Evaluation of Ruxolitinib versus Best Available Therapy in Treating Primary Myelofibrosis.

Objectives: This study aimed to evaluate the effectiveness of ruxolitinib and compare it with the best available therapy in patients with primary myelofibrosis. Ruxolitinib is a Janus kinase inhibitor that is approved for the treatment of myelofibrosis. The therapeutic protocol has changed after the introduction of ruxolitinib. Methods: In this retrospective study, 72 patients with primary myelofibrosis were scrutinised from January 2012 to January 2018 at Nanakali Hemato-Oncology Teaching Centre in Erbil, Iraqi Kurdistan. The patients were divided into two cohorts: 26 of them were treated with ruxolitinib and 46 received the best available therapy. The patients' characteristics, their response to the treatment and the outcomes were evaluated. The efficacy of the treatment in both groups was compared. Results: Most patients (n = 46; 63.9%) were in the high and intermediate-2 risk groups according to the international prognostic scoring system. At the time of diagnosis, no noticeable differences in the clinical characteristics and laboratory data were observed among the ruxolitinib and best available treatment groups. Ruxolitinib was found to be effective in reducing the size of the spleen and improving the overall survival rate when compared to the best available treatment group (P <0.001 and P = 0.008, respectively). The patients' performance status had a significant effect on the overall survival in both treatment groups (P = 0.003). Conclusion: Ruxolitinib appears to have a significant role in reducing the spleen size and potentially affect the survival outcomes in patients with myelofibrosis.

Potential Leukemic Cells Engraftment After Hematopoietic Stem Cell Transplantation From Unrelated Donors With Undiagnosed Chronic Leukemia.

BACKGROUND: Donor-related neoplasms are a potential complication of treatment strategies involving stem cell transplantation. Although mechanisms for detection of short-term complications after these procedures are well developed, complications with delayed onset, notably transmission of chronic diseases such as chronic myeloid leukemia (CML), have been difficult to assess. Consequently, we studied the potential of human CML cells to engraft hematopoietic tissues after intravenous implantation in mice. METHODS: Human peripheral blood cells, collected from CML patients presenting with moderately increased white blood cells count before treatment, were transplanted into sub-lethally irradiated, immunodeficient mice. Five weeks after transplantation the nuclear cells were isolated from the murine bone marrow, spleen, and peripheral blood and were used to quantitatively detect human CD45 antigen by flow cytometry; qRT-PCR was used to detect the BCR-ABL1 fusion gene, and the human or murine beta-glucuronidase housekeeping gene was used to examine human-murine chimerism. RESULTS: We found that all evaluated animals had donor chimerism at the selected interval after transplant and the presence of a specific BCR-ABL1 fusion gene transcript was also detected. CONCLUSIONS: Our results suggest that the risk of neoplasm transmission cannot be eliminated during hematopoietic stem cell transplantation from undiagnosed CML donors with borderline leukocytosis. The obtained data confirms the potential of leukemic cells to viably engraft the hematopoietic organs post-transplantation in an immunosuppressed recipient.

Clinical Aspects, Immunophenotypic Analysis and Survival Rate of Chronic Lymphocytic Leukaemia Patients in Erbil City, Iraq.

OBJECTIVES: Chronic lymphocytic leukaemia (CLL) is characterised by an accumulation of clonal B cells in the blood, bone marrow and lymphatic tissue. This study aimed to evaluate the clinical and immunophenotypic characteristics and survival rate of CLL patients. METHODS: This retrospective study was conducted at the Nanakaly Hospital for Blood Diseases & Oncology in Erbil, Iraq, between January 2011 and December 2017. A total of 105 CLL patients were assessed to determine clinical presentation and staging, immunophenotype and survival rate. RESULTS: The median age of the patients was 65 years and 63.8% were male. The main clinical presentations were splenomegaly (64.8%), pallor (61.9%) and lymphadenopathy (60%). More than half of the patients presented at an advanced clinical stage according to the Rai and Binet staging systems (59.1% and 55.2%, respectively). All CLL cases expressed both cluster of differentiation (CD)19 and CD5, 67.6% had monoclonal kappa light chains and 21% expressed CD38. The five-year overall survival (OS) rate was 61.3%. The mean duration of five-year survival was 41.3 months (95% confidence interval: 36.4-46.3 months). There were no correlations between survival and sociodemographic, clinical or laboratory characteristics. CONCLUSION: In comparison to the existing Western literature, Iraqi CLL patients more frequently presented with hepatosplenomegaly and at a more advanced clinical stage. In addition, the five-year OS rate was much lower.

Morphologic Changes in the Dermis After the Single Administration of Autologous Fibroblastic Cells: A Preliminary Study.

BACKGROUND: Aging is a multifactorial process defined by an accumulation of damage in all tissues and organs, including the skin, throughout the lifespan of an individual. The reduction of both cellular and extracellular matrix components of the dermis during the aging process is followed by the alteration of the morphology of the skin tissue. This study was conducted to assess skin morphology in men before and 3 months after the intradermal injection of autologous fibroblastic cells. METHODS: Tissue biopsies were surgically obtained before and 3 months after the treatment with autogenously harvested fibroblasts expanded in vitro, as well as after injection of phosphate-buffered saline. The thickness of collagen fiber bundles and number of fibroblasts in the dermis were analyzed in morphometric studies. The morphologic evaluation, using different methods of staining has been performed to analyze of extracellular matrix proteins, including collagen and reticular fibers, fibrillin-1-rich microfibrils, elastic fibers, and hyaluronic acid. RESULTS: After administration of the cells, we found a noticeable increase in the number of fibroblasts within the dermis, a significant enlargement in diameter of the collagen fiber bundles, and an improvement in the density of reticular fibers, fibrillin-1-rich microfibrils, and elastic fibers compared with the initial, steady-state condition. CONCLUSIONS: The administration of autogenous fibroblasts could be an effective and safe adjunctive therapy to conventional health care treatment to prevent and reduce the age-related accumulation of dermal tissue damage.

Effects of growth hormone therapeutic supplementation on hematopoietic stem/progenitor cells in children with growth hormone deficiency: focus on proliferation and differentiation capabilities.

We investigated the direct effects of growth hormone (GH) replacement therapy (GH-RT) on hematopoiesis in children with GH deficiency (GHD) with the special emphasis on proliferation and cell cycle regulation. Peripheral blood (PB) was collected from sixty control individuals and forty GHD children before GH-RT and in 3rd and 6th month of GH-RT to measure hematological parameters and isolate CD34(+)-enriched hematopoietic progenitor cells (HPCs). Selected parameters of PB were analyzed by hematological analyzer. Moreover, collected HPCs were used to analyze GH receptor (GHR) and IGF1 expression, clonogenicity, and cell cycle activity. Finally, global gene expression profile of collected HPCs was analyzed using genome-wide RNA microarrays. GHD resulted in a decrease in several hematological parameters related to RBCs and significantly diminished clonogenicity of erythroid progenies. In contrast, GH-RT stimulated increases in clonogenic growth of erythroid lineage and RBC counts as well as significant up-regulation of cell cycle-propagating genes, including MAP2K1, cyclins D1/E1, PCNA, and IGF1. Likewise, GH-RT significantly modified GHR expression in isolated HPCs and augmented systemic IGF1 levels. Global gene expression analysis revealed significantly higher expression of genes associated with cell cycle, proliferation, and differentiation in HPCs from GH-treated subjects. (i) GH-RT significantly augments cell cycle progression in HPCs and increases clonogenicity of erythroid progenitors; (ii) GHR expression in HPCs is modulated by GH status; (iii) molecular mechanisms by which GH influences hematopoiesis might provide a basis for designing therapeutic interventions for hematological complications related to GHD.

ß-Thalassemia Mutations among Transfusion-Dependent Thalassemia Major Patients in Northern Iraq.

Molecular defects responsible for ß-thalassemias (thal) were investigated among 254 chromosomes from 127 transfusion-dependent unrelated thalassemic patients from two provinces in Northern Iraq. Among fourteen identified mutations, the seven most common found in 88.2% of the thal chromosomes were: IVS-II-1 (G → A), IVS-I-1 (G → A), codon 8 (-AA), codon 39 (G → T), codon 8/9 (+G), codon 44 (-C), and codon 5 (-CT). There were some notable differences in frequencies of various mutations in comparison to other Eastern Mediterranean populations, as well as between the two provinces studied. The latter illustrates the relative heterogeneity of the mutations distribution in Iraq, and the need to screen other areas of the country, to ensure establishing an effective prenatal program.

Cells as vehicles for therapeutic genes to treat liver diseases.

Gene therapy involves the transfer of genetic sequences to tissues to obtain a curative effect. Effective gene transfer can be achieved by introducing the therapeutic gene into virus-like particles that facilitate the penetration of the transgene into the cells. However, direct injection of viral vectors may activate innate immunity leading to toxic effects. On the other hand, viral vectors frequently induce neutralizing antibodies, which limit the efficacy of repeated vector administration. Moreover, targeting of the transgene to the desired tissue is a goal that not always can be attained with current vectors. The use of cells as vehicles for therapeutic genes may offer solutions for these issues. Ex vivo transduction of specific cells with vectors encoding therapeutic genes followed by injection of the engineered cells to the patient will reduce the inherent toxicity of the vector while preventing the development of neutralizing antibodies. At the same time, this therapeutic approach can take advantage of the homing properties of the transduced cells to target transgene expression to the sites of interest. Thus, it has been shown that administration of dendritic cells engineered ex vivo with vectors encoding selected antigenic determinants or immunostimulatory molecules is an efficient means to elicit protective immune responses. Similarly, since endothelial progenitor cells (EPC) move to inflammed, ischemic or neoplastic tissues, the injection of EPC transduced ex vivo with appropriate therapeutic genes is an effective method to direct transgene expression to the lesions to be treated. Promising data in animal models of disease point to a future clinical application of this therapeutic strategy.

Preferences regarding end-of-life cancer care and associations with good-death concepts: a population-based survey in Japan.

BACKGROUND: The aims of this study were to clarify end-of-life cancer care preferences and associations with good-death concepts. METHODS: The general population was sampled using a stratified random sampling method (N = 2548; response rate = 51%) and bereaved families from 12 certified palliative care units ('PCU-bereaved families') were surveyed (N = 513; response rate = 70%). The respondents reported their end-of-life care preferences and good-death concepts. RESULTS: Regarding place of end-of-life care, approximately 50% of the general population preferred 'Home', while 73% of PCU-bereaved families preferred 'PCU'. The concepts of 'Maintaining hope and pleasure' and 'Dying in a favorite place' were associated with the preference for 'Home'. Regarding prognostic disclosure, approximately 50% of the participants preferred some level of negotiation with the physician. The concept of 'Control over the future' was associated with this preference. Regarding treatment of severe refractory physical distress, 75% of the general population and 85% of the PCU-bereaved families preferred palliative sedation therapy. The concepts of 'Physical and psychological comfort' and 'Unawareness of death' were associated with this preference. CONCLUSIONS: End-of-life care preferences were associated with good-death concepts. It would be useful for health-care workers to discuss patients' good-death concepts to support subsequent treatment decisions.

The influence of 3,3',5-triiodo-L-thyronine on human haematopoiesis.

OBJECTIVES: Thyroid hormones mediate many physiological and developmental functions in humans. The role of the 3,3',5-triiodo-L-thyronine (T3) in normal human haematopoiesis at the cellular and molecular levels has not been determined. In this study, it was revealed that the human haematopoietic system might be directly depended on T3 influence. MATERIALS AND METHODS: We detected the TRalpha1 and TRbeta1 gene expression at the mRNA level in human cord blood, peripheral blood and bone marrow CD34(+)-enriched progenitor cells, using the RT-PCR method. Furthermore, we performed Western blotting to prove TRalpha1 and TRbeta1 expression occurs at the protein level in human cord blood, peripheral blood and bone marrow CD34(+) cells. In addition, the examined populations of cells were exposed in serum-free conditions to increasing doses of T3 and were subsequently investigated for clonogenic growth of granulocyte-macrophage colony-forming unit and erythrocyte burst-forming unit in methylcellulose cultures, and for the level of apoptosis, by employing annexin V staining and the terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling method. We investigated expression levels of apoptosis-related Bax and antiapoptotic Bcl-2 and Bcl-x(L) genes in the examined cells. RESULTS: We found that exposure to higher and lower than normal concentration of thyroid hormone significantly influenced clonogenecity and induced apoptosis in human haematopoietic progenitor cells. CONCLUSIONS: This study expands the understanding of the role of thyroid disorders in normal human haematopoiesis and indicates a direct influence of T3 on this process.

An optimization of isolation of early hematopoietic cells from heparinized cadaveric organ donors.

Heparinized cadaveric organ donors are a potential source of hematopoietic cells for transplantation purposes. The aim of this study was to optimize the harvest of early hematopoietic cells from cadaver donors. Accordingly, we noticed that bone marrow cells harvested from cadaver donors should be resuspended in RPMI or Iscove's medium supplemented with heparin or ACD. Bags with harvested marrow should contain 20% atmosphere air during short-term storage/transportation. Finally, we noticed that BMMNC survive short-term storage better if the collected marrow is not depleted of erythrocytes.

Long-term follow up study of vertical HTLV-I infection in children breast-fed by seropositive mothers.

Infection of Human T-lymphotropic virus type-I (HTLV-I) was investigated by long-term follow up surveys of mother's milk-fed infants. HTLV-I infections of infants via seropositive mother's milk, that is, anti-HTLV-I antibody-positive infants, increased in number up to the age 2, but no infants became antibody-positive thereafter. Infants who had became antibody positive by age 2 remained so at age 11-12. HTLV-I infection via feeding with mother's milk was established by the age 2. While in epidemiologic surveys an increase of the anti-HTLV-I antibody-positive rate has been reported, this survey revealed that after acquisition of HTLV-I from breast feeding, there was no further horizontal transmission prior to puberty.

Long-term follow-up study of HTLV-I infection in bottle-fed children born to seropositive mothers.

Human T-lymphotropic virus Type-I (HTLV-I) infects children via mother's milk. Infection of Human T-lymphotropic virus Type-I (HTLV-I) was investigated by long-term follow-up surveys of modified milk-fed children. Our observations of modified milk-fed infants revealed that: 1 of 154 (0.6%) at year 1, 5 of 129 (3.9%) at 1.5 years, and 5 of 108 (4.6%) at year 2 were anti-HTLV-I antibody-positive. No infants or children became newly antibody-positive thereafter. Modified milk feeding could prevent the HTLV-I infection of infants from mothers in many cases, however the infants who had became anti-HTLV-I antibody-positive due to established infection by the age 2 remained positive at age 11-12 with persistent infections. Modified milk-fed infants who had been born from HTLV-I seropositive mothers did not show that they had complete protection from HTLV-I infection, but a low infection rate was seen, showing that modified milk feeding is useful to protect from HTLV-I infection.

XAFS studies of Tb or Eu cored dendrimer complexes with various properties of luminescence.

We studied the extended X-ray absorption fine structure (EXAFS) of Tb or Eu L3-edge of Tb(III) or Eu(III)-cored complexes of different luminescence to which different poly(benzyl ether) dendrons coordinated. The complexes were prepared via a ligand exchange reaction of dendritic carboxylic acid and acetate. Results show that (a) the Eu complex of the first generation of the 3,4- or 3,5-dibenzyloxyphenyl type had a coordination structure different from that of the 2,4-dibenzyloxyphenyl type; (b) the Eu complex of the first generation of 3,5-dinaphthylmethoxyphenyl type had a Eu-O coordination number higher than the total number of oxygen atoms in the carboxylate groups; (c) the Tb complex of the bulky fourth generation had a Th-O coordination number higher than that of the first generation. These results show that the different poly(benzyl ether) dendrons cause the different coordination, which might differently influence the luminescence properties of the complexes.

[Attitudes towards terminal care among the general population and medical practitioners in Japan].

OBJECTIVES: Assessment of attitudes held by the general population and medical practitioners in Japan regarding medical interventions in cases of painful terminal illness or a prolonged vegetative state. METHOD: A mail survey was conducted in 1998. The subjects were 5,000 persons randomly sampled members of the general population age 20 years or more, and 3,104 doctors and 6,059 nurses in hospitals, clinics, palliative units, and visiting nursing service stations randomly sampled. The response rates were 48% among general population, 51% among doctors, 56% among nurses. RESULTS: 1. 68-76% of the general population and medical practitioners expressed disapproval of life-extending medical treatment of terminal patients suffering pain. The application of euthanasia in certain cases was acceptable to 13% of the general population but only 1% of the medical practitioners. 2. Respondents in almost groups favored a home care setting for terminal patient in pain, and regarded relocation to a palliative unit as acceptable if necessary. 3. 46% of doctors and 22% of nurses indicated knowledge of the WHO method for cancer pain relief, and 45% of doctors and 25% of nurses showed that they were able to explain appropriate opioid administration. 4. 74-79% of general population and medical practitioners opposed life-extending medical intervention for patients in a vegetable state. 26% of the general population favored termination of all means of life support, while about 10% of the medical practitioners held this view. 5. Most medical practitioners felt that some medical treatments, such as bedsore care, should be continued in lieu of life support, but there were differences in opinion between practitioners at various types of medical facilities regarding the necessity of such specific measures as, for example, blood pressure monitoring by automatic sphygmomanometer among the medical facilities. CONCLUSION: Both the general population and the medical practitioners in Japan tended to oppose life-extending medical treatment for painful terminal cases and patients in a prolonged vegetable state. There are some differences in opinion between the general population and practitioners at various types of medical facilities regarding the extent of desirable medical care in such circumstances.

Hepatitis B virus and alcoholic liver damage in the Airin district. (Osaka's skid row area).

Healthy adults and non-liver disease patients in the Airin district gave a positivity rate for HBs antigen of approximately 2%, which is not so much different from the average of the entire Japanese population. In this district, the positive rate of HBs antigen 5% (11 out of 244) in chronic liver disease is much lower than that (31%) of control group patients at Osaka City University Hospital (O.C.U.H.). However the positive rate of anti-HBs antibody in this district is 46% which is extremely high compared with that (23.4%) in the rest of Japan. The positive for anti-HB antibodies (including of anti-HBs and anti-HBc antibody) accounted for 67% of non-liver disease patients and 68% of chronic liver disease patients in the Airin district. Corresponding values for patients at O.C.U.H. were 44% and 43% respectively. There were no significant differences in the histological picture between HBs antigen-negative patients with and those without demonstrable anti-HBs antibodies. Changes in the liver exhibited by heavy drinkers inhabitating the Airin district were primarily those of alcoholic liver damage.