Medium Corpuscular Volume Research Articles

Changes in echocardiographic and analytical parameters after iron replacement in stable heart failure patients. FER-STRAIN study

Abstract Background Iron deficiency replacement with intravenous ferric carboxymaltose (FCM) has demonstrated to improve clinical and prognostic parameters in patients with heart failure with reduced left ventricular ejection fraction (HFrEF). Improvement of myocardial contractility is proposed as possible hypothesis and strain could behave as an early and high sensitive parameter to assess this improvement in myocardial deformation due to increased iron availability after replacement of iron deficiency. Purpose Evaluate the effect of iron deficiency replacement with FCM in the strain of both ventricles in patients with stable HFrEF. Methods We performed an observational, analytical, longitudinal and prospective study that included 73 patients with stable HFrEF who presented iron deficiency without anemia defined as serum ferritin <100 ug/L or between 100 and 300 ug/L if the transferrin saturation was <20%. They were randomized 2:1 to iron replacement with FCM or control group. A central core lab, blinded to the treatment/control groups performed 3D echocardiograms and strain analysis. Analytical parameters and biomarkers were determined basal and four weeks after the randomization. Results Mean age of the study population was 70 years old, 77% were male, 75% had high blood pressure, 70% dyslipidemia and 41% diabetes. Of the total, 51 patients were assigned to the replacement group and 22 patients to the control group. Mean left ventricular ejection fraction (LVEF) was 33%, without basal differences between both groups. Four weeks after the randomization, a significant improvement of LVEF of 2.47±2.32% (p=0.018) was observed in the replacement group, as well as in the left ventricle global longitudinal strain (LV-GLS) of -0.96±2.08% (p=0.006). In blood tests, a significant improvement of ferritin (130.9±111.98mg/L, p<0.000), hemoglobin (0.3±0.7g/dL, p=0.027) and medium corpuscular volume (1.7±2.16fL, p=0.005), and also an increase of the biomarker myeloperoxidase (40.16±138.36pmol/L, p=0.02) were detected in the treatment group. Nevertheless, these changes were not observed in the control group. Conclusions In patients with stable HFrEF replacement of iron deficiency with FCM is associated with an improvement of myocardial contractility, of LVEF measured by 3D and LV-GLS.Variation of parametersLV-GLS before and after FCM

Circular Economy, Dairy Cow Feed Leftovers, and Withania somnifera Supplementation: Effects on Black Belly Ram's Libido, Sperm Quality, Sexual Behavior, and Hemogram Values.

Considering a circular economy perspective, this study evaluates the possible effect of targeted short-term supplementation with Withania somnifera L. (WS; Ashwagandha) on ram's seminal quality, socio-sexual behaviors, and blood constituents. Black Belly rams (n = 20) received a basal diet comprising feed-leftovers from dairy cows in the north-arid Mexico (i.e., Comarca Lagunera CL). The experimental units, with proven libido and fertility, were homogeneous in terms of age (3.41 ± 0.21 yr.), live weight (LW; 53.8 ± 3.3 kg), body condition (BC; 2.96 ± 0.01 units), initial sperm concentration (2387 ± 804 × 106), and viability (23.9 ± 15.6%). Rams were randomly assigned during the transition reproductive period (i.e., May to Jun; 25° NL) to three treatment groups: non-supplemented control group (CONT; n = 6), low WS-supplemented (LWS; i.e., 100 mg kg LW-1 d-1 × 40 d; n = 7), and high-WS-supplemented (HWS; i.e., 200 mg kg LW-1 d-1 × 40 d; n = 7). The basal leftover diet was offered twice daily (0700 and 1600 h); the experimental period (EP) lasted 47 d. No differences (p > 0.05) among treatments occurred regarding LW and BCS at the onset of the EP. Whereas the greater scrotal circumference (SCRC, cm) arose in the LWS and CONT rams, an increased ejaculated volume (VOLEJA, mL) occurred in the WS-rams. A total of 5/9 (i.e., 55%) appetitive and 3/3 (i.e., 100%) consummatory sexual behaviors favored (p < 0.05) the WS-rams, particularly the HWS rams, towards the final EP. The same was true (p < 0.05) regarding the hemogram variables white blood cell count (×109 cells L-1), hemoglobin concentration (g dL-1), and medium corpuscular volume (fL). This study, based on a rethink-reuse-reduce enquiry approach, enabled connectedness between two noteworthy animal systems in the CL: dairy cows and meat sheep schemes. Certainly, the use of dairy cow feed-leftovers aligned with the short-term supplementation with WS promoted enhanced testicular function, augmented seminal volume, and an increased sexual behavior in Black Belly rams in northern Mexico. Finally, while our research outcomes should enhance not only the resilience and sustainability of sheep production and the well-being of sheep-producers and their families, it may also embrace clinical translational applications.

Mean corpuscular haemoglobin concentration (MCHC): a new biomarker for high-altitude pulmonary edema in the Ecuadorian Andes

Ascent to high altitude (> 3000 m height above sea level or m.a.s.l) exposes people to hypobaric atmospheric pressure and hypoxemia, which provokes mountain sickness and whose symptoms vary from the mild acute mountain sickness to the life-threatening, high-altitude pulmonary edema (HAPE). This study analysed the risk factors underlying HAPE in dwellers and travellers of the Ecuadorian Andes after sojourning over 3000 m height. A group of HAPE patients (N = 58) was compared to a NO HAPE group (N = 713), through demographic (ethnicity, sex, and age), red blood cell parameters (erythrocytes counts, hematocrit, median corpuscular volume, median corpuscular haemoglobin, and median corpuscular haemoglobin concentration (MCHC)), altitude (threshold: 3000 m.a.s.l.), and health status (vital signs) variables. Analysis of Deviance for Generalised Linear Model Fits (logit regression) revealed patterns of significant associations. High-altitude dwellers, particularly children and elder people, were HAPE-prone, while women were more tolerant of HAPE than men. Interestingly, HAPE prevalence was strongly related to an increment of MCH. The residence at middle altitude was inversely related to the odds of suffering HAPE. Ethnicity did not have a significant influence in HAPE susceptibility. Elevated MCHC emerges like a blood adaptation of Andean highlanders to high altitude and biomarker of HAPE risk.

Oxidative Stress and DNA Damage Markers in Colorectal Cancer.

Oxidative stress (OS) and inflammation are known to play an important role in chronic diseases, including cancer, and specifically colorectal cancer (CRC). The main objective of this study was to explore the diagnostic potential of OS markers in patients with CRC, which may translate into an early diagnosis of the disease. To do this, we compared results with those in a group of healthy controls and assessed whether there were significant differences. In addition, we explored possible correlations with the presence of tumors and tumor stage, with anemia and with inflammatory markers used in clinical practice. The study included 80 patients with CRC and 60 healthy controls. The following OS markers were analyzed: catalase (CAT), reduced glutathione (GSH) and oxidized glutathione (GSSG) in serum; and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) and F2-isoprotanes in urine (F2-IsoPs). Tumor markers (CEA and CA 19.9), anemia markers (hemoglobin, hematocrit and medium corpuscular volume) and inflammatory markers (leukocytes, neutrophils, N/L index, platelets, fibrinogen, C-reactive protein, CRP and IL-6) were also determined. Comparison of means between patients and controls revealed highly significant differences for all OS markers, with an increase in the prooxidant markers GSSG, GSSG/GSH ratio, 8-oxodG and F2-IsoPs, and a decrease in the antioxidant markers CAT and GSH. Tumor and inflammatory markers (except CRP) correlated positively with GSSG, GSSG/GSH ratio, 8-oxodG and F2-IsoPs, and negatively with CAT and GSH. In view of the results obtained, OS markers may constitute a useful tool for the early diagnosis of CRC patients.

Comportamiento fisiológico de pollos de engorde usando dietas con Tithonia diversifolia y

Los probióticos permiten establecer y controlar la microflora beneficiosa y gradualmente reducen la microflora patógena. Tithonia diversifolia (TD) es una alternativa para reemplazar la proteína de otras fuentes más caras en las dietas para aves de corral; por lo tanto, los tratamientos usaron reemplazos de TD con 0, 5, 10 y 15% con (WP) y sin adición de probióticos (W/OP): Saccharomyces cerevisiae (SC), Lactobacillus Acidophilus (LA) and Bacillus subtilis (BS) y la mezcla de estos (MP) para estimar la digestibilidad in vivo. Se utilizaron 300 aves Cobb avian, con un peso promedio de 453.4 ± 24 g distribuidos aleatoriamente con un arreglo factorial 4x5x3x5. La digestibilidad de la materia seca (DM), proteína cruda (CP) y fibra (CF) se determinó mediante el método de recolección de heces totales; adicionalmente: se analizaron el hemograma (CBC) y la química sanguínea (BC). La digestibilidad de la DM aumentó con la proporción de TD (15%) en la dieta y alcanzó el 68% W/OP; mientras que esta variable aumentó a 70.67% con SC (P

Relationship between Obesity and Iron Deficiency in Healthy Adolescents.

The relationship between overweight/obesity (excess of weight [EW]) and iron deficiency (ID) is not well defined. Objective: To analyze the relationship between EW and ID in healthy adolescents, assessing the contribution of new diagnostic measures of iron status and erythropoietic activity. Method: A cross-sectional study was made of 405 healthy adolescents, 12-16 years of age. A total of 289 were normal weight (NW) and 116 were otherwise healthy EW. Epidemiological, socioeconomic, diet, BMI Z-score, CRP (C-reactive protein), hematological, iron status, and erythropoietic activity parameters were measured. Statistical tests were Student's, analysis of variance (ANOVA), Chi-square, Pearson's correlation, and odds ratio. Results: ID prevalence in the EW group was 22.6% vs. 29.5% in the NW group (p: 0.3). Greater body weight was associated with lower reticulocyte hemoglobin content (CHr) (NW: 31.3 ± 1.7 pg vs. OW: 30.2 ± 1.7 pg, p: 0.007) and greater CRP (NW: 0.1 ± 0.2 mg/dL vs. OW: 0.2 ± 0.18 mg/dL, p < 0.001), leukocytes (NW: 6.69 ± 1.57 × 103/L vs. OW: 7.43 ± 1.63 × 103/L, p < 0.02), platelets (NW: 265.6 ± 58.9 × 103/L vs. OW: 291.8 ± 54.4 × 103/L, p < 0.002), ferritin (NW: 32.1 ± 17.9 ng/mL vs. OW: 42.8 ± 20.3 ng/mL, p: 0.01), serum transferrin receptor (sTfR) (NW: 1.39 ± 0.4 mg/L vs. OW: 1.73 ± 0.45 mg/L, p: 0.008), sTfR-F index (sTfR/log Ferritin) (NW: 1.06 ± 0.7 vs. OW: 1.33 ± 0.85, p: 0.036) and reticulocyte count (NW: 54.2 ± 18 × 103/L vs. OW: 65.4 ± 24.2 × 103/L, p: 0.003). A positive correlation was observed between the BMI Z-score and CRP, ferritin, sTfR, leukocytes, platelets, and reticulocyte count, and a negative one between the BMI Z-score and CHr and medium corpuscular volume. Conclusions: The prevalence of ID in otherwise healthy EW adolescents was no higher than in NW. The effect of obesity on iron status was low. The findings reveal the concomitant low-grade inflammation, and probably the effect of adiposity on erythropoietic activity. Specific cutoff values for ID in adolescents with OW need to be defined.

Significance of Markers of Monocyte Activation (CD163 and sCD14) and Inflammation (IL-6) in Patients Admitted for Alcohol Use Disorder Treatment.

Monocyte activation and inflammation are prominent features of alcohol-related liver disease; however, they have not been thoroughly assessed in patients with alcohol use disorder (AUD) without overt liver disease. This study aimed to analyze associations among clinical and laboratory variables and markers of monocyte activation (CD163 and sCD14), and inflammation (interleukin [IL]-6) among AUD patients. We analyzed the aforementioned associations in the highest quartile in 289 patients (77.5% male; median age, 50years) consecutively admitted for alcohol detoxification in 2 tertiary hospitals in the Barcelona metropolitan area, Spain. Median alcohol intake was 142g/d; median glucose, albumin, creatinine, and bilirubin levels (mg/dl), 92, 40, 0.78, and 0.69, respectively; median AST, 41 U/l; median hemoglobin, median corpuscular volume, and platelet count, 14.1g/dl, 94.8fL, and 189×109 /l, respectively; median cholesterol, triglyceride, fibrinogen, and ferritin levels, 187mg/dl, 109.3mg/dl, 341mg/dl, and 177ng/ml, respectively. In addition, 36.7% patients had an erythrocyte sedimentation rate >20mm, 32.5% had a C-reactive protein (CRP) level of >5mg/l, and 10.9% were hepatitis C virus (HCV)-positive. Median CD163, sCD14, and IL-6 levels were 759, 1.68×106 , and 4.37pg/ml, respectively. On logistic regression analyses, glucose, AST, bilirubin, hemoglobin levels, and HCV infection (adjusted odds ratio [aORs]: 1.01, 1.02, 3.04, and 9.73, respectively) were associated with CD163. Glucose, AST, triglyceride, and CRP >5mg/l (aORs: 1.02, 1.01, 1.00, and 3.49, respectively) were associated with sCD14. Alcohol consumption upon admission, MCV, total cholesterol levels, and CRP >5mg/l (aORs: 0.99, 1.05, 0.99, and 2.56, respectively) were associated with IL-6. Monocyte activation and systemic inflammation are associated with higher glucose, liver enzyme, and lipid levels, HCV infections, and CRP of >5mg/l, thus potentially identifying patients with AUD at high risk of midterm poor outcomes.

Géophagie associée à une anémie sévère chez la femme non gravide : à propos de 12 cas

IntroductionGeophagy or soil eating is mostly described in pregnant women from Sub-Saharan Africa, South America. Here, we report 12 cases of geophagy associated with severe anemia in non-pregnant Nigerian women. Results/case reportsThe median age at diagnosis was 34.5 years. The socioeconomic level was average for all patients. The median hemoglobin level at admission was 6.9g/dL (3.3–8.6), median corpuscular volume was 78.3fL (63–106) and median serum ferritin was 9.2ng/mL (3.6–11.2). The reasons of this practice were “desire” (5/12) and tradition (4/12). All patients received psychotherapy and supplementation with intravenous iron. ConclusionGeophagy is an underestimated practice in developed countries and in non-pregnant women. It can be the cause of severe iron deficiency and must be discussed in patients with anemia, including non-pregnant patients, and in Africa as well as in migration areas, where the practice can be exported.

Alcohol Use in Patients After Liver Transplantation for Alcoholic Liver Cirrhosis Detected by the Questionnaire (Regional Experience)

Background: Alcoholic liver cirrhosis is a common indication for liver transplantation. Recurrence of alcohol use was retrospectively analyzed after liver transplantation. Methods: From X/1995 to II/2002, 42 out of 246 patients underwent liver transplantation for alcoholic liver cirrhosis. We administered a questionnaire to all patients surviving at least 1 year post liver transplantation for alcoholic liver cirrhosis. Liver biopsy was performed annually. The results of the questionnaires of both alcohol use and sobriety group were compared with laboratory parameters, graft histopathology, socioeconomic status and survival rate. Results: Sixteen patients admitted alcohol consumption in the questionnaire. Serious graft damage was diagnosed in 4 recipients, 2 of them died due to graft failure, one for another reason, 12 returned to alcohol use. We have found more steatosis, presence of Mallory hyaline and no difference in fibrosis between alcohol use and sobriety group. The alcohol use and sobriety group differed in body mass index, triacylglycerols, medium corpuscular volume and activity of � - glutamyltransferase. Cumulative survival rate (1, 3, 5 years) was significantly lower in alcohol use group than in sobriety group. Conclusion: Patients after liver transplantation for alcoholic liver cirrhosis with alcohol use had lower survival rate as compared with sobriety group. Considering the differences in histopathology and laboratory parameters in patients with and without alcohol use after liver transplantation, administration of questionnaires seems to be a valid tool for recurrence of alcohol use detection.

Hematologia de tilápia-do-nilo alimentada com suplemento à base de algas frente a desafios de estresse agudo e crônico

Avaliou-se o efeito da suplementação com produto à base de algas, ergosan, em dietas para tilápias Oreochromis niloticus, submetidas ao desafio de estresse agudo e crônico sobre as características hematológicas. O ensaio foi realizado com tilápias jovens (15g) em caixas de 500L de capacidade de estocagem, em sistema com recirculação da água, com fluxo constante, na densidade de 35 peixes por caixa. Os peixes foram alimentados com as dietas-teste durante 10 dias e, ao final deste período, voltaram a receber dieta isenta de ergosan, quando foram submetidos aos desafios de estresses agudo e crônico. Utilizou-se o delineamento inteiramente ao acaso (DIC), em esquema fatorial 4x2, sendo testados: quatro porcentagens do suplemento 0; 0,25; 0,5 e 1% de ergosan na ração em dois tipos de estresse, agudo e crônico. Os resultados permitem concluir que os estímulos de estresse aplicados foram capazes de provocar alterações fisiológicas nos peixes, incluindo redução no número de eritrócitos, eritroblastos e leucócitos e aumento do volume corpuscular médio dos eritrócitos. A administração oral de 1% de suplemento durante 10 dias para juvenis de tilápia-do-nilo, em condição de estresse agudo, induz o incremento de 69% no número de trombócitos circulantes.

A HPLC–MS method for the simultaneous quantification of fourteen antiretroviral agents in peripheral blood mononuclear cell of HIV infected patients optimized using medium corpuscular volume evaluation

A sensitive and accurate high performance liquid chromatography–mass spectrometric (HPLC–MS) method for the intracellular determination of 14 antiretroviral drugs in peripheral blood mononuclear cells (PBMCs) for HIV+ patients was validated. PBMCs are isolated by Ficoll density gradient centrifugation and cells count and the relative mean volume is performed with a Coulter® instrument. Extraction of drugs from PBMCs pellets was obtained with methanol:water (70:30, v/v), with quinoxaline added as internal standard, after a sonication step. Supernatant was dried and then dissolved in water/acetonitrile (60/40, v/v), before injection into a 2.1mm×150mm Atlantis® T3 3μ column. Chromatographic separations were performed using a gradient program with a mixture of water (0.05% formic acid), as mobile phase A and acetonitrile (0.05% formic acid), as mobile phase B. Analytes quantification was performed by electro-spray ionisation–single quadrupole mass spectrometry using the selected ion recording (SIR) detection mode. The positive ionization was used for the HIV protease inhibitors (PIs) indinavir, saquinavir, nelfinavir, nelfinavir M8 metabolite, amprenavir, darunavir, atazanavir, ritonavir, lopinavir, tipranavir, the integrase inhibitor (II) raltegravir and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine and etravirine, while the negative ionization is applied for efavirenz. The calibration curves were built using blank PBMCs spiked with antiretroviral drugs at concentrations ranging from 0.1 to 32ng/mL (1–320ng/mL for tipranavir) and fitted to a quadratic regression model weighted by 1/X. The mean extraction recovery for all PIs, II and NNRTIs was always above 82%. The method was precise, with a range of intra/inter-day percent standard deviation within 2.6–14.8%, and accurate with mean of percent coefficient of variation (CV%) from nominal values −7.85 to +9.7%. Each drug concentration evaluated was expressed in ng/mL and optimized using each patient medium corpuscolar volume and cell number. This analytical method is routinely used in our clinical research center for the assessment of intracellular levels of all PIs, raltegravir and NNRTIs commercially available at present.

Prevalencia y factores predictivos de la presencia de anticuerpos anticélula parietal gástrica en la enfermedad tiroidea autoinmune

Our aim was to evaluate the prevalence of parietal cell antibodies (PCA) in patients with autoimmune thyroid disease (ATD). We performed a descriptive, cross-sectional study of patients with ATD. The presence of PCA was determined. Elevated antithyroid antibodies (ATAs) were defined as those higher than the 75th percentile of distribution. Multivariate logistic regression models were built to assess the independent contribution of the following variables to PCA positivity: age, sex, hemoglobin, medium corpuscular volume (MCV), dose/Kg of levothyroxine (LT4), disease duration and elevated ATA levels. A total of 148 patients were included (137 females). The mean age was 45.7 (SD 15) years and disease duration was 4.5 (SD 4) years. Forty-three patients (29%) with Graves' disease and 105 (71%) with primary hypothyroidism were included. The 75th percentile of distribution was 420U/ml for anti-peroxidase antibodies and 200U/ml for anti-thyroglobulin antibodies. PCA positivity was found in 30 patients, with an overall prevalence of 20.3%. PCA positivity with titers higher than 1/640 was found in 19 patients (12.8%). The only independent predictive factor of PCA positivity was the presence of elevated levels of ATAs (odds ratio (OR)=3; 95% confidence interval (CI): 1.1-8.6; p=0.04). The only independent predictive factor of PCA positivity at titers >/=1/640 was also the presence of elevated levels of ATAS (OR=7.3; 95% CI: 1.6-32.7; p=0.009). The prevalence of PCA positivity in patients with ATD was 20%. Elevated levels of ATAs increase the risk of PCA positivity.

T lymphocyte subsets of the umbilical cord blood of dogs

The hematological parameters red blood cells (RBC) and total white blood cells (WBC) counts, hematocrit, hemoglobin concentration, and RBC indexes (median corpuscular volume and median corpuscular hemoglobin concentration) were determined and T CD5+ lymphocytes and CD4+ and CD8+ subpopulations of the umbilical cord blood (UCB) of dogs were quantified by the cytofluorimetric technique. Nine adult Beagles, from two do five-year old, were used as control. The umbilical cord blood (UCB) was collected from 20 neonate dogs. The method for the UCB collection was adequate to obtain sufficient quantity of blood for the accomplishment of the hematological analyses and lymphocyte quantification. Cytoscopic preparations of the UCB suggested high erythropoietic activity. There was no difference for the global leukocyte and lymphocyte counts between the groups. UCB T lymphocyte counts were lower than those obtained for adult dogs. The proportion of CD4:CD8 showed a great dominance of T CD4+ cells over T CD8+ lymphocytes in UCB.

A Case Report of Pernicious Anemia and Recurrent Aphthous Stomatitis

The aim of this report is to present the management of a patient with pernicious anemia afflicted with recurrent aphthous stomatitis (RAS). RAS is one of the most common lesions of the oral mucosa. Although the exact etiology of RAS is still unknown different hematinic deficiencies have been proposed. Painful recurrent ulcers covered with a grayish pseudomembrane surrounded by an erythematous margin were identified on the tongue and in the buccal mucosa of a 71-year-old woman. The patient also presented with depapilation of the tongue. The clinical diagnosis was RAS. Laboratory tests including a hemogram were ordered to determine existing levels of folic acid, iron, ferritin, and vitamins B2, B6, and B12. Levels of serum vitamin B12 and serum hemoglobin were low. The laboratory investigation also showed a medium corpuscular volume of 104.1 fl. A gastroduodenoscopy revealed no macroscopic abnormality. A gastric biopsy showed mucosal atrophy in the gastric corpus with evidence of intestinal metaplasia. Antibodies against an intrinsic factor were negative. The diagnosis pernicious anemia was made, with RAS caused by vitamin B12 malabsorption. Treatment consisted of the administration of 1.0 ml of hydroxocolabamin intramuscularly twice weekly over four weeks followed by 1.0 ml once weekly for four weeks. Clinical resolution was observed after two months. The association of RAS with vitamin B12 malabsorption is a rare event. However, along with conventional RAS clinical management, iron, folic acid, vitamin B deficiencies, and nutritional intolerance must be considered. Evaluation of the predisposing factors is imperative in treating patients with RAS including vitamin B12 malabsorption. Determination of the levels of vitamin B12 should be the basis for replacement therapy. Such therapy can be considered a benefit to the patients with RAS as its etiology remains unclear. Clinicians must be alert to the possibility this lesion could be a signal of systemic disease.

Sunitinib‐induced macrocytosis in patients with metastatic renal cell carcinoma

Sunitinib and sorafenib are small molecules that inhibit the vascular endothelial growth factor and related receptors with substantial clinical activity reported in metastatic renal cell carcinoma (RCC). Cytopenia and macrocytosis have been described in patients treated with these agents. A retrospective review of all patients with metastatic RCC who were treated with sunitinib or sorafenib for at least 3 months at the Cleveland Clinic Taussig Cancer Institute was undertaken. Complete blood count (CBC) data including red blood cell indices were recorded at baseline, after 3 months of therapy, and at the end of treatment. A total of 61 patients were treated with sunitinib and 37 patients were treated with sorafenib with available CBC data. In patients treated with sunitinib, the median corpuscular volume (MCV) increased significantly at 3 months compared with baseline (median increase of 5.1 femtoliters [fL]; P < .001) and continued to increase throughout treatment. Patients who developed hypothyroidism had a larger MCV increase at 3 months than patients who remained euthyroid (P = .06), although macrocytosis was observed in patients without hypothyroidism. Ten patients discontinued sunitinib therapy, and the MCV decreased in all patients within 2 to 4 months, without further intervention. Bone marrow analysis of 4 patients revealed a hypocellular bone marrow with trilineage hematopoiesis and no evidence of metastasis. There was no evidence of folate or vitamin B12 deficiency. In contrast to sunitinib, there was no change in the MCV for patients treated with sorafenib. Macrocytosis was a common occurrence after treatment with sunitinib but not sorafenib in patients with metastatic RCC. Sunitinib-induced macrocytosis is reversible with drug discontinuation.

JAK2 V617F Mutation Induces a Myeloproliferative Disorder in Mice.

Background: An activating mutation (V617F) in the JH2 domain of the Jak2 kinase is found in 74%–97% of patients with polycythemia vera (PV), 23%–57% of patients with essential thrombocythemia and 35%–57% of patients with idiopathic myelofibrosis. We studied the effects of this mutant in murine hematopoietic cells in vitro and in vivo.Materials and Methods: Murine wild type (Jak2-WT) and V617F mutant JAK2 (Jak2-V617F) were cloned into the MIGR1-IRES-GFP retroviral vector. BaF3 cells stably expressing mutant and wild type Jak2 were generated by electroporation followed by prolonged culture in IL-3 containing media and FACS selection for GFP-positive cells. Proliferation and viability assays were performed in graded concentrations of IL-3. Bone marrow from 5-FU treated Balb/c mice were infected with JAK2-V617F, JAK2-WT and empty vector retrovirus and injected into lethally irradiated recipients. The mice were monitored by observation and blood counts. Plasma levels of erythropoietin (EPO) were determined by ELISA on day 44 after transplantation. 1-way analysis of variance (ANOVA) was used to compare differences between cell line and hematologic parameters. The relationship between EPO levels and Hct/Hgb was analysed by Pearson correlation.Results: BaF3 cells expressing JAK2-V617F showed enhanced proliferation in response to murine IL-3 compared to cells expressing Jak2-WT (p<0.001) and parental cells (p= 0.009). Similarly, there was a trend for increased viability (p=0.071 vs. Jak-WT and p=0.059 vs. parental cells). Sudden complete IL-3 withdrawal induced >95% cell death, followed by outgrowth of IL-3-independent lines after 20 days.Mice transplanted with Jak2-V617F showed increased median hematocrit (Hct), hemoglobin (Hgb) and median corpuscular volume (MCV) compared to Jak2-WT and empty vector mice (table). There was a trend for increased white cell counts (WBC). Median EPO levels were 65 pg/ml in the Jak2-V617F mice compared to 88 pg/ml in the Jak2-WT and 131 pg/ml in the empty vector controls (p= 0.032). There was a significant correlation between EPO and Hct (p = 0.011, r = −0.677) and Hgb (p= 0.002, r= −0.770) across the three groups of mice. On day 44 post transplant one Jak2-V617F mouse developed trilineage MPD with a Hct of 56.8%, leukocytosis (22.6 x 103/ul) and thrombocytosis (1004 x 103/ul). The peripheral blood smear showed neutrophilia, elevated platelets and multiple nucleated red cells. Necropsy revealed hepatosplenomegaly.Conclusions: (i) JAK2-V617F induces hypersensitivity to IL-3 in BaF3 cells. Complete IL-3 independence was seen only after “crisis” and prolonged culture, suggesting additional mutations may be required. (ii) In a murine transduction-transplantation model JAK2-V617F induces MPD closely mimicking PV. Studies are in progress to determine whether additional phenotypes may be observed in a larger cohort of animals.JAK2-V617F (n= 4)JAK2- WT (n= 4)MIGR-1 vector-control (n= 5)P value (V617F vs. WT)P value (V617F vs. empty vector)Hct (%)51.1 (44.6–56.8)43.7 (42.8–45)43.6 (40.4–45.2)P = 0.015P= 0.011Hgb (g/dl)16.2 (14.6–17.6)14.6 (14.2–15.4)14.2 (13–15)P= 0.033P= 0.010MCV (fl)49.5 (47–54)46.2 (46–47)46.2 (45–48)P= 0.041P= 0.031WBC (103/ul)9.6 (3.8–22.6)5.5 (3.6–9.4)3.4 (2.2–4.6)nsP= 0.096EPO (pg/ml)65 (44–98)88 (80–112)131 (95–231)nsP= 0.032Median blood parameters (range) in groups of mice 44 days after transplant

An Itching and Excoriated Dermatosis During Intrahepatic Cholestasis of Pregnancy

A 35-year-old woman at 30th gestation week of her second pregnancy presented to our department with a 2-month history of intense and generalized pruritus. She had a spontaneous abortion 1 year earlier. Itching initially presented during nighttime and localized on lower limbs and after became continuous, diffuse, and associated with excoriations due to scratching. The patient was previously treated with oral corticosteroids (25 mg/d) in a gynecological department with temporary response. On our examination, she presented linear excoriations with hemorrhagic crusts localized on the trunk, buttocks, and upper and lower limbs. Biopsy specimen from the lesional area of the right buttock submitted for routine histology documented a mild perivascular and interstitial infiltrate of lymphocytes and monocytes with rare eosinophils on superficial dermis. Indirect and direct immunofluorescence (performed on perilesional skin) were negative. Laboratory investigations revealed microcytic anemia (hemoglobin 7.5 g/dL; medium corpuscular volume 61.7 fl), erythrocyte sedimentation rate (21 mm) and serum bile acid levels (18.3 nmol/L; normal values 1.00-8.90) increase. On the basis of clinical, serological, and histological findings, we diagnosed an itching dermatosis during an intrahepatic cholestasis of pregnancy. We treated the patient with ursodeoxycholic acid (600 mg) and topical corticosteroids with gradual resolution of itching. Furthermore, she delivered a healthy boy at 39th gestation week with normal birth weight and normal Apgar score.

A CASE OF SUBACUTE COMBINED DEGENERATION OF SPINAL CORD

Clerici R., De Riz M., Corrà B., Baron P.L., Scarpini E., Conti G., Scarlato G.Department of Neurological Sciences, Univ. of Milan, IRCCS Ospedale Maggiore, MilanoThe specific spinal cord lesion caused by vitamin B12 deficiency is known as subacute combined degeneration (SCD). This is a rare cause of demyelination of the dorsal and lateral columns of spinal cord and even more rarely of peripheral nerves, optic nerve and brain.We report a case of SCD in a vegetarian 55‐year‐old fe‐ male who presented with 6 weeks history of gradually progressive paresthesia involving thoracoabdominal skin below C5 dermatome and both lower limbs and postural instability due to a mild impairment of deep sensation. Hematological tests revealed abnormal Medium Corpuscular Volume (MCV) without anaemia and low vitamin B12 levels. The cause of this deficiency was secondary to atrophic gastritis. A spinal cord MRI demonstrated two T2‐weighted hyperintense signal alterations (C1‐C3 and C3‐C4). She was treated with parenteral vitamin B12 supplements and experienced gradual improvement in her clinical symptoms. Repeat MRI of cervical spinal cord after 5 months showed a relevant decrease in the areas of abnormal signal. In other documented cases reported in literature, there was a precocious clinical improvement, while the MRI lesions recovered with a delay. However, it is essential to recognize SCD among the different demyelinating diseases and treat it as soon as possible.

The First Findings on the Differences in Complete Blood Count Between Diploid and Triploid Tench, Tinca tinca L.

Svobodova. Z .. 1. Kolarova, M. Flajshans: The First Findings of the Differences in Complete Blood COltllt Between Diploid and Triploid Tench. Tinea tinca L. Acta vet. Bmo 1998, 67: 243-248. The aim of this work was to evaluate differences in complete blood count of diploid and triploid tench and concentration of total protein in blood plasma of these fish. Haematological examination of 78 specimens of three-year-old tench. consisting of 21 triploids and their 57 diploid siblings was performed in July 1996. Analysis of variance test confirmed a significantly lower erythrocyte count in triploids (P < 0.01). as well as a significantly lower haemoglobin content (P < 0.05), significantly higher medium corpuscular volume. higher medium corpuscular haemoglobin and lower medium corpuscular haemoglobin concentration (all at P < 0.01) compared to diploids. The haematocrit value of triploids was insignificantly lower compared to diploids. The differences of erythrocyte nuclear area. perimeter. average size, major axis and aspect ratio between diploids and triploids were found signiticant at P < 0.01 and the effect of ploidy level on these dimensions was considered highly significant (P < 0.000 I). No effect of ploidy level was proven for erythrocyte nuclear minor axis. The leukocyte count of triploids was insignificantly lesser compared to diploids. Triploid tench were found of significantly lower concentration of total protein in blood plasma compared to diploids (P < 0.01). Triploid tench could be hypothesized to have a lower oxygenation capacity of blood. as well as a lower non-specific immunity level compared to diploids. Insigniticantly higher erythrocyte count, haemoglobin content. haematocrit value. and significantly higher concentration of total protein in blood plasma (P < 0.0 I) was reported for diploid females compared to males.