mutation.Patient 1 was diagnosed as having a idiopathic myelofibrosis in Oct 1997. At that time a Ph-chromosome was not detected. The patient remained untreated until March 1999, when WBC started to increase up to 90.000/cmm with a parallel drop in platelet count and increase in spleen size. Hydroxiurea was started with partial hematological remission. In Jan 2000 the presence of a BCR/ABL transcript (p190 type) and a Ph-chromosome in 84% of the cells were observed. Imatinib treatment resulted in a initial improvement, soon followed by progressive increase in the WBC and platelet counts, in spite of a reduction of Ph+ cells to 4%. Hydroxyurea was added but hematological control remained unsatisfactory and imatinib treatment was stopped after 6 months. The patient continued on hydroxyurea for the following year. In 2002 he developed a myeloid blast crisis; karyotype was normal, 4.8% of the cells were Ph+ at FISH and the p190 transcript was still present at low levels.Patient 2 was diagnosed as essential throbocythemia in 1990; cytogenetic analysis showed a normal karyotype. A good control of the thrombocytosis was obtained with hydroxyurea for more than 10 years. In 2003 the hematological data showed leukocytosis (up to WBC 42.900/cmm), immature myeloid cells and 16% monocytes in PB. A masked Ph chromosome was observed by FISH in 47% of cells. RT-PCR was positive for a p190 transcript. The patient started therapy with imatinib which induced a rapid decrease in WBC count; however, the platelet count rised up to 977.000/cmm and hydroxiurea was added again. In subsequent bone marrow samples a progressive decrease of BCR/ABL positive cells to 2% was observed and a BCR-ABL/ABL ratio of 0.18 % was reached in January 2006. However, continuous hydroxyurea therapy was needed to partially control thrombocytosis.Both patients were tested for the presence of JAK2 mutation using the ABI Prims 3100 genetic analyzer and a JAK2-V617F mutation was present. Presence of two clones, possibly derived from a common abnormal stem cell, can be hypothesized in these cases. The possibility of a multistep patogenesis for CML has been widely discussed; in these patients, CML apparently developed as a late event, partially replacing a Ph-CMPD cell population characterized by the JAK2 mutation. Imatinib treatment allowed the reexpansion of the Ph- cells resulting in a overall poor hematological control. Both patients only displayed p190 BCR/ABL transcript, a quite unusual finding in chronic phase CML. It is not clear if any relationship may sussist between this molecular event and the unusual clinical history of these patients.

Pulmonary alveolar proteinosis with myeloproliferative syndrome with myelodysplasia: Bronchoalveolar lavage reduces white blood cell count

Pulmonary alveolar proteinosis (PAP) is a rare disorder characterized by surfactant component accumulation in the alveolar space. Primary PAP is likely an autoimmune disorder caused by antibodies to granulocyte-macrophage colony stimulating factor (GM-CSF). When an underlying disease causes PAP, this is called secondary PAP. Hematologic malignancies are an important cause of secondary PAP. As the pathogenesis of primary PAP has become more fully understood, improvements in diagnostic and therapeutic approaches have followed. However, when PAP is secondary to an underlying hematologic malignancy, much remains unclear. Here we describe for the first time a patient with hybrid myelodysplastic syndrome/myeloproliferative syndrome and PAP who had a marked decrease in her white blood cell count following a transbronchial biopsy accompanied by bronchoalveolar lavage (BAL). Similar significant decreases in WBC count accompanied clinical improvement following two unilateral BALs. Given that patients with pulmonary alveolar proteinosis frequently have elevated GM-CSF in bronchoalveolar fluid, this observation provides a unique vantage point to understand the pathophysiology of secondary PAP.

Thrombocytopenia and Cardiomyopathy (trac): A New Single Gene Mutation Resulting in Severe Platelet and Heart Abnormalities in Mice.

We have discovered a new spontaneous mouse mutation, named “ thrombocytopenia and cardiomyopathy” (trac), in a colony of A/J mice at The Jackson Laboratory. Homozygosity for this recessive mutation results in severe pathologic changes in platelets, dilated cardiomyopathy, and infertility. Although platelet counts in A/J-trac/trac mice are normal at 3 weeks of age, there is a precipitous drop in platelet numbers and increases in platelet volumes by 4 weeks of age. Quantitative analyses of peripheral blood in A/J-trac/trac mice at 10 weeks of age showed a 20-fold reduction in platelet numbers accompanied by a 3-fold increase in platelet volumes compared with non-mutant (+/?) sex-matched controls. Other abnormalities in trac/trac platelets include greatly increased size of the microtubule coil and increased CD41 expression. Thrombocytopenia in trac/trac mice was associated with greatly increased bleeding times. Homozygotes (trac/trac) showed other hematological changes that included mild microcytic anemia accompanied by a doubling of reticulocyte numbers and two-fold decrease in WBC counts. Histological examination of blood smears confirmed abnormally enlarged platelets as well as the presence of megakaryocytoid cells. Increased numbers of megakaryocytes were present in BM, spleen, and lungs of trac/trac mice. Ultrastructural studies showed a lack of well-defined demarcation channels in trac/trac megakaryocytes. In order to determine whether the trac/trac platelet abnormality is intrinsic to defects at the level of BM progenitor cells, reciprocal BM chimeras were examined. Lethally irradiated +/+ recipients of 5 x 106 trac/trac BM cells were reconstituted with normal platelet numbers and volumes. In contrast, lethally irradiated trac/trac recipients of 5 x 106 +/+ BM cells remained thrombocytopenic. Backcrossing of the trac mutation onto the BALB/cBy strain background facilitated the identification of homozygotes prior to weaning and indicated the presence of modifying genes. Our preliminary data has produced a fine structure map of a 5 megabase interval containing the trac locus on mouse Chromosome 17. The human synteneic region is Chr 2p21-p22. We hypothesize that identification of the molecular basis of this mutation will reveal a novel gene that plays a critical role in the homeostasis of platelets and cardiomyocytes.

Occurrence of Side Effects from Anti-Tuberculosis Drugs in Urban Nepalese Population under DOTS Treatment

Tuberculosis is one of the foremost public health problems causing an enormous burden of suffering and deaths. Chemotherapy is the basic approach to clinical tuberculosis control. 1Essential anti-tuberculosis drugs are Isoniazide (INH), Rifampicin (RMP), Pyrazinamide (PZA), Ethambutol (E) and Streptomycin (S). Department of Pharmacy, KU, KUTH, and Dhulikhel conducted this study in collaboration with German-Nepal Tuberculosis Project (GENETUP) and Korea-Thimi Friendship Hospital during March 2005 to July 2005. Total 70 newly diagnosed pulmonary or extra-pulmonary tuberculosis patients were included in the present study. Patients were kept under observation during whole treatment period. Clinical, hematological and biochemical monitoring form the basis of diagnosing side effects due to anti-tuberculosis therapy. Among the 70 cases studied, 70% were male and 30% were female. There was significant increase in the bilirubin total, bilirubin direct, AST, ALT, alkaline phosphatase, eosinophils, lymphocytes, monocytes after anti-tuberculosis treatment whereas there was significant decrease in WBC count, neutrophils, platelets count, and ESR after anti-tuberculosis treatment. Serum level of creatinine was normal before and after anti- tuberculosis treatment. In our study, slight increase in uric acid level was detected after anti- tuberculosis treatment when compared to pretreatment, though it was in the normal range. Of the 70 patients treated for tuberculosis, 80% of the total number of patients reported at least one type of side effects. 24 patients (34.29%) had experienced major side effects. 8 elderly patients (44.44%) and 16 younger patients (30.76%) had experienced major side effects. Female gender (38.10%), alcoholics (38.89%) and sputum-smear positive (32.14%) were associated with increased occurrence of major side effects. The overall side effects indicate that side effects are not to be neglected. It was experienced that timely detection and temporary withdrawal of the offending agent can completely cure anti-tuberculosis drugs- induced side effects. For this, careful patient monitoring clinically, hematologically and biochemically especially during first few weeks of chemotherapy and patient counseling regarding side effects of the anti-TB medications are necessary.