bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of HIF as a Growth and Survival Factor in Multiple Myeloma.
Authors: Feinman Rena , Abungu Billy , Barlos Dimitrios
Published: October 2019

Given that hypoxia and insulin-like growth factor-1 (IGF-1) induce hypoxia inducible factor (HIF-1)-dependent vascular endothelial growth factor (VEGF) expression in different cancer models, we began to explore the possibility that HIF-1α may act as a proliferative and survival factor in multiple myeloma cells. In this study, we demonstrate that MM cells express constitutive HIF-1α protein levels and that hypoxia and cobalt chloride, a hypoxia mimetic, increase HIF-1α and VEGF expression in several MM cell lines (ARP-1, MM1.S, RPMI8266). In the presence of IGF-1, maximal induction of HIF-1α expression in MM cells under normoxic conditions occurred after 12 hr of IGF-1 and declined thereafter. In contrast to hypoxia which inhibits the proteasomal degradation of HIF-1α, IGF-1 requires de novo protein synthesis of HIF-1α. Pharmacologic inhibition of PI3K/Akt or MAPK signaling pathways prevented IGF-1 induced HIF-1α and VEGF expression in ARP-1 cells. Since 2-methoxyestradiol (2-ME) has been shown to inhibit MM growth and associated angiogenesis, we examined whether 2-ME would inhibit the accumulation of IGF-1 induced HIF-1α expression. 2-ME markedly reduced constitutive and IGF-1-induced HIF-1α protein levels in ARP-1 cells. To further demonstrate that HIF-1α plays a role in the growth and survival of MM cells, RNA interference was used to inhibit HIF-1α expression. in ARP-1 cells. Nucleofection of ARP-1 cells with small interfering RNA (siRNA) targeting HIF-1α inhibited constitutive HIF-1α as well as IGF-1-induced HIF-1α protein levels. HIF-1β protein levels were unaffected. Consequently, the suppression of HIF-1α inhibited the proliferation of untreated and IGF-1 treated ARP-1 cells. More interestingly, IGF-1 did not fully overcome the growth inhibitory effect of dexamethasone in ARP-1 cells that were nucleofected with HIF-1 siRNA.

Taken together, our results suggest that HIF-1 plays a role in the pathophysiology of MM.
Hemin Prophylaxis for Patients with Frequently Recurring Acute Porphyric Attacks: Experience with Five Cases.
Authors: Griffith Harry S.
Published: October 2019

Background

The acute porphyrias are a group of four clinically indistinguishable diseases characterized by sudden, acute episodes of neurovisceral symptoms that can be life-threatening. Attacks are precipitated by a genetic deficiency in one of the enzymes involved in heme biosynthesis, acting in concert with one or more endogenous or exogenous triggers. The resultant stimulation of hepatic heme results in overproduction of highly reactive heme intermediaries - porphyrins and porphyrin precursors - which accumulate in the liver, blood, and bone marrow. A small percentage of patients have attacks that recur every several weeks, even after all known triggers are eliminated. Although hemin prophylaxis has been recommended for such patients (Anderson K, et al. Recommendations for the Diagnosis and Treatment of the Acute Porphyrias. Annals of Internal Medicine. 2005 Mar; 142(6):439–50), no adequate and well-controlled clinical studies have been conducted.

Methods

We report on five patients, all diagnosed with acute porphyria on the basis of elevated levels of urinary aminolevulinic acid and porphobilinogen (4 with acute intermittent porphyria, 1 with hereditary coproporphyria). All had a history of recurring acute porphyric attacks and received hemin prophylaxis for at least one year in an out-patient setting and were previously treated for a minimum of one year in a hospital setting. The dosage and frequency of prophylactic hemin was based on each patient’s clinical picture, and ranged from a one-day course of 1–4mg/kg/day every 10 to 40 days to a four day-course of 1–4 mg/kg/day every 30 days. The outcome of prophylaxis hemin treatment was determined on the basis of frequency of severe attacks requiring hospitalization and both physician and patient assessment of the severity of symptoms.

Results

Five patients (3 female, 2 male), ranging in age from 25–61 years old suffered from frequently recurring acute porphyric attacks, with symptoms that included abdominal pain (5), psychological disturbances (2), peripheral motor neuropathy (2), and nausea (2). Two patients had a significant reduction in the frequency of their acute attacks and three reported a moderate decrease with prophylactic hemin treatment. All five patients experienced a marked improvement in the reduction of the severity of their attacks. Overall, the occurrence of severe acute attacks requiring hospitalization declined by 66%. Three of the five patients did not require hospitalization over the 12 month period of time. One patient experienced phlebitis during one course of hemin treatment.

Conclusions

Hemin prophylaxis can be an effective disease management strategy in acute porphyria patients with frequently recurring acute attacks.
Association of Mild Anemia and Executive Cognitive Function Impairment in High Functioning Community-Dwelling Older Women.
Authors: Chaves Paulo H.M. , Fried Linda P. , Carlson Michelle
Published: October 2019

Background: Anemia in the elderly is common and may be associated with atypical clinical presentations. Although the impact of severe anemia on cognition has been studied in patients with cancer, less is known about the association of mild anemia on cognitive function in the elderly. Executive cognitive functioning (ECF) refers to the execution of complex activities that involve a set of higher-order cognitive abilities primarily dependent on the frontal lobes. ECF plays a major role in the maintenance of the functional status of older adults. Knowledge about potentially modifiable risk factors for ECF impairment in older adults is limited. The goal of this study was to test the hypothesis that mild anemia is associated with ECF impairment in high functioning, community-dwelling older women.

Methods: Cross-sectional analysis of baseline data from the Women’s Health and Aging Study (WHAS) II, 1994–1996, Baltimore, MD. WHAS II is a population-based study of 436 cognitively intact (Mini-Mental State Exam >24) and highly physically functioning women aged 70–80 years. The analytic sample included 364 subjects with Hb >10 g/dL and the following ECF test components: Trail Making Tests part B (TMTB) - primary outcome - and part A (TMTA), and the difference TMTB-TMTA. Multivariate polytomous logistic regression models were used to model the relationship between performance (based on tertiles) on each ECF test as a function of Hb, while controlling for age, education, cardiovascular disease, and a number of other potential confounders, such as hypertension, diabetes, depressive symptoms, calculated creatinine clearance, forced expiratory volume in the first second, thyroid stimulating hormone, total cholesterol levels, body mass index, walking speed, and prevalent mobility difficulty.

Results: Subjects with mild anemia (Hb 10–12 g/dL) performed worst in all ECF tests; i.e., the percentage of subjects in the best performance tertile for each test was lowest for those with Hb within 10–12 g/dL. Estimates from multivariate models revealed that, as compared to those with Hb≥12 g/dL, those with Hb within 10–12 g/dL were substantially less likely to be in the best performance tertile of the TMTB [odds ratio (OR).23, 95% confidence interval (CI):.06–.86; p=.029], TMTA (OR.27, 95%CI:.07–.98; p=.047), and TMTB-TMTA (OR.30, 95%CI:.10–.89; p=.031), even after comprehensive adjustment.

Conclusion: In this cross-sectional study, mild anemia was an independent risk factor for prevalent ECF impairment in high functioning community-dwelling older women. This raises the hypothesis that correction of mild anemia could potentially provide an opportunity for prevention of cognitive and functional decline in older adults. To test this hypothesis, prospective observational studies and clinical trials are warranted.
Essential Role for Activated Leukocyte Cell Adhesion Molecule in Transendothelial Migration.
Authors: Ofori-Acquah Solomon F.
Published: October 2019

Activated leukocyte cell adhesion molecule (ALCAM/CD166) is a member of the immunoglobulin cell adhesion super family, which has been implicated in diverse physiological and pathophysiological events involving cell migration. Hitherto, ALCAM’s role in inflammation has not been determined. In this study, we show ALCAM is involved in controlling migration of mononuclear leukocytes across the pulmonary endothelium. We demonstrated that ALCAM is localized at intercellular junctions in pulmonary microvascular endothelial cells in vitro and in vivo. ALCAM co-localized with multiple adherens junction molecules including cadherins, catenins and Dlg, as determined by confocal microscopy, and these observations were confirmed by co-immunoprecipitation and co-distribution assays. Treatment of endothelial cultures with EGTA and cytochalasin D translocated ALCAM from intercellular junctions to the cytosol indicating a requirement for homotypic cadherin adhesion and an intact endothelial cytoskeleton for maintaining ALCAM at endothelial cell junctions. Collectively, these data supports the conclusion that ALCAM contributes to the adherens junction complex in endothelial cells. To determine ALCAM’s role in leukocyte-endothelial cell interactions, adult Sprague Dawley rats were intratracheally instilled with macrophage inflammatory protein-1, and this treatment caused acute expression of ALCAM exclusively in newly recruited mononuclear but not polymorphonuclear leukocytes in the alveolar airway. Given that no ALCAM reactivity was observed in peripheral blood leukocytes, we concluded ALCAM is activated as part of the phenotypic switch by mononuclear leukocytes transitioning from circulation to interstitial tissue compartments. To determine the physiological relevance of this finding we examined whether ALCAM was required for transendothelial migration using monocyte chemoattractant protein 1 (MCP-1). MCP-1 dose- and time-dependently increased the number of transmigrated THP-1 monocytes across pulmonary microvascular endothelial monolayers. Recombinant soluble ALCAM dose-dependently reduced the number of transmigrated THP-1 monocytes, whereas in control experiments recombinant soluble vascular endothelial cadherin had no effect on transmigration. This study shows for the first time that ALCAM is located at endothelial cell junctions where it is intimately involved in controlling the number of monocytes that pass through endothelial barriers. ALCAM may therefore play an essential role in the response to inflammation by enhancing recruitment of mononuclear leukocytes by inflamed tissues.
A Study on the Etiology of 912 Anemia Patients in General Hospital.
Authors: Zhao Xiaoying , Zhang Ping , Huang Liansheng
Published: October 2019

Objectives To investigate incidence rate and the etiological factors of anemia in general hospital.

Methods We selected 912 patients with anemia from 4198 inpatients in our hospital from Jan.to Feb.2004,and analyzed their clinical data retrospectively. All inpatients had been detected Hb,RBC,MCV and MCHC. We collected 2ml venous blood in anticoagulant EDTA-k2 tube and measured them with Gen.STM System2 blood cell radiomete in 2 hours. some patients had been punctured bone marrow examination. According to diagnostic standard of anemia(male adult Hb<120g/L, female adult Hb<110g/L, gravida Hb<100g/L) and anemia degree(Mild degree anemia: Hb 90~120 g/L, Moderate degree anemia: Hb 60~90 g/L, Severe degree anemia: Hb30~60 g/L, Profound degree anemia: Hb<30g/L), we classified anemia according to clinical types and cell morphology. according to clinical types, anemia was resulted from primary hematologic disease and secondary anemia. Main causes of secondary anemia were chronic disease, blood loss and chronic systemic disease. According to red blood cell morphology, anemia was classified: microcytic hypochromic anemia (MCV<80fl,MCHC<32%), pure microcytic anemia(MCV<80fl,MCHC32%~36%),normocytic normochromic anemia (MCV80fl~100fl,MCHC32%~36%) and macrocytic anemia( MCV>100fl).

Results The incidence of anemia in inpatients in general hospital was 21.72%. 96.71% of patients were mild-moderate degree anemia and only 13.71% anemia patients showed symptom of anemia such as dizzying,drumming. But only 1.10% of patients had been diagnosed by doctors. 6.36% of patients were caused by hematologic disease and 89.74% of patients coexisted primary disease.53.62% of patient were secondary to chronic disease such as tumor,chronic infection, rheumatic disease. 27.85% of patients were secondary to blood loss. Then 8.00% of patients was caused by chronic systemic disease.

Conclusions Anemia existed in various clinical diseases. Misdiagnosis rate of anemia was high. Appropriate treatment of secondary anemia was beneficial for prognosis of primary diseases. All doctors should pay more attention to anemia in general hospital.
Effects of Zotarolimus (ABT-578), Sirolimus, Paclitaxel and Dexamethasone on Cytokine Production by Human Monocytes In Vitro.
Authors: Li Lanlan , Burke Sandra E. , Mack Matthew M.
Published: October 2019

Introduction: Percutaneous coronary intervention (PCI) and stenting are common therapies used to mitigate symptomatic coronary artery stenosis. These mechanical procedures are invariably associated with local and systemic inflammation, including activation and cytokine release from monocytes/macrophages, which are major stimuli for smooth muscle cell proliferation, migration and restenosis. Drug-eluting stents (DES) may potentially attenuate this inflammatory response, provided that the drug has antiinflammatory potency. This study was conducted to determine the effects of common DES agents on the generation of proinflammatory cytokines by stimulated human monocytes in vitro.

Methods: The production of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) by LPS-stimulated monocytes was determined by ELISA after exposure to either zotarolimus (ABT-578), sirolimus, dexamethasone or paclitaxel.

Results: The results showed that zotarolimus (ABT-578) and sirolimus were potent inhibitors of MCP-1 production (IC50 = 2.6 ± 0.2 and 5.9 ± 1.9 nM, respectively). Dexamethasone also inhibited MCP-1 production, but it was less potent (IC50 = 626.9 ± 60.8 nM). TNF-a and IL-6 production were dose-dependently inhibited by dexamethasone (IC50 = 5.5 ± 0.2 and 13.0 ± 3.7 nM, respectively). However, the production of neither cytokine was significantly inhibited by exposure to zotarolimus or sirolimus. Stimulated monocytes were not affected by paclitaxel; cytokine release after exposure to paclitaxel was no different from controls.

Conclusions: Zotarolimus and sirolimus are potent inhibitors of MCP-1 secretion by stimulated monocytes in culture. Dexamethasone is a less potent inhibitor of MCP-1 secretion, but effectively inhibits the release of TNF-a and IL-6. The anti-restenotic effect of these agents may, in part, be due to their potent antiinflammatory properties.
High Incidence of Ancestral HLA Haplotype 8.1 and Incomplete DH-JH Immunoglobulin Rearrangement in Persistent Polyclonal B-Cell Lymphocytosis.
Authors: Wolowiec Dariusz , Majewski Miroslaw , Nowak Jacek , Stella-Holowiecka Beata , Krawcewicz Angelika , Mika-Witkowska Renata , Makuch-Lasica Hanna , Solarska Iwona , Haus Olga , Duszenko Ewa , Kuliczkowski Kazimierz , Warzocha Krzysztof
Published: October 2019

Four women aged from 48 to 51 years with persistent polyclonal B-cell lymphocytosis (PPBCL) were studied for the clonality of B-cell population and HLA haplotype. All patients were smokers, one presented an immune deficiency syndrome due to decreased IgG and IgA levels, the others were asymptomatic. All patients presented serum increase of polyclonal IgM (5.55 – 8.12 g/L), absolute peripheral lymphocytosis ranging from 6100 to 10500/μL with elevated proportion of B cells, the percentage of CD5/CD19 lymphocytes < 5%, and binucleated/bilobulated lymphocytes accounting for 5–9% of nucleated cells on the blood smear. In one patient, cytogenetic study revealed trisomy 3 in 4 out of 40 metaphases, +18 in 2 cells, and i(18q), +15, t(13,14) in single cells. In the remaining patients no clonal chromosomal abnormalities were found. All patients were HLA typed by PCR based sequence specific primer amplification. HLA allele and haplotype incidence in patients’ group were compared to healthy population of 286 ethnically matched (Caucasian, Polish) controls. DRB1*07 allele was found in 2/4 patients, seems than to be elevated than in general Polish population but a small number of patients does not allow to perform relevant statistical tests. Moreover we found significantly more frequent B*08-DRB1*03 fragment of ancestral HLA 8.1 haplotype (8.1 AH) in PPBCL patients (3/4) than in control group (47/286) (OR=15.3; 95% CI 1.55–150; p=0.02). Peripheral blood mononuclear cells from all PPBCL patients were also studied for monoclonality in immunoglobulin (IGH VH-JH, incomplete DH-JH, IGK, IGL) and crosslineage TCR (TCRB, TCRG, TCRD) genes rearrangements according to BIOMED-2 protocol. The multiplex PCR reactions were followed with heteroduplex analysis of PCR products. We found polyclonality of lymphocytes population in all tests except incomplete DH7-JH IGH rearrangements in 2 patients. The described gene rearrangement was not related to other upstream DH or downstream JH gene segments. The HLA incidence we found is distinct from previously described elevated incidence HLA DRB1*07 allele and may suggest another genetic background of PPBCL pathogenesis. Moreover, our finding of high prevalence of 8.1 AH, known to be more frequent in autoimmune diseases, and to worsen the prognosis in some infectious diseases and non-Hodgkin lymphomas, as well as of the incomplete DJ monoclonality in 2/4 patients, warrant further studies aiming to determine the relationship between PPBCL, immune disorders and true lymphoproliferative diseases.
Vascular Endothelium Is Severely Perturbed and Undergoes Apoptosis in Experimental Heatstroke in Primates.
Authors: Roberts George T. , Chishti Muhammad A. , Al-Mohanna Fallah H. , El-Sayed Raafat M. , Bouchama Abderezak
Published: October 2019

Introduction: Ultrastructural evidence of endothelial cell (EC) injury has been associated with diffuse microvascular thrombosis in human heatstroke (HS). In vitro studies have also shown that heat stress accelerates apoptotic cell death. Using a recently described baboon model of heatstroke, we sought to examine pathological changes in the vascular endothelium and whether apoptosis is a mechanism of endothelial cell death.

Hypothesis: Major structural vascular endothelium alterations occur in HS and apoptosis is a mechanism of endothelial cell death in HS.

Methods: Anesthetized baboons (Papio hamadyras) were heat-stressed in a neonatal incubator maintained at 44 1.5 °C, until rectal temperature attained 42.5°C (moderate heatstroke; n =4) or systolic blood pressure fell to < 90 mm Hg (severe heatstroke n =4). Animals were resuscitated with normal saline and allowed to cool at room temperature. Four sham-heated animals served as control group. Spleen, liver, heart, kidney, gut, lung and adrenal tissue were obtained either by immediate autopsy in non-survivors or after euthanasia at 72-h for survivors. Vascular endothelium ultrastructure was evaluated by transmission electron microscopy (TEM) of ultra-thin tissue sections. Biological activity of EC was determined by light microscopy (LM) using a polyclonal antibody targeting von Willebrand Factor (vWF). Apoptosis was assessed, also in tissue sections, by deoxyuridine triphosphate nick end-labeling (TUNEL) procedure.

Results: In heatstroke animals, there were marked EC changes in lungs, spleen, jejunum, kidneys and liver, demonstrated by TEM, as increased cytoplasmic membrane convolutions that included formation of villi projecting into the vessel lumina, and increase in the width of the gaps between ECs. Migration of neutrophils, platelets and erythrocytes through these widened gaps was noted. Weibel-Palade bodies were increased both in size and number in EC of jejunum, lungs and kidneys. This increase correlated with increased endothelial expression of immunologically detectable vWF. TEM also showed that there was increased apoptosis manifested by nuclear chromatin condensation and karyorrhexis and formation of cytoplasmic myelin whorls. Increased EC apoptosis was also observed by TUNEL in the jejunum, lungs, liver and spleen. All these changes were greater in animals with severe HS than in animals with moderate HS, whereas sham heated control animals showed no significant changes.

Conclusion: Widespread EC injury with apoptotic cell death is consistent with the hypothesis that the endothelium may play a pathogenic role in heatstroke.
The Prognostic Value of Thrombocytosis Associated with Non Small Cell Lung Carcinoma.
Authors: Quispe Dolly , Hanbali Amr , Wollner Ira
Published: October 2019

Non small cell lung carcinoma (NSCLC) is the most common cause of cancer related death in US. During its clinical course patients may develop paraneoplastic syndromes such as thrombocytosis, which has been found in 17–21% patients with lung cancer. Thrombocytosis is associated with extensive disease and a poor prognosis. The objective of this study was to characterize the prognostic value of thrombocytosis in patients with NSCLC.

A retrospective study of inpatient and outpatient medical records at a large, midwestern, urban medical center was performed. Medical records from patients containing a new diagnosis of non small cell lung carcinoma based on ICD-9 codes from 1994 to 2000 were reviewed. Patients with a prior history of malignancy, less than 6 months of follow-up, or without an assessment of their platelet count were excluded. All records were reviewed for the following: TNM stage, histologic type by WHO criteria, and survival. Laboratory data was obtained prior to invasive diagnostic procedures, and thrombocytosis was defined as a platelet count > 400 K/uL. Survival was calculated by Kaplan Meier curves and the difference in survival curves was tested by log rank test.

Thrombocytosis was observed in 113/858 patients (13.2 %). The group with thrombocytosis in comparison with the group without thrombocytosis had a significantly lower mean age at diagnosis (63 y vs 67 y, p<0.001), hemoglobin (11.6 mg/dl vs 13.0 mg/dl, p<0.001) and advanced stages were more frequent, TNM III and IV (79.8% vs 66.2%, p=0.007). No difference was observed between histologic types. 5 year survival was lower in patients with thrombocytosis (4% vs 18%, p <0.001).

Thrombocytosis in patient with non small cell lung carcinoma is associated with advanced stages of the disease and poor survival.

5 Year survival non smell cell lung cancer 5 Year survival non smell cell lung cancer
Hyperhomocysteinemia and Heat Shock Proteins. Epidemiological Study in a General Population.
Authors: Dulin Elena , Guisasola Maria Concepcion , Desco Mª Mar , Suarez Antonio , Garcia-Barreno Pedro
Published: October 2019

Records and objectives: This study is planned with the objective of identifying new efficacious risk vascular markers. For this work the hyperhomocysteinemia as independent thrombotic vascular risk factor, atherosclerosis as inflammatory illness and Heat Shock Proteins (HSP70i) as molecules likely involved in the vascular damage are considered.

Subjects and methods: Workers of the HGGM have volunteered and given their consent. 53 male (M) and 54 female (F) were studied randomly. Anamnesis and epidemiological questionnaire were performed. Arterial pressure was measured and blood samples were taken for neutrophils leukocytes isolation by gradient, biochemical study and DNA extraction. We performed quantification of homocysteine (tHcy)

by HPLC; Reactive-C-Protein (RCP), serum and intraleukocitary HSP70i, and HSP70i antibodies by ELISA; molecular study of the polymorphism of C677T of the enzyme metilentetrahidropholatereductase (MTHFR) by PCR-RFLP. Task force of coronary risk was applied for the population classification.

Results: The studied population present [tHcy] ≥ 11 umol/L in the 12% of the cases. The mutation C677T appears in heterozygote subjects (CT) in the 45% of the cases and in homozygote subjects in the 17%.

Subjects were classified into 3 groups: G-0, without vascular risk factors, n=77 (32M and 45F); G-1, with a moderated risk factor (>10%) but without illness n=26 (18M and 8F) and G-2, with an evident atherosclerotic illness n=4 (3M and 1F)

Conclusions: The following correlations have been proved:

Inverse (p<0.05) between [tHcy] and [HSP70i] in neutrophils in females G-0 and males G1 wild type (CC) Inverse (p<0.05) between [tHcy] and serum [HSP70i] in males G-1 homozygotes (TT) Direct (p<0.05) between [tHcy] and [RCP] in females G-0 heterozygotes (CT) Direct (p<0.05) between [tHcy] and cholesterol in females G-0 homozygotes (TT) Direct (p<0.01) between [tHcy] and HTA in males G-0 heterozygotes (CT)

This study has been partially financed by FIS 03/1308
Acquired Risk Factors Play a Major Role in Venous Thrombosis in Chinese Population.
Authors: Feng Ying , Zhou X. , Xiong X. , Xiong Y. , Wang Y. , Jiang S. , Hu H. , Lu D. , Wang Z.
Published: October 2019

Acquired and genetic risk factors are associated with the occurrence of venous thrombotic embolism (VTE). In the Western population, 30 to 50% of the patients suffering from VTE are due to a genetic tendency to thrombosis. However, some common thrombotic risk factors in thrombophilia such as the factor V Leiden and the G20210A mutation of prothrombin are very rare in China and other Asiatic countries. Recently, we prospectively investigated the incidence of VTE among 768 hospitalized patients who had at least one of acquired risk factors such as prolonged immobilization, surgery, fracture, pregnancy, post-partum, cancer, diabetes mellitus, and some autoimmune diseases. The clinical VTE diagnosis was confirmed with Doppler ultrasonography, CT or/and venography. 17 patients (2.2%) developed VTE with one being complicated with pulmonary thromboembolism during their hospitalization. 90% of these cases had proximal deep vein thrombosis. It is notable that a high incidence of VTE (13.04%) has been found in patients in the intensive care unit, associated with many medical conditions including severe diseases, long lying in bed, multi-organ failure, infection and hypercoagulability. All these 17 VTE patients were extensively studied to identify and characterize the genetic anomalies predisposing possibly to thrombosis. Two were found to be of inherited antithrombin deficiency, and one of inherited protein C deficiency. Each of the three patients had the characteristics of recurrent VTE and/or familial history of VTE with episodes recognized at least in 2 family members. Our results suggest there might be a great variation according to the geographic origin of the population. The acquired risk factors play a much more important role than genetic risk factors in occurrence of VTE in Chinese population in comparison with Caucasians.
Drotrecogin Alfa (Xigris) Induced Coagulopathy as Measured by Routine Lab Tests in a Community Hospital Setting.
Authors: Maini Archana , Madangopal Chudaratna , Sheth Kishor
Published: October 2019

Drotrecogin alfa - Recombinant Human Activated Protein C (Xigris) is FDA approved for the reduction of mortality in adult patients with severe sepsis who have a high risk of death as determined by APACHE II. Patients with sepsis have decreased circulating levels of protein C and that is associated with increased mortality. Drotrecogin alfa presumably has three main mechanisms of action: anticoagulant, anti-inflammatory and pro-fibrinolytic. In the protein C pathway, protein C circulates as a zymogen which is converted to activated protein C (APC) when thrombin complexes with thrombomodulin, further augmented by Endothelial Protein C Receptor (EPCR). This APC, alongwith protein S, inactivates Factors Va and VIIIa, thereby limiting thrombin generation. Endogenous protein C activation is blunted in severe sepsis due to down-regulation of thrombomodulin and EPCR. PROWESS study was the first phase III landmark study to demonstrate an absolute 6.1% reduction in 28 day all cause mortality. The only significant toxicity was a 3.5% bleeding risk, especially with laboratory parameters of aPTT>120 secs, INR>3 and platelet count < 30,000/microliter as well as gastrointestinal ulceration and traumatic injury. Since then several other studies have been conducted with slightly varying results and as many as 19 different systemic biomarkers of thrombosis, coagulation, fibrinolysis and inflammation have been used. But practically, in the ICU setting, the anticoagulant activity of Drotrecogin alfa may be demonstrated by the prolongation of aPTT and PT during the drug infusion. No significant difference has been found in platelet counts in the major studies. The hematocrit is insignificantly changed in the absence of a bleeding complication. We report here a retrospective review of 10 adult patients with sepsis from our ICU who met the criteria to get drotrecogin alfa based on APACHE II. Their PT, INR, aPTT, Hb, Hct and platelet counts were recorded prior to drug infusion not exceeding 24 hours pre infusion. These counts were again looked at post infusion. The highest post infusion coagulation profile was selected and concordant Hb and Hct were recorded. We found the following mean changes in the selected parameters: Increase in aPTT of 58% (20.32 sec); increase in PT of 15% (2.61 sec); decrease in platelet count of 16% (26,000/microliter); decrease in Hb of 13% (1.44 gm/dl); decrease in Hct of 13% (4.09 %). Despite the seeming worsening of coagulopathy and some drop in Hb, Hct and platelets, patients did not have any major bleeding episodes attributable to Drotrecogin alfa. Out of our 10 patients, 7 have expired and 3 have survived for a mean of 130 days (range 118 days to 141 days) till today. Thus in our small retrospective sample, Drotrecogin alfa was found to be a safe and effective drug, based on routine blood tests and clinical observation alone in the ICU of a community hospital.
Clopidogrel Use in Children.
Authors: Finkelstein Yaron , Nurmohamed Laila , Avner Marina , Benson Lee N. , Koren Gideon
Published: October 2019

Background: Children with congenital heart problems have an increased incidence (up to 4%) of thrombotic vascular complications, particularly after interventional catheterization procedures that include device implantation. Clopidogrel, a thienopyridine derivative, is a specific platelet aggregation inhibitor. The preferred anti-thrombotic therapy after percutaneous coronary intervention (PCI) with stent placement in adults has evolved from aspirin and systemic anticoagulation with warfarin to dual antiplatelet therapy with aspirin and clopidogrel. Presently, no published data exists on clopidogrel use in children.

Objective: The aim of the present study was to report the first experience with clopidogrel therapy for primary and secondary prevention of thrombotic phenomena in children with complex heart disease after interventional cardiac catheterization, and to suggest a dose regimen for a paediatric population.

Methods: A retrospective chart review of all infants and children with complex heart disease treated with clopidogrel in the Hospital for Sick Children, Toronto between January 2001 and April 2004. Clopidogrel dosages, duration of therapy, complications and adverse effects in a paediatric population were explored.

Results: Fifteen infants and children with congenital and acquired heart disease were treated with clopidogrel (median age 3.5 years; range 6 weeks- 16 years). In 10 of them endovascular stents were inserted. Dosages ranged from 1 to 6 mg/kg/day, for periods ranging between 1 to 6 months. No thrombotic events were reported in these patients during clopidogrel therapy. One child had a bleeding complication (gastrointestinal) while on triple antithrombotic therapy. Other complications reported in adults, such as rash, were not noted in this paediatric series.

Conclusion: Clopidogrel was well tolerated, and there were no thrombotic events during treatment. We suggest a dose of 1mg/kg/day for children to be started.
The Effect of IL-3 in Ex Vivo Expansion of UCB CD34+ Cells.
Authors: Paek Jin-Young , Kim Do-Yeon
Published: October 2019

Background: Human umbilical cord blood (UCB) has been recognized as a source of hematopoietic stem cells for transplantation and the CD34+ cell dose is important factor predicting the hematologic recovery. However, the cell dose and CD34 (+) cells in one unit UCB are limited. This study compared the ex vivo culture conditions for expansion of total nucleated cells (TNC) and CD 34+ cells using isolated from UCB CD34 (+) and CD 133 (+) subset cells.

Material and method: The isolation of CD34 (+) and CD133 (+) cells from UCB was done by AutoMACS and the purity was more than 95%. We set three different cell culture conditions in stroma-free, serum-free medium. The culture conditions were 1) TPO, stem cell factor (SCF), Flt3-1 2) TPO/SCF/FLT3-1 and IL 3 10ng/dl, 3) TPO/SCF/FLT3-1 and IL 6 10ng/dl. After 7 days and 14days culture, we have been evaluated TNC and CD 34 (+) cell number for expansion potential.

Results: The TNC after 7 days and 14 days culture, in the CD 34(+) subset, the TPO/SCF/Flt3-1 condition was 15.6 fold and 8.0 fold and the TPO/SCF/Flt3-1 + IL-3 condition was 31.6 fold and 20.1 fold and the TPO/SCF/Flt3-1 + IL-6 condition was 25.5 fold and 12.0 fold, respectively. In the CD133 (+) subset, the TPO/SCF/Flt3-1 condition was 7.1 fold and 8.4 fold and in the TPO/SCF/Flt3-1 + IL-3 condition was 17.9 fold and 24.8 fold and in TPO/SCF/Flt3-1 + IL-6 condition was 15.9 fold and 19.8 fold, respectively. In terms of CD34 (+) cell population in the expanded cells, in the CD34 (+) subset, the TPO/SCF/Flt3-1 condition was 40%, the TPO/SCF/Flt3-1 + IL-3 condition was 64% and in the TPO/SCF/Flt3-1 + IL-6 condition was 40%, respectively. In the CD 133 (+) subset, the TPO/SCF/Flt3-1 condition was 83% and in TPO/SCF/Flt3-1 + IL-3 condition was 48% and in TPO/SCF/Flt3-1 + IL-6 condition was 62%, respectively.

Conclusions: To obtain more CD 34(+) cell population among ex vivo expanded cells, 7 days culture was more effective than 14 days culture and CD 133(+) subset was better than CD 34(+) subset. In terms of cell expansion ratio TPO/SCF/Flt3-1 + IL-3 was better than TPO/SCF/Flt3-1 and TPO/SCF/Flt3-1+ IL-6, which would be support the relevance of 7 days culture and CD133 (+) subset and IL-3 for expansion of hematopoietic stem cell ex vivo expansion.

Fig 1: The expansion ratio of both CD (34+)(a) and CD133 (+)(b) subset after 7 days and 14 days cultures in the 3 different cell culture conditions Fig 1:. The expansion ratio of both CD (34+)(a) and CD133 (+)(b) subset after 7 days and 14 days cultures in the 3 different cell culture conditions

Fig 2: The CD34(+) cell percentages in the different culture conditions both CD34(+) and CD133(+) subsets. Fig 2:. The CD34(+) cell percentages in the different culture conditions both CD34(+) and CD133(+) subsets.
Autologous Bone Marrow Stem Cells (ABMC) for Cardiormyoplasty in 15 Patients with Acute and Chronic Myocardial Disease (ACMD) in Colombia.
Authors: Cuéllar-Ambrosi Francisco , Senior-Sánchez Juan Manuel , Velásquez-Uribe Oscar , Navas-Rios Claudia , Londoño-Blair Juan Luis , Velásquez-Lopera Margarita
Published: October 2019

Introduction: ACMD disease accounts for 50% of all cardiovascular deaths and is the leading cause of congestive heart failure as well as premature permanent disability in workers. Unfortunately no medication or procedure has shown efficacy in replacing myocardial tissue or neovascularization. Some clinical human trials indicate that ABMC transplantation improves cardiac function and is feasible, safe and beneficial. We report the first experience in Colombia with this type of therapy

Methodology: This is the six months follow-up of 15 patients 23 - 60 years-old, 10M/5F with: A. an extensive, viability absence, acute myocardial infarction of anterior myocardial wall, 10 patients, with ejection fraction ≤ 40% that received standard postinfarction medical treatment, successful coronary percutaneous intervention and after intracoronary transfer of G-CSF mobilized ABMC, CD34 range 15–20 in millions. B. Five patients with chronic coronary heart disease and/or severe ischemic heart failure without option for standard revascularization therapies recibed epicardic implantation of G-SCF mobilized ABMC, CD34 range 14.5–19 in millions.

Results: It was demonstrated in both groups:

improvement in the mean ejection fraction from 29.44±3.36% before to 45±14,97% (p<0.001) after six months the exercise capacity increased in meaningful form evidenced by mean increase in the six minutes test from 295.07±66.04ms before to 648.4±93.15 ms after six months (p<0.001), the treadmill exercise time and the functional capacity in Mets from a mean of 4,8±1.02 before to 12.8±2.59 (p<0.001). there were significant changes in the myocardial perfusion at six months follow-up by nuclear images at diagnosis

Figure Figure

and 6 months later

Figure Figure

There were no presented complications related to the Cellular Transplant or the utilization of the G-CSF.

Conclusions: This is the first experience in Colombia with stem cells for intracoronary and epicardic cardiomyoplasty in acute and chronic coronary heart disease. It was observed functional recovery of the left ventricle, improvement in the exercise capacity without adverse effects or complications related to this therapy. In patients with ACMD, treatment with G-CSF to mobilize ABMC is feasible and safe and seems to be effective under clinical conditions. The therapeutic effect might be attributed to ABMC-associated promotion of myocardial regeneration and neovascularization.
Induction of Human Umbilical Cord Blood Mesenchymal Stem Cells into Nerve-Like Cells by Salvia Miltiorrhiza.
Authors: Ma Lian , Feng Xueyong , Yang Liye , Luo Minjie , Xie Qingdong , Huang Tianhua
Published: October 2019

Salvia miltiorrhiza is a traditional Chinese herb with thousands of years history. Composite salvia injection (CSI), a widely used administration in China nowadays, consists of Salvia miltiorrhiza, notoginseng, radix and borneo. It can promote blood flow, but also regulate qi to alleviate pain. So it is frequently used for angina, coronary heart disease and chest discomfort. Recently, some laboratory studies show that CSI may have the effect of cell induction.. In this study, we aim to investigate its neural induction.

Objective: To investigate the differentiation-inducing effects of salvia miltiorrhiza on umbilical cord blood mesenchymal stem cells.

Methods: The immunophenotypes of human umbilical cord blood-derived mesenchymal stem cells(MSCs) were determined by flow-cytometry. Salvia miltiorrhiza was adapted to induce the differentiation of the primary cultured cells of umbilical cord blood and MSCs. The results were compared with nerve growth factor(NGF) and ganglioside(GM1). The differentiated and undifferentiated cells were identified with immunocytochemistry.

Results: In primary culture, certain cells proliferated and formed cell clusters. These cells were either fibroblast-like or round in shape with various sizes. The positive rates for MSCs phenotypes CD29, CD44, CD59 of primary culture cells were 10.7%,37.27% and 66.67%, respectively. Salvia miltiorrhiza could induce the primary cultured cells of umbilical cord blood to express neural phenotypes. After passage, the MSCs could be maintained in vitro for extended periods with stable population doubling. The positive rates for CD29, CD44, CD59 of human umbilical cord blood-derived MSCs were 40.2%,70.5% and 95.4%, respectively. Salvia miltiorrhiza could induce the MSCs to express three kinds of proteins: nestin(the marker of neuronal precursor stem cells), β-TubulinIII and neurofilament (NF) (the marker of neuronal phenotypes) and glial fibrillary acidic protein(GFAP, the marker of astrocyte). Besides, NGF and GM1 could also induce the same MSCs to nerve-like cell, which could express the same markers: β-TubulinIII, NF and GFAP. In comparison NGF and GM1, the induction effect of salvia miltiorrhiza demonstrated the higher rate of expressing neuron phenotypes at a faster speed.

Conclusion: The human MSCs could be isolated from human umbilical cord blood. Salvia miltiorrhiza can induce the MSCs to differentiate into nerve-like cells, therefore, might act as a potential drug for neural induction.
Mesenchymal Stem Cells Modulates Differentiation into Hepatocyte Cell in VX2 Implanted Rabbit Liver Cancer Model.
Authors: Ren Jun , Wang Xiaodong , Di Lijun , Yang Renjie , Jiang Hanfang , Hu Weiheng
Published: October 2019

The issues of differentiation of mesenchymal stem cells still remain great increasing interests for the last recent years. For non-hemotological malignancies could these pluripotential stem cells be served as surrogates for the reconstruction of cell and organ function when solid carcinoma occurred or metastasized. New Zealand rabbits liver cancer model with squamous cells of VX2 implanted were used to explore the possibilities of mesenchymal stem cell into the hepatocytes. The results showed that there were abundant dispersed small roundness cells with mononuclear or binuclear were found in portal areas of normal liver lobes after mesenchymal stem cells had been implanted at 4 weeks, and those were originated from mesenchymal stem cells without expression of albumin, CK18 and CK19 by confocol laser microspopy; but the arrangement of partial hepatic cords was disordered near the portal areas of normal liver lobes, the cells was small like as renewal cells expressing albumin, Ck18 and CK19. It is hopefully firstly provided the evidence that mesenchymal stem cell could have the abilities and modulate the differentiation of hepatocytes which secreting albumin as well as CK18, CK19, but did not differentiate into cancer cells.
Amiodarone- Induced Bone Marrow Granulomas.
Authors: Mohamed Tarek , Sycheva Tatyana , Aish Leo , Schneider Douglas , Oo Thein H.
Published: October 2019

INTRODUCTION: Amiodarone is associated with several adverse effects, including hematologic side effects such as pancytopenia, hemolytic anemia and aplastic anemia. Very few cases of amiodarone-induced bone marrow granulomas have been reported.

METHODS: We report 2 cases of amiodarone-induced bone marrow granulomas. Patient 1: 81-year-old male presented with leukopenia, thrombocytopenia, and hepatosplenomegaly after 2 years of amiodarone. Patient 2: 80-year-old male presented with pancytopenia 2 ½ years after starting amiodarone. Examination was unrmarkable. Bone marrow biopsies on both patients showed non-caseating granulomas with negative stains for acid-fast bacilli and fungi. Both had normal CBC before amiodarone therapy. The literature was reviewed for amiodarone-induced bone marrow granulomas on MEDLINE.

RESULTS: There are 8 cases of amiodarone-induced bone marrow granulomas in the literature. Male: female - 5:3. One also had amiodarone-induced hepatic granulomas. Amiodarone was stopped in 5 cases with the improvement of granulomas in 3 cases. Patient characteristics and outcomes are shown below.

Ref Age/Sex CBC/Chemistry abnormalities Associated blood disorders Amiodarone stopped outcome/Note Boutros et al 67M none MGUS Yes fewer granulomas in bone marrow Boutros et al 77F platelet ↓ none Yes platelet count normalized and fewer granulomas in bone marrow Rosenbaum et al 53/F WBC ↑, Platelet ↑ myelofibrosis No also has amiodarone-induced hypothyroidism Rosenbaum et al 78/F RBC ↓ Yes Doing well. CBC unknown Mukhopadhyay et al 81/M RBC ↓ MDS No Erythropoetin improved anemia Mukhopadhyay et al 62/M pancytopenia none No Also has splenomegaly. CBC stable Yamreudeewong et al 63/M Unknown none Yes Bone marrow biopsy performed for fever of unknown origin. Symptoms improved. Moran et al 76/M RBC ↓, LFT ↑ MGUS Yes Bone marrow granulomas resolved. Liver biopsy showed granulomas. Our patient 1 81/M WBC ↓, Platelet ↓ none Yes Also has amiodarone-induced hyperthyroidism. Hepatosplenomegaly and CBC stable Our Patient 2 80/M Pancytopenia none No Erythropoetin improved anemia

CONCLUSION: Bone marrow granulomas, although rare, should be considered as a differential diagnosis in patients on amiodarone presenting with cytopenias.
Cellular Iron Metabolism Studies Demonstate Safety of Magnetic Tracking of Mesenchymal Stem Cells.
Authors: Pawelczyk Edyta , Arbab Ali S. , Pundit Sunil , Frank Joseph A.
Published: October 2019

Background: Ferumoxides-protamine sulfate (Fe-Pro) complexes are used for intracellular magnetic labeling of mesenchymal stem cells (MSCs) to non invasively monitor cell trafficking and tissue distribution by in vivo magnetic resonance imaging (MRI). It has been previously shown that there was no short or long- term toxic effects on cell’s viability, proliferation, reactive oxygen species and biological functions including differentiation capacity of MSCs, however the effects of dextran coated superparamagnetic iron oxide (SPIO) nanoparticles labeling on cellular iron metabolism and storage is unknown.

Objective: Evaluate the effect of using Fe-Pro complexes for magnetic cell labeling in MSCs on the expression of transferrin receptor (TfR-1) and ferritin, proteins involved in intracellular iron metabolism and storage.

Methods: Confluent MSCs were labeled with Fe-Pro complexes at the ratio of 100 ug/mL to 6 ug/mL. After labeling, cells were washed and cultured for 2 months. TfR-1 and ferritin expression were evaluated at the gene and protein levels in total cell lysates, on day 1, 3, 7, 14, 28 and 35 days post-labeling, by real-time polymerase chain reaction (RT-PCR) and Western blotting. MSCs were evaluated for the presence of iron by Prussian blue stain and light microscopy. In addition, TfR-1 and ferritin levels were determined for Fe-Pro labeled and unlabeled MSCs, allowed to slowly multiply, as a result of dividing the cells in half when they reached confluence.

Results: All cells were Prussian blue positive on light microscopy following labeling with Fe-Pro. Labeling of MSCs grown to confluence resulted in short-term increase of TfR-1 mRNA (day 1 and day 3, p= 0.0056) without changes in TfR-1 protein levels. Fe- Pro labeled MSC demonstrated an increase in ferritin gene expression on day 7 and 14 (p=0.0003) compared to unlabeled cells, while protein levels were higher compared to unlabeled MSC at each time point (p=0.005). However, ferritin protein levels from Fe-Pro labeled MSCs were not significantly increased after 7 passages.

Conclusions: Magnetic labeling of cells with FDA approved ferumoxides complexed to protamine sulfate elicited short-term changes in cellular iron metabolism and storage that was dependent on ability to divide. Fe-Pro labeling did not have any long-term implications validating the safety of Fe-Pro labeling technique for MSCs trafficking studies by MRI.