Changes In Blood Counts Research Articles

Radiothérapie métabolique des tumeurs endocrines gastro-entéro-pancréatiques par les analogues radio-marqués de la somatostatine

Surgery is the only curative treatment of gastro-entero-pancreatic endocrine tumors. In inoperable or metastasized forms, therapeutic options are limited. The metabolic or systemic radiotherapy, using radiolabeled somatostatin analogs, constitutes a new therapeutic alternative, currently in development which requires the presence of high affinity somatostatin receptors on tumoral cells. Using (111)In-pentetreotide, the main result is a symptomatic effect. With new somatostatin analogs coupled to B- emitters, such as Octreother or (177)Lu-DOTA-Octreotate, 10 to 30% of objective tumoral responses are observed in progressive patients, unresponsive to conventional treatments. Such results are explained by the high affinity for somatostatin receptors and the large emission diameter of these radiolabeled compounds. Renal toxicity is limited by amino-acid infusion whereas changes in blood count are usually moderate and transient. Multicentric prospective studies are necessary to identify the predictive factors of tumoral response and toxicity. The prospects are related to the development of new radiopharmaceuticals, even more specific of somatostatin receptors sub-types and to the use of other peptide analogues whose applications will overflow the framework of endocrine tumours.

A Multicenter Case-Control Study on Predictive Factors Distinguishing Childhood Leukemia From Juvenile Rheumatoid Arthritis

Acute lymphocytic leukemia (ALL) often presents with musculoskeletal concerns such as pain or swelling, even before appearance of blasts in the peripheral blood. Such presentation may lead to misdiagnosis of a child with juvenile rheumatoid arthritis (JRA). This study was designed to identify the predictive factors for leukemia using basic clinical and laboratory information. A retrospective chart review was performed using a simple questionnaire to compare the clinical and laboratory findings present during the initial visit to a pediatric rheumatology clinic for 277 children who were ultimately diagnosed with either JRA (n = 206) or ALL (n = 71). Sensitivity and specificity analysis of a variety of parameters, both singly and in combination, was performed to identify predictive value for ALL. The majority (75%) of children with ALL did not have blasts in the peripheral blood at the time of evaluation by pediatric rheumatologists. In children presenting with unexplained musculoskeletal complaints, the 3 most important factors that predicted a diagnosis of ALL were low white blood cell count (< 4 x 10(9)/L), low-normal platelet count (150-250 x 10(9)/L), and history of nighttime pain. In the presence of all 3, the sensitivity and specificity for a diagnosis of ALL were 100% and 85%, respectively. Other findings, including antinuclear antibody, rash, and objective signs of arthritis, were not helpful in differentiating between these diagnoses because they occurred at similar rates in both groups. When a child develops new-onset bone-joint complaints, the presence of subtle complete blood count changes combined with nighttime pain should lead to consideration of leukemia as the underlying cause.

Clinical applications of 188Re-labelled radiopharmaceuticals for radionuclide therapy

188Re is a radionuclide in which there is widespread interest for therapeutic purposes because of its favourable physical characteristics. Moreover, it can be eluted from an on-site installable 188W/188Re generator, which has a useful shelf-life of several months. Most of the clinical experiences gained with 188Re concern the use of 188Re-1,1-hydroxyethylidenediphosphonate (188Re-HEDP) for bone pain palliation in patients suffering prostate cancer. The maximum tolerated activity was 3.3 GBq 188Re-HEDP and if the platelet count exceeded 200 x 10(9) l(-1), the administration of 4.4 GBq appeared safe. Evidence for repeated administrations of 188Re-HEDP rather than single injections was established. In general, pain palliation occurs in 60-92% of patients with only moderate transient toxicity, mainly related to changes in blood counts. Also in haematology, radioimmunotherapy by means of 188Re might play a role by selectively targeting the bone marrow in patients undergoing conditioning prior to haematopoetic stem cell transplantation. The feasibility of such an approach was proven using a Re-labelled monoclonal antibody directed toward the CD66-antigen. More recently, encouraging safety data on locoregional treatment of primary liver tumours using 188Re-labelled lipiodol were reported. The normal organs at greatest risk for toxicity are the normal liver and the lungs. About 50% of the patients reported mild and transient side effects, mainly consisting of low grade fever, right hypochondrial discomfort or aggravation of pre-existing liver impairment. Besides the applications in oncology 188Re-based therapies have also been pioneered for benign condition such as prevention of re-stenosis following angioplasty and for radiosynovectomy in cases of refractory arthritis.

Long-Term Follow up of Healthy Donors Receiving G-CSF for Peripheral Blood Progenitor Cell Mobilization and Collection.

Background: Peripheral blood progenitor cells (PBPC) are a common source of hematopoietic stem cells for allogeneic transplantation. Information about long-term follow up and donor safety is, however, very limited.Study design and methods: A National Donor Registry was developed in Spain in 1999 directed to know the short and long-term effects of G-CSF administration on healthy donors for PBPC mobilization. A total of 1287 donors (590 M/697 F; median age, 37 years, range 1–74) have been registered. G-CSF, either filgrastim (1041 cases) or lenograstim (291 cases) was the only cytokine administered. Median (range) dose of G-CSF was 10 (5–23) mcg/kg/day, for a median of 4 days. Sixty-five donors underwent more than one mobilization procedure. A baseline investigation was performed in every donor 2–4 weeks before G-CSF administration and follow up investigations were planned at 4 weeks, and thereafter annually up to 5 years after mobilization.Results: In 588 donors at least one of the scheduled controls have been done, and 187 donors have been followed for more than two years. The peripheral white blood cell (WBC) count decreased significantly 4 weeks after leukaphereses, from 6.4 x109/L at baseline to 5.9 x109/L (P <0.0001). WBC were normal 1 year after mobilization and maintained similar values during the remaining follow up. Four weeks after PBPC collection, hemoglbin presented lower values than that observed before mobilization (14.3 vs. 13.8 g/dL; P <0.0001). As with WBC, hemoglobin one year after G-CSF administration was similar to baseline values. Regarding short-tem complications, 740 donors complained of some side-effect during G-CSF administration, being bone pain (89%) and headache (31%) the symptoms most commonly observed. One donor presented with a splenic rupture 12 hours after last leukapheresis that required splenectomy. During follow up, three donors (0.23%) have developed neoplastic diseases (colon carcinoma, lung cancer, and melanoma in choroidal region of the left eye) at 3, 5 and 6 years following G-CSF administration. Finally, one donor developed thrombocytopenia 1 month after mobilization. The bone marrow study excluded myelodysplastic syndrome and was diagnosed as having idiopatic thrombocytopenic purpura.Conclusion: Our analysis represents the larger study of follow up of donors receiving G-CSF for PBPC mobilization and reveals that clinically side-effects of G-CSF administration in healthy donors are generally mild. Changes in blood counts were minimal and restored normal values within one year after G-CSF. With longer follow up, isolated neoplastic diseases have been detected, although no hematologic malignancy has been reported and no other unusual diseases have been observed. Greater number of donors and longer follow-up are, however, needed to definitively guarantee the safety of the procedure.

Bone Marrow Connexin-43 Expression Is Critical for Hematopoietic Regeneration after Chemotherapy.

Contact between bone marrow (BM) hematopoietic stem cells (HSC) and osteoblast/stromal (OS) cells has been shown to be crucial in the regulation of hematopoiesis. However, very little is known about the regulatory mechanisms of direct cell-to-cell communication in the hematopoietic microenvironment. Gap junctions (GJs) represent the best described intercellular communication (IC) system, and they are characterized by the existence of plaques of narrow channels between contacting cells. Each cell contributes with one hemichannel, which is composed of six proteins, called connexins. Connexin 43 (Cx43) is expressed by BM OS cells. Cx43 has been associated with the cadherin/β-catenin signaling pathway, recently reported as relevant in the OS/HSC interaction at the stem cell niche. Multiple osteogenic defects have been reported in human Cx43 mutations and Cx43 has been shown to be essential in controlling osteoblast functions. BM Cx43 expression is up-regulated up to 100-fold by 5-fluorouracil (5-FU) treatment. Due to the perinatal death of Cx43 germline null mice, a conditional genetic approach was employed to study the role of Cx43 in stem cell proliferation and differentiation. The interferon-inducible Mx1 gene is expressed by both hematopoietic and stromal BM cells. Therefore, we crossed Cx43flox/+ mice with Mx1-Cre transgenic (Mx1-CreTg/−) mice. Cx43+/+:Mx1-CreTg/−and Cx43flox/flox:Mx1-CreTg/− littermates have been analyzed. Gene deletion was induced in vivo by injecting the interferon-inducer polyI:C (8 injections of 300 μg every other day), generating control (Cx43+) and Cx43-deficient (Cx43KO) mice. After one week, Cx43+ and Cx43KO mice were injected with 5-FU (150 mg/Kg i.v.). Cx43 expression in Cx43KO BM was markedly reduced (>80%) as analyzed on day +14 post-5-FU treatment. Cx43 deficiency did not induce a significant change in peripheral blood counts before 5-FU treatment, but the hematopoiesis recovery after 5-FU treatment was severely impaired as demonstrated by absence of recovery of peripheral blood counts, including profound neutropenia, anemia with reticulocytopenia, thrombocytopenia and a 5 to 8-fold decrease of cellularity and hematopoietic progenitor content (granulomacrophagic colony-forming-units -CFU-GM-, erythroid burst forming units (BFU-E) and mixed colony forming units -CFU-mix-) in BM and spleen on day +14 post-5-FU treatment. However, the femoral content of Lin−/c-kit+/Sca1+ cells in Cx43KO BM was maintained when compared to Cx43+ BM (139±19 vs 117±32 x 103 Lin−/c-kit+/Sca1+ cells per femur, respectively). Short-term competitive repopulation ability of Cx43KO BM cells was diminished as compared to Cx43+ mice (5.9±0.35% vs 22.2±4.6%, respectively, p<0.01), specifically for myeloid (9.2±1.4% vs 35.8±4.3%, respectively, p<0.001) and B lymphoid (0.4±0.2% vs 3.0±1.0%, respectively, p<0.01) cells, but showed spared long-term (6-month) competitive repopulation ability with roughly normal hematopoietic differentiation. Altogether, these data suggest that hematopoietic regeneration after cycle-specific chemotherapy is blocked in Cx43-deficient mice at the long-term HSC repopulating level. Cx43 expression within the BM appears to be crucial in the development of an efficient response to hematopoietic stress.

Outcomes in Patients with Splenic Marginal Zone Lymphoma or Marginal Zone Leukemia/Lymphoma Treated with Immunotherapy, Chemoimmunotherapy, or Chemotherapy.

Purpose: The optimal management of splenic marginal zone lymphoma (SMZL) or marginal-zone leukemia/lymphoma (MZL) is controversial. We retrospectively assessed the clinical outcome of patients with SMZL/MZL treated with systemic therapy.Patients and Methods: Patients were assessed by the time of their first treatment at U. T. M. D. Anderson Cancer Center, Dept. of Leukemia (5/95 to 10/04). Diagnosis was confirmed in 70 patients by slide review. The indications for treatment were the same as those used for patients with CLL.Results: The median age was 64 years (range, 33–88); and 61% of patients had monoclonal gammopathy. The median number of CD20 molecules/cell was 65.2 x 103 (16–260 x 103). Of the patients who required systemic therapy, 26 were treated with immunotherapy (rituximab, 25; alemtuzumab, 1); 6 with chemoimmunotherapy (CI/T; rituximab combined with a fludarabine-based regimen); and 11 with chemotherapy (C/T). Ten patients had splenectomy, and 17 were in the observation group. The overall response rates were 88% (CR, 31%) in the immunotherapy group, 83% (CR, 17%) in the CI/T group, and 55% (CR, 18%) in the C/T group. The median follow-up was 2.7 years. Patients treated with immunotherapy +/− C/T had higher rates of overall and failure-free survival compared with those treated with C/T. [Display omitted] In univariate analysis, the only factors predicting longer survival were age >60 years (p=0.01) and immunotherapy +/− chemotherapy (p=0.04). Seventeen (24%) of 70 patients had other malignancies prior to (n=8) or after (n=8) treatment of SMZL/MZL or both (n=1). Changes in bone marrow and blood counts in patients treated with rituximab (n=25) were compared with those of patients who had splenectomy as initial therapy (n=17, including 7 who had subsequent therapies). Rituximab resulted in the disappearance of a palpable spleen (median size, 6 cm; range 0–20cm) in 23 (92%) patients. Rituximab was superior to splenectomy in normalizing the white blood cell (WBC) counts (p<0.001) and absolute lymphocyte counts (ALC)(p<0.001). Splenectomy resulted in higher platelet counts compared with rituximab, but platelet counts remained within the normal range in all patients treated with rituximab. Hemoglobin levels and bone marrow cellularity did not reach statistical significance, but there was a trend towards a significantly lower proportion of lymphocytes in patients treated with rituximab (p=0.1).Conclusions: Rituximab with or without C/T induces durable remissions and prolongs survival in patients with SMZL/MZL, probably because CD20 molecules/cell are higher in SMZL/MZL than in CLL. Our data demonstrate that rituximab effectively controls SMZL, as evidenced by improvement in WBC and ALC and splenomegaly, and may be the treatment of choice, at least in older SMZL patients with comorbid diseases. Clinical trials of immunotherapy or CI/T are warranted.

Reproductive Parameters of Adult Birds and Morphophysiological Characteristics of Chicks in the Pied Flycatcher (Ficedula hypoleuca Pall.) in Technogenically Polluted Habitats

Effects of toxic emissions (heavy metals and sulfur dioxide) on reproductive parameters of adult pied flycatchers (Ficedula hypoleuca Pall.) and morphophysiological characteristics of their progeny have been studied in the vicinity of the Middle Ural Copper-Smelting Plant. In polluted areas, the fecundity of adult birds is lower, mortality among the progeny is higher, and changes in the physiological parameters of birds are observed. The effects of toxic load are better manifested in nestlings than in adult birds. They include an increase in the liver index, the reduction of hemoglobin concentration, and an increase in the proportion of immature erythrocytes in the peripheral blood. The proportion of anemic birds also increases. The weakening of nestlings facilitates their infestation by the larvae of parasitic flies. Changes in the differential blood count of nestlings reflect the combined impact of toxic pollutants and parasites.

Adverse Reactions to Anticoagulants and to Antiplatelet Drugs Recorded by the German Spontaneous Reporting System

According to their code of professional conduct, German physicians are obliged to report suspected cases of adverse drug reactions (ADRs) to the Drug Commission of the German Medical Association (AkdA). On the basis of an agreement between the German Medical Association and the Federal Institute for Drugs and Medical Devices (BfArM) a common pharmacovigilance database within the German spontaneous reporting system was created. A user-friendly application program developed in-house enables the user to conduct searches about reported ADRs covering a wide variety of questions within a short period of time. ADRs caused by anticoagulants and by antiplatelet drugs still belong to the most reported adverse events. The most frequently reported suspected drugs are heparins, followed by ticlopidine, phenprocoumon, acetylsalicylic acid, and clopidogrel. Bleeding complications are the most often described ADR symptoms of any anticoagulation therapy, especially of phenprocoumon and acetylsalicylic acid. Another serious ADR is heparin-induced thrombocytopenia or changes in blood counts (CBC) due to ticlopidine and clopidogrel. During the past few years a reduction in severe reactions, such as cerebral hemorrhage, especially with fatal outcome was detectable because of better clinical management of oral anticoagulant therapy and of adverse events concerning heparin.

Changes in blood counts from cycle 1 to cycle 2 appear to be associated with an increased risk of grade 3 or 4 toxicity

Changes in blood counts from cycle 1 to cycle 2 appear to be associated with an increased risk of grade 3 or 4 toxicity

Changes in blood counts from chemotherapy cycle 1 to cycle 2 appear to be associated with an increased risk of grade 3 or 4 toxicity

Changes in blood counts from chemotherapy cycle 1 to cycle 2 appear to be associated with an increased risk of grade 3 or 4 toxicity

High Doses of Taurolidine Inhibit Advanced Intraperitoneal Tumor Growth in Rats

The antitumor agent taurolidine (TRD) affects tumor growth in animals. Thus far, no animal studies have been published concerning the systemic or local toxicity and the effectiveness of long-term intraperitoneal (i.p.) and intravenous (i.v.) administration on advanced tumor growths. In a first experiment (A) the systemic toxicity of the liver and kidneys was examined only after i.v. treatment in 40 rats (BD IX). For local toxicity the superior vena cava (SVC) was histologically analyzed. In a second study (B) 20,000 colon adenocarcinoma cells (DHD/K12/TRb) were initially applied i.p. after laparotomy in 80 rats (BD IX). After 28 days a port catheter system was placed in the SVC and left for 1 week. The animals were randomized into eight groups (n = 10) and received a 7-day treatment (eight hourly, 1 ml): 1, 2, 3% TRD or Ringer's solution (control group) either i.p. or i.v. Total i.p. tumor weight was measured 4 weeks after the end of the therapy. Side effects on differential blood counts and animal weight changes were examined. No organ lesions were detected in liver, kidneys, and SVC in experiment A. The i.v. administration of 2% TRD (P = 0.034) and 3% TRD (P = 0.05) as well the i.p. application of 2% TRD (P = 0.05) decreased the development of advanced i.p. tumor lesions. No changes of differential blood count nor relevant animal weight changes resulted. Three port catheter-related infections were examined. TRD does not impair the liver tissue, kidneys, SVC, and leucopoiesis. The intravenous therapy of 2% TRD is safe and anti-tumorigenic in advanced local tumor growth in rats.

696. Tumor Vaccine Combined with Cytokines and Suicide Gene Therapy for Canine Spontaneous Melanoma

Canine malignant melanoma, a spontaneous, highly aggressive and metastatic tumor that usually appears in the oral cavity, nail bed and mucocutaneous junction, is a useful animal model of human malignant melanoma. As an adjuvant treatment of surgery, we evaluated the safety, efficacy, and antitumoral effects of a vaccine containing both melanoma cells and genetically modified cells secreting human granulocyte-macrophage colony-stimulating factor (hGM-CSF) and human interleukin-2 (hIL-2), simultaneous to herpes simplex thymidine kinase (HSVtk) suicide gene therapy. Treated canine patients (T) were subjected to: (i) surgery followed by multiple injections of lipid-complexed plasmid DNA encoding HSVtk co-administrated with ganciclovir (GCV) in the areas surrounding the excised tumor; (ii) periodic subcutaneous injection of living irradiated xenogenic cells secreting hGM-CSF and hIL-2 mixed with formolized autologous tumor cells concurrent with (iii) periodic multiple intra and/or peritumoral injections of HSVtk-carrying lipoplexes and GCV, in cases of partial tumor resection or local relapse. The study was intended as an initial weekly treatment for 5-weeks, followed by a biweekly, a monthly and finally a bimonthly treatment according to the patient's response. Control patients (C) were subjected only to surgical tumor excision. This treatment did neither produce significant changes in complete blood counts or serum biochemical parameters nor induce systemic toxicity, organ dysfunction or fever in any patient. Some patients developed transient peritumoral edema after injection and/or induration at the vaccination sites. Treatment diminished local relapse from 95 % (C) to 38 % (T) and regional metastases (linfonodules) from 32 % (C) to 8 % (T), increasing survival free of distant metastases (as evidenced by X-rays and ultrasound) from 50 (C) to 71 (T) %. These effects suggest systemic antitumor immunity. In addition, there was a 3-fold (T/C) rise of mean survival rate of dead patients (p<0.001, ANOVA), and a 4-fold (T/C) rise of median survival rate (p<0.001, Kaplan-Meier analysis). The complete treatment induced local antitumor activity, as evidenced by tumor regression and the development of pronounced infiltration of immune cells in cases of partial surgery or relapse. The antitumor efficacy of the suicide gene treatment was verified by complete remissions in 57 % (4/7) of the patients with local relapse, while partial resection of the tumor resulted in 100 % (6/6) of a long-lasting local stable disease that slowly led to a progressive disease. In conclusion, the use of this treatment after surgical removal of the tumor could delay or prevent post-surgical recurrence and metastases, with the consequent quality of life and survival rate improvement. (SEe Table)

Disturbances of electrolytes and blood chemistry in acute alcohol intoxication

Prevalence of electrolyte disturbances and biochemical changes were determined in patients admitted to the emergency room of the Department of Internal Medicine in Innsbruck, Austria during a six-month period. The value of biochemical parameters for the detection of chronic alcohol abuse was also investigated. The most frequent electrolyte disturbances found were hypernatremia (41%), hyperchloremia (21%), hypermagnesemia (17%) and hypocalcemia (15%), whereas hypokalemia and hypophosphatemia were observed quite rarely (5% and 3.4%, respectively). The most frequent biochemical changes observed were consistent with signs of cellular toxicity i.e. increased liver enzymes (elevated gamma-glutamyltransferase (GGT), aspartate aminotransferase, alanine aminotransferase and lactic dehydrogenase) as well as signs of pancreatitis (elevated serum lipase and amylase) and muscle damage (elevated creatine kinase). The most frequent changes in blood counts were leucocytosis (23%), thrombocytopenia (14%), and anemia (12%). C-reactive protein showed only minimal elevation. Male sex and level of blood alcohol were detected as major risk factors for the diagnosis of chronic alcohol abuse in the patient sample investigated. When testing the value of routinely measured parameters for predicting the presence of chronic alcohol abuse, GGT and mean corpuscular volume of red blood cells (MCV) appeared to be of equal value. A combination of elevated blood alcohol with an increase in either of these markers may be interpreted as high risk for chronic alcohol abuse in this particular group of patients.

Change in Cycle 1 to Cycle 2 Haematological Counts Predicts Toxicity in Older Patients with Breast Cancer Receiving Adjuvant Chemotherapy

To determine the association between changes in complete blood counts and grade 3 or 4 toxicities from cycle 1 to cycle 2 during adjuvant chemotherapy in women > or =65 years of age with breast cancer. A retrospective review was performed on 1405 patients > or =65 years of age who were treated for primary invasive breast cancer at Memorial Sloan-Kettering Cancer Center between January 1998 and December 2000. From this cohort, we identified patients with stage I-III breast cancer who received adjuvant chemotherapy: cyclophosphamide, methotrexate and fluorouracil (CMF) or the anthracycline-based regimens doxorubicin and cyclophosphamide (AC) or AC followed by paclitaxel (AC-T). Patients were excluded from the analysis if they had a prior history of breast cancer or chemotherapy, or if they had no baseline blood counts available for review. Toxicities, dose modification and causality were recorded. The 104 patients who met our criteria had received either CMF (n = 58; mean age 70.6 years, range 65-78) or an anthracycline-based regimen (n = 46; mean age 68.9 years, range 65-77). Of these patients, 50% experienced treatment delay or treatment-related grade 3 or 4 toxicity. A decrease in white blood cell count and absolute neutrophil count from cycle 1 to cycle 2 was associated with grade 3 or 4 haematological toxicity, febrile neutropenia, hospitalisation and initiation of filgrastim for secondary prophylaxis. A decrease in haemoglobin was associated with febrile neutropenia and hospitalisation. Advanced age was not associated with a significant change in complete blood counts, other than a decline in absolute neutrophil count in patients receiving CMF. In this cohort of older patients who received chemotherapy for breast cancer, changes in blood counts from cycle 1 to cycle 2 were associated with increased risk of treatment-related grade 3 or 4 toxicity.

Bone Marrow Connexin-43 Expression Is Critical for Hematopoietic Regeneration After Chemotherapy

Contact between bone marrow (BM) hematopoietic stem cells (HSC) and osteoblast/stromal (OS) cells has been shown to be crucial in the regulation of hematopoiesis. However, very little is known about the regulatory mechanisms of direct cell-to-cell communication in the hematopoietic microenvironment. Gap junction channels (connexons) are formed by polypeptides (connexins) arranged in hexamers and represent the best described intercellular communication system. Connexin-43 (Cx43) is expressed by BM OS cells and has been associated with the cadherin/beta -catenin signaling pathway, recently reported as relevant in the OS/HSC interaction at the stem cell niche. Here, we employed an inducible gene-targeted murine approach to study the role of Cx43 in HSC proliferation and differentiation in vivo. Mx-Cre/Cx43+/+ and Mx-Cre/Cx43flox/flox littermates have been analyzed after gene deletion induced in vivo by the interferon-inducer poly (I)-poly (C), generating control (Cx43+) and Cx43-deficient (Cx43-/-) mice. After one week, Cx43+ and Cx43-/- mice were treated with 5-fluorouracil (5-FU). Cx43 expression in Cx43-/- BM was markedly reduced (> 90%) as analyzed on day +14 post-5-FU treatment. Cx43 deficiency did not induce a significant change in peripheral blood counts before 5-FU treatment, but the hematopoiesis recovery after 5-FU treatment was severely impaired as demonstrated by absence of recovery of peripheral blood counts, including profound neutropenia, anemia with reticulocytopenia, thrombocytopenia and a 5- to 8-fold decrease of cellularity and hematopoietic progenitor content (granulomacrophagic colony-forming-units (CFU-GM-), erythroid burst forming units (BFU-E) and mixed colony forming units (CFU-mix-) in BM and spleen on day +14 post-5-FU treatment. However, the femoral content of Lin-/c-kit+/Sca1+ cells in Cx43-/- BM was maintained when compared to Cx43+ BM. Short-term competitive repopulation ability of Cx43-/- BM cells was diminished as compared to Cx43+ mice, specifically for myeloid and B lymphoid cells, but showed spared long-term competitive repopulation ability with roughly normal hematopoietic differentiation. These data suggest that hematopoietic regeneration after cycle-specific chemotherapy is blocked in Cx43-deficient mice at the long-term HSC repopulating level. Cx43 expression within the BM appears to be crucial in the development of an efficient response to hematopoietic stress.

Sudden Blastic Transformation (SBT) in Patients (pts) with Chronic Myeloid Leukemia (CML) Treated with Imatinib Mesylate.

Imatinib has become standard therapy for pts with CML in chronic phase (CP). SBT has been reported in pts receiving IFN-α, at a rate of 0.5% to 2.5% during the first 3 years of therapy. There is little information on the occurrence of SBT among pts treated with imatinib. Here we report 3 pts who developed SBT, defined as occurring after being found in complete cytogenetic remission (CG CR). These 3 pts represent 0.5% of 557 pts who received imatinib at MD Anderson for chronic phase CML. The total population has been followed for a median of 33 months (range, 1 to 56). Pt #1 was a 54 yr old female with a variant Ph translocation, t(9;22;19;10) (q34;q11.2;p13.1;q22). Sokal risk was low. She started imatinib 400mg/day within 4 mos from diagnosis, achieved a CG CR within 3 mos and was still in CG CR after 9 mos with BCR-ABL/ABL 0.66%. On routine follow-up at 12 mos she was found in myeloid blast phase with 48,XX,+8,t(8;21)(q22;q22),t(9;22;19;10) (q34;q11.2;p13.1;q22), +der(22)t(9;22;19;10) in all 20 metaphases. She received standard induction chemotherapy followed by bone marrow transplantation and is alive and in CR after 24+ months. Pt #2 was a 51 yr old female initially treated with IFN-α. She achieved a CG CR but discontinued therapy because of toxicity after 18 mos. Fourteen mos after stopping IFN-α she had a CG recurrence (35% Ph-positive) and was started on imatinib 400mg/day. Three mos later she was in CG CR and BCR-ABL was undetectable by nested PCR; a similar result was found after 9 mos of therapy. 9 mos later (18 mos after start of imatinib) she had a lymphoid SBT (precursor B-cell). CG showed 45,XX,-8, t(9;22) (q34;q11.2), −20,+mar in one metaphase, and 11 metaphases were diploid. No ABL mutations were identified. She achieved CR with HCVAD + imatinib and is currently receiving a BMT. Patient #3 was a 27 yr old pt who received imatinib 800 mg/d as first line of therapy for CML in CP with intermediate Sokal risk score. Three mos after start of therapy he was in CG CR and repeated analyses showed continued CG CR up to mo 15, with the lowest BCR-ABL/ABL 0.12 at mo 6. On mo 18 he had a lymphoid SBT (precursor B phenotype) with 43–46,XY, −7, −8,i(9)(q10),t(9;22)(q34;q11.2), −13, −17, −21,+der(22)t(9;22),+mar in 13 metaphases and one diploid metaphase. No ABL mutations were identified. He was treated with HCVAD + imatinib and achieve a CR, becoming undetectable for BCR-ABL by nested PCR. He is undergoing a BMT. In pts 2 and 3, transformation was associated with a sudden drop in platelet counts. These isolated events should alert the physicians to continued monitoring of pts on imatinib and consider marrow studies in the event of late, unexpected peripheral blood count changes. Still, SBT is rare, probably less common that seen with IFN-α therapy.

Investigation on the clinical efficacy and safety of 0.1% tacrolimus ointment (Protopic®) in canine atopic dermatitis: a randomized, double‐blinded, placebo‐controlled, cross‐over study

Topical tacrolimus is successfully used in people with atopic dermatitis. Preliminary studies in dogs with atopic dermatitis using tacrolimus in a compounded lotion formulation indicated that tacrolimus significantly decreased erythema and pruritus according to investigator, but no significant improvement was reported by the dog owners. The objectives of this study were to evaluate the clinical efficacy and safety of the commercially available 0.1% tacrolimus ointment (Protopic) in dogs with atopic dermatitis. The study was designed as a double-blinded, placebo-controlled, cross-over study. Selected dogs were allocated to either tacrolimus or placebo for 4 weeks. After 4 weeks there was a wash-out period of 2 weeks and treatments were switched. Twelve dogs completed the study. Clinical signs were scored. Blood samples were collected for complete blood count, chemistry panels and tacrolimus levels at week 0 and 4 of each treatment. Tacrolimus ointment significantly decreased severity of symptoms for both owners and investigators at the end of the trial. When the same dogs received the placebo, there were no differences between week 0 and week 4 scores. Dogs with localized disease responded better than dogs with generalized disease. Tacrolimus was detected in the blood of animals receiving the active ingredient. Levels were below the level of toxicity and no adverse effects were reported in any of the dogs. No changes in complete blood count and chemistry parameters were detected between groups or within groups. In conclusion, tacrolimus appears to be a safe alternative treatment in dogs with atopic dermatitis, especially in those with localized disease.

Repetitive propofol administration in dogs and cats.

A bolus of propofol was administered to 10 dogs (6 mg/kg intravenously [IV]) and 10 cats (10 mg/kg IV) on three consecutive days. The occurrence of apnea, heart and respiratory rates, blood pressure, time to movement, and changes in a complete blood count and biochemical profile were recorded. Apnea was not seen in the dogs but was seen in three cats. Slight increases in the number of Heinz bodies were seen in six cats, but the increases were not considered clinically significant. No apparent cumulative adverse effects were seen from a bolus of bisulfite-containing propofol, administered on three consecutive days.

Lipopolysaccharide dose response in baboons.

A lipopolysaccharide (LPS) dose-response study in an experimental baboon endotoxemia model is presented to define the relevance of this model compared with human endotoxemia. We describe acute and subacute endotoxemic models in baboons, the first evoked by bolus injection of LPS (1 mg, 0.1 mg, or 4 ng per kg of Escherichia coli LPS), and the second evoked by infusion of 1.5 mg/kg of E. coli LPS over 30 min. We report the analysis of LPS clearance, the kinetics of tumor necrosis factor, interleukin (IL) 6, and IL-8 expression on the protein as well as on the mRNA level, change in blood counts (white and red blood cells and circulating platelets), and several hemodynamic parameters such as temperature, cardiac index, heart rate, and mean arterial pressure via multiple sampling. The resulting data are compared with previously published human data. Our results show that the LPS-induced kinetics of cytokine release, as well as of hemodynamic and hematologic changes in baboons, were similar to those observed in humans, even though baboons required a approximately 104-fold higher initial LPS dose to develop these manifestations. Hence, we demonstrate that endotoxemia in baboons qualitatively, yet not quantitatively, resembles endotoxemia in humans and, therefore, proves to constitute a useful model for studying the pathogenic mechanisms of sepsis in relation to humans.

Treatment of canine cyclic neutropenia by lentivirus-mediated G-CSF delivery

AbstractCyclic neutropenia is a rare disease that occurs both in humans and gray collie dogs and is characterized by recurrent severe neutropenia leading to bacterial infections and shortened life expectancy. Daily injections of recombinant granulocyte colony-stimulating factor (rG-CSF) are effective in shortening the period of severe neutropenia and reducing infections. After demonstrating that rG-CSF induced elevated neutrophil production in an affected dog, cytokine administration was stopped and 109 infectious units (IUs) of a lentivirus pseudotyped with vesicular stomatitis virus G protein (VSV-G) encoding canine G-CSF cDNA was administered intramuscularly. Serial blood cell counts showed elevated neutrophil production for longer than 17 months. Although neutrophil counts continued to cycle, the range at nadirs was from 3710 to 5300 cells/μL, well above the nadirs before lentivirus administration. After the injection of lentivirus, mean neutrophil counts ± SD were 12 460 ± 4240 cells/μL, significantly increased over both pretreatment values of 3040 ± 2540 cells/μL (P < .0001) and neutrophil counts during G-CSF administration of 10 290 ± 4860 cells/μL(P < .007). The changes in blood counts from lentivirus injection were associated with absence of clinical signs of infection and fever. The gray collie continued to gain weight and was no longer housed in a pathogen-free environment. Genomic DNA from muscle at injection sites was positive for provirus, whereas gonad, lung, spleen, heart, liver, kidney, leukocytes, and noninjected muscle samples were all negative for provirus. Thus, intramuscular administration of lentivirus encoding G-CSF provided sustained therapeutic levels of neutrophils, suggesting this approach may be applied for long-term treatment of patients with cyclic and other neutropenias.