Preclinical Study of Cell Therapy for Osteonecrosis of the Femoral Head with Allogenic Peripheral Blood-Derived Mesenchymal Stem Cells

PURPOSE: To explore the value of transplanting peripheral blood-derived mesenchymal stem cells from allogenic rabbits (rPBMSCs) to treat osteonecrosis of the femoral head (ONFH). MATERIALS AND METHODS: rPBMSCs were separated/cultured from peripheral blood after granulocyte colony-stimulating factor mobilization. Afterwards, mobilized rPBMSCs from a second passage labeled with PKH26 were transplanted into rabbit ONFH models, which were established by liquid nitrogen freezing, to observe the effect of rPBMSCs on ONFH repair. Then, the mRNA expressions of BMP-2 and PPAR-γ in the femoral head were assessed by RT-PCR. RESULTS: After mobilization, the cultured rPBMSCs expressed mesenchymal markers of CD90, CD44, CD29, and CD105, but failed to express CD45, CD14, and CD34. The colony forming efficiency of mobilized rPBMSCs ranged from 2.8 to 10.8 per million peripheral mononuclear cells. After local transplantation, survival of the engrafted cells reached at least 8 weeks. Therein, BMP-2 was up-regulated, while PPAR-γ mRNA was down-regulated. Additionally, bone density and bone trabeculae tended to increase gradually. CONCLUSION: We confirmed that local transplantation of rPBMSCs benefits ONFH treatment and that the beneficial effects are related to the up-regulation of BMP-2 expression and the down-regulation of PPAR-γ expression.

The New Sysmex XN-2000 Automated Blood Cell Analyzer More Accurately Measures the Absolute Number and the Proportion of Hematopoietic Stem and Progenitor Cells Than XE-2100 When Compared to Flow Cytometric Enumeration of CD34+ Cells

No abstract available.

Hematology of Swiss mice (Mus musculus) of both genders and different ages

PURPOSE: To describe the hematologic values of male and female, young and adult, Swiss mice (Mus musculus). METHODS: Mus musculus (n=14) were randomly selected and separated by gender. The male and female, young and adult animals were sedation to obtain a blood sample, by intracardiac route at 30, 45, 60, 75, 90, 105 and 120 days after birth. RESULTS: The Swiss mouse hemogram values obtained, in relation to total eosinophils, basophils, and number of platelets, there was no statistical differences according to the genders or the age of the animals. Regarding the erythrocyte, hemoglobin and hematocrit values obtained, these were higher in females. The RDW-CD and MPV values were higher in the females than in the males. CONCLUSIONS: Lymphocytes are the predominant cells in the peripheral blood. The collection of 800 µL of blood by intracardiac route, every 15 days, did not affect the health of the animals. Analyses of the blood samples contribute to the experimental models provided by the Central Animal Facility of UFMS and used by professors. .

HTLV 1 associated adult T cell lymphoma/leukemia a clinicopathologic, immunophenotypic tale of three cases from non-endemic region of south India.

Adult T cell lymphoma/leukemia is a peripheral T-cell neoplasm caused by human T-cell lymphotrophic virus-1, affects mostly adults with systemic involvement and poor prognosis. Diagnosis of adult T-Cell leukemia/Lymphoma is challenging. The clinico-pathologic and immuno-phenotypic features of the three cases will be presented.

Morphological spectrum of leukemic mantle cell lymphoma.

Background: Leukemic involvement in mantle cell lymphoma (MCL) is common, and can be secondary to nodal or extranodal disease or can be de-novo. There is paucity of literature that describes the morphological spectrum. Aim: This study was aimed at studying the morphological spectrum of leukemic MCL and to correlate the morphology with other features. Materials and Methods: Twenty six such cases diagnosed over a period of four years were studied. Peripheral blood and bone marrow aspiration smears stained with Wrights stain were examined by three hematopathologists. Immunophenotyping was done using multicolor flow cytometry. Fluorescence in situ hybridization (FISH) done in 12 cases showed t(11;14)(q13:q32). Results: Six cases had de-novo leukemic involvement; while 20 cases had secondary involvement. Morphologically, the cells were small (less than twice the size of red blood cell) or large. Small cell morphology in turn showed irregular nuclear border (n=13) or round nuclear contour (n=6). Large cells had blastic morphology (n=5) or had central prominent nucleoli resembling prolymhphocytes (n=2). Twenty cases showed characteristic immunophenotype of CD5+/CD19+/CD20+/FMC7+/CD10-/CD23- and light chain restrictions. Three cases expressed CD23 and two cases were negative for FMC7. Five out of 12 cases, where FISH was done, showed cytogenetic abnormalities in addition to t(11;14)(q13;q32). Conclusion: Morphological spectrum of leukemic MCL ranges from small cells resembling chronic lymphocytic leukemia (CLL) or follicular lymphoma (FL) to large cell mimicking prolymphocytic leukemia (PLL) or acute leukemia. Large cell morphology was associated with more frequent additional cytogenetic abnormality as well as a poorer outcome.

Chronic myelogenous leukemia in sickle cell/beta0-thalassemia.

Sickle cell/beta 0 -thalassemia (S/β0 -thal) is a compound heterozygous state for βS and β0 thalassemia. There are rare reported cases of patients with sickle cell disease who developed hematological neoplasms including myeloid and lymphoid conditions; however, to the best of our knowledge, chronic myelogenous leukemia (CML) occurring in S/β0 -thal has been reported in one case and this is the second such report.

Clinico-hematological profile of Chediak-Higashi syndrome: Experience from a tertiary care center in south India.

Introduction: Chediak-Higashi syndrome (CHS) is a rare autosomal recessive disorder characterized by partial ocular and cutaneous albinism, increased susceptibility to pyogenic infections, the presence of large lysosomal-like organelles in most granule-containing cells and a bleeding tendency. The abnormal granules are most readily seen in blood and marrow leukocytes, especially granulocytes; and in melanocytes. Other clinical features include silvery hair, photophobia, horizontal and rotatory nystagmus and hepatosplenomegaly. Materials and Methods: The clinico-hematological profile of a series of 5 cases of CHS encountered at JIPMER Hospital with diagnostic work-up done in the Department of Pathology over the last 6 years is presented. The diagnostic work-up included complete hemogram with peripheral smear, bone marrow examination, skin and liver biopsies. Results: The age of the patients ranged from 5 months to 3 years. All patients had silvery hair and partial albinism and presented with fever and recurrent chest infection. Two patients were stable. Three patients were in accelerated phase; of them, 1 patient with associated hemophagocytic syndrome had a rapidly fulminant course. Peripheral blood smear showed anomalously large granules in the leukocytes. Skin biopsy showed sparse, coarse melanin pigment in the epidermis, and liver biopsy done in 2 patients with accelerated phase showed portal lymphohistiocytic aggregates. Conclusions: The diagnostic hallmark of CHS is the occurrence of giant inclusion bodies (granules) in the peripheral leukocyte and their bone marrow precursors. The case series is being presented because of the rarity of CHS and varied spectrum of clinical and hematological presentation.

Hematología y citoquímica de las células sanguíneas de Rhinella fernandezae (Anura: Bufonidae) en Espinal y Delta-Islas del río Paraná, Argentina

Hematology and blood cell cytochemistry of Rhinella fernandezae (Amphibia: Anura) from Espinal and Delta-Islands of Paraná River, Argentina. The description of amphibian hematology is scarce and most of these studies have been done in species from North America, Asia and Europe. With the purpose to obtain basic hematological information of Rhinella fernandezae, 23 blood samples from Santa Fe and Entre Ríos natural reserves were studied. Blood of each individual was extracted by cardiac puncture and hemograms were carried out. Morphological and cytochemical description of blood cells were analyzed in slides and were inspected for extra and intra cellular parasites. Five leucocytes types were observed, being lymphocytes the predominant ones followed by basophiles. Heterophils and eosinophils were positive to PAS, Sudan B and peroxidase. The erythrocytes and its precursors were negative for cytochemical reactions. Micronuclei and nuclear alterations frequencies were scarce. No significant differences (p>0.05) were observed between sexes neither in hemograms nor in blood cells morphology. Microfilarias were the only hemoparasites found with a relative low prevalence and infection intensity. The hematological characteristics studied were similar to those reported for other amphibians, suggesting that R. fernandezae individuals present optimal nutritional and immunological status. Rev. Biol. Trop. 59 (1): 17-28. Epub 2011 March 01.

La descripción de la hematología de los anfibios anuros es escasa, habiéndose realizado la mayoría de los trabajos en especies de Norteamérica, Asia y Europa. Con el propósito de obtener datos hematológicos para Rhinella fernandezae, fueron estudiados 23 especímenes provenientes de zonas protegidas de las provincias de Santa Fe y Entre Ríos. Se les extrajo sangre por punción cardíaca y se realizaron hemogramas. En los extendidos sanguíneos, se efectuaron la descripción morfológica y citoquímica de las células sanguíneas y búsqueda de parásitos. Se observaron cinco tipos de leucocitos, donde predominaron los linfocitos pequeños. Heterófilos y eosinófilos resultaron positivos para PAS, Sudan B y peroxidasa; contrariamente, los eritrocitos y sus precursores fueron negativos. Los puntajes de las reacciones citoquímicas fueron variables para basófilos, linfocitos, monocitos y trombocitos. Las frecuencias de micronúcleos y de alteraciones nucleares fueron escasas. No se observaron diferencias significativas (p>0.05) entre sexos en el hemograma ni en la morfología sanguínea. Los únicos hemoparásitos encontrados fueron microfilarias (Nematoda: Filaroidea), cuya prevalencia e intensidad de infección fueron bajas. Las características hematológicas estudiadas fueron semejantes a los valores reportados para otros anfibios, pudiendo inferir que los individuos de R. fernandezae estudiados se encuentran en un buen estado nutricional e inmunológico.

Effect of instrumentation using curettes, piezoelectric ultrasonic scaler and Er,Cr:YSGG laser on the morphology and adhesion of blood components on root surfaces: a SEM study

This study used scanning electron microscopy (SEM) to evaluate the morphology and adhesion of blood components on root surfaces instrumented by curettes, piezoelectric ultrasonic scaler and Er,Cr:YSGG laser. One hundred samples from 25 teeth were divided into 5 groups: 1) Curettes; 2) Piezoelectric ultrasonic scaler; 3) Curettes plus piezoelectric ultrasonic scaler; 4) Er,Cr:YSGG laser; 5) Curettes plus Er,Cr:YSGG laser. Ten samples from each group were used for analysis of root morphology and the other 10 were used for analysis of adhesion of blood components on root surface. The results were analyzed statistically by the Kruskall-Wallis and Mann-Whitney tests with a significance level of 5 percent. The group treated with curettes showed smoother surfaces when compared to the groups were instrumented with piezoelectric ultrasonic scaler and the Er,Cr:YSGG laser. The surfaces instrumented with piezoelectric ultrasonic scaler and Er,Cr:YSGG laser, alone or in combination with hand scaling and root planing, did not differ significantly (p>0.05) among themselves. No statistically significant differences (p>0.05) among groups were found as to the adhesion of blood components on root surface. Ultrasonic instrumentation and Er,Cr:YSGG irradiation produced rougher root surfaces than the use of curettes, but there were no differences among treatments with respect to the adhesion of blood components.

Esse estudo utilizou microscopia eletrônica de varredura (MEV) para avaliar a morfologia e a adesão de elementos sanguíneos em superfícies radiculares instrumentadas com curetas, ultrassom piezoelétrico e laser de Er,Cr:YSGG. Foram utilizadas no presente estudo 100 amostras provenientes de 25 dentes que foram divididas em 5 grupos: 1) Raspagem manual com curetas; 2) Raspagem com ultrassom; 3) Associação instrumento manual e ultrassom; 4)Irradiação do laser de Er,Cr:YSGG;5)Associação raspagem manual com irradiação com laser de Er,Cr:YSGG. Dez amostras de cada grupo foram utilizadas para análise da morfologia e as outras 10 foram utilizadas para a análise de adesão de elementos sanguíneos. As eletromicrografias foram analisadas através dos escores de adesão de elementos sanguíneos e pelo índice de morfologia radicular e os resultados foram analisados estatisticamente através dos testes de Kruskall-Wallis e de Mann-Whitney com nível de significância de 5 por cento. O grupo que foi tratado com instrumentos manuais apresentou superfície mais lisa em relação aos grupos que foram instrumentados com ultrassom e com o laser de Er,Cr:YSGG. As superfícies instrumentadas com ultrassom e com o laser de Er,Cr:YSGG de forma isolada ou associada a raspagem manual não apresentaram diferenças estatísticas entre si (p>0,05). Não houve diferenças estatísticas entre os grupos em relação a adesão de elementos sanguíneos(p>0,05). A instrumentação ultrassônica e a irradiação com o laser de Er,Cr:YSGG produziram superfícies radiculares mais rugosas em relação a raspagem com curetas, porém não houve diferenças entre os tratamentos com relação à adesão de elementos sanguíneos.

Plasma cell leukemia presenting as acute renal failure: a case report.

Plasma cell leukemia is a rare form of malignant plasma cell dyscrasia. It can occur as a primary form without prior evidence of multiple myeloma or as a secondary form which is a terminal event in multiple myeloma. A case of primary plasma cell leukemia presenting as acute renal failure is reported here.

Hematology, cytochemistry and ultrastructure of blood cells in fishing cat (Felis viverrina)

Hematological, cytochemical and ultrastructural features of blood cells in fishing cat (Felis viverrina) were evaluated using complete blood cell counts with routine and cytochemical blood stains, and scanning and transmission electron microscopy. No statistically significant difference was found in different genders of this animal. Unique features of blood cells in this animal were identified in hematological, cytochemical and ultrastructural studies. This study contributes to broaden hematological resources in wildlife animals and provides a guideline for identification of blood cells in the fishing cat.

A new method for isolation of leukocytes from the peripheral blood of amphibians [Bufo himalayanus, (Gunther)] and study of their surface morphology by scanning electron microscopy.

Using percoll as the density gradient, a new single step method to isolate leukocytes from the peripheral blood of amphibians (B. himalayanus) has been described. Isolated leukocytes were photographed under the scanning electron microscope and an attempt has been made to characterize the leukocyte population on the basis of surface morphology. Apart from regular blood cell types, B. himalayanus have slender, elongated and slightly curved leukocyte type cells in their peripheral blood. Such slender elongated cells were absent in the blood of a related species B. stomnaticus and hence could not be categorized under the known blood cell types.

Haemopoietic organs and effect of their ablation on total haemocyte count in lemon-butterfly, Papilio demoleus L.

The haemopoietic organs in Vth instar larvae of P. demoleus are in the form of thin transparent cellular sheets, closely wrapped around the base of each wing-pad. Three cell types viz; prohaemocytes, plasmatocytes and oenocytoids appear to be derived from these organs and their ablation caused a reduction in cell number which, in turn, revealed that the haemocytes in general are derived both from the haemopoietic organs as well as from the circulating blood cells.

[Morphological studies on the blood of healthy native north Iranian green-head ducks]

Diagnosis of diseases is mainly possible using previous available knowledge of healthy conditions such as normal physiological parameters and values. There is no information on the morphology of the various blood cells of the native ducks of the north of Iran. Therefore blood samples were collected from 100 apparently healthy adult green head Iranian ducks of both sexes. The smears were prepared and stained with Giemsa and new methylen blue technique. The stained smears were studied by light microscopy and the size of blood cells were determined by micrometry. Results of this study showed blood cell morphology in native ducks of north of Iran were similar to chicken and other birds. The size of red blood cells in these ducks were smaller than Indian ducks. Azurophilic granules were not seen in native duck's monocytes

Ultramicrometodo inmunocitoquimico. Titulacion de anticuerpos utilizados para el inmunofenotipaje celular

Se describe el procedimiento para la titulacion de los anticuerpos primarios y los antisueros conjugados, que hizo posible la introduccion en el Instituto de Hematologia Inmunologia, de un ultrametodo inmunocitoquimico para el estudio del inmunofenotipo en enfermedades inmunologicas y en las hemopatias malignas. Este metodo emplea celulas no deshidratadas adheridas a laminas portaobjetostratadas con poli-lisina. Al determinar la dilucion de trabajo de 32 anticuerpos monoclonales y 3 policlonales, se observo que con la misma cantidad de estos reactivos se pueden realizar entre 5 y 100 veces mas determinaciones con respecto al metodo de inmunofluorescencia indirecta. Entre otras ventajas, este metodo permite realizar 21 determinaciones en una misma lamina portaobjeto, requiere una menor cantidad de celulas, los resultados se evaluan en un microscopio de luz visible y las colaboraciones no se desvanecen con el tiempo, lo que permite realizar estudios retrospectivos

Hairy cell leukaemia: a clinicopathological study on 29 Iraqi patients

A total of twenty-nine Iraqi cases diagnosed as Hairy Cell Leukaemia [HCL] in the 3 between 1985-1992 at Baghdad Teaching Hospital were studied. They had ages ranging from 29 to 80 years [median of 50 years] with MF ratio of 5:1 The main clinical features at presentation included constitutional symptoms [86 pyrexia [37.9%]. weight loss [20.7%], bleeding tendency [13.8%] palpable sp1enomegaly [89.7%] and palpable hepatomegaly [48.3%]. Haematologically, pancytopenia was seen in the majority [75.9%]. Bone marrow biopsy showed complete replacement pattern in 69%, large confluent areas of infilteration in 27.6% and small multiple areas of infilteration in 3.4%. Cytoloickally, the hairy cells were of the ovoid type in 58.6%. convoluted type in 27.6%, and indented cell type in 13.8%. The application of the clinical staging system of Jansen and Hermans [1982] showed that the majority were at the advanced stage 111[65.5%] at presentation. This study revealed that the clinical and cytological features of HCL in Iraqi patients are similar to those reported from the West, but they appear to be at a more advanced stage at presentation [as shown by prognostic clinical staging and histological infilteration pattern] as compared to their Western counterparts