High molecular risk variants, severe thrombocytopenia and large unstained cells count affect the outcome in primary myelofibrosis.

Apart from the driver mutations, high molecular risk (HMR) variants and other factors have been reported to influence the prognosis of primary myelofibrosis (PMF). The aim of our study was to investigate the impact of laboratory and molecular characteristics at the time of diagnosis (TOD) on the PMF outcome. The study group consisted of 82 patients recruited from three Polish university centers. Among the driver mutations, only CALR type 1 positively influenced the overall survival (OS). The risk of progression to accelerated or blastic disease phase (AP/BP) did not depend on the driver mutation type, but was closely associated with the presence of HMR variants (p = 0.0062). The risk of death (ROD) was higher in patients with HMR variants (OR[95%CI] = 4.33[1.52;12.34], p = 0.0044) and in patients with a platelet count at the TOD between 50-100 G/L (HR[95%CI] = 2.66[1.11;6.35]) and < 50 G/L (HR[95%CI] = 8.44[2.50;28.44]). Median survival time was 7.8, 2.2 and 1.4 years in patients with large unstained cells (LUC) count of [0.0-0.2], (0.2-0.4] and > 0.4 G/L at the TOD, respectively. We found an unexpected, hitherto undescribed, association between LUC count at the TOD and PMF prognosis. Our analysis led to the following conclusions: in PMF patients at the TOD 1) the presence of HMR variants, especially combined, is associated with an increased risk of progression to the AP and BP, and shorter OS, 2) severe thrombocytopenia confers worse prognosis than the moderate one, 3) LUC count is closely related with the disease phase, and associated with the ROD and OS.

Large unstained cell (LUC) count as a predictor of carotid artery occlusion.

BACKGROUND: Carotid artery stenosis is often considered a stable clinical condition, and the underlying atherosclerosis is thought to have an inflammatory background. OBJECTIVES: The aim of the study was to assess the value of different parameters obtained from whole blood counts for the prediction of advanced carotid artery atherosclerosis, including vessel occlusion, irrespective of symptom occurrence. MATERIAL AND METHODS: The study group comprised 290 patients (84 (29%) females and 206 (71%) males) with a mean age of 68 ±8 years, who were admitted to the Vascular Surgery Department due to significant carotid artery disease. Patients were retrospectively divided into 2 subgroups regarding the presence or absence of artery occlusion. The demographic, clinical and laboratory preoperative data were compared between both groups. RESULTS: We found significant differences in preoperative large unstained cell (LUC) counts between patients with and without carotid artery occlusion (p = 0.003), when analyzed with the Mann-Whitney test for independent samples. The receiver operating characteristic (ROC) curve showed that LUC count has prognostic properties for carotid artery occlusion, with an area under the curve (AUC) of 0.637 (p = 0.033), yielding a 69.70% sensitivity and a 51.75% specificity. CONCLUSIONS: Large unstained cells represent an acute inflammatory state related to artery occlusion. An LUC count below the cutoff value of 0.16×109/L may be a predictor of carotid artery occlusion. Therefore, carotid artery occlusion should not be regarded as a chronic state, but as a clinical challenge being promoted by active inflammatory processes.

Role of large unstained cells in predicting successful stem cell collection in autologous stem cell transplantation.

INTRODUCTION: Sufficient stem cell collection is mandatory for Autologous stem cell transplantation (ASCT). Peripheral CD34+ stem CD34 + stem cell counting by flow cytometry is the gold standard method in both the predicting and timing of successful stem cell collection. Large unstained cells (LUC) are large peroxidase-negative cells that are displayed on certain automatic cell counters and present large lymphocytes, virocytes, blasts, abnormal cells and hematopoietic stem cells. In this study, we evaluated the role of LUC parameters in the timing and prediction of successful stem cell collection. METHODS: Patients with a diagnosis of multiple myeloma, lymphoma and testis tumor who proceed to ASCT were included in this study. Preapheresis LUC parameters were analyzed with Siemens ADVIA® 2120i system., Kruskal Wallis, Mann-Whitney U, Spearman Rho and receiver-operator curve (ROC) tests were used for analyses. RESULTS: Ninety patients were evaluated. Peripheral CD34 + cell count was positively correlated with both LUC count (p = 0014) and LUC percentage (p = 0,01). LUC percentage in peripheral blood was positively correlated with mobilized stem cell count in the yield (p = 0.003). We found a LUC count of > 0.485 × 109/L as a cut-off value for detecting > 20 × 106/L CD34 +cells in the peripheral blood with a sensitivity of 64.6% and specificity of 75%. We defined > 2.15% as a cut-off value for LUC percentage to collect > 5 × 106/kg of stem cells with a sensitivity of 64% and specificity of 63%. Additionally, total nucleated cell (TNC) count was negatively correlated with LUC percentage (p = 0.014) and positively correlated with LUC count (p = 0.001). CONCLUSION: LUC parameters are readily available, simple and cheap tools that can be useful in both timing of CD34 count by flow cytometry in peripheral blood and in the prediction of successful mobilization. LUCs can also be an indicator of graft composition.

Repeated dose (90 days) oral toxicity study of ursolic acid in Han-Wistar rats.

BACKGROUND: Ursolic acid (UA) has been used in alternative medicine for decades, and there has been a great interest in its medicinal properties. Despite this increased interest, a detailed long-term toxicity study has not been performed. The objective of this study was to determine the long-term toxic effect of UA on clinical chemistry, haematology, coagulation, pathology/morphology, behaviour and motor skills in rats. METHODS: A solution was made by dissolving UA in a mixture of 0.1% Tween 80 and 0.5% hydroxypropyl methylcellulose in Milli-Q Water. The control group received the vehicle, and the test groups received a dose up to 1000 mg/kg/day via oral gavage. The solution was administered to both male and female (Han-Wistar) rats for 90 consecutive days. RESULTS: UA did not cause any deaths, abnormal body weights or abnormal pathology at all test doses. In addition to that, no toxicological changes were observed in behaviour, neurotoxicity, coagulation, haematology or clinical chemistry that are related to the administration of UA. CONCLUSION: This study indicates that oral dosing of UA for 90 consecutive days does not lead to toxic effects at any of the doses. Therefore, the NOAEL for UA is likely to be higher than 1000 mg/kg/day.

Analysis of Separation of White Blood Cells in Peripheral Blood as an Indicator of MPO Deficiency.

The purpose of this study was to analyze the white blood cell (WBC) percentage pattern of patients with myeloperoxidase disorder. During the 18 months of routine work, 36 blood samples were found with disorders of myeloperoxidase activity: 12 cases of total myeloperoxidase (MPOt) deficiency and 24 cases of partial myeloperoxidase (MPOp) deficiency. In the group with MPOp, according to the results, monocytes (MONO) were the dominant population 33.2%±21.3; however, the microscopic evaluation of leucocytes showed the dominance of neutrophil (NEUT). The average NEUT value was 66.63%±12.31; LYMPH 23.33%±10.08; MONO 6.00%±3.20; EOS 2.04%±2.20; BASO 0.29%±0.62; ATYP 0.83%±1.09. In the group with MPOt, the results of automated leukocyte analysis showed that the dominant group consisted of large unstained cells (LUC) 72.6%±8.64. LUC category reflects large immature cells such as blusts. In the microscopic evaluation: NEUT 67% ±11.40; LYMPH 23%±8.94; MONO 6.17%±3.47; EOS 1.25%±1.06; BASO 0.08%±0.29; ATYP 0.92%±1.38. During microscopic verification, no LUC cells were found. Results of the evaluation of automatic WBC separation according to morphology and functionality of cells led to the conclusion that monocyte dominance in the differential WBC count is associated with a high likelihood of MPOp, and the domination of large unstained cells with MPOt.

The Usefulness of Hematological Parameters for Assessing Disease Progression in Patients with HIV Infection

BACKGROUND: In patients with HIV, CD4+ T cell count and viral load are the main laboratory tests performed to assess clinical management. However, they require extensive resources. In this study, we aimed to determine whether hematological parameters measured using a hematology analyzer are useful as surrogate markers of CD4+ T cell count and viral load in HIV-infected patients. METHODS: Peripheral blood samples were obtained from 14 HIV-naïve, 105 HIV-treated, and 103 uninfected individuals. Hematological parameters were measured using the ADVIA 2120i hematology analyzer (Siemens Healthcare Diagnostics, USA). RESULTS: In HIV-naïve and -treated patients, the percentage of large unstained cells (%LUCs) was 2.5±1.6% and 1.9±0.7%, respectively, compared to 1.6±0.5% in HIV-uninfected controls. The %LUCs was higher in HIV patients with low CD4⁺ T cell count below 200/μL (2.4±1.0%) or high viral load ≥200 copies/mL (2.4±0.8%) than in other infected groups. Significant differences in lymphocyte count were observed between the HIV-naïve (1.5±0.6×10⁹/L) and uninfected (2.0±0.6×10⁹/L) groups as well as between HIV patients with CD4⁺ T cells ≥500/μL (2.5±0.6×10⁹/L) and other infected groups. Neutrophil count varied between high viral load (3.0±1.4×10⁹/L) and low viral load (3.7±1.3×10⁹/L) groups. The CD4⁺ T cell count correlated with lymphocyte count (r=0.642, P<0.0001) and %LUCs (r=-0.287, P=0.002). CONCLUSIONS: %LUCs, lymphocyte count, and neutrophil count are probable surrogate markers of CD4⁺ T cells and viral load.

Increased large unstained cells value in varicella patients: A valuable parameter to aid rapid diagnosis of varicella infection.

Varicella is a highly contagious infection caused by varicella zoster virus. Sometimes it is difficult to differentiate between other viral infections such as Kaposi's varicelliform eruption (KVE) and disseminated herpes zoster (HZ). The large unstained cells (LUC) value is a differential count parameter reported by routing hematology analysis. LUC have been studied previously, but never been reported in the context of varicella or in dermatological published work. The aim of this study was to compare the LUC values in varicella patients with that in KVE and disseminated HZ patients. Sixty-nine varicella patients, 30 KVE patients and 11 disseminated HZ patients were included in this retrospective study. All data were analyzed using SPSS version 17.0 or GraphPad Prism version 5.0. The mean percentage of LUC (%LUC) in varicella patients was higher than the upper limit of normal reference range and it was increased compared to %LUC of both KVE (P < 0.0001) and disseminated HZ (P = 0.0051) patients. %LUC of varicella patients significantly decreased with clinical improvements (P = 0.0017). %LUC was significantly increased in varicella patients and corresponded with clinical improvements. Patients with %LUC of 3.55 or more favor the diagnosis of varicella over both KVE and disseminated HZ with 71.01% sensitivity and 84.44% specificity. We suggest that %LUC can assist in making a precise diagnosis of varicella in confusing cases.

Safety assessment of consumption of glabrous canary seed (Phalaris canariensis L.) in rats.

Canary seed is a nutrient-rich cereal grain; however, it has not been used in human food in part due to concerns regarding safety of consumption. Glabrous or hairless canary seed has potential human food use as trichomes are absent. The objective of the oral feeding studies reported here was to assess the safety of yellow and brown glabrous canary seed cultivars as human cereal foods. The first study was a 90-day rat oral toxicity study, which compared the effects of diets containing 50% of either brown dehulled glabrous, brown hulled glabrous, or brown hulled pubescent (hairy) hulled canary seed to a diet containing 50% wheat. No significant adverse effects were observed. In a 28-day and a 90-day study rats were fed yellow or brown glabrous canary seed groats in the AIN-76 diet at concentrations levels of 2.5%, 5% and 10%. The NOAELs in 90-day study were 5.15 g/kg/d and 5.23 g/kg/d for yellow and brown canary seed groats. Consumption of canary seed was associated with reduced incidence and severity of liver lipidosis as compared to controls. The combined results of these studies clearly demonstrate the safety of consumption of glabrous canary seed, and support its use as a human cereal grain.

Absence of systemic toxicity changes following intramuscular administration of novel pSG2.HIVconsv DNA, ChAdV63.HIVconsv and MVA.HIVconsv vaccines to BALB/c mice.

BACKGROUND: The systemic toxicity of three candidate HIV-1 vaccines plasmid pSG2.HIVconsv DNA (D), ChAdV63.HIVconsv (C) and MVA.HIVconsv (M) expressing chimeric immunogen derived from the most conserved regions of the HIV-1 proteome was evaluated in two repeat-dose studies in the male and female BALB/c mice. METHODS: In study UNO011, mice received three doses of 2×10(7) plaque-forming units of MVA.HIVconsv vaccine (MMM). In study UNO012, mice received 3 doses of 50µg of pSG2.HIVconsv DNA followed by a single dose of 5.95×10(9) virus particles of ChAdV63.HIVconsv vaccine (DDDC). Similarly constituted control groups received the vehicle alone (phosphate buffered saline) at the same volume-dose. All vaccines were administered by intramuscular needle injection into the right hind limb at 14-day intervals and animals were sacrificed 7 days after the last dose. Assessment of local and systemic toxicity was made. Induction of HIV-1-specific responses was confirmed. Parameters assessed included clinical condition, body weight, food consumption, ophthalmoscopy, haematology, blood chemistry, organ weight and macroscopic and microscopic pathology. RESULTS: In both studies, treatment with the candidate vaccines elicited strong HIV-1-specific T-cell responses. The vaccine treatment was well-tolerated without any adverse systemic toxicological changes. The local toxicity findings observed in these studies were consistent with the predicted response to a vaccine/substance administration by intramuscular injection. CONCLUSIONS: The three novel anti-HIV-1 vaccines were well tolerated when administered by intramuscular injection to BALB/c mice. These results supported an application for authorisation by the Medicines and Healthcare Products Regulatory Agency of the UK to test these vaccines for the first time in phase I clinical trials in healthy both uninfected subjects and HIV-1-infected patients stable on antiretroviral treatment.