Search results for: intramuscular

Authors: Firjeeth C. Paramba, Vamanjore A. Naushad, Nishan K. Purayil, Osama H. Mohammed, Prem Chandra

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Background: Fever is a common problem in adults visiting the emergency department. Extensive studies have been done in children comparing the efficacy of various antipyretics. However, studies on the efficacy of antipyretic drugs in adults are very scarce. To the best of our knowledge, no controlled trial has been carried out comparing the antipyretic efficacy of paracetamol (oral and intravenous) and intramuscular diclofenac in adults. Methods: In this parallel-group, open-label trial, participants aged 14–75 years presenting with fever who had a temperature of more than 38.5°C were enrolled and treated. Participants were randomly allocated to receive treatment with 1,000 mg oral paracetamol (n=145), 1,000 mg intravenous paracetamol (n=139), or 75 mg intramuscular diclofenac (n=150). The primary outcome was degree of reduction in mean oral temperature at 90 minutes. The efficacy of diclofenac versus oral and intravenous paracetamol was assessed by superiority comparison. Analysis was done using intention to treat principles. Results: After 90 minutes, all three groups showed a significant reduction in mean temperature, with intramuscular diclofenac showing the greatest reduction (−1.44 ± 0.43, 95% confidence interval [CI] −1.4 to −2.5) and oral paracetamol the least (−1.08 ± 0.51, 95% CI −0.99 to −2.2). After 120 minutes, there was a significant difference observed in the mean change from baseline temperature between the three treatment groups (P, 0.0001). Significant changes in temperature were observed in favor of intramuscular diclofenac over oral and intravenous paracetamol at each time point from 60 minutes through 120 minutes inclusive. Conclusion: Both intramuscular diclofenac and intravenous paracetamol showed superior antipyretic activity than oral paracetamol. However, in view of its ease of administration, intramuscular diclofenac can be used as a first-choice antipyretic in febrile adults in the emergency department.

Keywords: antipyretic, intramuscular, intravenous, paracetamol, diclofenac, emergency department

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Authors: Seong Gu Hwang, Young Kyoon Oh, Joseph F. dela Cruz

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Marbling, or intramuscular fat, has been consistently identified as one of the top beef quality problems. Intramuscular adipocytes distribute throughout the perimysial connective tissue of skeletal muscle and are the major site for the deposition of intramuscular fat, which is essential for the eating quality of meat. The stromal vascular fraction of the skeletal muscle contains progenitor cells that can be enhanced to differentiate to adipocytes and increase intramuscular fat. Primary cultures of bovine intramuscular stromal vascular cells were used in this study to elucidate the effects of extracellular calcium and retinoic acid concentration on adipocyte differentiation. Cell viability assay revealed that even at different concentrations of calcium and retinoic acid, there was no significant difference on cell viability. Monitoring of the adipocyte differentiation showed that bovine intramuscular stromal vascular cells cultured in a low concentration of extracellular calcium and retinoic acid had a better degree of fat accumulation. The mRNA and protein expressions of PPARγ, C/EBPα, SREBP-1c and aP2 were analyzed and showed a significant upregulation upon the reduction in the level of extracellular calcium and retinoic acid. The upregulation of these adipogenic related genes means that the decreasing concentration of calcium and retinoic acid is able to stimulate the adipogenic differentiation of bovine intramuscular stromal vascular cells. To further elucidate the effect of calcium, the expression level of calreticulin was measured. Calreticulin which is known to be an inhibitor of PPARγ was down regulated by the decreased level of calcium and retinoic acid in the culture media. The same tendency was observed on retinoic acid receptors RARα and CRABP-II. These receptors are recognized as adipogenic inhibitors, and the downregulation of their expression allowed a better level of differentiation in bovine intramuscular stromal vascular cells. In conclusion, data show that decreasing the level of extracellular calcium and retinoic acid can significantly promote adipogenesis in intramuscular stromal vascular cells of Hanwoo beef cattle. These findings may provide new insights in enhancing intramuscular adipogenesis and marbling in beef cattle.

Keywords: calcium, calreticulin, hanwoo beef, retinoic acid

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Authors: Amer Elgerwi, Abdelrazzag El-Magdoub, Abubakr El-Mahmoudy

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There are many bacterial infections affecting sheep that necessitates antibiotic intervention. Amoxicillin is among commonly used antibiotics in such case for its broad spectrum of activity. However, the side alterations in blood and organ function that may be associated during or after treatment are questionable. Therefore, the aim of the present study was to assess the possible alterations in blood parameters and organ function bio markers of sheep that may occur following intramuscular injection of amoxicillin. Amoxicillin has been administered intramuscularly to 10 sheep at a dosage regimen of 7 mg/kg of body weight for 5 successive days. Two types of blood samples (with and without anticoagulant) were collected from the jugular vein pre- and post-administration of the drug. Amoxicillin significantly (P < 0.001) increased total leukocyte count and (P < 0.05) absolute eosinophilic count when compared with those of the control samples. Aspartate aminotransferase, alkaline phosphatase and cholesterol were significantly (P < 0.05) higher than the corresponding control values. In addition, amoxicillin significantly (P < 0.05) increased blood urea nitrogen and creatinine but decreased phosphorus level when compared with those of prior-administration samples. These data may indicate that although the side changes caused by amoxicillin are minor in sheep, yet the liver and kidney functions should be monitored during its usage in therapy and it should be used with care for treatment of sheep with renal and/or hepatic impairments.

Keywords: amoxicillin, biogram, haemogram, sheep

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Authors: Zahid Iqbal, Ijaz Javed, Riaz Hussain, Ibadullah Jan, Amir Ali Khan

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This study was conducted to investigate the disposition kinetics of ciprofloxacin and calculate its optimal dosage in Pakistani sheep of Lohi breed. Injectable preparation of ciprofloxacin was given intramuscularly to eight sheep at a dose of 5 mg/Kg. Before administration of drug blood sample was drawn from each animal. Post drug administration, blood samples were also drawn at various predetermined time periods. Drug concentration in the blood samples was assessed through high performance liquid chromatograph (HPLC). Data were best described by two compartment open model and different pharmacokinetic (PK) parameters were calculated. Cmax of 1.97 ± 0.15 µg/ml was reached at Tmax of 0.88 ± 0.09 hours. Half life of absorption (t1/2 abs) was observed to be 0.63 ± 0.16 hours while t1/2 α (distribution half life) and t1/2 ß (elimination half life) were found to be 0.46 ± 0.05 and 2.93 ± 0.45 hours, respectively. Vd (apparent volume of distribution) was calculated as 2.89 ± 0.30 L/kg while AUC (area under the curve) was 7.19 ± 0.38 µg.hr/mL and CL (total body clearance) was 0.75 ± 0.04 L/hr/kg. Using these parameters, an optimal intramuscular dosage of ciprofloxacin in adult Lohi sheep was calculated as 21.43 mg/kg, advised to be repeated after 24 hours. From this, we came to the conclusion that calculated dose was much higher than the dose advised by the foreign manufacturer and to avoid antimicrobial resistance, it is advised that this locally investigated dosage regimen should be strictly followed in local sheep.

Keywords: pharmacokinetics, dosage regimen, ciprofloxacin, HPLC, sheep

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Authors: Silvia Grassi, Stefano Schiavon, Ernestina Casiraghi, Cristina Alamprese

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Dry-cured ham is an uncooked meat product particularly appreciated for its peculiar sensory traits among which lipid component plays a key role in defining quality and, consequently, consumers’ acceptability. Usually, fat content and distribution are chemically determined by expensive, time-consuming, and destructive analyses. Moreover, different sensory techniques are applied to assess product conformity to desired standards. In this context, visual systems are getting a foothold in the meat market envisioning more reliable and time-saving assessment of food quality traits. The present work aims at developing a simple but systematic and objective visual methodology to assess the fat amount of dry-cured ham slices, in terms of total, intermuscular and intramuscular fractions. To the aim, 160 slices from 80 PDO dry-cured hams were evaluated by digital image analysis and Soxhlet extraction. RGB images were captured by a flatbed scanner, converted in grey-scale images, and segmented based on intensity histograms as well as on a multi-stage algorithm aimed at edge enhancement. The latter was performed applying the Canny algorithm, which consists of image noise reduction, calculation of the intensity gradient for each image, spurious response removal, actual thresholding on corrected images, and confirmation of strong edge boundaries. The approach allowed for the automatic calculation of total, intermuscular and intramuscular fat fractions as percentages of the total slice area. Linear regression models were run to estimate the relationships between the image analysis results and the chemical data, thus allowing for the prediction of the total, intermuscular and intramuscular fat content by the dry-cured ham images. The goodness of fit of the obtained models was confirmed in terms of coefficient of determination (R²), hypothesis testing and pattern of residuals. Good regression models have been found being 0.73, 0.82, and 0.73 the R2 values for the total fat, the sum of intermuscular and intramuscular fat and the intermuscular fraction, respectively. In conclusion, the edge enhancement visual procedure brought to a good fat segmentation making the simple visual approach for the quantification of the different fat fractions in dry-cured ham slices sufficiently simple, accurate and precise. The presented image analysis approach steers towards the development of instruments that can overcome destructive, tedious and time-consuming chemical determinations. As future perspectives, the results of the proposed image analysis methodology will be compared with those of sensory tests in order to develop a fast grading method of dry-cured hams based on fat distribution. Therefore, the system will be able not only to predict the actual fat content but it will also reflect the visual appearance of samples as perceived by consumers.

Keywords: dry-cured ham, edge detection algorithm, fat content, image analysis

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Authors: Ghibeche Abderrahmane

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To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n=7) and age-matched (n=5) and young (n=9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of β-hyroxyacyl-CoA dehydrogenase (β-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50,P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both β-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) ∼70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.

Keywords: euglycemic-hyperinsulinemic, HSP72, mRNA, diabete

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Authors: Samad Lotfollahzadeh

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To evaluate the effect of intramuscular administration of vitamin A during dry period in pregnant dairy cows, which already received it in their daily ration, on vitamin A status of neonatal calves, a total of 30 cows were randomly selected and divided to two main groups; treatment and control group. Animals in the treatment group were subdivided into two groups. Single intramuscular injection of 2000000 IU vitamin A; was carried in 10 dairy cows at 7 months of pregnancy (group 1). In the second group of treated animals (10 cows) the injection was performed in 8 months of pregnancy (group 2). Ten pregnant dairy cows were received saline injection as placebo and selected as the control group. Blood samples were collected from experimental dairy cows at 7 and 8 months of pregnancy as well as their newborn calves’ pre and after colostrum intake. There was no significant difference between vitamin A and β- carotene concentration of dairy cows of three groups in two last months of pregnancy (P > 0.05). Vitamin A concentration of calves of two treatment groups before and after receiving of colostrum were significantly higher than that in the control group (P < 0.05). There was no significant difference between serum concentrations of vitamin A in calves of two treated groups (P > 0.05). β- Carotene concentration of serum samples of dairy cows and neonatal calves of three groups were not significantly different as compared with together. From results of the present study it can be concluded that daily supplementation of vitamin A in late pregnancy in dairy cows may not compensate the calves need for vitamin A and single injection of this vitamin A during dry either in 7 or 8 months of pregnancy can significantly increase level of vitamin A in their colostrum and neonatal calves.

Keywords: dry cow, beta carotene, newborn calves, vitamin A, dry cows

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Authors: Balakrishna Shetty

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Introduction: Stem Cell Imaging is a challenging field since the advent of Stem Cell treatment in humans. Series of research on tagging and tracking the stem cells has not been very effective. The present study is an effort by the authors to track the stem cells injected into calf muscles by Magnetic Resonance Diffusion Weighted Imaging. Materials and methods: Stem Cell injection deep into the calf muscles of patients with peripheral vascular disease is one of the recent treatment modalities followed in our institution. 5 patients who underwent deep intramuscular injection of stem cells as treatment were included for this study. Pre and two hours Post injection MRI of bilateral calf regions was done using 1.5 T Philips Achieva, 16 channel system using 16 channel torso coils. Axial STIR, Axial Diffusion weighted images with b=0 and b=1000 values with back ground suppression (DWIBS sequence of Philips MR Imaging Systems) were obtained at 5 mm interval covering the entire calf. The invert images were obtained for better visualization. 120ml of autologous bone marrow derived stem cells were processed and enriched under c-GMP conditions and reduced to 40ml solution containing mixture of above stem cells. Approximately 40 to 50 injections, each containing 0.75ml of processed stem cells, was injected with marked grids over the calf region. Around 40 injections, each of 1ml normal saline, is injected into contralateral leg as control. Results: Significant Diffusion hyper intensity is noted at the site of injected stem cells. No hyper intensity noted before the injection and also in the control side where saline was injected conclusion: This is one of the earliest studies in literature showing diffusion hyper intensity in intramuscularly injected stem cells. The advantages and deficiencies in this study will be discussed during the presentation.

Keywords: stem cells, imaging, DWI, peripheral vascular disease

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Authors: Brid Dinnee, Jessica Taylor, Joseph Hartland, Michael Natarajan

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Background:The third stage of labour (TSL) can be complicated by Post-Partum Haemorrhage (PPH), which can have a significant impact on maternal mortality and morbidity. In Africa, 33.9% of maternal deaths are attributable to PPH1. In order to minimise this figure, current recommendations for the developing world are that all women have active management of the third stage of labour (AMTSL). The aim of this project was to examine TSL practice in a rural Ugandan Hospital, highlight any deviation from best practice and identify barriers to change in resource limited settings as part of a 4th year medical student External Student Selected Component field trip. Method: Five key elements from the current World Health Organisation (WHO) guidelines on AMTSL were used to develop an audit tool. All daytime vaginal deliveries over a two week period in July 2016 were audited. In addition to this, a retrospective comparison of PPH rates, between 2006 (when ubiquitous use of intramuscular oxytocin for management of TSL was introduced) and 2015 was performed. Results: Eight vaginal deliveries were observed; at all of which intramuscular oxytocin was administered and controlled cord traction used. Against WHO recommendation, all umbilical cords were clamped within one minute, and no infants received early skin-to-skin contact. In only one case was uterine massage performed after placental delivery. A retrospective comparison of data rates identified a 40% reduction in total number of PPHs from November 2006 to November 2015. Maternal deaths per delivery reduced from 2% to 0.5%. Discussion: Maternal mortality and PPH are still major issues in developing countries. Maternal mortality due to PPH can be reduced by good practices regarding TSL, but not all of these are used in low-resource settings. There is a notable difference in outcomes between the developed and developing world. At Kitovu Hospital, there has been a reduction in maternal mortality and number of PPHs following introduction of IM Oxytocin administration. In order to further improve these rates, staff education and further government funding is key.

Keywords: post-partum haemorrhage, PPH, third stage labour, Uganda

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Authors: M. A. Akulov, V. O. Zaharov, A. A. Tomskij

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Introduction: Scapulohumeral periarthrosis, resulting as a reaction to mechanical injury of shoulder tendons and muscles, is associated with high incidence of temporal and permanent disability. There is a strong need for investigation of treatment of that patient group. Severe pain leads to limitation of movements range, which result in secondary alterations of joint capsule and ligamentous apparatus. Muscle tension and edema, swelling of fascial and fibrous structures result in nerve and vascular compression in intramuscular and osseo-muscular-fibrous spaces. Botulinum toxin injection leads to decrease of muscle tone, increase of movements range and associated pain alleviation. Study aim: Optimization of rehabilitation process in scapolohumeral periarthrosis using Xeomin. Patients and methods: 40 patients aged 37-56 years with scapulohumeral periarthrosis were evaluated. Patients were divided into two groups according to treatment regimen. The first (main) group included 21 patients, receiving intramuscular Xeomin 150-200 U in the area of brachio-scapular joint and trigger points (inducing motion range limitation and pain). Treatment procedures were combined with physical therapy and osteopathic procedures. The second (control) group included 19 patients, receiving conventional physical therapy and osteopathic procedures. The evaluation and efficacy comparison was carried out using McGill pain questionnaire, Clinical Global Impression scale (CGI), and patient-reported increase of brachio-scapular joint movement range and pain decrease at 1, 3 and 6 months of treatment. Results. The study demonstrated a significant improvement in the main group after one month of treatment, which persisted during months of treatment. At baseline, rank pain index on McGill pain questionnaire was 18,4±4,9 and 17,8±5,1 in the main and control group, respectively (p > 0,05). At 1 month of treatment we observed a significant decrease of pain syndrome (no pain or modest pain) and increase of movement range in angular degrees in the main group (р < 0,05). In the control group significant improvements were observed only on the 3 month of treatment (р < 0,05), but at 6 months of treatment the improvement in pain syndrome and motion range in brachio-scapular joint was significantly smaller, than in the main group. Rank pain index on McGill pain scale was 5,2±1,8 in the main group compared to 12,0±2,6 in the control group (р < 0,05). At 6 months of treatment patients in the first group reported a significant/highly significant improvement of general health on CGI, whereas in the second group most patients reported a minimal improvement. We observed a sustained and persistent improvement of motion range in brachio-scapular joint in the main group. Conclusion: Xeomin injections as a part of rehabilitation process in scapulohumeral periarthrosis lead to reduced time and increased quality of rehabilitation.

Keywords: botulinum toxin, rehabilitation, scapulohumeral periarthrosis

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Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

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Authors: Hamid Ghasemzadeh- Nava, Maziar Kaveh Baghbadorani, Amin Tamadon

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Considering response of uterus to ecbolic effect of oxytocin near the time of parturition, this study was done for investigating the effect of prophylactic administration of this hormone on duration of fetal membrane retention, time interval to first detectable estrus, time interval to first service, and conception rate at first service in cases of both normal parturition and dystocia. For this reason cows with (n=18) and without (n=18) dystocia assigned randomly to treatment (n=12) or control (n=6) groups and received intramuscular injection of 100 IU of oxytocin or 10 mL of normal saline respectively. Further observations and investigations indicate that duration of fetal retention is significantly shorter in treatment group cows compared to control groups, regardless of having dystocia (P=0.002) or normal spontaneous calving (P=0.001). The same trend exists for conception rate at first service in which cows in treatment groups had significantly higher conception rate (CR) in comparison to cows in control groups with (P=0.0003) or without dystocia (P=0.017). The time interval to first detected heat and first service didn’t show any difference between groups.

Keywords: conception rate, oxytocin, RFM, time to first service

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Authors: Jelena Maletkovic

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Background: Endogenous Cushing’s syndrome is a rare disease, but iatrogenic or drug related Cushing syndrome from glucocorticoid products is commonly seen in clinical practice. With high dose and long term use of glucocorticoids, patients can develop isolated hypothalamic-pituitary-adrenal (HPA) suppression, or HPA axis suppression can be accompanied by overt iatrogenic Cushing’s syndrome. This is a rare case where severe Cushing’s syndrome developed from an unknown medication and was followed by severe and prolonged adrenal insufficiency and multiple potentially fatal complications. Case: This is a 37-year-old woman who is presented to Emergency Room (ER) with shortness of breath and chest pain. Four months prior to this presentation the patient was a generally healthy woman who was looking for improvement in her appearance and visited local Rejuvenation Clinic. After initial consultation with a nurse, she was contacted by a physician over the phone and was advised to start taking multiple injectable medications that will arrive by mail. Medications without any labels on bottles were delivered and the patient started daily intramuscular injections. Over the next two months, she noticed rounding of her face and swelling around her eyes. She gained 20 pounds, mostly abdominal fat and became extremely fatigued. Her muscles on legs were visibly decreasing in size and she felt significant muscle weakness. Unexplained bruising occurred. She started growing hair on face and developed secondary amenorrhea. New severe back pain started. She developed depression and headaches. Finally, over a few days, a number of red-purple stretch marks that were sensitive and painful appeared over her abdomen, upper part of arms and legs. She then became suspicious that these dramatic symptoms are caused by injectable medication and she discontinued injections. Over the next few days she presented to ER with low blood pressure and oxygen saturation of 75%. Studies revealed extensive pneumonia as well as multiple pulmonary emboli. Her white blood count was elevated with 32 000 and she also had acute kidney failure on admission. She was treated for sepsis and was also given stress dose steroids. Steroids were tapered over 48 hours and discontinued. After being discharged to home, on her first visit to endocrinology clinic she had undetectable ACTH of < 2pg/mL and undetectable 8am cortisol of < 0.2mcg/dL. She did not respond to an intramuscular injection of cosyntropin 250mcg and her repeated cortisol after 60 minutes was only 1mcg/dL. The patient was diagnosed with adrenal insufficiency and was started on hydrocortisone 20mg+10mg. It took close to 2 years of slow tapering for recovery of this patient’s HPA axis and resolve all the sequelae from Cushing’s syndrome. Conclusion: Misuse and abuse of glucocorticoids have been present almost since these medications were discovered. This is a rare case where not only severe Cushing’s syndrome in full clinical picture developed but also the patient suffered multiple potentially fatal complications and prolonged adrenal insufficiency. Visits to herbal, rejuvenation, esthetic, and similar clinics are becoming more and more popular and physicians need to be aware of possible non-benign nature of medications that their patients may be using.

Keywords: iatrogenic, Cushing's syndrome, adrenal crisis, steroid abuse

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Authors: Ranjeeta Kumari, Makesh M, Gayatri Tripathi, K V Rajendran, Megha Bedekar

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A comparative study on DNA immunization with recombinant glyceraldehyde-3-phosphate dehydrogenase (GAPDH) construct of Edwardsiella tarda (pGPD group) and a bicistronic construct expressing GAPDH plus IFN-γ of Labeo rohita as adjuvant (pGPD+IFN group) was undertaken in Labeo rohita along with the control animals. Successful co-expression of two genes that is GAPDH and IFN-γ was confirmed in SSN-1 cells line by RT-qPCR and western blot. The protective immune response of host to DNA vaccine construct was determined by RPS and specific antibody production. Fishes immunized with plasmids via intramuscular injection (I/M) exhibited a considerable relative percentage survivability of 66.66% in pGPD+IFN immunized group and 53.34% in pGPD immunized group after challenge with E. tarda. Antibody response was also significantly high in pGPD+IFN group at all time points under study. This was analysed by competitive ELISA, using anti GAPDH monoclonal antibodies. The experiment revealed that the GAPDH gene of E. tarda is one of the ideal candidates for generating protective immune response in L. rohita. Further addition of Interferon gamma to DNA vaccine construct can enhance the immune response in host.

Keywords: DNA vaccine, Edwardsiella tarda, Labeo rohita, zoonosis, immune response

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Authors: P. O. Ughachukwu, P. O. Okonkwo, P. C. Unekwe, J. O. Ogamba

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Background: The prevalence of hepatitis B viral infection is high worldwide with liver cirrhosis and hepatocellular carcinoma as important complications. Cases of poor antibody response to hepatitis B vaccination abound. Immunosuppression, especially from glucocorticoids, is often cited as a cause of poor antibody response and there are documented evidences of irrational administration of glucocorticoids to children and adults. The study was, therefore, designed to find out if administration of glucocorticoids affects immune response to vaccination against hepatitis B in mice. Methods: Mice of both sexes were randomly divided into 2 groups. Daily intramuscular methylprednisolone injections, (15 mg kg-1), were given to the test group while sterile deionized water (0.1ml) was given to control mice for 30 days. On day 6 all mice were given 2 μg (0.1ml) hepatitis B vaccine and a booster dose on day 27. On day 34, blood samples were collected and analyzed for anti-HBs titres using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was done using Graph Pad Prism 5.0 and the results taken as statistically significant at p value < 0.05. Results: There were positive serum anti-HBs responses in all mice groups but the differences in titres were not statistically significant. Conclusions: At the dosages and length of exposure used in this study, methylprednisolone injection did not significantly inhibit anti-HBs response in mice following immunization against hepatitis B virus. By extrapolation, methylprednisolone, when used in the usual clinical doses and duration of therapy, is not likely to inhibit immune response to hepatitis B vaccinations in man.

Keywords: anti-HBs, hepatitis B vaccine, immune response, methylprednisolone, mice

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Authors: Jeong Woo Kang, MiYoung Baek, Ki-Suk Kim, Kwang-Jick Lee, ByungJae So

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Introduction: A fast, easy and sensitive detection method was developed and validated by liquid chromatography tandem mass spectrometry for the determination of marbofloxacin in pig plasma which was further applied to study the pharmacokinetics of marbofloxacin. Materials and Methods: The plasma sample (500 μL) was mixed with 1.5 ml of 0.1% formic acid in MeCN to precipitate plasma proteins. After shaking for 20 min, The mixture was centrifuged at 5,000 × g for 30 min. It was dried under a nitrogen flow at 50℃. 500 μL aliquot of the sample was injected into the LC-MS/MS system. Chromatographic analysis was carried out mobile phase gradient consisting 0.1% formic acid in D.W. (A) and 0.1% formic acid in MeCN (B) with C18 reverse phase column. Mass spectrometry was performed using the positive ion mode and the selected ion monitoring (MRM). Results and Conclusions: The method validation was performed in the sample matrix. Good linearities (R2>0.999) were observed and the quantified average recoveries of marbofloxacin were 87 - 92% at level of 10 ng g-1 -100 ng g-1. The percent of coefficient of variation (CV) for the described method was less than 10 % over the range of concentrations studied. The limits of detection (LOD) and quantification (LOQ) were 2 and 5 ng g-1, respectively. This method has also been applied successfully to pharmacokinetic analysis of marbofloxacin after intravenous (IV), intramuscular (IM) and oral administration (PO). The mean peak plasma concentration (Cmax) was 2,597 ng g-1at 0.25 h, 2,587 ng g-1at 0.44 h and 2,355 ng g-1at 1.58 h for IV, IM and PO, respectively. The area under the plasma concentration-time curve (AUC0–t) was 24.8, 29.0 and 25.2 h μg/mL for IV, IM and PO, respectively. The elimination half-life (T1/2) was 8.6, 13.1 and 9.5 for IV, IM and PO, respectively. Bioavailability (F) of the marbofloxacin in pig was 117 and 101 % for IM and PO, respectively. Based on these result, marbofloxacin does not have any obstacles as therapeutics to develop the oral formulations such as tablets and capsules.

Keywords: marbofloxacin, LC-MS/MS, pharmacokinetics, chromatographic

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Authors: Seyyed Javad Ziaolhagh

Abstract:

Background: The purpose of present study was to investigate, the effects of anabolic steroid Boldenone (BOL) with eight weeks of resistance training on structural changes in rat liver. Method: 21 Male adult Wistar rats, 12 weeks old and 228/53±7/94 g initial body weight were randomly assigned to three groups: group1: Control+ Placebo (C), group2: training+ Placebo (T), group3: Boldenone intramuscular injections 5mg/kg (B). The endurance training protocol consisted three exercise sessions weekly started by a 30-minute run with the speed of 12 m/min and lasted by 60min run with the speed of 30 m/min in 8 weeks. At the end of the experiment, for light microscopic study Slides were prepared. Results: Sections stained of rat's livers showed no any cell degeneration and cytoplasmic lipid vacuoles in all groups, but few samples were seen. Indeed, congested blood sinusoids, cell infiltration and degeneration were seen in the Boldenone-treated group. Hepatotoxic effects were severe in group treatment received 5 mg/kg and directly depended on the doses. Indeed, training group was no any hepatocyte degeneration, inflammation and congestion. Conclusion: The present results showed that BOL has a marked adverse effect on the liver tissue, even with low– dose and endurance training. As a result, athletes should aware of Boldenone dosage consumption.

Keywords: anabolic androgenic steroids, Boldenone, blood congestion, cellular inflammation, cellular degeneration, lipid vocuolations, endurance training

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Authors: Karissa Graham, Andrew Campbell-Lloyd

Abstract:

The deep inferior epigastric artery perforator flap (DIEP) is used to reconstruct large volumes of tissue. The DIEP flap is based on the deep inferior epigastric artery (DIEA) and vein. Accurate knowledge of the anatomy of these vessels allows for efficient dissection of the flap, minimal damage to surrounding tissue, and a well vascularized flap. A 54 year old lady was assessed for bilateral delayed autologous reconstruction with DIEP free flaps. The right DIEA was consistent with the described anatomy. The left DIEA had a vessel branching shortly after leaving the external iliac artery and before entering the muscle. This independent branch entered the muscle and had a long intramuscular course to the largest perforator. The main DIEA vessel demonstrated a type II branching pattern but had perforators that were too small to have a viable DIEP flap. There were no communicating arterial branches between the independent vessel and DIEA, however, there was one venous communication between them. A muscle sparing transverse rectus abdominis muscle flap was raised using the main periumbilical perforator from the independent vessel. Our case report demonstrated an unreported anatomical variant of the DIEA. A few anatomical variants have been described in the literature, including a unilateral absent DIEA and peritoneal-cutaneous perforators that had no connection to the DIEA. Doing a pre-operative CTA helps to identify these rare anatomical variations, which leads to safer, more efficient, and effective operating.

Keywords: aberrant anatomy, CT angiography, DIEP anatomy, free flap

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Authors: A. M. Prasad

Abstract:

Knowledge of variations of nerves of gluteal region is important for clinicians administering intramuscular injections, for orthopedic surgeons dealing with the hip surgeries, possibly for physiotherapists managing the painful conditions and paralysis of this region. Herein, we report multiple variations of the nerves of gluteal region. In the current case, the sciatic nerve was absent. The common peroneal and tibial nerves arose from sacral plexus and reached the gluteal region through greater sciatic foramen above and below piriformis respectively. The common peroneal nerve gave a muscular branch to the gluteus maximus. The inferior gluteal nerve and posterior cutaneous nerve of the thigh arose from a common trunk. The common trunk was formed by three roots. Upper and middle roots arose from sacral plexus and entered gluteal region through greater sciatic foramen respectively above and below piriformis. The lower root arose from the pudendal nerve and joined the common trunk. These variations were seen in the right gluteal region of an adult male cadaver aged approximately 70 years. Innervation of gluteus maximus by common peroneal nerve and presence of a common trunk of inferior gluteal nerve and posterior cutaneous nerve of the thigh make this case unique. The variant nerves may be subjected to iatrogenic injuries during surgical approach to the hip. They may also get compressed if there is a hypertrophy of the piriformis syndrome. Hence, the knowledge of these variations is of importance to clinicians, orthopedic surgeons and possibly for physiotherapists.

Keywords: gluteal region, multiple variations, nerve injury, sciatic nerve

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Authors: Changde Zhang, Qiang Zhang, Shilong Zheng, Jiawang Liu, Shanchun Guo, Qiu Zhong, Guangdi Wang

Abstract:

Fulvestrant was approved by FDA to treat breast cancer as a selective estrogen receptor downregulator (SERD) with intramuscular injection administration. ZB716, a fulvestarnt-3 boronic acid, is an SERD with comparable anticancer effect to fulvestrant, but could produce good pharmacokinetic properties under oral administration with mice or rat models. To understand why ZB716 produced much better oral bioavailability, it was proposed that the boronic acid blocked the phase II direct biotransformation with the hydroxyl group on the 3 position of the aromatic ring on fulvestrant. In this study, ZB716 or fulvestrant was incubated with human liver microsome and oxidation cofactor NADPH in vitro. Their metabolites after oxidation were profiled with the Q-Exactive, a high-resolution mass spectrometer. The result showed that ZB716 blocked the forming of hydroxyl groups on its benzene ring except for the oxidation of C-B bond forming fulvestrant in its metabolites, and the concentration of fulvestrant with one more hydroxyl group found in the metabolites from incubation with fulvestrant was about 34 fold high as that formed from incubation with ZB716. Compared to fulvestrant, ZB716 is expected to be much difficult to be further bio-transformed into more hydrophilic compounds, to be difficult excreted out of blood system, and to have longer residence time in blood, which can lead to higher oral bioavailability. This study provided evidence to explain the high bioavailability of ZB716 after oral administration from the perspective of its difficulty of oxidation, a phase I biotransformation, on positions on its aromatic ring.

Keywords: biotransformation, fulvestrant, metabolite profiling, ZB716

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Authors: Ntalamagka N., Sanchis-Benlloch P. J., Mayoral-Serrano R., Tena-Medialdea J., García-March J. R.

Abstract:

The pen shell Pinna nobilis population has declined dramatically since 2016 due to die-off events observed in the whole extent of the Mediterranean Sea associated with the protozoan Haplosporidium pinnae. As of 2019, it is considered a critically endangered species. Due to its ecological importance and its endangered status, several initiatives have been developed for its salvation and recovery. This research is an effort to understand and control its reproduction under captivity. As a limited number of Pinna nobilis individuals could be used for experimentation, the possibility of using the Pinna rudis as a model animal was explored. The molecular mechanism that regulates the reproduction of both species is unknown; consequently, transcriptomic analysis was performed to identify neuropeptides that are expressed in the key regulatory tissues of the visceral ganglia and gonads of both species. Neuropeptides form an important group of signaling peptides that regulate reproductive, behavioral and physiological functions in molluscs. In total, 17 neuropeptide precursors were identified in P. nobilis and 14 in P. rudis transcriptomes; 14 of them were identical in both species. This affinity verified the genetic similarity of these species at the reproduction level. APGWamide, buccalin, ELH and GnRH were tested in P. rudis and demonstrated their capacity to advance gonadal maturation and trigger spawning while spawning was recorded in P. nobilis after the usage of APGWamide and buccalin. The neuropeptides were administered using intramuscular injection and cholesterol implants following relative literature as well as a new method was developed for external administration without the use of anesthesia using a mathematical model. The know-how of this research will not only lead to the survival of the species but also will narrow the horizons of broodstock conditioning of other similar species.

Keywords: neuropeptides, Pinna nobilis, reproduction, transcriptomics

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Authors: Catrin Morgan, Atholl Johnston

Abstract:

Creatine supplementation should theoretically increase total muscle creatine and so enhance the generation of intramuscular phosphocreatine and subsequent ATP formation. The use of creatine as a potential ergogenic aid in sport has been an area of significant scientific research for a number of years. However the effect of creatine supplementation and swimming performance is a relatively new area of research and is the subject of this review. In swimming creatine supplementation could help maintain maximal power output, aid recovery and increase lean body mass. After investigating the underlying theory and science behind creatine supplementation, a literature review was conducted to identify the best evidence looking at the effect of creatine supplementation on swimming performance. The search identified 27 potential studies, and of these 17 were selected for review. The studies were then categorised into single sprint performance, which involves swimming a short distance race, or repeated interval performance, which involves swimming a series of sprints with intervals of rest between them. None of the studies on the effect of creatine controlled for the multiple confounding factors associated with measurement of swimming performance. The sample size in the studies was limited and this reduced the reliability of the studies and introduced the possibility of bias. The studies reviewed provided insufficient evidence to determine if creatine supplementation is beneficial to swimming performance. However, what data there was supported the use of creatine supplementation in repeated interval swimming rather than in single sprint swimming. From a review of the studies, it was calculated on average, there was a 1.37% increase in swimming performance with the use of creatine for repeated intervals and a 0.86% increase in performance for single sprint. While this may seem minor, it should be remembered that swimming races are often won by much smaller margins. In the 2012 London Olympics the Men’s 100 metres freestyle race was won by a margin of only 0.01 of a second. Therefore any potential benefit could make a dramatic difference to the final outcome of the race. Overall more research is warranted before the benefits of creatine supplementation in swimming performance can be further clarified.

Keywords: creatine supplementation, repeated interval, single sprint, swimming performance

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Authors: Olga Shapovalova, Vladimir Vinogradov

Abstract:

Among many different sol – gel matrices only alumina can be successfully parenteral injected in the human body. And this is not surprising, because boehmite (aluminium oxyhydroxide) is the metal oxide approved by FDA and EMA for intravenous and intramuscular administrations, and also has been using for a longtime as adjuvant for producing of many modern vaccines. In our earlier study, it has been shown, that denaturation temperature of enzymes entrapped in sol-gel boehmite matrix increases for 30 – 60 °С with preserving of initial activity. It makes such matrices more attractive for long-term storage of non-stable drugs. In current work we present ultrasound-assisted sol-gel synthesis of nano-boehmite. This method provides bio-friendly, very stable, highly homogeneous alumina sol with using only water and aluminium isopropoxide as a precursor. Many parameters of the synthesis were studied in details: time of ultrasound treatment, US frequency, surface area, pore and nanoparticle size, zeta potential and others. Here we investigated the dependence of stability of colloidal sols and textural properties of the final composites as a function of the time of ultrasonic treatment. Chosen ultrasonic treatment time was between 30 and 180 minutes. Surface area, average pore diameter and total pore volume of the final composites were measured by surface and pore size analyzer Nova 1200 Quntachrome. It was shown that the matrices with ultrasonic treatment time equal to 90 minutes have the biggest surface area 431 ± 24 m2/g. On the other had such matrices have a smaller stability in comparison with the samples with ultrasonic treatment time equal to 120 minutes that have the surface area 390 ± 21 m2/g. It was shown that the stable sols could be formed only after 120 minutes of ultrasonic treatment, otherwise the white precipitate of boehmite is formed. We conclude that the optimal ultrasonic treatment time is 120 minutes.

Keywords: boehmite matrix, stabilisation, ultrasound-assisted sol-gel synthesis

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Authors: M. Soltani, K. Pakzad, A. Haghigh-Khiyabani, M. Alavi, R. Naderi, M. Castex

Abstract:

Effect of probiotic Bactocell (Pediococcus acidilactici) as a supplementary diet was studied on post-larvae 12-15 of white leg shrimp (Litopenaeus vannamei) (150000 PL/0.5 h pond, average body weight=0.02 g) growth performance under farm condition for 102 days at water quality parameters consisting of temperature at 30.5-36οC, dissolved oxygen 4.1-6.6 mg/l, salinity 40-54 g/l, turbidity 35-110 cm, ammonia 0.1-0.8 mg/l and nitrite 0.1-0.9 mg/l. Also, the resistance level of the treated shrimps was assessed against a virulent strain of Vibrio paraheamolyticus as intramuscular injection route at 1.4 x 106 cells/shrimp. Significantly higher growth rate (11.3±1.54 g) and lower feed conversion ratio (1.1) were obtained in shrimps fed diets supplemented with Bactocell at 350 g/ tone feed compared to other treatments of 250 g Bactocell/ton feed (10.8±2 g, 1.3), 500 g Bactocell/ton feed (10.3±1.7 g, 1.3) and untreated control (10.1±2 g, 1.4). Also, thermal growth coefficient (0.057%) and protein efficiency ratio (2.13) were significantly improved in shrimps fed diets supplemented with Bactocell at 350 g/ton feed compare to other groups. Shrimps fed diet supplemented with Bactocell at 350 g/tone feed showed significantly higher protein content (72.56%) in their carcass composition than treatments of 250 g/ton feed (65.9%), 500 g/ton feed (67.5%) and control group (65.9%), while the carcass contents of moisture, lipid and ash in all shrimp groups were not significantly affected by different concentrations of Bactocell. No mortality occurred in the experimentally infected shrimps fed with Bactocell at 500 g/tone feed after 7 hours post-challenge with V. parahemolyticus. The mortality levels of 100%, 40%, 50% and 70% were obtained in shrimps fed with 0.0, 500 g/tone feed, 350 g/ton feed and 250 g/ton feed, respectively 14 hours post-infection. Also, the cumulative mortalities were achieved in 100%, 92% and 81% in shrimps few with Bactocell at 500 g/ton feed, 250 g/ton feed and 350 g/ton feed, respectively.

Keywords: litopenaeus vannamei, vibrio paraheamolyticus, pediococcus acidilactici, growth performance, bactocell

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Authors: Prataksitorn Chonlakan, Tiparat Wongsilarat

Abstract:

The traditional clinical practice guideline used at the emergency room at Songkhla Hospital in caring for patients who come in contact with jellyfish venom took a long time for the pain to reduce to the level that patients can cope with. To investigate the effectiveness of clinical practice guidelines by comparing the effectiveness of a newly developed clinical practice guideline with the traditional clinical practice guideline in the following aspects: 1) pain reduction, 2) length of pain, 3) the rate of patient’s re-visit, 4) the rate of severe complications such as anaphylactic shock, and cardiac arrest, and death, and 5) patient satisfaction. This study employed a quasi-experimental research design. Thirty subjects were selected with purposive sampling from jellyfish-sting patients who came for treatment at the Emergency Room of Songkhla Hospital. The subjects were divided using random assignment into two groups of 15 each: an experimental group, and the control group. The control group was treated using the traditional clinical practice guideline consisting of rinsing the affected area with 0.9% normal saline, using a cloth soaked with vinegar to press against the affected area, and controlling pain using tramadol or diclofenac intramuscular injection. The data were analyzed using descriptive statistics and paired t-test at the significance level p < 0.05. The results of the study revealed the following. The pain level in the experimental group was significantly lower than that of the control group (the average pain score of the experimental group was 3.46 while that of the control group was 6.33) (p < 0.05).The length of pain in the experimental group was significantly lower than that of the control group (the average length of pain in the experimental group was 48.67 minutes while that of the control group was 105.35 minutes) (p < 0.05). The rate of re-visit within 12 hours in the experimental group was significantly lower than that of the control group (the rate of re-visit within 12 hours of the experimental group was 0.07 while that of the control group was 0.00) (p < 0.05).No severe complications such as anaphylactic shock, and cardiac arrest were found in the two groups of subjects.The rate of satisfaction among the subjects in the experimental group was significantly higher than that of the control group (the rate of satisfaction among the subjects of the experimental group was 90.00 percent while that among the control group was 66.33 percent) (p < 0.05). The newly develop clinical practice guideline could reduce pain and increase satisfaction among jellyfish-sting patients better than the traditional clinical practice guideline.

Keywords: effectiveness, clinical practice guideline, jellyfish-sting patients, cardiac arrest

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Authors: Angela Cristina Da F. De Oliveira, Salma E. Asmar, Norbert P. Battlori, Yaz Vera, Uriel R. Valencia, Tâmara D. Borges, Antoni D. Bueno, Leandro B. Costa

Abstract:

The present study aimed to evaluate the replacement of 5% of starch per 5% of sunflower oil (SO) on meat quality and animal welfare of growing and finishing pigs (Iberic x Duroc), exposed to a heat stress environment. The experiment lasted 90 days, and it was carried out in a randomized block design, in a 2 x 2 factorial, composed of two diets (starch or sunflower oil (with or without) and two feed intake management (ad libitum and restriction). Seventy-two crossbred males (51± 6,29 kg body weight - BW) were housed in climate-controlled rooms, in collective pens and exposed to heat stress environment (32°C; 35% to 50% humidity). The treatments studies were: 1) control diet (5% starch x 0% SO) with ad libitum intake (n = 18); 2) SO diet (replacement of 5% of starch per 5% of SO) with ad libitum intake (n = 18); 3) control diet with restriction feed intake (n = 18); or 4) SO diet with restriction feed intake (n = 18). Feed were provided in two phases, 50-100 Kg BW for growing and 100-140 Kg BW for finishing, respectively. Within welfare evaluations, dirtiness score was evaluated all morning during ninety days of the experiment. The presence of manure was individually measured based on one side of the pig´s body and scored according to: 0 (less than 20% of the body surface); 1 (more than 20% but less than 50% of the body surface); 2 (over 50% of the body surface). After the experimental period, when animals reach 130-140 kg BW, they were slaughtered using carbon dioxide (CO2) stunning. Carcass weight, leanness and fat content, measured at the last rib, were recorded within 20 min post-mortem (PM). At 24h PM, pH, electrical conductivity and color measures (L, a*, b*) were recorded in the Longissimus thoracis and Semimembranosus muscles. Data shown no interaction between diet (control x SO) and management feed intake (ad libitum x restriction) on the meat quality parameters. Animals in ad libitum management presented an increase (p < 0.05) on BW, carcass weight (CW), back fat thickness (BT), and intramuscular fat content (IM) when compared with animals in restriction management. In contrast, animals in restriction management showing a higher (p < 0.05) carcass yield, percentage of lean and loin thickness. To welfare evaluations, the interaction between diet and management feed intake did not influence the degree of dirtiness. Although, the animals that received SO diet, independently of the management, were cleaner than animals in control group (p < 0,05), which, for pigs, demonstrate an important strategy to reduce body temperature. Based in our results, the diet and management feed intake had a significant influence on meat quality and animal welfare being considered efficient nutritional strategies to reduce heat stress and improved meat quality.

Keywords: dirtiness, environment, meat, pig

Procedia PDF Downloads 244

Authors: Mohamad F. Jamiluddin, Emeline Sarry, Ana Bejanariu, Cécile Bauche

Abstract:

Introduction: Over 360 million people are chronically infected with hepatitis B virus (HBV), of whom 1 million die each year from HBV-associated liver cirrhosis or hepatocellular carcinoma. Current treatment options for chronic hepatitis B depend on interferon-α (IFNα) or nucleos(t)ide analogs, which control virus replication but rarely eliminate the virus. Treatment with PEG-IFNα leads to a sustained antiviral response in only one third of patients. After withdrawal of the drugs, the rebound of viremia is observed in the majority of patients. Furthermore, the long-term treatment is subsequently associated with the appearance of drug resistant HBV strains that is often the cause of the therapy failure. Among the new therapeutic avenues being developed, therapeutic vaccine aimed at inducing immune responses similar to those found in resolvers is of growing interest. The high prevalence of chronic hepatitis B necessitates the design of better vaccination strategies capable of eliciting broad-spectrum of cell-mediated immunity(CMI) and humoral immune response that can control chronic hepatitis B. Induction of HBV-specific T cells and B cells by therapeutic vaccination may be an innovative strategy to overcome virus persistence. Lentiviral vectors developed and optimized by THERAVECTYS, due to their ability to transduce non-dividing cells, including dendritic cells, and induce CMI response, have demonstrated their effectiveness as vaccination tools. Method: To develop a HBV therapeutic vaccine that can induce a broad but specific immune response, we generated recombinant lentiviral vector carrying IRES(Internal Ribosome Entry Site)-containing bicistronic constructs which allow the coexpression of two vaccine products, namely HBV T- cell epitope vaccine and HBV virus like particle (VLP) vaccine. HBV T-cell epitope vaccine consists of immunodominant cluster of CD4 and CD8 epitopes with spacer in between them and epitopes are derived from HBV surface protein, HBV core, HBV X and polymerase. While HBV VLP vaccine is a HBV core protein based chimeric VLP with surface protein B-cell epitopes displayed. In order to evaluate the immunogenicity, mice were immunized with lentiviral constructs by intramuscular injection. The T cell and antibody immune responses of the two vaccine products were analyzed using IFN-γ ELISpot assay and ELISA respectively to quantify the adaptive response to HBV antigens. Results: Following a single administration in mice, lentiviral construct elicited robust antigen-specific IFN-γ responses to the encoded antigens. The HBV T- cell epitope vaccine demonstrated significantly higher T cell immunogenicity than HBV VLP vaccine. Importantly, we demonstrated by ELISA that antibodies are induced against both HBV surface protein and HBV core protein when mice injected with vaccine construct (p < 0.05). Conclusion: Our results highlight that THERAVECTYS lentiviral vectors may represent a powerful platform for immunization strategy against chronic hepatitis B. Our data suggests the likely importance of Lentiviral vector based novel bicistronic construct for further study, in combination with drugs or as standalone antigens, as a therapeutic lentiviral based HBV vaccines. THERAVECTYS bicistronic HBV vaccine will be further evaluated in animal efficacy studies.

Keywords: chronic hepatitis B, lentiviral vectors, therapeutic vaccine, virus-like particle

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