Influence of height on endothelial maintenance activity: a narrative review

Recent studies have revealed an inverse association between height and cardiovascular disease. However, the background mechanism of this association has not yet been clarified. Height has also been reported to be positively associated with cancer. Therefore, well-known cardiovascular risk factors, such as increased oxidative stress and chronic inflammation, are not the best explanations for this inverse association because these risk factors are also related to cancer. However, impaired blood flow is the main pathological problem in cardiovascular disease, while glowing feeding vessels (angiogenesis) are the main characteristic of cancer pathologies. Therefore, endothelial maintenance activity, especially for the productivity of hematopoietic stem cells such as CD34-positive cells, could be associated with the height of an individual because this cell contributes not only to the progression of atherosclerosis but also to the development of angiogenesis. In addition, recent studies have also revealed a close connection between bone marrow activity and endothelial maintenance; bone marrow-derived hematopoietic stem cells contribute towards endothelial maintenance. Since the absolute volume of bone marrow is positively associated with height, height could influence endothelial maintenance activity. Based on these hypotheses, we performed several studies. The aim of this review is not only to discuss the association between height and bone marrow activity, but also to describe the potential mechanism underlying endothelial maintenance. In addition, this review also aims to explain some of the reasons that implicate hypertension as a major risk factor for stroke among the Japanese population. The review also aims to clarify the anthropological reasons behind the high risk of atherosclerosis progression in Japanese individuals with acquired genetic characteristics.

Expression of non-neuronal cholinergic system in maxilla of rat in vivo

BACKGROUND: Acetylcholine (ACh) is known to be a key neurotransmitter in the central and peripheral nervous systems, which is also produced in a variety of non-neuronal tissues and cell. The existence of ACh in maxilla in vivo and potential regulation role for osteogenesis need further study. RESULTS: Components of the cholinergic system (ACh, esterase, choline acetyltransferase, high-affinity choline uptake, n- and mAChRs) were determined in maxilla of rat in vivo, by means of Real-Time PCR and immunohistochemistry. Results showed RNA for CarAT, carnitine/acylcarnitine translocase member 20 (Slc25a20), VAChT, OCTN2, OCT1, OCT3, organic cation transporter member 4 (Slc22a4), AChE, BChE, nAChR subunits α1, α2, α3, α5, α7, α10, ß1, ß2, ß4, γ and mAChR subunits M1, M2, M3, M4, M5 were detected in rat's maxilla. RNA of VAChT, AChE, nAChR subunits α2, ß1, ß4 and mAChR subunits M4 had abundant expression (2(-ΔCt) > 0.03). Immunohistochemical staining was conducted for ACh, VAChT, nAChRα7 and AChE. ACh was expressed in mesenchymal cells, chondroblast, bone and cartilage matrix and bone marrow cells, The VAChT expression was very extensively while ACh receptor α7 was strongly expressed in newly formed bone matrix of endochondral and bone marrow ossification, AchE was found only in mesenchymal stem cells, cartilage and bone marrow cells. CONCLUSIONS: ACh might exert its effect on the endochondral and bone marrow ossification, and bone matrix mineralization in maxilla.

Potencial de transdiferenciação neural das células-tronco mesenquimais da medula óssea de equino / Potential of neural transdifferentiation of mesenchymal stem cells from equine bone marrow

Os primeiros estudos demonstrando o potencial de trandiferenciação neural das células-tronco mesenquimais (CTMs) provenientes da medula óssea (MO) foram conduzidos em camundogos e humanos no início da década de 2000. Após esse período, o número de pesquisas e publicações com o mesmo propósito tem aumentado, mas com raros ou escassos estudos na espécie equina. Nesse sentindo, o objetivo desse trabalho foi avaliar o potencial in vitro da transdiferenciação neural das CTMs provenientes da MO de equinos utilizando-se dois protocolos: P1 (forksolin e ácido retinóico) e P2 (2-βmecarptoetanol). Após a confirmação das linhagens mesenquimais, pela positividade para o marcador CD90 (X=97,94%), negatividade para o marcador CD34 e resposta positiva a diferenciação osteogênica, as CTMs foram submetidas a transdiferenciação neural (P1 e P2) para avaliação morfológica e expressão dos marcadores neurais GFAP e β3 tubulina por citometria de fluxo. Os resultados revelaram mudanças morfológicas em graus variados entre os protocolos testados. No protocolo 1, vinte quatro horas após a incubação com o meio de diferenciação neural, grande proporção de células (>80%) apresentaram morfologia semelhante a células neurais, caracterizadas por retração do corpo celular e grande número de projeções protoplasmáticas (filopodia). Por outro lado, de forma comparativa, já nos primeiros 30 minutos após a exposição ao antioxidante β-mercaptoetanol (P2) as CTMs apresentaram rápida mudança morfológica caracterizada principalmente por retração do corpo celular e menor número de projeções protoplasmáticas. Também ficou evidenciado com o uso deste protocolo, menor aderência das células após tempo de exposição ao meio de diferenciação, quando comparado ao P1. Com relação a análise imunofenotípica foi observado uma maior (P<0,001) expressão dos marcadores GFAP e β3 tubulina ao término do P2 quando comparado ao P1. A habilidade das CTMs em gerar tipos celulares relacionados a linhagem neural é complexa e multifatorial, dependendo não só dos agentes indutores, mas também do ambiente no qual estas células são cultivadas. Desta forma um maior número de estudos é necessário para o melhor entendimento do processo de transdiferenciação neural a partir de CTMs de equinos.

The first studies showing the potential of neural transdifferentiation of mesenchymal stem cells (MSCs) from bone marrow (BM) were conducted in camundogos and humans in the early 2000s. After this period, the number of research and publications with the same purpose increased, but with rare or scarce studies in horses. The aim of this study was to evaluate in vitro neuronal transdifferentiation potential of MSCs from equine BM using two protocols: P1 (forksolin and retinoic acid) and P2 (2-βmecarptoetanol). After confirming the mesenchymal lineages, by positivity for the marker CD90 (X=97.94%), negative for the marker CD34 and positive response for osteogenic differentiation, MSCs were subjected to neural transdifferentiation (P1 and P2) for morphological analysis and expression of neural markers GFAP and β3 tubulin by flow cytometry. The results revealed morphological changes in varying degrees between the tested protocols. In protocol 1, twenty four hours after incubation with the media of neural differentiation, a large proportion of cells (>80%) had similar morphology to neural cells, characterized by retraction of cellular body and a large number of cytoplasmic extension (filopodia). However, comparatively, within the first 30 minutes after exposure to the antioxidant β-mercaptoethanol (P2) MSCs showed rapid morphological changes characterized mainly by retraction of cellular body and less cytoplasmic extension. It was also evidenced with the use of this protocol, lower cellular adhesion after exposure to media when compared to P1. Regarding the immunophenotyping analysis it was observed a higher (P<0.001) expression of the markers GFAP and β3 tubulin at the end of P2 compared to P1. The ability of MSCs to generate cell types related to neural lineage is complex and multifactorial, depending not only of inducing agents, but also the environment in which these cells will be cultivated. Thus a greater number of studies are necessary to better understand the process of neural transdifferentiation of MSCs from equine.

Diferenciação in vitro de células-tronco mesenquimais da medula óssea de cães em precursores osteogênicos / In vitro differentiation of mesenchimal stem cells of dogs into osteogenic precursors

O objetivo principal da nossa pesquisa foi avaliar o potencial de diferenciação osteogênica de células-tronco mesenquimais (MSC) obtidas da medula óssea do cão. As MSC foram separadas pelo método Ficoll e cultivadas sob duas condições distintas: DMEM baixa glicose ou DMEM/F12, ambos contendo L-glutamina, 20% de SFB e antibióticos. Marcadores de MSC foram testados, confirmando células CD44+ e CD34- através da citometria de fluxo. Para a diferenciação osteogênica, as células foram submetidas a quatro diferentes condições: Grupo 1, as mesmas condições utilizadas para a cultura de células primárias com os meios DMEM baixa glicose suplementado; Grupo 2, as mesmas condições do Grupo 1, mais os indutores de diferenciação dexametasona, ácido ascórbico e b-glicerolfosfato; Grupo 3, células cultivadas com meios DMEM/F12 suplementado; e Grupo 4, nas mesmas condições que no Grupo 3, mais indutores de diferenciação de dexametasona, ácido ascórbico e b-glicerolfosfato. A diferenciação celular foi confirmada através da coloração com alizarin red e da imunomarcação com o anticorpo SP7/Osterix. Nós observamos através da coloração com alizarin red que o depósito de cálcio foi mais evidente nas células cultivadas em DMEM/F12. Além disso, usando a imunomarcação com o anticorpo SP/7Osterix obtivemos positividade em 1:6 células para o Meio DMEM/F12 comparada com 1:12 para o meio DMEM-baixa glicose. Com base nos nossos resultados concluímos que o meio DMEM/F12 é mais eficiente para a indução da diferenciação de células-tronco mesenquimais caninas em promotores osteogênicos. Este efeito provavelmente ocorre em decorrência da maior quantidade de glicose neste meio, bem como da presença de diversos aminoácidos.

The aim of our research was to evaluate the potential for osteogenic differentiation of mesenchimal stem cells (MSC) obtained from dog bone marrow. The MSC were separated using the Ficoll method and cultured under two different conditions: DMEM low glucose or DMEM/F12, both containing L-glutamine, 20% of FBS and antibiotics. MSC markers were tested, confirming CD44+ and CD34- cells with flow cytometry. For osteogenic differentiation, cells were submitted to four different conditions: Group 1, same conditions used for primary cell culture with DMEM supplemented media; Group 2, same conditions of Group 1 plus differentiation inductors Dexametazone, ascorbic acid and β-glicerolphosphate. Group 3, Cells cultured with supplemented DMEM/F12 media, and Group 4, same conditions as in Group 3 plus differentiation inductors Dexametazone, ascorbic acid and β-glicerolphosphate. The cellular differentiation was confirmed using alizarin red and imunostaining with SP7/Osterix antibody. We observed by alizarin staining that calcium deposit was more evident in cells cultivated in DMEM/F12.Furthermore, by SP/7Osterix antibody immunostaining we obtained 1:6 positive cells when using DMEM/F12 compared with 1:12 for low-glucose DMEM. Based on our results, we conclude that the medium DMEM/F12 is more efficient for induction of differentiation of mesenchymal stem cells in canine osteogenic progenitors. This effect is probably due to the greater amount of glucose in the medium and the presence of various amino acids.

Effect of titanium dioxide [TiO2] as a color additive on the testes, sperms and chromosomes of male albino rats

Titanium dioxide [TiO2] is one of the top 50 produced substances for use around the world. 70% of all [TiO2] produced is used as pigment in consumer products such as plastics, health, beauty aids and other personal care product that we use. Toothpaste products use [TiO2] to get that desirable bright white color as do many other products such as lotions, creams, shave foam, cosmetics, sunscreen lotions and more. Food products such as sour cream, cottage cheese [via the cheese dressing] ice cream and other dairy products use a small quantity of the pigment to attain that familiar brightwhite coloration. The aim of this study was to evaluate the histopathological effect of 1/20 of LD50 of [Ti02] on the testes, sperms and chromosomes of albino rats and its relation to the duration of its adminstration. Forty male albino rats had been divided into four groups, ten rats for each. The first was served as a control group, the second was gavaged by [TiO2] 600mg/kg daily for 4weeks. The third was gavaged with same dose of [TiO2] for 8 weeks and the forth group was gavaged by same dose of [Ti02] for 12 weeks. Each rat's group were sacrificed after each duration, testes specimens were taken and stained with Haematoxylin and Eosin. The sperms were examined for number, viability, motility and shape abnormalities. For chromosomal study, rats from each group were anaesthetized and the bone marrow cells were obtained by Rabello-Gay and Ahmed method. Microscopic examination of the testicular specimens, revealed disorganized germinal epithelium with abnormal mitotic figures and apoptotic cells. Sperm analysis showed that sperm count, viability and motility were decreased and the sperm anomalies were increased. Chromosomal analysis of bone marrow cells showed many aberrations as, chromatid deletions, ring chromosomes, chromosomal gaps, dicentric chromosomes, clumping of the chromosomes and polyploidy. All the former revealed that the histopathological changes and abnormalities caused by [TiO2] had been aggravated by prolonged duration of administration

Intrathoracic and pelvic extramedullary haematopoiesis in sickle cell disease: a case report

Intrathoracic extramedullary haematopoiesis is a rare entity encountered in patients with long standing anaemias such as thalassaemia and congenital spherocytosis. It is rare in patients with homozygous sickle cell disease; only 11 cases of intrathoracic and two cases of pelvic extramedullary haematopoiesis have been documented in the literature. We report the case of a 30-year old man with homozygous sickle cell disease with intrathoracic and pelvic extramedullary haematopoiesis, the first case to be documented from the Caribbean.

La hematopoyesis extramedular intratorácica es una entidad que raras veces se encuentra en pacientescon anemias de larga duración tales como la talasemia y la esferocitosis congénita. También es rara en pacientes que padecen la enfermedad de células falciformes homocigóticas. En la literatura se han documentado sólo 11 casos de hematopoyesis extramedular intratorácica y dos casos de hematopoyesis extramedular pélvica. Reportamos el caso de un hombre de 30 años de edad con la enfermedad decélulas falciformes homocigóticas con hematopoyesis extramedular intratorácica y pélvica – el primercaso que se documenta en el Caribe.

Age-related marrow conversion and developing epiphysis in the proximal femur: evaluation with STIR MR imaging / 华中科技大学学报（医学）（英德文版）

In order to observe the feature of age-related marrow conversion and maturation of epiphyseal cartilage and analyze the distribution of red and yellow marrow in the proximal femur at STIR MR imaging, STIR and T(1) weighted MR imaging of the proximal femur in 52 subjects, aged 4 months to 25 years old, were retrospectively analyzed for the distribution and appearance of red and yellow marrow. The subjects with no known bone marrow abnormalities were divided into 6 age groups. The signal intensity of the marrow in the proximal epiphysis, proximal metaphysis, proximal diaphysis, distal diaphysis and greater trochanter was compared with the signal intensity and homogeneity of surrounding muscle and fat and graded by two observers. The results showed that the conversion of hematopoietic marrow in the proximal femur followed a well-defined sequence, occurring first in the proximal epiphysis, followed by the distal diaphysis, and then greater trochanter and metaphysis. STIR in combination with T(1)-weighted imaging could display clearly the origin of ossification center and the course of conversion from red to yellow marrow in proximal epiphysis and greater trochanter. STIR imaging showed that the marrow conversion in proximal metaphysic began below epiphyseal plate and intertrochanter. The site of red yellow was distributed in weight-bearing axis by 20 years of age. The marrow conversion of diaphysis was from distal end to proximal end, and the consequence of conversion was that distal diaphysis contained yellow marrow but proximal diaphysis partly red marrow connected with the red marrow of metaphysic. The epiphyseal cartilage had different characters of signal-intensity with age in STIR sequence. The distribution of red marrow in STIR imaging was more close to that of anatomy than T(1)-weighted imaging. It was concluded that STIR could dynamically display the feature of marrow conversion and the development of epiphyseal cartilage and accurately reveal the age-related distribution of red and yellow marrow on STIR imaging in the proximal femur.

Proportions of Cells Expressing CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, or CD13,33+/CD34+ in the Regenerating Bone Marrows During Complete Remission of Acute Leukemia or After Bone Marrow Transplantation / 대한진단검사의학회지

BACKGROUND: The hemopoietic stem cells increase in number during the regeneration after chemotherapy or bone marrow transplantation (BMT). Although the proportion of hemopoietic stem cells and their differentiation have been studied by immunophenotyping using the flow cytometry, no substantial research efforts have been directed toward the regenerating marrow. We attempted to discover the proportions of undifferentiated stem cells, committed stem cells, B cell precursors, and myeloid precursors in the regenerating bone marrows during complete remission (CR) and after engraftment of BMT. METHODS: Bone marrow samples from 82 patients with acute leukemia in CR and from 25 patients after BMT engraftment, along with 22 control samples, were used to find the numbers of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells in the large lymphocyte gate by flow cytometry. We cross-analyzed our results in terms of groups: CR, BMT, and initial diagnosis groups. We performed significance tests on age, relapse, chromosomal abnormalities, clinical outcomes, and initial immunophenotypes of the leukemic cells. RESULTS: The proportions of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells are more highly distributed in acute B-lymphoblastic leukemia than the normal group and also in the CR than the BMT group. CD19+/CD34+ cells were increased in the relapse group and CD38+/ CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells were increased in the group with chromosomal abnormality. The results were irrelevant to the initial immunophenotype of the leukemic blasts. CONCLUSIONS: The increases of the markers spanned too widely to apply one specific cutoff value to analyze them. They seemed to be the results of normal regeneration, irrelevant to relapse or initial immunophenotype of leukemic blasts.

Role of macrophage-colony stimulating factor and osteoclast differentiation factor in osteoclastogenesis of bone marrow derived stem cells.

Macrophage colony stimulating factor (M-CSF) and osteoclast differentiation factor (ODF) regulate osteoclastogenesis in vivo. Regulation of osteoclast development in vitro by these cytokines has been reported in the present study. Simultaneous addition of ODF and M-CSF during initiation of bone marrow culture inhibited osteoclastogenesis. However, delayed addition of ODF (three days after initiation of the culture) resulted in dramatic increase in phenotypically and functionally mature osteoclast cells. Delayed addition of ODF beyond day three decreased osteoclastogenesis. Further, removal of M-CSF as early as day three inhibited ODF-induced osteoclastogenesis. These studies provided evidence for the importance of co-ordinated regulation of osteoclastogenesis by M-CSF and ODF.

Defective Erythropoiesis in Bone Marrow is a Mechanism of Anemia in Children with Cancer

Evaluation of the mechanism of anemia in cancer patients might help to select patients for the more efficient use of erythropoietin (EPO, a growth factor for erythroid precursor cells). For this, we investigated whether the production of EPO responds to anemia and the bone marrow responds to EPO appropriately, and whether chronic inflammation is inhibitory to erythropoiesis in anemic cancer children. Serum levels of EPO, soluble transferrin receptor (sTfR), tumor necrosis factor (TNF)-alpha, and erythrocyte sedimentation rate (ESR) in anemic cancer children were measured by enzyme-linked immunosorbent assay and then the correlation coefficients between those parameters and hemoglobin (Hb) were determined. Both in leukemia and in solid tumor patients, there were significant inverse correlations between Hb and EPO (leukemia: tau=-0.547, p<0.0001; solid tumor: tau=-0.591, p<0.0001), and between sTfR and EPO (leukemia: tau=-0.223, p<0.05; solid tumor: tau=-0.401, p<0.05). In contrast, sTfR showed a correlation with Hb in leukemia (tau=0.216, p<0.05) but not in solid tumor patients. sTfR was suppressed in 53% of anemic episodes of leukemia and 78% of those of solid tumor patients. Our results suggest that in cancer children, the EPO production is not defective and chronic inflammation is not inhibitory to erythropoiesis. Rather, the defective erythropoiesis itself is thought to be responsible for the anemia.

In vivo radioprotection of mouse bone marrow: protection from apoptotic death by WR-2721

Apoptotic cell death is an important phenomenon in radiation bone marrow injury and the compound WR-2721 can protect hematopoietic tissue against such injury. In this study, we assessed the ability of WR-2721 to prevent radiation-induced apoptosis in bone marrow cells. Femoral bone marrow from C57BL male mice was used. The marrow was studied 4 h, 12 h, 24 h and 10 days after a single whole-body radiation dose of 7 Gy. Mice which received WR-2721 (400 mg/kg, i.p.) 30 min before irradiation were compared with unprotected mice. Less injury and a significant reduction in the number of apoptotic cells were observed 4 h (49.6 per cent less) and 12 h (40.8 per cent less) after irradiation in the group that received WR-2721 compared to the unprotected mice. An earlier than expected recovery was also observed 10 days after irradiation in the protected group. This is the first study in vivo to report the protection by WR-2721 of bone marrow cells from apoptosis following exposure to radiation.

Review of bone marrow examinations at sandman teaching hospital, Quetta

Balochistan is the largest province of Pakistan and has only one teaching hospital in Quetta, which serves people from all over the province, and also from Afghanistan. As yet there is no reported incidence of prevalence of "Bone Marrow disorder" in this part of the country. The present analysis is of 191 bone marrow examinations done in 1 1/2 year in this hospital, included 128 males [67%] and 63 females [33%]. Out of these 113 [59%] were adults and 78[41%] childen. Leukemias were the most frequent diagnosis in our patients. The total number was 40 [21%] which included 20 cases of ALL [L1=6, L2=11, L3=3] and 7 cases of AML [M1=1, M2=1, M3=1, M4=3 M6=1]. There was no case of M5 and M7. All the 27 cases of acute leukaemia presented with anemia; 25 showed thrombocytopenia and in only 2 cases platelets were normal. Leucocytosis was found in 8 cases, leucopenia in 12 and TLC was normal in 7 cases. Twelve [44%] patients presented with pancytopenia. Out of 40 there were 11 cases of CML and 2 cases of CLL. Two patients of CML later went into blast crisis. In all the cases of malaria, Gametocytes of P.falciparum were found in Bone marrow and out of them, 5 had negative P/Smear for MP. All cases of Kala-azar presented with pancytopenia. In Aplastic anemia, myelofibrosis and metastatic disease of Bone marrow, diagnosis was made after doing bone marrow Trephine. Metastasis was from Ca prostate. Both patients of essential thrombocythaemia had platelet count of more than 1 million. Of the 2 cases of myelodysplastic syndrome one was RAEBt and other was RAEB

Autologous bone marrow injection for delayed union: a preliminary report

Bone marrow injection is a useful semi-invasive method of bone healing enhancement in cases of delayed union. It is indicated in particular in cases where an open procedure is contraindicated or not accepted by the patient. We present the method and preliminary results of treatment of delayed union with this method

Putative pathways involved in cardiovascular responses evoked from the caudal pressor area

1. The caudal pressor area (CPA) is a recently identified site within the ventrolateral medulla which is involved in cardiovascular regulation. CPA chemical stimulation by L-glutamate produces an increase in arterial blood pressure (ABP) while its inhibition by GABA or glycine evokes marked hypotension. In the present study, we sought to determine the potential neural pathways underlyng these responses. 2. In urethane-anesthetized, paralyzed, artificially ventilated rats, CPA inhibition by bilateral microinjection of the inhibitory amino acid glycine (Gly, 100 nmol 200 nl-1 site-1) produced an average decrease of -38 + or - 4.3 mmHg in ABP (n = 6). Ten min after bilateral microinjection of the broad-spectrum glutamate antagonist kynurenic acid (KYN, 2 nmol 200 nl-1 site-1) into the cauldal ventrolateral medulla (CVLM) depressor responses to CPA inhibition were virtually abolished (-3 + or - 1.7 mmHg, P<0.05). Similar microinjection of KYN into the rostral ventrolateral medulla (RVLM) or into the CPA itself did not modify depressor responses to CPA inhibiton by glycine. 3. CPA stimulation by bilateral microinjection of the excitatory amino acid L-glutamate (L-glu, 50 nmol 200 nl-1 site-1) produced an increase in ABP (+43 + or - 5.4 mmHg, N= 6). Bilateral microinjection of the GABA A antagonist bicuculline methiodide (BIC, 200 pmol 200 nl-1 site-1) into the CVLM markedly reduced pressor responses to CPA stimulation (+6 + or - 2.7 mmHg, P<0.05). Similar application of BIC into the RVLM or CPA did not modify pressor responses to CPA stimulation by glutamic acid

Enhancement of haemopoietic stem cell adherence to bone-marrow derived stroma by phyto-haemagglutinin treated leucocyte conditioned medium (PHA-LCM).

Stem cell adhesion to bone-marrow derived stroma, plays a crucial role in haemopoiesis. However, there is very little information as to the nature of the adhesion molecule. In this paper we have shown that human bone-marrow derived stroma can be established in tissue culture. This stroma is able to adhere human bone-marrow mononuclear cells including the multipotent stem cell, viz. CFU-GEMM. Their adherence increases when the stroma is treated with lymphokines in the form of PHA-treated leucocyte conditioned medium (PHA-LCM). Triton X-100 extracts of the untreated and PHA-LCM treated stroma were analysed on single dimension PAGE. It was observed that PHA-LCM treated stromal extracts showed two extra bands and an increase in the density of a band of approximately 14 kDa. Whether these changes have anything to do with the increased adhesion of stem cell is not yet known.