Histological grading is a good parameter to stratify tumors according to their biological aggressiveness. The Elston and Ellis grading method in humans, invasive ductal breast carcinomas and other invasive tumors are routinely used. The aims of this study were classification of mammary gland tumors and also application of a human grading method in canine mammary carcinoma. The samples included 37 tumors of mammary glands. Mammary tumors were carcinomas (n = 32) and sarcomas (n = 5). The carcinomas were classified as simple carcinoma 56.8% (n = 21), complex carcinoma 13.5% (n = 5), carcinoma arising from benign tumor 10.8% (n= 4) and special type of carcinoma 5.4% (n = 2). Out of 32 carcinomas studied, 37.5% (n = 12) grade I, 46.9% (n = 15) grade II and 15.6% (n = 5) grade III. This study demonstrated that the Elston and Ellis method of histological grading in canine mammary tumor is a reliable prognostic factor which is correlated with histopathological classification. Simple and complex carcinomas are recorded as the most common type of malignant CMGTs.6 Canine mammary sarcomas often have multi-differentiation  which is not unusual in human mammary sarcomas. Sarcomas resembling  cystosarcoma phyllodes in women appear to be very rare in the dog. There is need for further studies on the histogenesis and biological behavior of mammary sarcomas.7Canine mammary gland tumors (CMGTs) are one of the most common neoplasms of bitches.8Domestic pets are particularly valuable models, as possible sentinels for human environmental and life-style risks. Experimental models, principally genetically engineered mice (GEMS), have been useful for investigating the role of steroid hormones, hormone receptors, and other growth factors in the pathogenesis of breast cancer. However, rodents differ considerably in mammary gland development and types of breast cancer from women. Furthermore, many mammary gland cancers in mice are viral- or toxin- induced, so the validity of these models has been questioned. The high prevalence of spontaneous mammary cancer in domestic dogs and cats closely mimics the disease in women, making these species more suitable comparative models.9,10 The Elston and Ellis grading method in humans, invasive ductal breast carcinomas and other invasive tumors are routinely graded.11 The “Elston and Ellis method” is the most common histological grading method for invasive carcinomas and shows a strong correlation with prognosis.11,12Histological grading is a good parameter to stratify tumors according to their biological aggressiveness.2,11-13 Morphological criteria alone may be insufficient for a proper diagnosis because when only histologically determined, benign tumors may incidentally give rise to metastasis. Despite benign biological behavior, canine complex adenomas and mixed tumors often show histomorphological evidence of malignancy (carcinoma or sarcoma in benign tumor).14 Tumor grade and degree of invasion (stage) are also of prognostic signiﬁcance.13Surgery remains the basic treatment for dogs and cats with most type of mammary gland tumors. The exceptions are inoperable disease  and distant (organ) metastasis. One of the major problems in veterinary oncology is accurate prognosis for post-surgical mammary cancer cases. In human breast cancer and in canine mammary tumor, histological type, histological grade and lymph node involvement are standard prognostic features.et al., used this method to grade these tumors.15The aim of this study was classification and also application of a human grading method in canine mammary carcinoma. The incidence of canine mammary gland carcinoma is common in Iran, however, only Rezaie Specimens. Thirty seven spontaneous tumors of the mammary gland from bitches aged from 4 to 15 years (average 8.5 years), of various pure or mixed breed during September 2009 to September 2011 for diagnostic purposes were obtained. They were selected from cases treated surgically or from archive of Pathology Department of Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran. In surgically removed samples, the mammary tumors were excised by simple mastectomy or regional mastectomy, with or without the superﬁcial inguinal lymph nodes. Tissues, submitted in 10% neutral buffered formalin were embedded in paraffin wax. Sections (5 µm thick) were cut and routinely stained with hematoxylin and eosin (H & E) for histological examination. Histopathological evaluation. Tumors were classiﬁed according to the World Health Organization (WHO) criteria for canine mammary lesions6 based on the most pronounced histological pattern observed in more than 50.0% of the tumor mass. Whenever tumors displayed multiple morphological patterns, without more prominent growth pattern present in 50.0% of the tumor mass, lesions were classiﬁed as tumors with mixed morphology tumor.Tumor grade. Grade was determined according to the Elston and Ellis scoring system12 based on the assessment of three morphological features: (1) the degree of glandular differentiation assessed by tubular formation; (2) nuclear pleomorphism, and (3) mitotic activity. Each parameter was graded into three categories, to which a score of 1-3 was assigned as follows: (1) tubule formation ; (2) nuclear pleomorphism , and (3) number of mitosis per 10 high power ﬁelds  . The scores of all three components were added together to give a total of 3-9 points. Grade was allocated by an arbitrary division as follows: (1) grade I, well differentiated or low grade: 3-5 points; (2) grade II, moderately differentiated or intermediate grade: 6-7 points, and (3) grade III, poorly differentiated or high grade: 8-9 points  and 86.5% (n = 32) and 13.5% (n = 5) of mammary tumors were carcinomas and sarcomas, respectively. The most frequently represented tumor type were simple carcinoma 56.8% (n = 21) followed by complex carcinoma 13.5% (n = 5) sarcoma 13.5% (n = 5) carcinoma arising from benign tumor 10.8% (n = 4) and special type of carcinoma 5.4% (n = 2). Simple carcinomas n = 21) were sub classified to 80.9% (n = 17) tubulopapillary carcinoma, 4.8% (n = 1) solid carcinoma and 14.3% (n =3) cribriform carcinoma , 2 and 31 were suThe carcinomas of this study were as follow: Out of 32 carcinomas studied, 37.5% (n = 12) grade I, 46.9% (n = 15) grade II and 15.6% (n = 5) grade III with high mitotic index . Tumors 9One of the major challenges for a veterinary oncologist is to identify the prognostic variables which allowed disease behavior to be predicted.6In canine mammary neoplasm, tumor type was an important factor and an increasing range was observed in malignancy of complex carcinoma  to simple carcinoma  to sarcoma.16 Meuten reported that about 20.0-40.0% of bitches with mammary tumors developed malignant kinds.17 Although Simeonov and Stoikov reported that only 19.0% benign and 81.0% mammary tumors were malignant.18 In the present study all samples (n = 37) were malignant. Rezaie et al. found that 70.6% of bitches had tubulopapillary carcinoma, 23.5%- solid carcinoma, and 5.9% - cribriform carcinoma.15 Ežerskytė et al. showed that the most common tumor types of mammary glands in bitches were simple carcinoma, complex carcinoma and carcinosarcoma 46.0%, 27.0% and 13.0%, respectively.19Some studies showed that half (42.0-55.0%) of the surgically removed mammary tumors in bitches were malignant.In the present study, 86.5 % (n = 32), and 13.5 % (n = 5) of mammary tumors were carcinomas and sarcomas, respectively. The most frequently represented tumor type was simple carcinoma 56.8% (n = 21), followed by complex carcinoma 13.5% (n = 5), sarcoma 13.5% (n = 5), carcinoma arising from benign tumor 10.8% (n= 4) and special type of carcinoma 5.4% (n = 2).In sub classification of simple carcinomas: 80.9% (n = 17) tubulopapillary carcinoma, 4.8% (n = 1) solid carcinoma and 14.3% (n = 3) cribriform carcinoma. These results were in agreement with above researches.10,20 The Elston and Ellis grading method has been used previously to study canine mammary carcinoma10,15,21,22 and feline mammary carcinomas.23-25Most grading systems of mammary carcinomas in dogs are a modification of the numeric method of Elston and Ellis.11In human, the combination of histological type and grade produce a more accurate assessment of prognosis of breast cancer than histological type alone. Histological grade may also provide useful information to predict the response to chemotherapy and, therefore, be a predictive factor.2,6 These results are in agreement with other studies.10,15In this study, most carcinomas were graded II and I, with just 15.6 % of lesions being classiﬁed as grade III and also we found complex carcinoma and carcinoma arising from benign mixed tumor were usually of grade I or II. The presence of myo-epithelial cells in complex carcinomas is associated with a less aggressive biological behavior and a better prognosis than with simple carcinomas.26 In this study we described 3 simple cribriform carcinoma (two grade II and one grade III). This type of tumor has worse prognoses than tubulopapillary and solid carcinoma.Ten samples out of 12 samples of grade I (83.3%) were tubulopapillary carcinoma, that were consisted of more than 75.0% tubules with a clear central lumina, cells with regular outline and uniform and also less than 7 mitosis per 10 HPF. On the other hands, simple solid carcinoma  was grade III. Cribriform carcinoma, which is uncommon, is characterized by the proliferation of a population of neoplastic epithelial cells forming a sieve-like arrangement.27,28 It is an uncommon mammary gland neoplasm, but it most often presents as a carcinoma and osteosarcoma. The epithelial component metastasizes via lymphatic vessels to regional lymph nodes and the lungs, and the mesenchymal component, via the hematogenous route to the lungs.26The malignant mixed mammary tumor (carcinosarcoma) is composed partly of cells morphologically resembling the epithelial component and partly of cells morphologically resembling connective tissue elements, both types of which are malignant.29 There are limited reports of canine primary mammary chondrosarcoma in the literature. Present paper described a primary mammary chondrosarcoma. According to authors’ knowledge, this is the first report of this kind of mammary tumor in Iran.Matrix-producing carcinoma is a very rare breast neoplasm accounting for less than 0.1% of all human breast malignancies.30The prognosis is based on multiple factors. The type of tumor is important in determining the prognosis. Sarcomas are associated with shorter survival times than carcinomas. Other factors for poor prognosis are size of tumor, lymph node involvement, ulcerated tumor surface, rapid growth of tumor, tumor adherence to deeper tissues and nuclear differentiation.et al. found significant differences in survival between cases with different tumor grades.10 Survival was worse in dogs with grade III carcinomas than in those with grade I or grade II. Dogs with simple carcinomas had a worse prognosis than with other carcinomas; there was no significant difference in survival between grade II and grade III cases, with both having a very poor prognosis. Undifferentiated carcinomas (grade III) had an increased risk of death when compared with differentiated carcinomas (grade I and II). As noted in many of the studies, lymphatic/vascular invasion and lymph node metastasis are, as expected, associated with a poor prognosis.26 In feline mammary carcinoma there was a signiﬁcantly shorter overall survival in queens with grade III tumors, which was proved in other reports.23,25 In this study, cases graded III had significant invasion to blood and lymphatic vessels.Karayannopoulou This study demonstrated that the Elston and Ellis method of histological grading in canine mammary tumor is a reliable prognostic factor. That is correlated with histopathological classification. Grading should be a usual procedure in the evaluation of biological aggressiveness of canine mammary carcinomas. Its routine use should be helpful in indicating appropriate post-surgical treatment.


Several studies have reported the favorable effect of leucine supplementation on insulin resistance or insulin sensitivity. However, whether or not leucine supplementation improves leptin sensitivity remains unclear.Forty-eight male Sprague-Dawley rats were fed with either a high-fat diet (HFD) or HFD supplemented with 1.5, 3.0, and 4.5% leucine for 16 weeks. At the end of the experiment, serum leptin level was measured by ELISA, and leptin receptor (ObR) in the hypothalamus was examined by immunohistochemistry. The protein expressions of ObR and leptin-signaling pathway in adipose tissues were detected by western blot.No significant differences in body weight and food/energy intake existed among the four groups. Serum leptin levels were significantly lower, and ObR expression in the hypothalamus and adipose tissues was significantly higher in the three leucine groups than in the control group. These phenomena suggested that leptin sensitivity was improved in the leucine groups. Furthermore, the expressions of JAK2 and STAT3 (activated by ObR) were significantly higher, and that of SOCS3  was significantly lower in the three leucine groups than in the control group.Leucine supplementation improves leptin sensitivity in rats on HFD likely by promoting leptin signaling. The discovery of leptin has led to astonishing advances in understanding energy homeostasis in rodents and humans . Leptin Leucine is an essential branched chain amino acid (BCAA). Many studies have demonstrated that leucine plays an important role in regulating energy balance and improving insulin resistance by directly affecting the central nervous system and peripheral tissues –10. SomeForty-eight 5-week-old male Sprague-Dawley rats  were individually housed in stainless steel wire-mesh cages in an air-conditioned room (22–25°C) with a 12 h light/dark cycle and 55±5% humidity in compliance with the Guide for the Care and Use of Laboratory Animals in Soochow University. After 10 days of acclimatization, the rats were randomly assigned to one of four groups (12 rats per group) fed with HFD (HFD group) or HFD supplemented with different leucine concentrations by weight % . Leucine was purchased from Shandong Luzhou Amino Acid Co., Ltd. The base diet was modified AIN-93. HFD was made according to feed formation of Teklad Custom Research Diet (TD 06414), and fat sources were lard and soybean oil. HFD contained 5.10 kcal/g with 60.3, 18.4, and 21.3% calories from fat, protein, and carbohydrates, respectively. The animals had free access to food and water. The experiment lasted for 16 weeks.During the experiment, body weights were recorded weekly. Food was dispensed in a glass container  and renewed every day. Food intake was measured, and energy intake was calculated once a week. At 16 weeks after the experiment, the rats were deprived of food for 12 h and blood was collected from the femoral artery under ether anesthesia. Serum was separated and stored at −80°C. Peritoneal adipose tissues (except mesenteric one) were quickly removed and weighed. Serum leptin level was determined using commercial ELISA kits .2O2. The sections were counterstained with hematoxylin and then sliced. The slices were observed under a microscope at 40× objective. Five areas of section were randomly selected and quantified as one animal by Image pro-Plus program, and four animals in each group were counted and averaged.Four animals in each group were perfused transcardially with 4% paraformaldehyde. Brain was carefully removed from the skull, post fixed with the same fixative solution for 24 h, and then dehydrated and embedded in paraffin. Tissue blocks were transversally cut from the head into 3-µm-thick sections and incubated in a blocking solution containing 10% goat serum. The sections were incubated overnight with polyclonal antibodies directed against ObR  and then with biotinylated goat anti-rabbit IgG  and streptavidin–biotin complex for 1 h each. The sections were washed three times by PBS for 5 min after each incubation. Immunolabeling was revealed with 0.05% diaminobenzidine  and 0.03% HPerirenal fat was homogenized in lysis buffer (1:10 wt/vol). Protein concentration was determined using the BCA protein assay. Equal amounts of protein (30–50 µg) were loaded into a 12% sodium dodecyl sulfate –polyacrylamide gel and then transferred to a polyvinylidene difluoride membrane by electrophoretic transfer. After being blocked, the membrane was incubated overnight at 4°C with the following primary antibodies: ObR , JAK2 , STAT3 , SOCS3 , and β-actin . After being washed three times, the antigen–antibody complexes were visualized with peroxidase affinipure goat anti-mouse or anti-rabbit IgG antibody  at room temperature for 1 h. Antibody reactivity was detected by chemiluminescence ECL Detection Systems. Blots were performed at least three times to confirm the reproducibility of the results. The intensity of the bands was normalized using each corresponding β-actin density as an internal control. The bands were quantified using Gbox Chemi-XR 5 .post hoc test. Differences were considered significant at P<0.05. All the analyses were performed using SPSS version 17.0 statistical analysis package .Data are reported as means±standard deviation (SD). One-way ANOVA was used to determine differences among groups, followed by the Tukey The body weight in the four groups was similar at the baseline and constantly increased over time . At the P<0.01; At the end of the experiment, the serum levels of leptin were 2.31±0.23 ng/mL, 1.69±0.26 ng/mL, 1.87±0.27 ng/mL, and 1.91±0.29 ng/mL in the HFD, HFD+1.5% Leu, HFD+3.0% Leu, and HFD+4.5% Leu groups, respectively. The serum leptin level was significantly lower in the three leucine groups than in the HFD group . Furthermore, the expression was significantly higher in the HFD+1.5% Leu group than in the HFD+3.0% Leu or HFD+4.5% Leu group (both P<0.05)  b.P<0.01). However, SOCS3 expression was significantly lower in the HFD+1.5% Leu and HFD+4.5% Leu groups than in the HFD group (P<0.01). Notably, the expression of ObR, JAK2, and STAT3 was the highest, and the expression of SOCS3 was the lowest in HFD+1.5% Leu group .The effects of supplemental leucine on the leptin-signaling pathway in adipose tissues were shown in In the present study, animals were exposed to HFD with or without leucine. Chronic leucine supplementation significantly decreased the circulating leptin level and increased the hypothalamic ObR. Accordingly, the protein expression of ObR, JAK2, and STAT3 increased, whereas that of SOCS3 decreased in adipose tissues.Most animal studies that investigated the effect of leucine supplementation observed the weight/fat loss in diet-induced obese rats. Zhang found that leucine supplementation in drinking water can reduce the weight gain of HFD-fed mice by up to 32% . HoweverAt present, convincing evidence suggests that leucine benefits protein synthesis and improves insulin resistance and insulin sensitivity mainly through the mammalian target of rapamycin (mTOR) pathway , 18, 19.in vitro experiment confirmed that the ObR expression is elevated in mouse C2C12 myotubes cultured with leucine for 2 h  showed a significant effect among the three leucine groups. Our result was similar to that of Li, who found that 1.5% leucine significantly affects the insulin sensitivity of rats on HFD . LeucineIn conclusion, the present study is the first to demonstrate that chronic leucine supplementation can improve HFD-induced leptin resistance by decreasing the circulating leptin level and increasing the ObR. This improvement was ascribed to leptin signaling in adipose tissues, which increased the expression of JAK2 and STAT3 but decreased that of SOCS3.


Ovarian masses represent a range of pathology from benign cyst to highly aggressive malignant tumors. It has been estimated that gynecologic malignancy account for approximately 2% of all types of cancer in children, 60-70% of these lesions arise in the ovary.All ovarian masses which were resected or biopsied in Mofid Children's Hospital from 2002 to 2012 were reviewed retrospectively. Patient’s age, presenting symptoms, surgical procedures, pathological diagnosis, postoperative treatment, and outcome were obtained from medical records.Fifty-seven girls (aged 40.2±57months with the range of 1 day to 15 years) underwent different types of ovarian operations . 50 children had unilateral ovarian mass . The most common presenting symptoms were acute abdominal pain in 46%.Twenty one (37%) of our patients had ovarian torsion. Four (7%) patients had benign tumors, and 8 (14%) had malignant tumors. There were no age differences between those with benign type (8.2±2.6years) and malignant tumors (6.1±5.3years) (P=0.683). Ovarian tumors are rare in children. Most are benign, in children presenting with acute abdominal pain, ovarian mass particularly neoplastic tumors should be suspected. An important proportion of these patients may require postoperative chemotherapy. Ovarian tumors are uncommon in children, and the clinical presentation and pathology are different from adult patients . The cauHowever, in patients requiring emergency operation, presumptive preoperative diagnosis is helpful for surgical or postoperative management. Data in this context particularly in children are scarce. The present study was designed to determine the cause of ovarian mass in children presented to Mofid Children’s Hospital from 2002-2012.Fifty-seven children with ovarian mass confirmed by ultrasonography and CT scan admitted to Mofid Children's Hospital from March 10, 2002 to April 2012 were studied. All cases with ovarian samples submitted for pathologic review were included for further evaluation data. Data regarding demographic data, clinical presentation, treatment, tumor histology, and outcome were collected from medical records. This retrospective analysis was performed with the approval of the hospital institutional review board (IRB). All data were recorded and evaluated using SPSS software Version 18.The mean age of the study patients was 40.2±57 months, (range 1 day to 15 years). Eight (14%) girls had no symptoms on admission. The presenting symptoms were acute abdominal pain in 26 (45.6%), palpable abdominal mass 20 (35%), fever 3 (5.3%), nausea and vomiting 3 (5.3%) patients. Laboratory findings showed leukocytosis in 42.1% and anemia in 8.8% patients. The patients underwent different types of ovarian surgeries included; Salpingo-oophorectomy in 24 (42%) cases, oophorectomy in 10 (17.5%), ovarian cystectomy in 21 (36.8%), and ovarian biopsies in 2 (3.5%) patients. The total number of mass was 64 in which 50 children had unilateral ovarian masses; right side 49.1% and 38.6% in left side, and 12.3% had bilateral mass respectively. Intra-operative findings were as followes: torsion 32.8%, hemorrhagis 12.5%, and necrosis in 14.1%. Pathology types in our patients were seen in The mean age of patients with ovarian cyst, benign and malignant tumors were 2.2±4.6, 8.2±2.6 and 6.1±5.3 years, respectively. There was a statistical significant difference between the mean of age in cases with ovarian cyst and benign (P=0.019) and malignant tumors (P=0.035), but there was no mean age difference between benign and malignant tumors in our study patients (P=0.683). All malignant masses received chemotherapy performed by a pediatric oncologist.The mean size of patient masses with ovarian cyst, benign and malignant tumors were 60.6±21.4, 104.3±53.6 and 75.9±39.6 centimeters, respectively. There was statistical significant difference between the average mass size in cases with ovarian cyst and benign tumors (P=0.006), but there was no difference in the mean size of masses in patients with malignant tumors, benign tumors (P=0.373) and ovarian cyst (P=0.158) in our study patients.Ovarian tissue arises from the mesenchyme of the urogenital ridge, the germinal epithelium that covers the urogenital ridge, the germ cells which arise in the yolk sac, and follicles in the ovary develop from all three of these embryonic cell lines . OvarianOvarian masses may represent physiologic cyst, benign type of neoplasm, or malignant neoplasms. Presenting symptoms may be pain or asymptomatic mass. Pelvic sonography is a very useful tool in the diagnosis of ovarian cyst, and torsion in children . Acute abdominal pain (47%) was the most common presenting symptom in our study, but in Zhang et al.’s study series  it was 3Detorsion and preservation of all ovaries, even in necrotic cases has been recently recommended, also, aspiration of simple benign appearing cyst or contralateral ovary may be helpful . In our In conclusion ovarian tumors are rare in children. Abdominal pain was the most common complaint. The most masses are non-neoplastic (cystic). Surgical treatment is conservative for both benign and malignant lesions; however, postoperative chemotherapy is needed in most malignant lesions.Limitation: We had limitations in tumor marker evaluation and small sample size.


Chemical analysis indicated that PE was high in crude protein (11.30%) and low in neutral detergent fiber (26.20%). Total phenols, total tannins, condensed tannins and hydrolysable tannins contents in PE were 8.29%, 4.48%, 0.49% and 3.79%, respectively. Ruminal dry matter and crude protein degradation after 48 hr incubation were 75.21% and 82.52%, respectively. The gas production volume at 48 hr for PE was 122.47 mL g-1DM. As a whole, adding polyethylene glycol (PEG) to PE increased (p < 0.05) gas production volumes, organic matter digestibility and the metabolizable energy that illustrated inhibitory effect of phenolics on rumen microbial fermentation and the positive influence of PEG on digestion PE. The results showed that PE possessed potentials to being used as feed supplements.The nutritive value of pistachio epicarp (PE) was evaluated by  The crude protein (CP) content of PB varies from 92 to 120 g kg-1 on a dry matter (DM) basis and the metabolizable energy (ME) content of PB may vary from 7.10 to 7.50 MJ kg-1 DM.2 However, PE have been reported to contain high levels of tannins in both hydrolysable and condensed forms.3 Therefore, the value of PE for ruminants is offset by their potentially negative effects on protein utilization, and the risk of toxicity when intake is high.4 Polyethylene glycol (PEG), a non-nutritive synthetic polymer, has a high affinity to tannins and can make them inert by forming tannin-PEG complexes.5 Polyethylene glycol can also liberate protein from tannin–protein complexes and/or prevent their formation.6 Therefore, PEG has been used to mitigate adverse effects of tannins on rumen fermentation.Due to shortage of water and locally produced feeds in many developing and underdeveloped countries, livestock often suffer from under-feeding and malnutrition. Thus, better utilization of non-agricultural by-products which do not compete with human foods is imperative. One of the agro-industrial co-products is pistachio epicarp (PE) which offers some promises of overcoming this problem. In Iran, production pistachio by-products (PB) exceeds 400,000 tons per year.in situ and in vitro gas production techniques.The aim of the current study was to assess the chemical composition and nutritive value of PE using Pistachio epicarp. Fresh PE (Pistacia vera) was collected from pistachio plant cleaning factories of Ghazvin, Iran. Samples were oven-dried at 50˚C for 48 hr and stored in sealed plastic bags. Samples were milled through a 2 mm screen for in situ analysis, in vitro gas production and chemical analysis.Chemical composition. Pistachio epicarp dry matter , ash (Method ID 942.05), ether extract  and crude protein (Method ID 984.13) were determined by procedures of Association of Official Analytical Chemists (AOAC).7 The NDFand ADF concentrations were determined using the methods of Van Soest et al. without sodium sulphite.8 The NDF was analyzed without amylase with ash included. Total phenolics (TP) were measured using Folin Ciocalteau method according to Makkar et al. 5 Total tannin (TT) was determined after adding insoluble PVPP and reacting with Folin Ciocalteau reagent.9 Tannic acid was used to express TP and TT. The condensed tannins (CT) were measured by the HCL-butanol method.12 Hydrolysable tannins were analyzed using the Rhodanine assay,9 and results are expressed as gallotannin.In situ DM and protein degradability analysis. Rumen degradation characteristics of feeds were calculated after the incubation of 5 g sample of PE in nylon bags.10 The bag size was 12 cm × 6 cm with a pore size of 50 µm. There were four bags for each incubation time. Bags were incubated at 0, 2, 4, 6, 8, 12, 16, 24, 36, 48, 72, and 96 hr. The sheep equipped with ruminal fistulae that were fed daily containing of 400 g lucerne, 300 g barley and 300 g soybean meal. In each sheep two bags were used for each time interval. Animals had free access to water. After removal, bags were washed under tap water for at least 15 min until the washing water was clear. After hand squeezing, bags were dried for 48 hr at 105 ˚C then weighed. Feed residues were recovered from each bag and stored pending analysis for Kjeldahl nitrogen. The value of degradability at time 0 was obtained by washing four bags under tap water for at least 15 min. For each bag, the residue was analyzed for DM and nitrogen. The percentage of degradability (Y) of DM and nitrogen at time (t) was obtained from an exponential curve of the type: -ct)Y = a + b  per time. The effective degradability of DM and protein for PE were then estimated by the following equation: which was fitted to the experimental data by iterative regression analysis.-1 DM) = a + bc / (c + k)Effective degradability  were fed a daily ration 400 g lucerne, 300 g barley and 300 g soybean meal twice daily at 09:00 and 17:00 hours.12 About 300 mg dried PE alone was incubated in duplicate with 20 mL rumen liquor- artificial saliva mixture (1:2) in calibrated glass syringes at 39 ˚C. A second assay was carried out with 300 mg PE incubated with 600 mg polyethylene glycol (PEG 6000) as described by Makkar et al.5 Readings were recorded after 2, 4, 6, 8, 12, 16, 24, 36, 48, 72, and 96 hr of incubation and cumulative gas production data was fitted into an exponential equation as described in previous section.11Gas production was completed as described by Menke and Steingass.-1 DM) and metabolizable energy (ME) in MJ kg-1 DM were estimated by equations of Menke and Steingass,12 based on 24 hr gas production  and CP content (g kg-1DM) as: Feed OMD (g kg-1 DM) = 148.8 + 8.89 Gas + 4.5 CP + 0. 651 XAOMD (g kgand -1 DM) = 2.20 + 0.136 Gas + 0.057 CP + 0.0029 CPME .Statistical analysis. Data obtained from in situ and in vitro gas production was subjected to ANOVA as a completely randomized design with four replicates by the GLM procedure, and treatment means were compared by the Duncan test. The relationship between in situ CP degradability (ISCPD) with in situ dry matter degradability (ISDM), determined by linear regression using the PROC REG procedure of SAS . Chemical composition. The chemical composition of PE is presented in In situ DM and protein degradability. in situ CP and DM degradability of PE at incubation times. The disappearance of DM and CP increased with increasing time of incubation. After 96 hr of incubation, the CP and DM degradability for PE were 84.14% and 85.83%, respectively.in situ degradation of PE are given in in situ DM degradability and CP degradability is shown in 2 = 0.926; p < 0.05) in all incubation times.Characteristics of the DM and CP In vitro gas production. The cumulative gas production is shown in -1DM. The addition of PEG increased gas production at all times of incubation (p < 0.05).cubation . Kineticet al. while the NDF and ADF contents were lower than that reported by these authors.3 The amount of total phenol and total tannins for PB  reported by Bagheripour et al. were higher than obtained values for PE in this study.1 Generally, differences can be depended on the maturity stage, species or variety,13 drying methods, the condition environment,14 and soil type.15 Nitrogen, NDF and phenolic concentration suggest that PE have the potential to be used as feed supplements.The CP and TP contents of PE were similar to that reported by Mahdavi et al. showed The values of a, b and c for DM dried PE were 44.69%, 44.31% and 0.04%, respectively.3 The DM degradabilities of PE at 48 hr were higher than those reported by Mahdavi et al.3 Variation in protein degradability is believed to be related to the proportion of structural and non-structural protein and carbohydrate fractions, which in turn affects their solubility and bio-availability.16Mahdavi et al. reported the gas production of PB at 24 hr of incubation about 35.00 mL g-1 OM which was lower than the value reported for PE in this study.1 Tacon et al.16 reported that OMD feeds and gas production was reduced with increasing of phenolic contents. The phenolic contents in study of Bagheripour et al. were higher than the values for PE in this study (9.13% vs 8.29). 1Bagheripour et al. who found PEG utilization increased gas production in PB about 14%.1Inclusion of PEG increased the gas production in PE, the increasing in gas production at 24 hr incubation was about 39.46%. The similar results have been reported by Bagheripour in vitro suggests a negative influence of tannins on digestibility.et al. when using PEG as the phenolic binding agent in conjunction with an in vitro gas technique for assessment of secondary compounds in browse.18Increased gas production and OMD due to addition of PEG p < 0.05, r2 =0.82),  correlation between in situ degradability and in vitro gas production data suggests that either method could be used to estimate nutritive value of such feeds. Previous experiments have shown a positive relationship between in situ degradability and both voluntary intake and in vivo digestibility10,22 as well as 24 hr in vitro gas production and metabolizable energy for ruminants from various forages.23 Khazaal et al. reported that in situ method should be used with caution when estimating the nutritive value of high phenolic feeds.24 The potential negative effect of phenolic compounds on rumen microbial fermentation is unlikely to be detected by in situ method. In this respect in vitro methods are more reliable in detecting inhibitory compounds in feeds. The in vitro gas production technique is a closed system with limited supply of rumen liquor where if there is any anti-nutritive compound, it is likely to affect the activity of rumen microbes. There was a positive correlation between 48 hr gas production and 24 hr DMD data ( =0.82), . This isin situ degradability data suggest that PE have a high nutritive value. In vitro gas production of PE was increased when that combined with PEG. In order to further optimize PE use as a feed supplement, more in vivo studies are warranted. In conclusion, our results showed that the PE evaluated in the present study can be used for supplying part of protein requirements. The


They arRetrospective case-note review and database interrogation of all patients starting anti-retroviral therapy (ART) from 1st March 2011 to 31st March 2014.215 patients started ART. 58% (125/215) were black-African and 47% (100/215) were female. Median CD4 at baseline was 272 cells/µL (range 1-1044 cells/µL). 69 (32%) had a viral load (VL) >100,000 copies/mL. 7 individuals had evidence of transmitted drug resistance. 88 patients started an STR. Two tested positive for HLAB5701. None had a 10 year CVS risk score of >20% and 36% had a baseline VL >100,000 copies/mL. 127 patients started a non-STR regimen, 29% had a VL >100 000 copies/mL and none had an elevated CVS risk. Information regarding baseline renal function and HLAB5701 will be provided at the conference. The use of STRs increased over the three years . There was no difference in STR prescribing between men and women. In men, heterosexual male patients were more likely to be prescribed an STR than MSM males . In 43% (38/88) patients had indicated a preference for an STR and 32/88 expressed a preference for a particular drug. 2% (2/88) requested to be on the same treatment as a partner. In those patients who expressed an interest in a particular drug, four had received information from a friend or partner. In 33% (29/88) cases an STR was felt by the clinician to be the best option, largely based on concerns around pill-burden and adherence (31%), viral load (16%) and renal or cardiac risk (7%).Use of STRs is increasing. This is not driven on in our cohort by cardiovascular risk of HLAB5701 carriage but by patient and clinician preference, and to a lesser extent by higher baseline viral loads.


Many instruments have been developed to evaluate hospital mortality, but less attention has been paid to long-term mortality of traumatic brain injury patients.To analyze the relationship between type of injury used computerized tomography (CT) by Marshall score on admission and mortality at four years in traumatic brain injury critical patients admitted to intensive care.Prospective cohort study of traumatic brain injury patients admitted in the University Clinical Hospital  between 2004 to 2008. Data were collected to calculate different prognostic scores and cranial CT at admission by Marshall score.2 and multiple logistic regression. P < 0.05 was considered significant.The results were analyzed at discharge, at one and four years. T-test, X531 adult patients. Mean age 40.35 ± 19.75 years. At first year, 171 patients had died (32.2%): 133 in intensive care (25%), 19 in hospital  and 19 out of the hospital to follow up; 35 missing cases (6.6%). At 4 years, 181 had died (34.1%); 86 cases were lost to follow up (16.2%).The patients dead at four years to follow up were older , more severity by APACHE-II  and higher admission coma degree .Mortality at 4 years is associated with type of injury used computerized tomography (CT) by Marshall score on admission. Mortality at diffuse injury type I was 9.1% vs 68.2% in injury type IV.By logistic regression the mortality at 4 years was associated with APACHE-II , age , Injury Severity Score , Glasgow coma scale at admission , tracheostomy  and Marshall classification: diffuse injury type I (OR: 1), type II , type III  and type IV ; evacuated mass , not evacuated mass .The type of injury at admission CT in traumatic brain injury patients admitted to ICU and evaluated by Marshall score is related to mortality at 4 years. Mortality after the first year is very low.


Introduction of new vaccines in low- and lower middle-income countries has accelerated since Gavi, the Vaccine Alliance was established in 2000. This study sought to (i) estimate the costs of introducing pneumococcal conjugate vaccine, rotavirus vaccine and a second dose of measles vaccine in Zambia; and (ii) assess affordability of the new vaccines in relation to Gavi’s co-financing and eligibility policies.Data on ‘one-time’ costs of cold storage expansions, training and social mobilisation were collected from the government and development partners. A detailed economic cost study of routine immunisation based on a representative sample of 51 health facilities provided information on labour and vaccine transport costs. Gavi co-financing payments and immunisation programme costs were projected until 2022 when Zambia is expected to transition from Gavi support. The ability of Zambia to self-finance both new and traditional vaccines was assessed by comparing these with projected government health expenditures.‘One-time’ costs of introducing the three vaccines amounted to US$ 0.28 per capita. The new vaccines increased annual immunisation programme costs by 38%, resulting in economic cost per fully immunised child of US$ 102. Co-financing payments on average increased by 10% during 2008–2017, but must increase 49% annually between 2017 and 2022. In 2014, the government spent approximately 6% of its health expenditures on immunisation. Assuming no real budget increases, immunisation would account for around 10% in 2022. Vaccines represented 1% of government, non-personnel expenditures for health in 2014, and would be 6% in 2022, assuming no real budget increases.While the introduction of new vaccines is justified by expected positive health impacts, long-term affordability will be challenging in light of the current economic climate in Zambia. The government needs to both allocate more resources to the health sector and seek efficiency gains within service provision. This was switched to DTP-hepatitis B-Hib  vaccine in 2005. In 2009, a proposal was submitted for pneumococcal conjugate vaccine (PCV), rotavirus vaccine (RV) and a second dose of measles (MSD). Gavi approved PCV and MSD in 2010 and RV in 2011, following evidence of plans for cold chain expansions. However, PCV and MSD were only introduced in July 2013 and RV in November 2013. Delays were due to a measles outbreak in 2012, relocation of the Child Health Unit from the Ministry of Health to the new Ministry of Community Development, Mother and Child Health in 2011, delays in receiving the Gavi vaccine introduction grant, and delays in disbursements to subnational levels Zambia has introduced four new vaccines with Gavi support. The combined diphteria-tetanus-pertussis (DTP)-A comprehensive study on the economic and fiscal costs of Zambia’s routine immunisation services was undertaken in 2012–13, before introduction of the three new vaccines Our study objectives were to estimate the incremental costs of introducing PCV, MSD and RV and evaluate affordability after cessation of Gavi support. Although ‘one-time’ vaccine introduction costs were calculated, the primary objective was to examine the longer-term economic costs.1.1Gavi’s current eligibility criteria, established in 2011, is Gross National Income (GNI) per capita of less than US$ 1500, which is adjusted annually for inflation to remain constant in real terms. In 2015, the threshold was US$ 1580 Gavi’s co-financing policy requires countries to co-procure a portion of their new vaccines and injection equipment. MSD is exempted from co-financing, but after five years countries must take on the full costs 22.1Economic costs were estimated in 2014 values, using an exchange rate of 6.18 Zambian Kwacha for one US$ The number of fully immunised children was approximated by the number reported to receive three doses of pentavalent vaccine (penta3). With 602,000 surviving infants and 86% penta3 coverage in 2014, this was 517,720 children 2.1.1Vaccine costs were calculated by multiplying price per dose, coverage rate of the first dose, target population size, number of doses per child in the schedule and the vaccine wastage factor 2.1.23 per dose for PCV, 17.1 cm3 for RV and 2.13 cm3 for measles 3 to 138 cm3 per penta3 child Cold storage expansions were undertaken at national, provincial, district and health facility levels Power consumption for equipment was collected from WHO Product Information Sheets 2.1.3The EPIC study found that annualised costs of vehicles, vehicle maintenance and fuel comprised 12.5% of total immunisation programme costs, amounting to US$ 4.35 million in 2014 values, or US$ 8.39 per penta3 child 2.1.4As PCV and MSD were introduced concurrently, training and social mobilisation were done at the same time. Based on advice from key informants, we allocated 80% of these costs to PCV and 20% to MSD. Costs were annualised over two years, reflecting the usual frequency of training.New vaccination cards and health facility registers were printed, the Health Management Information System was modified, and a post introduction evaluation was conducted in July 2014 2.1.5Introduction of new vaccines did not lead to additional employments, but staff opportunity costs need to be accounted for. In the EPIC study, clinical staff at 51 randomly selected health facilities in nine districts were asked to allocate their working hours among all activities during one month The EPIC study took place before introduction of the new vaccines, but facility staff were asked if they thought introductions would impact operating costs, and 73% responded that they did not think so 2.2Zambia’s co-financing contributions were provided by the Gavi secretariat, and the Gavi website gave values of its vaccine and injection supplies donations Total government immunisation programme costs were the sum of systems costs, procurement of traditional vaccines and injection supplies, and government expenditure to co-procure vaccines for Gavi co-financing. The EPIC study found that immunisation systems costs amounted to US$ 31.8 million in 2011, equivalent to US$ 61 per penta3 child When assessing affordability, we compared immunisation programme costs with total government health expenditures. For 2008–2014, government health expenditures were collected from the Accountant General’s Office For 2018–2022, we assumed a nominal growth in health care expenditures of 10% per year, which was the average increase during 2014–2016, and equivalent to 5% real, annual increase with predicted inflation rates of around 5% 33.13.1.1‘One-time’, non-annualised, incremental costs amounted to US$ 4.21 million, equivalent to costs per infant in the birth cohort of US$ 0.96 for MSD, US$ 2.02 for PCV and US$ 3.58 for RV . Cold stCold storage expansions were financed by JICA (36%), ELMA Foundation (34%), Zambian government (10%), Canadian International Development Agency (9%), ARK (7%), WHO (4%), and Boston University (0.3%). Gavi, through the vaccine introduction grant, funded 64% of the social mobilisation, training and monitoring activities. The remainder was paid by the government (29%), GSK (3%), WHO (3%) and Absolute Returns for Kids (1%).3.1.2Annual costs of vaccines and injection equipment increased from US$ 6.8 million to US$ 7.0 million after introduction of MSD, to US$ 14.0 million after PCV, and to US$ 16.9 million after RV . CompareWhen ‘one-time’ costs were annualised, training and social mobilisation were the most important capital costs, each comprising 2% of total costs . Cold stThe EPIC study concluded that economic costs of routine immunisation in Zambia totalled US$ 38.2 million in 2011, equivalent to US$ 66 per penta3 child 3.23.2.1Zambia introduced pentavalent vaccine in 2005. Gavi’s first co-financing policy required countries to start co-procuring six years after introducing the first new vaccine; thus 2011 for Zambia. With GNI per capita of US$ 490 in 2005, Zambia belonged to the poorest co-financing country group, entailing flat levels of US$ 0.20 per dose for the first vaccine When Gavi’s second co-financing policy came into effect in 2012, Zambia’s GNI had increased to US$ 1,650 per capita, placing it in the intermediate group, requiring a 15% annual increase in the per dose amount. During 2012, Zambia procured the equivalent of US$ 0.27 per dose in co-financing for pentavalent vaccine and US$ 0.20 for PCV. During 2013, co-financing of RV was added, starting at US$ 0.20 per dose. During 2015, approximately 12% of pentavalent vaccines, 8% of PCV and 9% of RV were co-financed.Gavi’s new co-financing policy starts in 2016, but the first year is a grace year with the old policy still applying. Zambia is in the accelerated transition group, as its GNI per capita has exceeded the Gavi eligibility threshold for three consecutive years . Hence, Zambia is predicted to fully finance all vaccines in 2022.3.2.2During 2008–2016, per capita government health expenditures fluctuated between US$ 14 in 2009 and US$ 45 in 2013 . BetweenRoutine immunisation programme costs were US$ 52.9 million in 2014, equivalent to 0.2% of GDP. Gavi funded approximately 20% of these costs. Immunisation programme expenditures paid for by the government comprised approximately 6% of government health expenditures. Vaccine expenditures comprised 1% of government health expenditures excluding personnel . Per capAssuming a 5% annual, inflation-adjusted increase in government health expenditures during 2017–2022, immunisation programme costs are projected to be 5% of total health expenditures in 2022. When assuming 0% increase in government health expenditures, immunisation costs would account for 10% in 2022.Considering vaccine costs alone and assuming 5% real increase in government health expenditures, these will account for 3% in 2022 exclusive personnel. When assuming 0% annual increase until 2022, the proportion is 6%. For vaccine costs to remain at 1% of government health expenditures, excluding human resources, the nominal budget needs to increase by 30% each year between 2017 and 2022.4Incremental, annual systems costs of introducing the three new vaccines were US$ 4.9 million, equivalent to US$ 7.69 per child in the birth cohort and US$ 0.31 per capita. With a total of 2.8 million doses delivered, the systems costs per vaccine dose delivered of all three vaccines were US$ 1.78. The systems costs were US$ 1.33 for PCV, US$ 2.15 for RV and US$ 3.43 for MSD. Costs of simultaneous introduction of PCV, RV and MSD in Ghana in 2012 totalled US$ 2.42 per dose When including costs of vaccines, annual incremental costs of the three introductions amounted to US$ 23 per child in the birth cohort. It led to a cost increase of 38%, resulting in total immunisation programme costs of US$ 52.9 million in 2014, equivalent to US$ 102 per penta3 child. This is comparable to US$ 132 per penta3 child in Honduras, where PCV, RV and MSD have also been introduced Cold chain investments may be essential for new vaccines, and have been a major focus of cost studies of earlier new vaccine introductions Government health expenditures have fluctuated since 2008. Zambia experienced a serious economic situation in 2015. The annual economic growth rate declined from an average of 7% since 2010 to 3.4% in 2015 Immunisation is projected to comprise 5–10% of government health expenditures in 2022, compared to 6% in 2014. The proportion depends on the extent real government health expenditures increases and assuming no further deterioration in the exchange rate. Even though there are no benchmarks for optimal spending on immunisation, these proportions must be considered relatively high. Projected vaccine and injection equipment costs of 3–6% of non-personnel health spending are also higher proportions than those reported as potentially challenging in other countries Numerous studies from sub-Saharan Africa have shown substantial health impacts and cost-effectiveness of PCV, RV and MSD


Human immunodeficiency virus (HIV) associated anemia is common and it is a challenge for prognosis of HIV positive patients. It is stated in different literature that the prevalence of anemia due to HIV before highly active antiretroviral therapy (HAART) initiation is more prevalent than after HAART initiation. Thus this study aimed to assess anemia prevalence before and after initiation of HAART among HIV patients attending university of Gondar hospital from 2008 – 2013.A retrospective study was conducted by collecting data from antiretroviral clinic of University of Gondar Hospital from January 1, 2008 to December 30, 2013. Data was collected by using data collection sheet which contains age, sex, regimen type, hematological parameters and CD4 cell count. Data were analyzed using SPSS version 20. In order to compare means paired t-test was used. P- Value <0.05 was considered as significant.Prevalence of anemia before and after HAART initiation was 21.2% and 11.5% respectively. There is a significance difference in CD4 cell count, hemoglobin and hematocrit values on patients before and after HAART initiation (P <0.001). Opportunistic infection and CD4 cell count were associated with prevalence of anemia before HAART initiation.There was a decline in the prevalence of anemia and increment of mean CD4 cell count among HIV infected patients after HAART initiation. HIV patients are recommended checking up their CD4 cell counts regularly and starting HAART when it is appropriate with regular follow-up. Anemia is the most common hematological abnormality in HIV positive patients. It has been shown to be the risk factor for early death in patients with AIDS. Anemia is not only associated with decreased quality of life but it is also independently associated with decreased survival with HIV . It is dPrevious studies have found that the incidence of anemia increases with progression of HIV infection. Several other etiologic factors may also be involved in the development of HIV-associated anemia, including micronutrient deficiencies, immunological myelosuppression, impaired erythropoietin production and blood loss from intestinal opportunistic disease .Anemia is a condition in which there is a decrease in the red blood cell count, hemoglobin and/or hematocrit values as compared to normal reference range for age, sex, race and altitude which decreases oxygen-carrying capacity of red blood cells to tissues . It is dAnemia is observed in both antiretroviral therapy treated and untreated individuals, but severity varies due to the immune status or CD4 cell levels and also the prevalence increases as the disease advanced and left untreated . If an iMost studies described the prevalence of anemia in HIV infected individuals before and after initiation of highly active antiretroviral therapy (HAART), however few studies were conducted in Ethiopia on the assessment of anemia among HIV positive patients , 12, 16.This retrospective study was conducted in the ART clinic of the University of Gondar Hospital which is found in Northwest of Ethiopia. Patients’ data with full information of sex, age, regimen type, hematological and immunological parameters, WHO clinical stage, type of regimen, opportunistic infections and cotrimoxazole were collected from follow up charts and registration books of patients who started HAART at GUH ART clinic from January 1, 2008 to December 30, 2013. Hematological and immunological parameters were collected at baseline when the HIV positive patients adhere to ART and after six month of HAART initiation. The quality of data were controlled at different levels for completeness and consistency. Whenever an error was found at any level, it was corrected by rechecking from the registration book.The data were cleaned, edited, checked for completeness and entered into SPSS version 20 for analysis. Descriptive statistics was used to get a clear picture of dependent and independent variables. In order to compare means paired t-test was used and also chi square was computed to determine association between dependent and independent variables. P- Value <0.05 was considered as significant.The study was conducted after ethical letters obtained from School of Biomedical and Laboratory Science Ethical Committee. Then permission was taken from hospital higher management. After permission was obtained, data were collected from ART clinic. To keep confidentiality codes were used and unauthorized person didn’t have access to the data.A total of 349 HIV positive patients, 218 (62.5%) females and 131 (37.5%) males, were involved in this study. The overall mean age was 34.6 ± 8.5 years, within the range of 15 – 60 years old. Majority of the patients [154 (44.1%)] were within 26 – 35 years of age. About 226 (64.8%) of participants were under WHO clinical stage I & II, 87.7% of them were taking cotrimoxazole prophylaxis therapy and about 4.3% of them were TB co-infected   before HAART initiation and 11.5% (40/114) after HAART initiation. About 40 (11.5%) had mild anemia, 27 (7.7%) had moderate anemia, and 7 (2%) had severe anemia before HAART initiation. However, the prevalence of anemia after HAART initiation was significantly decreased by 9.7%  were males and 22% (48/118) were females. Similarly 30.4% (14/46) patients at the age of <25 years old and 35.2% (50/142) of patients with CD4 cell count <200 cells/μl were developed anemia. There was a significant association between anemia with CD4 cell count and TB before HAART initiation. HIV patients with CD4 cell count <200 cells/μl before HAART initiation had higher prevalence of anemia . Similarly about 53.3% (P =0.002) patients with TB had developed anemia before HAART initiation. However, there was no association of anemia with sex, age, and WHO clinical stage  were males and 13.8% (30/118) were females. Similarly 12.3% 14/146) patients at the age of 36–45 years old and 18% (18/100) patients with CD4 cell count <200 cells/μl were developed anemia. Although, HIV patients with CD4 cell count <200 cells/μl had higher prevalence of anemia (18%) after HAART initiation, there was no significance association between anemia and CD4 cell count (P =0.051). Similarly sex, age regimen type and WHO clinical stage had no association with anemia  and 37.3%) , Hawassa% and  in this study was lower than study done in different countries, such as Germany from 2001–2002 (61%) , India iPrevalence of anemia after HAART initiation (11.5%) was lower study done in in studies from Minillik II hospital, Addis Ababa 200 (37.3%)  and HawaThe prevalence of anemia observed across sex groups before HAART initiation was 19.8% (26/131) for males and 22% (48/218) for females similarly after HAART initiation 7.6% (10/131) and 13.8% (30/218) were male and female patients were anemic respectively. This was slightly lower than a report from Minillik II hospital ART clinic, Addis Ababa Ethiopia, 70.25% vs. 29.75% in males and females before HAART initiation and 69.23% vs. 30.77% in males and females after HAART initiation respectively . PrevaleIn this study the minimum and maximum CD4 cell count before HAART initiation was 1 cell/μl and 795 cells/μl and after HAART initiation were 14 cells/μl and 969 cells/μl. Patients with CD4 cell count <200 cells/μl before HAART initiation was 142 (40.7%) while after HAART initiation was 100 (28.7%). The risk of having CD4 cell count <200 cells/μl was increased before HAART initiation, because before HAART initiation patients have less immunity towards the HIV and the virus attacks CD4 cells easily.There was a decline in the prevalence of anemia and increment of mean CD4+ T cell count among HIV infected patients after HAART initiation. Before HAART initiation, HIV patients have significantly higher prevalence of anemia than those after HAART initiation. The prevalence of anemia was significantly higher in patients with CD4 cell count <200 cell/μl. There was a significant association between prevalence of anemia with CD4 cell count and opportunistic infections before HAART initiation. But there was no association of prevalence of anemia with sex, age, regimen type and WHO clinical stage. Based on the present finding, HIV patients are recommended to check up their CD4 cell counts regularly and start HAART when it is appropriate in order to decrease the prevalence of anemia.


Introduction. The risk of gastrointestinal (GI) bleeding with rivaroxaban has not been studied extensively. The aim of our study was to assess this risk in comparison to warfarin. Methods. We examined the medical records for patients who were started on rivaroxaban or warfarin from April 2011 to April 2013. Results. We identified 300 patients (147 on rivaroxaban versus 153 on warfarin). GI bleeding occurred in 4.8% patients with rivaroxaban when compared to 9.8% patients in warfarin group (p = 0.094). GI bleeding occurred in 8% with therapeutic doses of rivaroxaban (>10 mg/d) compared to 9.8% with warfarin (p = 0.65). Multivariate analysis showed that patients who were on rivaroxaban for ≤40 days had a higher incidence of GI bleeding than those who were on it for >40 days . Concomitant use of dual antiplatelet agents was associated with increased risk of GI bleeding in the rivaroxaban group . Prior GI bleeding was also a risk factor for GI bleeding in rivaroxaban group (OR = 15.5). Conclusion. The incidence of GI bleeding was similar between rivaroxaban and warfarin. The risk factors for GI bleeding with rivaroxaban were the first 40 days of taking the drug, concomitant dual antiplatelet agents, and prior GI bleeding. Rivaroxaban is the first direct factor Xa inhibitor that has been approved by the US Food and Drug Administration (FDA) for the prevention of stroke and systemic embolism in patients with nonvalvular atrial fibrillation (AF), for the prevention and the treatment of venous thromboembolic events (VTE) –7. It ha2) .Rivaroxaban selectively inhibits the active site of factor Xa (both free and prothrombinase-bound forms) without a requirement for cofactors such as antithrombin III . The oraDespite these advantages of rivaroxaban over warfarin, the risk of bleeding, including gastrointestinal (GI) bleeding, is a concern with its use. Our hypothesis was that GI bleeding events occur more often in patients on rivaroxaban when compared to patients on warfarin. The primary aim of our study was to assess the risk of GI bleeding with rivaroxaban and compare to that of warfarin in a community hospitals setting and to identify the risk factors for developing GI bleeding in patients on rivaroxaban.In this retrospective cohort study, we examined the medical records of all patients who were started on rivaroxaban from April 2011 to April 2013 and compared to a control group of consecutive patients who were started on warfarin during the same period in 1 : 1 fashion. The study was conducted in two hospitals  after obtaining the Institutional Review Board (IRB) approval from each institution. This study was conducted in collaboration with Georgia Regents University, Augusta, GA.2 or end-stage renal disease on dialysis).Demographic details, laboratory studies, information on the concomitant use of antiplatelet agents or nonsteroidal anti-inflammatory drugs (NSAIDs), duration of rivaroxaban and warfarin (≤ or >40 days), dose of rivaroxaban (10 mg/day (prophylactic dose), >10 mg/day (therapeutic dose)), indication for anticoagulation, GI bleeding events and the site of GI bleeding, major bleeding events and the site of bleeding, acute coronary syndrome (ACS), venous thromboembolic events (VTE), cerebrovascular events, and all-cause deaths while on an anticoagulation agent were collected. The primary aim was to assess the risk of GI bleeding in patients on rivaroxaban when compared to patients on warfarin during the study period and to identify any risk factors for developing GI bleeding. Gastrointestinal bleeding was defined as any bleeding in the GI tract that required hospitalization. The secondary aims were the risk of major bleeding events , ACS that required angioplasty (with or without stenting) or heart surgery, VTE events (deep venous thrombosis or pulmonary embolism), cerebrovascular events (stroke or transient ischemic attack), and all-cause deaths while on an anticoagulation agent. Inclusion criteria were patients who were on rivaroxaban for ≥4 days during the study period and were compared to age and gender-matched patients who were on warfarin for ≥4 days. Exclusion criteria were an unknown duration of rivaroxaban or warfarin, lack of follow-up, age <18 years, pregnancy, mechanical valve replacement, and advanced kidney disease (glomerular filtration rate (GFR) < 15 mL/min/1.73 mp value of < 0.05 was considered statistically significant. Parametric two-sample t-tests were conducted to compare the means of the two groups involved, but the assumptions associated with t-tests  were not satisfied. Hence, the nonparametric Wilcoxon's rank sum test was conducted to compare the means of the two groups involved. A chi-square analysis, Fisher's exact test, and a Pearson correlation were conducted for the other variables. A logistic regression was carried out in multivariate analyses. Odds ratios (OR) were generated between group comparisons. A chi-square analysis was used for parameters in secondary aims.Patients' data were entered into Microsoft Excel spreadsheet in a coded format which was locked with a password. All analyses were performed with the use of SAS software . A 2-sided p = 0.0573) (p = 0.0738). The majority of people in both groups were White (76.87% versus 86.93%) and female (52.38% versus 54.25%) in rivaroxaban and warfarin groups, respectively. Atrial fibrillation was the indication for anticoagulation in 46.94% in rivaroxaban group versus 60.78% in warfarin group (p < 0.0001). Prophylactic dose (10 mg/day) was used for VTE prophylaxis in 60 patients (40.82%) in rivaroxaban group, whereas therapeutic doses (>10 mg/day) were used in 87 patients (59.18%). The mean duration of drug use was significantly different between rivaroxaban and warfarin groups  with 55.1% of patients in rivaroxaban group being on the drug for ≤40 days when compared to only 10.46% of patients in warfarin group (p < 0.0001). Concomitant use of aspirin, thienopyridines , dual antiplatelet agents (aspirin and thienopyridine), and NSAIDs was found in 38.10%, 8.16%, 5.44%, and 9.52%, respectively, in the rivaroxaban group when compared to 41.18%, 17.65%, 9.80%, and 4.58%, respectively, in the warfarin group . History of prior GI bleeding was present in 5 patients (3.40%) in the rivaroxaban group and 15 patients (9.80%) in the warfarin group (p = 0.0356). Laboratory studies  and BMI  were not significantly different between both groups . We also separated the patients on therapeutic doses (>10 mg/day) of rivaroxaban from prophylactic dose group and compared them separately to the patients in warfarin group, to minimize the heterogeneity between both groups. The characteristics of patients on therapeutic doses are shown in A total of 300 patients were identified, of whom 147 patients were on rivaroxaban (mean age 68.3 ± 14.5 years) and 153 patients were on warfarin (mean age 71.4 ± 13.1 years) ( 0.0573) . Eighty-Rivaroxaban was discontinued prematurely in 24 patients (16.33%) for various reasons (GI bleeding (7), major bleeding events (6), switching to warfarin for financial issue (6), high risk for bleeding (1), intolerance due to intractable vomiting (1), switch to dabigatran (1), switching to apixaban (1), and hospice (1)), whereas warfarin was discontinued prematurely in 28 patients (18.30%) for the following reasons (GI bleeding (15), major bleeding events (4), high risk for bleeding (4), switching to dabigatran (4), and switching to rivaroxaban (1)).p = 0.094) (p = 0.65). The mean age for GI bleeders in rivaroxaban group was 72.14 ± 15.40 years when compared to 75.80 ± 11.38 years in warfarin group (p = 0.4801). The mean duration of being on the drug was 29.00 ± 38.03 days in rivaroxaban group as compared to 163.87 ± 143.5 days in warfarin group (p = 0.0239). Concomitant use of antiplatelet agents or NSAIDs, laboratory parameters, BMI, and prior GI bleeding were not statistically different between GI bleeders in both groups .GI bleeding occurred in 7 patients (4.76%) in the rivaroxaban group when compared to 15 patients (9.80%) in the warfarin group (= 0.094) . All casp = 0.023). Concomitant use of dual antiplatelet agents (aspirin and thienopyridine) was associated with an increased risk of GI bleeding in rivaroxaban group . A history of prior GI bleeding was a risk factor for GI bleeding in the rivaroxaban group (OR = 15.5). Age, gender, ethnicity, BMI, concomitant use of aspirin , thienopyridines , or NSAIDs, hemoglobin <12 g/dL, creatinine >1.5 mg/dL, GFR ≤ 30 mL/min/1.73 m2, and alanine aminotransferase >40 IU/L were not risk factors. The site of GI bleeding in rivaroxaban group was the lower GI tract in 57.14% and upper GI tract in 42.86% when compared to 33.33% and 40%, respectively, in the warfarin group with 26.67% without obvious site of GI bleeding (p > 0.05). Multivariate analysis carried out using a logistic regression showed that patients who were on rivaroxaban for ≤40 days had a higher incidence of GI bleeding than those who were on the drug for >40 days . ACS that required angioplasty with or without stenting or heart surgery occurred in 3 cases (2.04%) in rivaroxaban group, compared to 8 cases (5.23%) in warfarin group (p = 0.2189). Symptomatic VTE events occurred in 2 cases in each group, and cerebrovascular events (stroke or TIA) occurred in one case of each group (p > 0.05 for both). There was no incidence of intracranial hemorrhage in warfarin group but there was one case in rivaroxaban group (p = 0.49). All-cause deaths while on the anticoagulation agent occurred in 1.36% in rivaroxaban group (Sepsis (2)) and in 5.23% in warfarin group (Sepsis (3), retroperitoneal bleeding (1), respiratory failure (1), congestive heart failure (1), lung cancer (1), and vulvar cancer (1)) (p = 0.1042). All the cases of secondary aim parameters occurred in patients on the therapeutic doses of rivaroxaban except 2 cases (one VTE and the case of postoperative knee bleeding) which occurred on the prophylactic dose.Major bleeding events, defined as the bleeding events that required hospitalization and cessation of the anticoagulation agent, occurred in 6 patients (4.08%) in rivaroxaban group (hematuria (2), epistaxis (2), ICH (1), and postoperative knee {surgical site} bleeding that required reoperation (1)), when compared to 4 patients in warfarin group , hemarthrosis (1), and soft tissue bleeding due to valvular carcinoma (1))  of rivaroxaban but all the events occurred on therapeutic doses (>10 mg/day). In the RECORD 1 trial, major GI bleeding occurred in 2 patients (0.09%) treated with prophylactic dose (10 mg/day) of rivaroxaban when compared to 1 0.05%) in the enoxaparin group during the study period of 5 weeks for the thromboprophylaxis after hip arthroplasty . This pa% in the As mentioned above, all cases of GI bleeding in our study occurred in patients on the therapeutic doses of rivaroxaban. It has been shown that major and nonmajor bleeding events, including GI bleeding, with rivaroxaban are dose-dependent as demonstrated in the phase II trials , 28. In p < 0.0001) [2) when compared to those with preserved renal function, despite the dose reduction of rivaroxaban to 15 mg once daily instead of 20 mg once daily  [2 instead of 15 mg once daily) to warfarin, major GI bleeding events occurred in 8 cases (1.25%) in the rivaroxaban group and 15 cases in the warfarin group (2.35%) [2 [Major GI bleeding events were more common in rivaroxaban group when compared to warfarin group in ROCKET AF study  . In J-RO (2.35%) . Majorit (2.35%) . Of thos.35%) [2 .The number of GI bleeding events observed in our study was higher than the other studies, which may be attributed, in part, to the difference in the definitions of GI bleeding that was used in our study. We compared the incidence of all GI bleeding that required hospitalization and cessation of the active drug, as opposed to ROCKET AF trial which separated major GI bleeding (which was associated with dropping Hb by 2 g/dL or requiring transfusion of 2 units of blood) from nonmajor bleeding. By using our definition, it is possible that we included all GI bleeding events (major and nonmajor).In our study, the first 40 days of rivaroxaban use was a risk factor for developing GI bleeding. The odds of GI bleeding for patients on rivaroxaban for ≤ 40 days were 2.8 times higher than those on the drug for >40 days. This is possibly because rivaroxaban may achieve the anticoagulant effect earlier than warfarin. The time in therapeutic range (TTR) for international normalized ratio (INR) for warfarin is low in the first months of starting the drug. For example, in EINSTEIN-DVT trial, the TTR was 54.1% in the first month when compared to 66.4% in the tenth month [The concomitant use of NSAIDs and antiplatelet agents with anticoagulants is important because of the potential increased risk of GI bleeding with their use; however, our study did not show any increased risk of GI bleeding with concomitant use of NSAIDs or aspirin , or thienopyridines . It has been shown that neither NSAIDs nor aspirin has a clinically relevant interaction with rivaroxaban , 34. In 2, and alanine aminotransferase >40 IU/L were not the risk factors for the development of GI bleeding in rivaroxaban group. However, the advanced age, especially the age over 75, was a risk factor for GI bleeding in patients on rivaroxaban in a recent study [In our study, age, gender, ethnicity, BMI, hemoglobin of <12 g/dL, creatinine >1.5 mg/dL, GFR ≤ 30 mL/min/1.73 mnt study . Similarnt study . The sitThis study has several limitations. First, the size of the study was small when compared to other studies in this field. Second, since our study was retrospective, it carries its own bias such as missing information. Third, there was heterogeneity between both the groups such as age and previous GI bleeding which was minimized when only the therapeutic group was compared to the control group (compare Tables In conclusion, our study provides data that support the similarity of GI bleeding risks between rivaroxaban and warfarin. Rivaroxaban is more convenient than warfarin for both patients and health care providers. The risk factors for the development of GI bleeding in patients on rivaroxaban were the first 40 days of drug use, concomitant use of dual antiplatelet agents, and a history of previous GI bleeding. Our study is another step toward bid adieu to warfarin.


Background: Cardiovascular disease (CVD) is the leading cause of mortality and morbidity in the elderly and the ambient concentration of PM2.5 has been associated with several cardiovascular diseases. Methods: We describe the present state of planetary air pollution, analyze epidemiological studies linking PM2.5 and CVD, and discuss multiple pathophysiological mechanisms linking PM2.5 and CVD. Results: A few epidemiological studies show that the elderly appear specifically susceptible to adverse cardiovascular effects triggered by PM2.5 exposure. Plausible pathophysiological mechanisms include inflammatory dysfunction, oxidative stress, abnormal activation of the hemostatic system and disturbance of the autonomic nervous system. Conclusions: An in-depth knowledge of the chemical compounds, pathophysiological mechanisms, and epidemiological studies of PM2.5 are recommended to understand this important and modifiable factor contributing to geriatric CVD burden. We offer public health recommendations to reduce this preventable cause of disease and death. Elderly people have the highest rates of CVD and thus are the most susceptible population [2.5-related pathophysiological mechanisms of CVD and epidemiological studies linking PM2.5 and CVD, especially in susceptible people, to make recommendations for future public health and reduce this avoidable cause of disease and death.Cardiovascular diseases (CVD) include disorders of the heart  and blood vessels , particular those supplying the brain (ischemic and hemorrhagic stroke). Together, these disorders constitute the leading cause of death across the globe, with low- and middle-income countries most heavily affected. According to the World Health Organization (WHO), each year about 17.3 million people die of cardiovascular disease, accounting for 30% of all deaths  have pro2.5 refers to atmospheric particles with diameters less than or equal to 2.5 micrometers. The influence of PM on humans depends on particle size, which is linked with its aerodynamic diameter (AD)  is classified into four categories according to aerodynamic diameter: Total suspended particulate (TSP ≤ 100 μm); particulate matter (≤10 μm); fine particulate matter ≤2.5 μm), and ultrafine particles (≤0.1 μm). PMter (AD) . The ran effects ,5,6. Com μm, and ace area . The smaace area .The major sources of particulate matter are broadly divided into two parts, human activities and natural phenomena, including wildfires, volcanoes, and land dust . In addi2.5 is considered the main culprit for the adverse effects of air pollution on human health [2.5 pollution affects the whole planet, particularly densely populated metropolitan areas of eastern and southern China, northern India, and the emerging countries of South-East Asia. Parts of Europe and America are not spared as well  Part of the particulate matter causes the above reaction through other routes, such as entering into the circulatory system via the digestive tract. Animal studies have shown that PM2.5 can be devoured by macrophages and endothelial cells, indicating that air pollution may have direct effects on the vascular system [Air particulate matter enters the human body mainly in two ways: (1) PMr system ,5,6,18.The relationship between air pollution and respiratory diseases, such as asthma and chronic obstructive pulmonary disease, is well established. Nevertheless, the relationship between air pollution and CVD remains unclear. The plausible pathophysiological mechanisms can be divided into the following several aspects .2.5 inhalation can lead to the occurrence of systemic inflammation, increasing the risk of cardiovascular stress. PM2.5 can enter the alveolar epithelium, cause local inflammation and oxidative stress, resulting in the release from lung cells into bronchial fluid and the blood stream of several inflammatory mediators, such as IL-6, IL-8, TNF-α and interferon-γ [2.5 for six hours, there is increased expression of inflammation-related genes such as mRNA of TNF-α, TNF-β and IL-6, IL-8. A recent study finds that IL-6 can make a rapid response to air pollution and increase the production of C-reactive protein (CRP) [2.5; for every increase of 100 μg/m3, blood CRP increases 8.1 mg/L [2.5 exposure in older individuals [Both animal and human studies ,22,23,24rferon-γ . These sin (CRP) . CRP is 8.1 mg/L . Interes8.1 mg/L . Higher ividuals .2.5 exposure is an abnormal activation of the hemostatic system. Epidemiological studies have associated PM2.5 inhalation with venous thrombosis and a shorter prothrombin plasma-clotting time. The diffuse pulmonary inflammation caused by PM2.5 may spread to the circulatory system, causing abnormal activation of the hemostatic system [in vitro cell studies show that PM2.5 increases fibrinogen and tissue factors [2.5 exposure may increase the risk of acute thrombosis, as in myocardial infarction and ischemic stroke. At the same time, fibrinogen is also an important risk factor for stroke [Another probably detrimental effect of PMc system . Animal  factors , and finr stroke .2.5 can stimulate the ANS and increase the risk of arrhythmia and acute cardiovascular events, with serious impact on the elderly [2.5 is linked with HRV change in the elderly. Compared with exposure to clean air for two hours, HRV drops 35.7% after inhalation of 21.2~80.3 μg/m³ PM2.5 [2.5 was found to have a strong effect on the decrement in HRV [2.5 on subjects with hypertension were larger than on the subjects without hypertension [PM particles disturb the autonomic nervous system (ANS) . Under n elderly . A numbe elderly . Heart rm³ PM2.5 . Moreovet in HRV . Additiortension . The dec2.5 is one possible CVD mechanism. Cardiovascular endothelial cells treated in vitro with PM2.5 for 24 hours display adhesion molecules and apoptosis [2.5, suggesting that PM2.5 may damage the vascular endothelium, thereby causing CVDs. Recently, some studies suggest that specific metals may be important components responsible for PM2.5-induced cardiovascular effects and that the reduced capacity of endothelial repair may play a critical role [Because vascular endothelial injury is an important pathological basis of cardiovascular disease, the damage of vascular endothelial cells by PMpoptosis . Cell mocal role .2.5 can increase risk of CVD [et al. found that risk of CVD exposed to PM10 increased 0.26%, compared with people 5–44 years of age or 45–64 years of age[2.5 adversely affected cardiac autonomic function of the elderly people with heart diseases [10 and congestive heart failure, a 0.72% increase in risk per 10 μg/m3 elevation in same-day PM10 concentration [3 elevation), a marker of traffic-related PM, was associated with a 1.1% increase in carotid intima media thickness (CIMT) in a cohort of elderly men living in the Boston area [2.5 can increase the risk of CVD, it is noteworthy that some studies had not find relationships between PM2.5 and risk of CVD [Though several studies have shown exposure to PMk of CVD ,55,56,57rs of age,59. Anotdiseases . Dominicntration . This iston area . Short-tton area . In an eton area . While tk of CVD ,64,65,662.5 and a few epidemiological studies showed more risk of CVD increased in the elderly exposed to the same levels of PM2.5. Due to increasing of PM2.5 level and aged populations, more attention should be paid to CVD risk and PM2.5 exposure in the elderly. In addition, the relationship between PM2.5 concentration and CVD risk appears not to have a “safe” threshold. The “alarm” threshold periodically claimed by regulatory agencies during certain seasons should therefore be considered advisory, and efforts should be made to keep level of pollutants as low as possible [2.5 pollution and societal and personal measures to reduce other risk factors should be recommended for the elderly [Walking or cycling in parks or country roads instead of busy streets during rush hours.Using more public transport rather than private cars and motorbikes.2.5 .Avoiding outdoor exercise at times of high PMUsing an air purifier to optimize indoor air quality.Choosing appropriate respirators that fit snugly on the face if air pollution is severe.Reducing other known cardiovascular risk factors, such as smoking and alcohol.Warranting more aggressive use of primary and secondary preventive therapies, including antiplatelet agents, lipid-lowering agents, and treatments for hypertension or diabetes.From a point of pathophysiology, the elderly is a susceptible population to cardiovascular injury by PM elderly ,68:Walk


Ten bio-oil compounds were measured via pyrolysis/gas chromatography-mass spectrometry (Py/GC-MS) in maize cobs. 184 recombinant inbred lines (RILs) of the intermated B73 x Mo17 (IBM) Syn4 population were analyzed in two environments, using 1339 markers, for quantitative trait locus (QTL) mapping. QTL mapping was performed using composite interval mapping with significance thresholds established by 1000 permutations at α = 0.05. 50 QTL were found in total across those ten traits with R Corn cobs and stover have received attention for use as alternative energy platforms. It is estimated that more than 280 million tons of these residues are produced each year in the United States , 2. JansWhile there have been several quantitative trait locus (QTL) mapping experiments completed in maize in regards to ethanol conversion , 7, therWater content for bio-oil can vary from 15–30% and heating values range from 16–19 Megajoules (MJ)/kg, while petroleum oils typically contain very little water (0.1%) and have a higher heating value of 40 MJ/kg . By contet al. [brown midrib hybrids of maize. In this study, measurements for 26 compounds were available. However, we focused on ten of those compounds for QTL analysis. Two compounds each were chosen that derive from cellulose, hemicelluloses, and the hydroxyphenyl, guaiacyl, and syringyl lignin subunits. Levoglucosan and hydroxyacetaldehyde derive from cellulose [brown midrib3 mutant, for pyrolysis of neutral detergent fiber [Jeffrey et al.  demonstrellulose –21, and ellulose , 22. Hydellulose , and is ellulose . Hydroxyellulose  and cellellulose . Phenoliellulose . In addiellulose –27. We hnt fiber . Phenol,2 generation and has an overall map distance almost four times greater than previously used recombinant inbred lines (RILs) [The intermated B73 x Mo17 (IBM) Syn4 population is an advanced intercross lines (AILs) population that was developed in order to provide increased QTL resolution. The population was developed through four generations of random mating after the creation of the Fs (RILs) .Our objectives were to (1) determine whether there is significant genetic variation for 10 pyrolysis compounds in the IBM Syn4 population and to calculate phenotypic correlations among these compounds, (2) identify QTL for those 10 compounds, and (3) compare our results with previous QTL studies on maize cell wall traits and discuss implications for breeding dual purpose maize.et al. [et al. [et al. [et al. [Plant materials used in this study have been previously described by Lorenz et al.  and Jans [et al. . Briefly [et al.  analyzed [et al.  analyzedField trials were performed in 2006 and 2007 in Madison  and Arlington (43°18'13.57"N 89°23'16.10"W), WI and were planted in a randomized complete block design with two replications per location (i.e. one replication is one complete block). B73 (parent), Mo17 (parent), and 206 recombinant inbred lines (RILs) of the IBM Syn4 per se populations were planted at a density of 79,040 plants per hectare in single row plots that were 6.08m long and 0.76m apart. Trials were planted in Plano silt loam soil on May 21, 2006 in Madison and June 2, 2006 in Arlington. After most of the plots at a location reached physiological maturity, all plots at that location were harvested. In 2006, all ears were harvested by hand 125 days after planting in Madison and 114 days after planting in Arlington. Cobs were then shelled and dried in a forced-air dryer for one week at 55°C. Cobs were ground in a hammer mill to pass a 1mm screen. Ground cobs were further ground in a ball mill  to reduce particle size. Each parent (B73 and Mo17) was planted in two plots per block. Phenotypic data for the parents were not used in QTL analysis, but included for comparison to each other and the RILs.In this study, we used cob materials from 2006: one field replication from Madison and two field replications from Arlington, WI. 184 RILs from the IBM Syn4 per se population were phenotyped for 26 bio-oil compounds through pyrolysis/gas chromatography-mass spectrometry (Py/GC-MS). Field permits were not required, as trials were performed on University of Wisconsin-Madison field plots dedicated to experimentation. Field trials did not involve endangered or protected species.et al. [The method and instruments used are the same described by Jeffrey et al. . BrieflyLeast square means (lsmeans) were calculated for each RIL over all 3 environments in SAS PROC GLM , using RIL and environment (three environments: the two replications from Arlington were considered as distinct environments) as fixed effects. Least square means for each RIL were used as phenotypic data for correlation and QTL analysis. Phenotypic correlations were calculated as Pearson product-moment coefficients using SAS PROC CORR. Coefficients of variation (CV) were calculated by dividing square root of the mean square error by mean (multiplied by 100). Phenotypic data for B73 and Mo17 were compared against each other using SAS PROC GLM, with genotype and environment as fixed effects.et al. [Heritabilities were calculated according to Holland et al.  on an enMaizeGDB.org . This map provides a length of 6242.7centimorgans (cM), which conveys an average distance between markers of 4.66cM. This map does not convey cM in the traditional definition, thus we will refer to positions on the map in IBM centimorgans (IcM) [Genotypic data for 1339 markers were obtained for 184 RILs from ns (IcM) .2 effects, and sum of R2 effects over all QTL of a trait were calculated using MIM.Composite interval mapping (CIM) was performed using WinQTL Cartographer version 2.5 . Ten cofMeans, standard errors, heritabilities, and other summary statistics for the ten compounds are shown in Overall, loose trait correlations were found , with thQTL analysis resulted in the identification of 50 QTL across eight maize chromosomes, as no QTL for these traits were found on chromosomes 9 or 10 . All 50 The 50 QTL were spread across 15 different (greater than 10IcM separating QTL positions) chromosome regions, with 6 regions containing 4 or more QTL . The firFive QTL were found for levoglucosan that accounted for 18.5% of the phenotypic variation (17.4% under the MIM model): on chromosome 1 at position 892.8IcM, chromosome 3 at 157.9IcM, chromosome 4 at 560.9IcM, chromosome 5 at 192.3IcM, and chromosome 7 at 206.5IcM. Four of these QTL mapped to the same region as the other cellulose derived compound  and were located on chromosomes 1, 3, 4, and 5, respectively. The other two QTL found for hydroxyacetaldehyde were on chromosome 2 at 298.3IcM and on chromosome 6 at 412.3IcM. Together these six QTL accounted for 21.6% of the phenotypic variation (19.8% under the MIM model).Five QTL were found for acetic acid that accounted for 23.2% of the phenotypic variation (17.0% under the MIM model): on chromosomes 2 (299.4IcM), 5 (267.3IcM), 6 (414.3 and 484.5IcM), and 7 (288.9IcM). One of those QTL locations was also found for the other hemicellulose derived compound (hydroxyacetone) on chromosome 2. The second QTL for hydroxyacetone was identified on chromosome 4 at 557.1IcM, and the two QTL together accounted for 9.0% of the phenotypic variation (9.3% under the MIM model).Six QTL were found for phenol that accounted for 24.8% of the phenotypic variation (19.4% under the MIM model): chromosome 2 at 46.3IcM and 299.4IcM, chromosome 3 at 406.5IcM, chromosome 4 at 560.9IcM, and chromosome 6 at 414.3IcM and 477.9IcM. For the other hydroxyphenyl derived compound, 4-methylphenol, four QTL mapped to the same region as those for phenol: both QTL on chromosome 2, chromosome 3 at 407.0IcM, and on chromosome 4. The other two QTL found for 4-methylphenol were found on chromosome 3 at 156.9IcM and 438.9IcM. The six QTL for 4-methylphenol accounted for 23.3% of the phenotypic variation (16.9% under the MIM model).Five QTL were found for 2-methoxyphenol that accounted for 18.1% of the phenotypic variation (18.7% under the MIM model): chromosome 2 at 46.3IcM and 300.4IcM, chromosome 4 at 560.9IcM, and chromosome 6 at 413.3IcM and 477.9IcM. Three of those QTL were also found in similar regions in the other guaiacyl derived compound, vanillin: chromosome 2 at 297.3IcM, chromosome 4 at 557.1IcM, and chromosome 6 at 414.3IcM. One other QTL was found for vanillin on chromosome 3 at 155.9IcM and the four QTL accounted for 17.2% of the phenotypic variation (14.9% under the MIM model).The most QTL we found for a compound (7) were for acetosyringone and accounted for 29.8% of the phenotypic variance (28.2% under the MIM model). These occurred on chromosome 2 at position 47.3IcM, chromosome 4 at 558.1IcM, chromosome 5 at 267.3IcM, chromosome 6 at 118.4IcM and 411.3IcM and 477.9IcM, and chromosome 8 at 502.9IcM. Two of these QTL were found in similar regions for the other syringyl derived compound, 2,6-dimethoxyphenol: chromosome 2 at 44.8IcM and chromosome 4 at 560.9IcM. Two other QTL were found for 2,6-dimethoxyphenol: chromosome 2 at 297.3IcM and chromosome 5 at 192.3IcM. These four QTL accounted for 19.7% of the phenotypic variation (17.4% under the MIM model).et al. [et al. [2 by heritability for each trait, R2/h2. Explained heritability ranged from 13.5% to 95.5% under the CIM model with an average of 39.8%, and from 13.9% to 69.3% with an average of 33.6% under the MIM model. On average, 60.2% of heritability went unexplained.Due to exclusively finding QTL with minor genetic effects, we conclude that compounds resulting from pyrolysis can be considered as quantitative traits in the IBM Syn4 population. Jansen et al.  found an [et al.  found anHeritability is measured as a proportion of variance explained by genetic effects in relation to overall phenotypic variance. Sources of genetic variance include additive, dominance, and epistatic effects. Phenotypic variance includes all genetic variance plus environmental variance, genotype x environment interaction variance, and twice the covariance between genotype and environment. Most analyses, including ours, ignore epistasis because it is too complex to model and estimate for a large number of genes or QTL. However, this can lead to an overestimation of heritability, and therefore, an underestimation of explained heritability . In addiWe found the closest correlations among compounds derived from the same cell wall polymers. Consistent with these findings, QTL co-located for these compounds. Different compounds with close correlations, such as the guaiacyl and syringyl derived compounds, also shared several QTL. Of the six distinct chromosomal regions that contained QTL for guaiacyl derived compounds, five also had QTL for a syringyl derived compound. While this could be due to either pleiotropy or close linkage of QTL, it is notable that we found 50 QTL in only 15 different regions. In consequence, we identified a limited number of regions with co-locating QTL.For lignin related compounds, there are six regions with co-locating QTL. On chromosome 2, five QTL for lignin derived compounds were located between 44.8 and 47.3IcM. All five of these QTL had higher area % values for the Mo17 allele. Five additional QTL for lignin derived compounds were found on chromosome 2 between 297.3 and 300.4IcM. The favorable allele for maximizing lignin derived compounds was contributed by Mo17 for all five. QTL for hydroxyacetaldehyde, hydroxyacetone, and acetic acid were found in this region as well, with the trait increasing allele also coming from Mo17. Increased acetic acid content is likely to be undesirable as the low pH of bio-oil can cause usage and storage issues . If the While no QTL mapping studies have been previously performed for pyrolysis related traits, it is reasonable to compare our results to QTL studies for cell wall traits, ethanol conversion, or forage and digestibility traits, since all of these traits relate to cell wall composition.et al. [et al. [Barrière et al.  found 80et al. , 38. In et al. , which g [et al. , there aet al. [p-coumaric acid (PCA), syringaldehyde, and diferulic acids. The closest marker to these QTL was umc1425, located at 165IcM in our map, which is close to four QTL identified in our study , as compared to Py/GC-MS, are available. Because all of these studies found areas where QTL for multiple cell wall related traits co-located, it is highly likely that separate breeding programs will be required to produce maize for cellulosic ethanol conversion and maize for bio-oil conversion. Common QTL between our study and Lorenzana et al. , would iWe found genetic variation for 10 bio-oil compounds in the IBM Syn4 population, and exclusively minor QTL, each of which explained a small amount of phenotypic variance. This information, taken together with relatively high (> 0.62) heritabilities for seven compounds, suggests that favorable maize varieties for improved bio-oil composition can be developed. Due to finding exclusively minor QTL, we expect that genomic selection, rather than marker assisted selection, would be the best strategy for a breeding program to improve maize for bio-oil conversion. By using all available markers, genome wide selection can capture more genetic variation compared to marker-assisted selection , and, th


Premonitory symptoms are symptoms which precede the migraine attack by 2–48h. Little attention has been paid to the variety of PS and how severely PS are affecting a patient's life.Determine frequency of PS and investigation of a connection between PS and MIDAS scores.Unsponsored, prospective study lasted for 8 months. In order to be included in the study, subjects had to be above 18 years old, and meet IHS criteria for migraine (without or with aura). Patients have filled in a questionnaire, containing a total of 11 symptoms that have been described as possible migraine PS. Also they have done MIDAS test as a general measure of migraine severity.A total of 200 migraine patients (F:M=169:31), mean age 40.6 years (range19-71) participated in this survey. 145 were diagnosed as having migraine without aura and 55 migraine with aura. 69 had a MIDAS score ≤10, 54 between 11 and 20 and 77 scored ≥21. 73 patients took different antimigraine preventive treatments. Most frequent individual premonitory symptoms were fatigue(62.5%) and unhappiness(62.0%) followed by stiff neck(58.5%), photophobia(57.0%), phonophobia(54.5%), concentracion difficulties(50.5%), nausea(48.5%) and osmophobia(45.0%), food craving(35.5%), yawning(34.5%) and water craving(30.0%). Premonitory symptoms fatigue and stiff neck were related with higher MIDAS scores,p<.001).Gender and age did not influence the frequency or the profile of PS. Taking prophylactic therapy for migraine was protective only for nausea. Fatigue and stiff neck were related with MIDAS scores, as they were more frequent, higher were the scores on the test.No conflict of interest.