Search results for: protein metabolism
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2727

Search results for: protein metabolism

747 Vitamin D Levels of Patients with Rheumatoid Arthritis in Kosova

Authors: Mjellma Rexhepi, Blerta Rexhepi Kelmendi, Blana Krasniqi, Shaip Krasniqi

Abstract:

Rheumatoid arthritis is a chronic disease that causes inflammation of the joints which can be so severe that can cause not only deformities but also impairment of function that limits movement. This also contributes to the pain that accompanies this disease. This remains a problematic and challenging disease of modern medicine because treatment is still symptomatic. The main purpose of drug treatment is to reduce the activity of the disease, achieve remission, avoid disability and death. The etiology of the disease is idiopathic, but can also be linked to genetic, nongenetic factors such as hormonal, environmental or infectious. Current scientific evidence shows that vitamin D plays an important role in immune regulation mechanisms. Lack of this vitamin has been linked to loss of immune tolerance and the appearance of autoimmune processes, including rheumatoid arthritis. The purpose of the work was to define Vitamin D in patients hospitalized with rheumatoid arthritis in University Clinical Center of Kosova, as a basis of their connection with lifestyle and physical inactivity. The sample for the work was selected from patients with criteria met for rheumatoid arthritis who were hospitalized at the tertiary level of health care in Kosova. During the work have been investigated 100 consecutive patients fulfilling diagnostic criteria for rheumatoid arthritis, whereas in addition to the general characteristics are also determined the values of vitamin D at the beginning of hospitalization. The average age of the sample analyzed was 50.9±5.7 years old, with an average duration of rheumatoid arthritis disease 7.8±3.4 years. At the beginning of hospitalization, before treatment was initiated, the average value of vitamin D was 15.86±3.43, which according to current reference values is classified into the category of insufficient values. Correlating the duration of the disease, from the time of diagnosis to the day of hospitalization, on one side and the level of vitamin D on the other side, the negative correlation of a lower degree derived (r =-0.1). Physical activity affects the concentration of vitamin D in the blood through increased metabolism of fat and the release of vitamin D and its metabolites from adipose tissue. To now it is evident that physical activity is also accompanied by higher levels of vitamin D. In patients with rheumatoid arthritis, vitamin D levels were low compared to normal. Future works should be oriented toward investigating in detail the bone structure, quality of life and pain in patients with rheumatoid arthritis. More detailed scientific projects, with larger numbers of participants, should be designed for the future to clarify more possible mechanisms as factors related to this phenomenon such as inactivity, lifestyle and the duration of the disease, as well as the importance of keeping vitamin D values at normal limits.

Keywords: hospitalization, lifestyle, rheumatoid arthritis, vitamin D

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746 Designing of Food Products Enriched With Phytonutrients Assigned for Hypertension Suffering Consumers

Authors: Anna Gramza-Michałowska, Dominik Kmiecik, Justyna Bilon, Joanna Skręty, Joanna Kobus-Cisowska, Józef Korczak, Andrzej Sidor

Abstract:

Background: Hypertension is one of the civilization diseases with a global scope. Many research showed that every day diet influences significantly our health, helping with the prophylaxis and diseases treatment. The key factor here is the presence of plant origin natural bio active components. Aim: The following research describes snack health-oriented products for hypertension sufferers enriched with selected plant ingredients. Various analytical methods have been applied to determine product’s basic composition and their antioxidant activity. Methods: Snack products was formulated from a composition of different flours, oil, yeast, plant particles and extracts. Basic composition of a product was evaluated as content of protein, lipids, fiber, ash and caloricity. Antioxidant capacity of snacks was evaluated with use radical scavenging methods (DPPH, ABTS) and ORAC value. Proposed snacks as new product was also characterized with sensory analysis. Results and discussion: Results showed that addition of phyto nutrients allowed to improve nutritional and antioxidative value of examined products. Also the anti radical potential was significantly increased, with no loss of sensory value of a snacks. Conclusions: Designed snack is rich in polyphenolics, that express high antioxidant activity, helpful in hypertension and as low calories product obesity prophylaxis.

Keywords: antioxidant, well-being, hypertension, bioactive compounds

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745 Factors Associated with Weight Loss Maintenance after an Intervention Program

Authors: Filipa Cortez, Vanessa Pereira

Abstract:

Introduction: The main challenge of obesity treatment is long-term weight loss maintenance. The 3 phases method is a weight loss program that combines a low carb and moderately high-protein diet, food supplements and a weekly one-to-one consultation with a certified nutritionist. Sustained weight control is the ultimate goal of phase 3. Success criterion was the minimum loss of 10% of initial weight and its maintenance after 12 months. Objective: The aim of this study was to identify factors associated with successful weight loss maintenance after 12 months at the end of 3 phases method. Methods: The study included 199 subjects that achieved their weight loss goal (phase 3). Weight and body mass index (BMI) were obtained at the baseline and every week until the end of the program. Therapeutic adherence was measured weekly on a Likert scale from 1 to 5. Subjects were considered in compliance with nutritional recommendation and supplementation when their classification was ≥ 4. After 12 months of the method, the current weight and number of previous weight-loss attempts were collected by telephone interview. The statistical significance was assumed at p-values < 0.05. Statistical analyses were performed using SPSS TM software v.21. Results: 65.3% of subjects met the success criterion. The factors which displayed a significant weight loss maintenance prediction were: greater initial percentage weight loss (OR=1.44) during the weight loss intervention and a higher number of consultations in phase 3 (OR=1.10). Conclusion: These findings suggest that the percentage weight loss during the weight loss intervention and the number of consultations in phase 3 may facilitate maintenance of weight loss after the 3 phases method.

Keywords: obesity, weight maintenance, low-carbohydrate diet, dietary supplements

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744 Monocytic Paraoxonase 2 (PON 2) Lactonase Activity Is Related to Myocardial Infarction

Authors: Mukund Ramchandra Mogarekar, Pankaj Kumar, Shraddha V. More

Abstract:

Background: Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), very low-density lipoprotein cholesterol (VLDL-C), Apo B, and lipoprotein(a) was found as atherogenic factors while high-density lipoprotein cholesterol (HDL-C) was anti-atherogenic. Methods and Results: The study group consists of 40 MI subjects as cases and 40 healthy as controls. Monocytic PON 2 Lactonase (LACT) activity was measured by using Dihydrocoumarine (DHC) as substrate. Phenotyping was done by method of Mogarekar MR et al, serum AOPP by modified method of Witko-Sarsat V et al and Apo B by Turbidimetric immunoassay. PON 2 LACT activities were significantly lower (p< 0.05) and AOPPs & Apo B were higher in MI subjects (p> 0.05). Trimodal distribution of QQ, QR & RR phenotypes of study population showed no significant difference among cases and controls (p> 0.05). Univariate binary logistic regression analysis showed independent association of TC, HDL, LDL, AOPP, Apo B, and PON 2 LACT activity with MI and multiple forward binary logistic regression showed PON 2 LACT activity and serum Apo B as an independent predictor of MI. Conclusions- Decrease in PON 2 LACT activity in MI subjects than in controls suggests increased oxidative stress in MI which is reflected by significantly increased AOPP and Apo B. PON 1 polymorphism of QQ, QR and RR showed no significant difference in protection against MI. Univariate and multiple forward binary logistic regression showed PON 2 LACT activity and serum Apo B as an independent predictor of MI.

Keywords: advanced oxidation protein products, apolipoprotein-B, myocardial infarction, paraoxonase 2 lactonase

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743 Chemopreventive Efficacy Of Cdcl2(C14H21N3O2) in Rat Colon Carcinogenesis Model Using Aberrant Crypt Foci (ACF) as Endpoint Marker

Authors: Maryam Hajrezaie, Mahmood Ameen Abdulla, Nazia AbdulMajid, Maryam Zahedifard

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Colon cancer is one of the most prevalent cancers in the world. Cancer chemoprevention is defined as the use of natural or synthetic compounds capable of inducing biological mechanisms necessary to preserve genomic fidelity. New schiff based compounds are reported to exhibit a wide spectrum of biological activities of therapeutic importance. To evaluate inhibitory properties of CdCl2(C14H21N3O2) complex on colonic aberrant crypt foci, five groups of 7-week-old male rats were used. Control group was fed with 10% Tween 20 once a day, cancer control group was intra-peritoneally injected with 15 mg/kg Azoxymethan, drug control group was injected with 15 mg/kg azoxymethan and 5-Flourouracil, experimental groups were fed with 2.5 and 5 mg/kg CdCl2(C14H21N3O2) compound each once a day. Administration of compound were found to be effectively chemoprotective. Andrographolide suppressed total colonic ACF formation up to 72% to 74%, respectively, when compared with control group. The results also showed a significant increase in glutathione peroxidase, superoxide dismutase, catalase activities and a decrease in malondialdehyde level. Immunohistochemical staining demonstrated down-regulation of PCNA protein. According to the Western blot comparison analysis, COX-2 and Bcl2 is up-regulated whilst the Bax is down-regulated. according to these data, this compound plays promising chemoprotective activity, in a model of AOM-induced in ACF.

Keywords: chemopreventive, Schiff based compound, aberrant crypt foci (ACF), immunohistochemical staining

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742 Evaluation of Immune Checkpoint Inhibitors in Cancer Therapy

Authors: Mir Mohammad Reza Hosseini

Abstract:

In new years immune checkpoint inhibitors have gathered care as being one of the greatest talented kinds of immunotherapy on the prospect. There has been a specific emphasis on the immune checkpoint molecules, cytotoxic T-lymphocyte antigen-4 (CTLA-4) and programmed cell death protein 1 (PD-1). In 2011, ipilimumab, the primary antibody obstructive an immune checkpoint (CTLA4) was authorized. It is now documented that recognized tumors have many devices of overpowering the antitumor immune response, counting manufacture of repressive cytokines, staffing of immunosuppressive immune cells, and upregulation of coinhibitory receptors recognized as immune checkpoints. This was fast followed by the growth of monoclonal antibodies directing PD1 (pembrolizumab and nivolumab) and PDL1 (atezolizumab and durvalumab). Anti-PD1/PDL1 antibodies have developed some of the greatest extensively set anticancer therapies. We also compare and difference their present place in cancer therapy and designs of immune-related toxicities and deliberate the role of dual immune checkpoint inhibition and plans for the organization of immune-related opposing proceedings. In this review, the employed code and present growth of numerous immune checkpoint inhibitors are abridged, while the communicating device and new development of Immune checkpoint inhibitors in cancer therapy-based synergistic therapies with additional immunotherapy, chemotherapy, phototherapy, and radiotherapy in important and clinical educations in the historical 5 years are portrayed and tinted. Lastly, we disapprovingly measure these methods and effort to find their fortes and faintness based on pre-clinical and clinical information.

Keywords: checkpoint, cancer therapy, PD-1, PDL-1, CTLA4, immunosuppressive

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741 Serum Neurotrophins in Different Metabolic Types of Obesity

Authors: Irina M. Kolesnikova, Andrey M. Gaponov, Sergey A. Roumiantsev, Tatiana V. Grigoryeva, Alexander V. Laikov, Alexander V. Shestopalov

Abstract:

Background. Neuropathy is a common complication of obesity. In this regard, the content of neurotrophins in such patients is of particular interest. Neurotrophins are the proteins that regulate neuron survival and neuroplasticity and include brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). However, the risk of complications depends on the metabolic type of obesity. Metabolically unhealthy obesity (MUHO) is associated with a high risk of complications, while this is not the case with metabolically healthy obesity (MHO). Therefore, the aim of our work was to study the effect of the obesity metabolic type on serum neurotrophins levels. Patients, materials, methods. The study included 134 healthy donors and 104 obese patients. Depending on the metabolic type of obesity, the obese patients were divided into subgroups with MHO (n=40) and MUHO (n=55). In the blood serum, the concentration of BDNF and NGF was determined. In addition, the content of adipokines (leptin, asprosin, resistin, adiponectin), myokines (irisin, myostatin, osteocrin), indicators of carbohydrate, and lipid metabolism were measured. Correlation analysis revealed the relationship between the studied parameters. Results. We found that serum BDNF concentration was not different between obese patients and healthy donors, regardless of obesity metabolic type. At the same time, in obese patients, there was a decrease in serum NGF level versus control. A similar trend was characteristic of both MHO and MUHO. However, MUHO patients had a higher NGF level than MHO patients. The literature indicates that obesity is associated with an increase in the plasma concentration of NGF. It can be assumed that in obesity, there is a violation of NGF storage in platelets, which accelerates neurotrophin degradation. We found that BDNF concentration correlated with irisin levels in MUHO patients. Healthy donors had a weak association between NGF and VEGF levels. No such association was found in obese patients, but there was an association between NGF and leptin concentrations. In MHO, the concentration of NHF correlated with the content of leptin, irisin, osteocrin, insulin, and the HOMA-IR index. But in MUHO patients, we found only the relationship between NGF and adipokines (leptin, asprosin). It can be assumed that in patients with MHO, the replenishment of serum NGF occurs under the influence of muscle and adipose tissue. In the MUHO patients only the effect of adipose tissue on NGF was observed. Conclusion. Obesity, regardless of metabolic type, is associated with a decrease in serum NGF concentration. We showed that muscle and adipose tissues make a significant contribution to the serum NGF pool in the MHO patients. In MUHO there is no effect of muscle on the NGF level, but the effect of adipose tissue remains.

Keywords: neurotrophins, nerve growth factor, NGF, brain-derived neurotrophic factor, BDNF, obesity, metabolically healthy obesity, metabolically unhealthy obesity

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740 5-HT2CR Deficiency Causes Affective Disorders by Impairing E/I Balance through Augmenting Hippocampal nNOS-CAPON Coupling

Authors: Hu-Jiang Shi, Li-Juan Zhu

Abstract:

The implication of 5-hydroxytryptamine 2C receptor (5-HT2CR) in affective behaviors is a topic of debate, and the underlying mechanisms remain largely unclear. Here, we elucidate that the interaction between hippocampal neuronal nitric oxide synthase (nNOS) and carboxy-terminal PDZ ligand of nNOS (CAPON) contributes to the disruption of hippocampal excitation-inhibition (E/I) balance, which is responsible for the anxiety- and depressive-like behaviors caused by chronic stress-related 5-HT2CR signaling deficiency. In detail, activation or inhibition of 5-HT2CR by CP809101 or SB242084 modulates nNOS-CAPON interaction by influencing intracellular Ca²⁺ release. Notably, the dissociation of nNOS-CAPON abolishes SB242084-induced anxiety- and depressive-like behaviors, as well as the reduction in extracellular signal-regulated kinase (ERK)/cAMP-response element binding protein (CREB)/synapsin signaling and SNARE complex assembly. Furthermore, nNOS-CAPON blockers restore the impairments caused by SB242084, including the reduction in SNARE assembly-mediated γ-aminobutyric acid (GABA) vesicle release and a consequent shift of the E/I balance toward excitation in CA3 pyramidal neurons. Conclusively, our findings disclose the regulatory role of 5-HT2CR in anxiety- and depressive-like behaviors and highlight the hippocampal nNOS-CAPON coupling-triggered E/I imbalance as a pivotal cellular event underpinning the behavioral consequences of 5-HT2CR inhibition.

Keywords: 5-HT2CR, anxiety, depression, nNOS-CAPON coupling, excitation-inhibition balance, neurotransmitter release

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739 Contamination with Heavy Metals of Frozen Fish Sold in Open Markets in Ondo City, Southwest Nigeria

Authors: Adebisi M. Tiamiyu, Adewale F. Adeyemi, Olu-Ayobamikale V. Irewunmi

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Fish consumption has increased in recent years in both developing and advanced countries, owing to increased awareness of its nutritional and therapeutic benefits and its availability and affordability relative to other animal protein sources. Fish and fish products, however, are extremely prone to contamination by a wide range of hazardous organic and inorganic substances. This study assessed the levels of three heavy metals, copper (Cu), iron (Fe), and zinc (Zn), in frozen fish imported into Nigeria and sold in Ondo City for their safety for human consumption as recommended by WHO and FEPA. Three species of frozen fish (Scombrus scombrus, Merluccius merluccius, and Clupea harengus) were purchased, and the wet tissues (gills, muscles, and liver) were digested using a 3:1 mixture of nitric acid (HNO3) and hydrochloric acid (HCL). An atomic absorption spectrophotometer (AAS) was used to detect the amount of metal in the tissues. The levels of heavy metals in different fish species' organs varied. The fish had Zn > Fe > Cu heavy metal concentrations in that order. While the concentration of Cu and Fe in the tissues of all three fish species studied were within the WHO and FEPA prescribed limits for food fish, the concentration of Zn in the muscles of M. merluccius (0.262±0.052), C. harengus harengus (0.327±0.099), and S. scombrus (0.362±0.119) was above the prescribed limit (0.075 ppm) set by FEPA. An excessive amount of zinc in the body can cause nausea, headaches, decreased immunity, and appetite loss.

Keywords: heavy metal, atomic absorption spectrophotometer, fish, agencies

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738 New Targets Promoting Oncolytic Virotherapy

Authors: Felicia Segeth, Florian G. Klein, Lea Berger, Andreas Kolk, Per S. Holm

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The entry of oncolytic viruses (OVs) into clinical application opens groundbreaking changes in current and future treatment regimens. However, despite their potent anti-cancer activity in vitro, clinical studies revealed limitations of OVs as monotherapy. The same applies to CDK 4/6 inhibitors (CDK4/6i) targeting cell cycle as well as bromodomain and extra-terminal domain inhibitors (BETi) targeting gene expression. In this study, the anti-tumoral effect of XVir-N-31, an YB-1 dependent oncolytic adenovirus, was evaluated in combination with Ribociclib, a CDK4/6i, and JQ1, a BETi. The head and neck squamous cell carcinoma (HNSCC) cell lines Fadu, SAS, and Cal-33 were used. DNA replication and gene expression of XVir-N-31 was measured by RT-qPCR, protein expression by western blotting, and cell lysis by SRB assays. Treatment with CDK4/6i and BETi increased viral gene expression, viral DNA replication, and viral particle formation. The data show that the combination of oncolytic adenovirus XVir-N-31 with CDK4/6i & BETi acts highly synergistic in cancer cell lysis. Furthermore, additional molecular analyses on this subject demonstrate that the positive transcription elongation factor P-TEFb plays a decisive role in this regard, indicating an influence of the combinational therapy on gene transcription control. The combination of CDK4/6i & BETi and XVir-N-31 is an attractive strategy to achieve substantial cancer cell killing and is highly suitable for clinical testing.

Keywords: adenovirus, BET, CDK4/6, HNSCC, P-TEFb, YB-1

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737 Functionalized Titanium Dioxide Nanoparticles for Targeting and Disrupting Amyloid Fibrils

Authors: Elad Arad, Raz Jelinek, Hanna Rapaport

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Amyloidoses are a family of diseases characterized by abnormal protein folding that leads to aggregation. They accumulate to form fibrillar plaques which are implicated in the pathogenesis of Alzheimer, prion, diabetes type II and other diseases. To the best of our knowledge, despite extensive research efforts devoted to plaque aggregates inhibition, there is yet no cure for this phenomenon. Titanium and its alloys are found in growing interest for biomedical applications. Variety of surface modifications enable porous, adhesive, bioactive coatings for its surface. Titanium oxides (titania) are also being developed for photothermal and photodynamic treatments. Inspired by this, we set to explore the effect of functionalized titania nanoparticles in combination with external stimuli, as potential photothermal ablating agents against amyloids. Titania nanoparticles were coated with bi-functional catechol derivatives (dihydroxy-phenylalanine propanoic acid, noted DPA) to gain targeting properties. In conjunction with UV-radiation, these nanoparticles may selectively destroy the vicinity of their target. Titania modified 5 nm nanoparticles coated with DPA were further conjugated to the amyloid-targeting Congo Red (CR). These Titania-DPA-CR nanoparticles were found to target mature amyloid fibril of both amyloid-β (Aβ 1-42 a.a). Moreover, irradiation of the peptides in presence of the modified nanoparticles decreased the aggregate content and oligomer fraction. This work provides insights into the use of modified titania nanoparticles for amyloid plaque targeting and photothermal destruction. It may shed light on future modifications and functionalization of titania nanoparticles for different applications.

Keywords: titanium dioxide, amyloids, photothermal treatment, catechol, Congo-red

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736 Calcium Biochemical Indicators in a Group of Schoolchildren with Low Socioeconomic Status from Barranquilla, Colombia

Authors: Carmiña L. Vargas-Zapata, María A. Conde-Sarmiento, Maria Consuelo Maestre-Vargas

Abstract:

Calcium is an essential element for good growth and development of the organism, and its requirement is increased at school age. Low socio-economic populations of developing countries such as Colombia may have food deficiency of this mineral in schoolchildren that could be reflected in calcium biochemical indicators, bone alterations and anthropometric indicators. The objective of this investigation was to evaluate some calcium biochemical indicators in a group of schoolchildren of low socioeconomic level from Barranquilla city and to correlate with body mass index. 60 schoolchildren aged 7 to 15 years were selected from Jesus’s Heart Educational Institution in Barranquilla-Atlántico, apparently healthy, without suffering from infectious or gastrointestinal diseases, without habits of drinking alcohol or smoking another hallucinogenic substance and without taking supplementation with calcium in the last six months or another substance that compromises bone metabolism. The research was approved by the ethics committee at Universidad del Atlántico. The selected children were invited to donate a blood and urine sample in a fasting time of 12 hours, the serum was separated by centrifugation and frozen at ˗20 ℃ until analyzed and the same was done with the urine sample. On the day of the biological collections, the weight and height of the students were measured to determine the nutritional status by BMI using the WHO tables. Calcium concentrations in serum and urine (SCa, UCa), alkaline phosphatase activity total and of bone origin (SAPT, SBAP) and urinary creatinine (UCr) were determined by spectrophotometric methods using commercial kits. Osteocalcin and Cross-linked N-telopeptides of type I collagen (NTx-1) in serum were measured with an enzyme-linked inmunosorbent assay. For statistical analysis the Statgraphics software Centurium XVII was used. 63% (n = 38) and 37% (n = 22) of the participants were male and female, respectively. 78% (n = 47), 5% (n = 3) and 17% (n = 10) had a normal, malnutrition and high nutritional status, respectively. The averages of evaluated indicators levels were (mean ± SD): 9.50 ± 1.06 mg/dL for SCa; 181.3 ± 64.3 U/L for SAPT, 143.8 ± 73.9 U/L for SBAP; 9.0 ± 3.48 ng/mL for osteocalcin and 101.3 ± 12.8 ng/mL for NTx-1. UCa level was 12.8 ± 7.7 mg/dL that adjusted with creatinine ranged from 0.005 to 0.395 mg/mg. Considering serum calcium values, approximately 7% of school children were hypocalcemic, 16% hypercalcemic and 77% normocalcemic. The indicators evaluated did not correlate with the BMI. Low values ​​were observed in calcium urinary excretion and high in NTx-1, suggesting that mechanisms such as increase in renal retention of calcium and in bone remodeling may be contributing to calcium homeostasis.

Keywords: calcium, calcium biochemical, indicators, school children, low socioeconomic status

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735 Antioxidant Defense Mechanisms in Murine Epidermis and Dermis and Their Responses to Ultraviolet Light

Authors: Ben Abderrahmane Ayoub El Fateh, Bnina Rachid

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A comprehensive comparison of antioxidant defenses in the dermis and epidermis and their response to exposure to ultraviolet (UV) irradiation has not previously been attempted. In this study, enzymic and non-enzymic antioxidants in epidermis and dermis of hairless mice were compared. Enzyme activities are presented both as units/gram of skin and units/milligram of protein; arguments are presented for the superiority of skin wet weight as a reference base. Catalase, glutathione peroxidase, and glutathione reductase (units/gram of skin) were higher in the epidermis than dermis by 49%, 86%, and 74%, respectively. Superoxide dismutase did not follow this pattern. Lipophilic antioxidants ( -tocopherol, ubiquinol 9, and ubiquinone 9) and hydrophilic antioxidants (ascorbic acid, dehydroascorbic acid, and glutathione) were 24–95% higher in the epidermis than in dermis. In contrast, oxidized glutathione was 60% lower in the epidermis than in dermis. Mice were irradiated with solar light to examine the response of these cutaneous layers to UV irradiation. After irradiation with 25 J/cm2 (UVA + UVB, from a solar simulator), 10 times the minimum erythemal dose, epidermal and dermal catalase and superoxide dismutase activities were greatly decreased. Tocopherol, ubiquinol 9, ubiquinone 9, ascorbic acid, dehydroascorbic acid, and reduced glutathione decreased in both epidermis and dermis by 26-93%. Oxidizedgiutathione showed a slight, non-significant increase. Because the reduction in total ascorbate and catalase was much more severe in the epidermis than dermis, it can be concluded that UV light is more damaging to the antioxidant defenses in the epidermis than in the dermis.

Keywords: antioxidant defenses, enzymic, epidermis, oxidizedgiutathione

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734 Production of Soy Yoghurt Using Soymilk-Based Lactic Acid Bacteria as Starter Culture

Authors: Ayobami Solomon Popoola, Victor N. Enujiugha

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Production of soy-yogurt by fermentation of soymilk with lactic acid bacteria isolated from soymilk was studied. Soymilk was extracted from dehulled soybean seeds and pasteurized at 95 °C for 15 min. The soymilk was left to naturally ferment (temperature 40 °C; time 8 h) and lactic acid bacteria were isolated, screened and selected for yogurt production. Freshly prepared soymilk was pasteurized (95 °C, 15 min), inoculated with the lactic acid bacteria isolated (3% w/v starter culture) and incubated at 40 °C for 8 h. The yogurt produced was stored at 4 °C. Investigations were carried out with the aim of improving the sensory qualities and acceptability of soy yogurt. Commercial yogurt was used as a control. The percentage of soymilk inoculated was 70% of the broth. Soy-yoghurt samples produced were subsequently subjected to biochemical and microbiological assays which included total viable counts of fresh milk and soy-based yoghurt; proximate composition of functional soy-based yoghurt fermented with Lactobacillus plantarum; changes in pH, Titratable acidity, and lactic acid bacteria during a 14 day period of storage; as well as morphological and biochemical characteristics of lactic acid bacteria isolated. The results demonstrated that using Lactobacillus plantarum to inoculate soy milk for yogurt production takes about 8 h. The overall acceptability of the soy-based yogurt produced was not significantly different from that of the control sample. The use of isolate from soymilk had the added advantage of reducing the cost of yogurt starter culture, thereby making soy-yogurt, a good source of much desired good quality protein. However, more experiments are needed to improve the sensory qualities such as beany or astringent flavor and color.

Keywords: soy, soymilk, yoghurt, starter culture

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733 Effect of Seasonal Variation on Two Introduced Columbiformes in Awba Dam Tourism Centre, University of Ibadan, Ibadan

Authors: Kolawole F. Farinloye, Samson O. Ojo

Abstract:

Two Columbiformes species were recently introduced to the newly established Awba Dam Tourism Centre [ADTC], hence there is need to investigate the effect of seasonal variation on these species with respect to hematological composition. Blood samples were obtained from superficial ulna vein of the 128 apparently healthy C. livia and C. guinea into tubes containing EDTA as anticoagulant. Thin blood smears (TBS) were prepared, stained and viewed under microscope. Values of Red Blood Cell (RBC) count, White Blood Cell (WBC) count, cholesterol (CH), Uric Acid (UA), Protein (PR), Mean Corpuscular Volume (MCV), Haemoglobin Content (HB), Blood Volume (BV), Plasma Glucose (PG) and Length/Width (L/W) ratio of red blood cells were assessed. The procedure was carried out on a seasonal basis (wet and dry seasons of 2013-2014). Data was analyzed using descriptive and inferential statistics. Lymphocyte count for C. livia was F3, 161 = 13.15, while for C. guinea was F3, 178 = 13.15. Heterophil, H/L ratio and Muscle score values for both species were (rs = -0.38, rs = -0.44), (rs = 0.51, rs = 0.31) (4, 3) respectively. Analyses also demonstrated a low WBC to RBC ratio (0.004: 25.3) in both species during the wet season compared to dry season, respectively. L/W varied significantly among sampling seasons i.e. wet (19.1% of BV, 12.6% of BV, 0.1% of BV) and dry (18.9% of BV, 12.7% of BV, 0.08% of BV). The level of HB in wet season (19.20±8.46108) is lower compared to dry season (19.70±8.48762). T-test also showed (wet=15.625, 0.111), (dry=12.125, 0.146) respectively, hence there is no association between species and haematological parameters. Species introduced were found to be haematologically stable. Although there were slight differences in seasonal composition, however this can be attributed to seasonal variation; suggesting little or no effect of seasons on their blood composition.

Keywords: seasonal variation, Columbiformes, Awba Dam tourism centre, University of Ibadan, Ibadan

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732 Use of Chlorophyll Meters to Assess In-Season Wheat Nitrogen Fertilizer Requirements in the Southern San Joaquin Valley

Authors: Brian Marsh

Abstract:

Nitrogen fertilizer is the most used and often the most mismanaged nutrient input. Nitrogen management has tremendous implications on crop productivity, quality and environmental stewardship. Sufficient nitrogen is needed to optimum yield and quality. Soil and in-season plant tissue testing for nitrogen status are a time consuming and expensive process. Real time sensing of plant nitrogen status can be a useful tool in managing nitrogen inputs. The objectives of this project were to assess the reliability of remotely sensed non-destructive plant nitrogen measurements compared to wet chemistry data from sampled plant tissue, develop in-season nitrogen recommendations based on remotely sensed data for improved nitrogen use efficiency and assess the potential for determining yield and quality from remotely sensed data. Very good correlations were observed between early-season remotely sensed crop nitrogen status and plant nitrogen concentrations and subsequent in-season fertilizer recommendations. The transmittance/absorbance type meters gave the most accurate readings. Early in-season fertilizer recommendation would be to apply 40 kg nitrogen per hectare plus 16 kg nitrogen per hectare for each unit difference measured with the SPAD meter between the crop and reference area or 25 kg plus 13 kg per hectare for each unit difference measured with the CCM 200. Once the crop was sufficiently fertilized meter readings became inconclusive and were of no benefit for determining nitrogen status, silage yield and quality and grain yield and protein.

Keywords: wheat, nitrogen fertilization, chlorophyll meter

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731 Total Longitudinal Displacement (tLoD) of the Common Carotid Artery (CCA) Does Not Differ between Patients with Moderate or High Cardiovascular Risk (CV) and Patients after Acute Myocardial Infarction (AMI)

Authors: P. Serpytis, K. Azukaitis, U. Gargalskaite, R. Navickas, J. Badariene, V. Dzenkeviciute

Abstract:

Purpose: Total longitudinal displacement (tLoD) of the common carotid artery (CCA) wall is a novel ultrasound marker of vascular function that can be evaluated using modified speckle tracking techniques. Decreased CCA tLoD has already been shown to be associated with diabetes and was shown to predict one year cardiovascular outcome in patients with suspected coronary artery disease (CAD) . The aim of our study was to evaluate if CCA tLoD differ between patients with moderate or high cardiovascular (CV) risk and patients after recent acute myocardial infarction (AMI). Methods: 49 patients (54±6 years) with moderate or high CV risk and 42 patients (58±7 years) after recent AMI were included. All patients were non-diabetic. CCA tLoD was evaluated using GE EchoPAC speckle tracking software and expressed as mean of both sides. Data on systolic blood pressure, total and high density lipoprotein (HDL) cholesterol levels, high sensitivity C-reactive protein (hsCRP) level, smoking status and family history of early CV events was evaluated and assessed for association with CCA tLoD. Results: tLoD of CCA did not differ between patients with moderate or high CV risk and patients with very high CV risk after MI (0.265±0.128 mm vs. 0.237±0.103 mm, p>0.05). Lower tLoD was associated with lower HDL cholesterol levels (r=0.211, p=0.04) and male sex (0.228±0.1 vs. 0.297±0.134, p=0.01). Conclusions: tLoD of CCA did not differ between patients with moderate or high CV risk and patients with very high CV risk after AMI. However, lower CCA tLoD was significantly associated with low HDL cholesterol levels and male sex.

Keywords: total longitudinal displacement, carotid artery, cardiovascular risk, acute myocardial infarction

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730 Characteristics of Serum Exosomes after Burn Injury and Dermal Fibroblast Regulation by Exosomes in Vitro

Authors: Jie Ding, Yingying Pan, Shammy Raj, Lindy Schaffrick, Jolene Wong, Antoinette Nguyen, Sharada Manchikanti, Larry Unsworth, Peter Kwan, Edward E. Tredget

Abstract:

Background: Exosomes (EXOs) have been considered a new target that is thought to be involved in and treat wound healing. More research is needed to fully understand the EXO characteristics and mechanisms of EXO-mediated wound healing, especially wound healing after burn injury. Methods: Total EXOs were isolated from 85 serum samples of 29 burn patients and 13 healthy individuals. We characterized the EXOs for morphology and density, serum concentration, protein level, marker expression, size distribution, and cytokine content. After confirmation of EXO uptake by dermal fibroblasts, we also explored functional regulation of primary human normal skin and hypertrophic scar fibroblast cell lines by the EXOs in vitro, including cell proliferation and apoptosis. Results: EXOs dynamically changed their morphology, density, size, and cytokine level during wound healing in burn patients, which were correlated with burn severity and the stages of wound healing. EXOs from both burn patients and healthy individuals stimulated dermal fibroblast proliferation and apoptosis. Conclusion: EXO features may be important signals that influence wound healing after burn injury; however, to understand the mechanisms by which EXOs regulated the fibroblasts in healing wounds, further studies will be required in the future.

Keywords: exosome, burn, wound healing, hypertrophic scarring, cytokines

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729 Orange Leaves and Rice Straw on Methane Emission and Milk Production in Murciano-Granadina Dairy Goat Diet

Authors: Tamara Romero, Manuel Romero-Huelva, Jose V. Segarra, Jose Castro, Carlos Fernandez

Abstract:

Many foods resulting from processing and manufacturing end up as waste, most of which is burned, dumped into landfills or used as compost, which leads to wasted resources, and environmental problems due to unsuitable disposal. Using residues of the crop and food processing industries to feed livestock has the advantage to obviating the need for costly waste management programs. The main residue generated in citrus cultivations and rice crop are pruning waste and rice straw, respectively. Within Spain, the Valencian Community is one of the world's oldest citrus and rice production areas. The objective of this experiment found out the effects of including orange leaves and rice straw as ingredients in the concentrate diets of goats, on milk production and methane (CH₄) emissions. Ten Murciano-Granadina dairy goats (45 kg of body weight, on average) in mid-lactation were selected in a crossover design experiment, where each goat received two treatments in 2 periods. Both groups were fed with 1.7 kg pelleted mixed ration; one group (n= 5) was a control (C) and the other group (n= 5) used orange leaves and rice straw (OR). The forage was alfalfa hay, and it was the same for the two groups (1 kg of alfalfa was offered by goat and day). The diets employed to achieve the requirements during lactation period for caprine livestock. The goats were allocated to individual metabolism cages. After 14 days of adaptation, feed intake and milk yield were recorded daily over a 5 days period. Physico-chemical parameters and somatic cell count in milk samples were determined. Then, gas exchange measurements were recorded individually by an open-circuit indirect calorimetry system using a head box. The data were analyzed by mixed model with diet and digestibility as fixed effect and goat as random effect. No differences were found for dry matter intake (2.23 kg/d, on average). Higher milk yield was found for C diet than OR (2.3 vs. 2.1 kg/goat and day, respectively) and, greater milk fat content was observed for OR than C (6.5 vs. 5.5%, respectively). The cheese extract was also greater in OR than C (10.7 vs. 9.6%). Goats fed OR diet produced significantly fewer CH₄ emissions than C diet (27 vs. 30 g/d, respectively). These preliminary results (LIFE Project LOWCARBON FEED LIFE/CCM/ES/000088) suggested that the use of these waste by-products was effective in reducing CH₄ emission without detrimental effect on milk yield.

Keywords: agricultural waste, goat, milk production, methane emission

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728 Expression of Gro-El under Phloem-Specific Promoter Protects Transgenic Plants against Diverse Begomovirus-Beta Satellite Complex

Authors: Muhammad Yousaf Ali, Shahid Mansoor, Javeria Qazi, Imran Amin, Musarrat Shaheen

Abstract:

Cotton leaf curl disease (CLCuD) is the major threat to the cotton crop and is transmitted by whitefly (Bemisia tabaci). Since multiple begomoviruses and associated satellites are involved in CLCuD, approaches based on the concept of broad-spectrum resistance are essential for effective disease control. Gro-El and G5 are two proteins from whitefly endosymbiont and M13 bacteriophage origin, respectively. Gro-El encapsulates the virus particle when it enters the whitefly and protects the virus from the immune system of the whitefly as well as prevents viral expression in it. This characteristic of Gro-El can be exploited to get resistance against viruses if expressed in plants. G5 is a single-stranded DNA binding protein, expression of which in transgenic plants will stop viral expression on its binding with ssDNA. The use of tissue-specific promoters is more efficient than constitutive promoters. Transgenics of Nicotiana benthamiana for Gro-El under constitutive promoter and Gro-El under phloem specific promoter were made. In comparison to non-transgenic plants, transgenic plants with Gro-El under NSP promoter showed promising results when challenged against cotton leaf curl Multan virus (CLCuMuV) along with cotton leaf curl Multan beta satellite (CLCuMB), cotton leaf curl Khokhran virus (CLCuKoV) along with cotton leaf curl Multan beta satellite (CLCuMB) and Pedilenthus leaf curl virus (PedLCV) along with Tobacco leaf curl beta satellite (TbLCB).

Keywords: cotton leaf curl disease, whitefly, endosymbionts, transgenic, resistance

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727 Protective Effect of L-Carnitine against Gentamicin-Induced Nephrotoxicity in Rats

Authors: Mohamed F. Ahmed, Mabruka S. Elashheb, Fatma M. Ben Rabha

Abstract:

This study aimed to determine the possible protective effects of L‐carnitine against gentamicin‐induced nephrotoxicity. Forty male albino rats were divided into 4 groups (10 rats each); Group 1: normal control, group 2: induced nephrotoxicity (gentamicin 50 mg/kg/day S.C; 8 days) , group 3: treated with L‐carnitine (40 mg/kg/d SC for 12 days) and group 4: treated with L‐carnitine 4 days before and for 8 days in concomitant with gentamicin. Gentamicin‐induced nephrotoxicity (group 2): caused significant increase in serum urea, creatinine, urinary N‐acetyl‐B‐D‐glucosaminidase (NAG), gamma glutamyl transpeptidase (GGT), urinary total protein and kidney tissue malondialdehyde (MDA) with significant decrease in serum superoxide dismutase (SOD), serum catalase and creatinine clearance and marked tubular necrosis in the proximal convoluted tubules with interruption in the basement membrane around the necrotic tubule compared to the normal control group. L‐carnitine 4 days before and for 8 days in concomitant with gentamicin (group 4) offered marked decrease in serum urea, serum creatinine, urinary NAG, urinary GGT, urinary proteins and kidney tissue MDA, with marked increase in serum SOD, serum catalase and creatinine clearance with marked improvement in the tubular damage compared to gentamicin‐induced nephrotoxicity group. L‐carnitine administered for 12 days produced no change in the above-mentioned parameters as compared to the normal control group. In conclusion: L‐carnitine could reduce most of the biochemical parameters and also improve the histopathological features of the kidney associated with gentamicin-induced nephrotoxicity.

Keywords: gentamicin, nephrotoxicity, L‐carnitine, kidney disease

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726 Effect of Chronic Exposure to Diazinon on Glucose Homeostasis and Oxidative Stress in Pancreas of Rats and the Potential Role of Mesna in Ameliorating This Effect

Authors: Azza El-Medany, Jamila El-Medany

Abstract:

Residential and agricultural pesticide use is widespread in the world. Their extensive and indiscriminative use, in addition with their ability to interact with biological systems other than their primary targets constitute a health hazards to both humans and animals. The toxic effects of pesticides include alterations in metabolism; there is a lack of knowledge that organophosphates can cause pancreatic toxicity. The primary goal of this work is to study the effects of chronic exposure to Diazinon an organophosphate used in agriculture on pancreatic tissues and evaluate the ameliorating effect of Mesna as antioxidant on the toxicity of Diazinon on pancreatic tissues.40 adult male rats, their weight ranged between 300-350 g. The rats were classified into three groups; control (10 rats) was received corn oil at a dose of 1 0 mg/kg/day by gavage once a day for 2 months. Diazinon (15 rats) was received Diazinon at a dose of 10 mg/kg/day dissolved in corn oil by gavage once a day for 2 months. Treated group (15 rats), were received Mesna 180mg/kg once a week by gavage 15 minutes before administration of Diazinon for 2 months. At the end of the experiment, animals were anesthetized, blood samples were taken by cardiac puncture for glucose and insulin assays and pancreas was removed and divided into 3 portions; first portion for histopathological study; second portion for ultrastructural study; third portion for biochemical study using Elisa Kits including determination of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), myeloperoxidase activity (MPO), interleukin 1β (IL-1β). A significant increase in the levels of MDA, TNF-α, MPO activity, IL-1β, serum glucose levels in the toxicated group with Diazinon were observed, while a significant reduction was noticed in GSH in serum insulin levels. After treatment with Mesna a significant reduction was observed in the previously mentioned parameters except that there was a significant rise in GSH in insulin levels. Histopathological and ultra-structural studies showed destruction in pancreatic tissues and β cells were the most affected cells among the injured islets as compared with the control group. The current study try to spot light about the effects of chronic exposure to pesticides on vital organs as pancreas also the role of oxidative stress that may be induced by them in evoking their toxicity. This study shows the role of antioxidant drugs in ameliorating or preventing the toxicity. This appears to be a promising approach that may be considered as a complementary treatment of pesticide toxicity.

Keywords: Diazinon, reduced glutathione, myeloperoxidase activity, tumor necrosis factor α, Mesna

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725 Effect of Satureja khuzestanica Jamzad Supplementation on Inflammatory and Antioxidant Indicators in Type 2 Diabetes Patients: A Randomized Controlled Clinical Trial Study

Authors: Maryam Bordbar, Yaser Mokhayeri, Sajjad Roosta, Fatemeh Ghasemi, Saeed Choobkar, Hamidreza Nikbakht, Ebrahim Falahi

Abstract:

Objective: Diabetes mellitus type 2 is the most common metabolic disorder that is growing exponentially worldwide. Satureja Khuzestanica Jamzad is a native plant of Iran that grows widely in the south of Iran. Its antimicrobial, antioxidant, anti-inflammatory and pain-relieving effects have been documented in animal studies. The purpose of this study is to investigate the effect of consumption daily S. khuzestanica on inflammatory and antioxidant indicators in type 2 diabetic patients. Methods and Materials: In a double-blind, placebo-controlled clinical trial, 67 patients with type 2 diabetes were included and divided into two groups. One group received S. khuzestanica (capsule containing 500 mg) and the other group received placebo (500 mg talcum powder) once a day for 12 weeks. After the intervention, the inflammatory and antioxidant indicators of the two groups were compared. Results: In comparison to placebo groups, there was a significant difference in levels of total antioxidant capacity, superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase; these antioxidant indicators were higher in the intervention group (P<0.05). Moreover, a considerable decrease in weight, CRP and IL-6 levels were observed in patients in the S.Khuzestanica group. Conclusion: Our findings may provide novel complementary treatments without adverse effects for diabetes complications.

Keywords: Satureja khuzestanica Jamzad, diabetes mellitus, antioxidant indicators, IL-6, C-reactive protein

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724 Isolation and Identification Fibrinolytic Protease Endophytic Fungi from Hibiscus Leaves in Shah Alam

Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

Abstract:

Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

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723 Association of Human Immunodeficiency Virus with Incident Autoimmune Hemolytic Anemia: A Population-Based Cohort Study in Taiwan

Authors: Yung-Feng Yen, I-an Jen, Yi-Ming Arthur Chen

Abstract:

The molecular mimicry between human immunodeficiency virus (HIV) protein and red blood cell (RBC) antigens could induce the production of anti-RBC autoantibodies. However, the association between HIV infection and subsequent development of autoimmune hemolytic anemia (AIHA) remains unclear. This nationwide population-based cohort study aimed to determine the association between incident AIHA and HIV in Taiwan. From 2000–2012, we identified adult people living with HIV/AIDS (PLWHA) from the Taiwan centers for disease control HIV Surveillance System. HIV-infected individuals were defined by positive HIV-1 western blot. Age- and sex-matched controls without HIV infection were selected from the Taiwan National Health Insurance Research Database for comparison. All patients were followed until Dec. 31, 2012, and observed for occurrence of AIHA. Of 171,468 subjects (19,052 PLWHA, 152,416 controls), 30 (0.02%) had incident AIHA during a mean follow-up of 5.45 years, including 23 (0.12%) PLWHA and 7 (0.01%) controls. After adjusting for potential confounders, HIV infection was found to be an independent risk factor of incident AIHA (adjusted hazard ratio [AHR], 20.9; 95% confidence interval [CI], 8.34-52.3). Moreover, PLWHA receiving HAART were more likely to develop AIHA than those not receiving HAART (AHR, 10.8; 95% CI, 2.90-40.1). Additionally, the risk of AIHA was significantly increased in those taking efavirenz (AHR, 3.15; 95% CI, 1.18-8.43) or atazanavir (AHR, 6.58; 95% CI, 1.88-22.9) component of the HAART. In conclusion, HIV infection is an independent risk factor for incident AIHA. Clinicians need to be aware of the higher risk of AIHA in PLWHA.

Keywords: autoimmune disease , hemolytic anemia, HIV, highly active antiretroviral treatment

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722 Targeting NLRP3 Inflammasome Activation: A New Mechanism Underlying the Protective Effects of Nafamostat Against Acute Pancreatitis

Authors: Jiandong Ren, Lijun Zhao, Peng Chen

Abstract:

Nafamostat (NA), a synthetic broad-spectrum serine protease inhibitor, has been routinely employed for the treatment of acute pancreatitis (AP) and other inflammatory-associated diseases in some East Asia countries. Although the potent inhibitory activity against inflammation-related proteases such as thrombin, trypsin, kallikrein, plasmin, coagulation factors and complement factors is generally considered to be responsible for the anti-inflammatory effects of NA, precise target and molecular mechanism underlying the anti-inflammatory activity in the treatment of AP remain largely unknown yet. As an intracellular inflammatory signaling platform, the NOD-like receptor protein 3 (NLRP3) inflammasome is recently identified to be involved in the development of AP. In present study, we have revealed that NA alleviated pancreatic injury in a caerulein-induced AP model by inhibiting the NLRP3 inflammasome activation in pancreas. Mechanistically, NA interacted with HDAC6, a cytoplasmic deacetylase implicated in the NLRP3 inflammasome pathway, and efficiently abrogated the function of HDAC6. This property enabled NA to influence HDAC6 dependent NF-κB transcriptional activity and thus block NF-κB-driven transcriptional priming of NLRP3 inflammasome. Moreover, NA exerted the potential to interfere HDAC6-mediated intracellular transport of NLRP3, thereby leading to the failure of NLRP3 inflammasome activation. Our current work has provided valuable insight into the molecular mechanism underlying the immunomodulatory effect of NA in treatment of AP, highlighting its promising application in prevention of NLRP3 inflammasome-associated inflammatory pathological damage.

Keywords: acute pancreatitis, HDAC6, nafamostat, NLRP3 inflammasome

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721 The Instablity of TetM Gene Encode Tetracycline Resistance Gene in Lactobacillus casei FNCC 0090

Authors: Sarah Devi Silvian, Hanna Shobrina Iqomatul Haq, Fara Cholidatun Nabila, Agustin Krisna Wardani

Abstract:

Bacteria ability to survive in antibiotic is controlled by the presence of gene that encodes the antibiotic resistance protein. The instability of the antibiotic resistance gene can be observed by exposing the bacteria under the lethal dose of antibiotic. Low concentration of antibiotic can induce mutation, which may take a role in bacterial adaptation through the antibiotic concentration. Lactobacillus casei FNCC 0090 is one of the probiotic bacteria that has an ability to survive in tetracycline by expressing the tetM gene. The aims of this study are to observe the possibilities of mutation happened in L.casei FNCC 0090 by exposing in sub-lethal dose of tetracycline and also observing the instability of the tetM gene by comparing the sequence between the wild type and mutant. L.casei FNCC 0090 has a lethal dose in 60 µg/ml, low concentration is applied to induce the mutation, the range from 10 µg/ml, 15 µg/ml, 30 µg/ml, 45 µg/ml, and 50 µg/ml. L.casei FNCC 0090 is exposed to the low concentration from lowest to the highest concentration to induce the adaptation. Plasmid is isolated from the highest concentration culture which is 50 µg/ml by using modified alkali lysis method with the addition of lysozyme. The tetM gene is isolated by using PCR (Polymerase Chain Reaction) method, then PCR amplicon is purified and sequenced. Sequencing is done on both samples, wild type and mutant. Both sequences are compared and the mutations can be traced in the presence of nucleotides changes. The changing of the nucleotides means that the tetM gene is instable.

Keywords: L. casei FNCC 0090, probiotic, tetM, tetracycline

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720 Network Analysis of Genes Involved in the Biosynthesis of Medicinally Important Naphthodianthrone Derivatives of Hypericum perforatum

Authors: Nafiseh Noormohammadi, Ahmad Sobhani Najafabadi

Abstract:

Hypericins (hypericin and pseudohypericin) are natural napthodianthrone derivatives produced by Hypericum perforatum (St. John’s Wort), which have many medicinal properties such as antitumor, antineoplastic, antiviral, and antidepressant activities. Production and accumulation of hypericin in the plant are influenced by both genetic and environmental conditions. Despite the existence of different high-throughput data on the plant, genetic dimensions of hypericin biosynthesis have not yet been completely understood. In this research, 21 high-quality RNA-seq data on different parts of the plant were integrated into metabolic data to reconstruct a coexpression network. Results showed that a cluster of 30 transcripts was correlated with total hypericin. The identified transcripts were divided into three main groups based on their functions, including hypericin biosynthesis genes, transporters, detoxification genes, and transcription factors (TFs). In the biosynthetic group, different isoforms of polyketide synthase (PKSs) and phenolic oxidative coupling proteins (POCPs) were identified. Phylogenetic analysis of protein sequences integrated into gene expression analysis showed that some of the POCPs seem to be very important in the biosynthetic pathway of hypericin. In the TFs group, six TFs were correlated with total hypericin. qPCR analysis of these six TFs confirmed that three of them were highly correlated. The identified genes in this research are a rich resource for further studies on the molecular breeding of H. perforatum in order to obtain varieties with high hypericin production.

Keywords: hypericin, St. John’s Wort, data mining, transcription factors, secondary metabolites

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719 In-silico Antimicrobial Activity of Bioactive Compounds of Ricinus communis against DNA Gyrase of Staphylococcus aureus as Molecular Target

Authors: S. Rajeswari

Abstract:

Medicinal Plant extracts and their bioactive compounds have been used for antimicrobial activities and have significant remedial properties. In the recent years, a wide range of investigations have been carried out throughout the world to confirm antimicrobial properties of different medicinally important plants. A number of plants showed efficient antimicrobial activities, which were comparable to that of synthetic standard drugs or antimicrobial agents. The large family Euphorbiaceae contains nearly about 300 genera and 7,500 speciesand one among is Ricinus communis or castor plant which has high traditional and medicinal value for disease free healthy life. Traditionally the plant is used as laxative, purgative, fertilizer and fungicide etc. whereas the plant possess beneficial effects such as anti-oxidant, antihistamine, antinociceptive, antiasthmatic, antiulcer, immunomodulatory anti diabetic, hepatoprotective, anti inflammatory, antimicrobial, and many other medicinal properties. This activity of the plant possess due to the important phytochemical constituents like flavonoids, saponins, glycosides, alkaloids and steroids. The presents study includes the phytochemical properties of Ricinus communis and to prediction of the anti-microbial activity of Ricinus communis using DNA gyrase of Staphylococcus aureus as molecular target. Docking results of varies chemicals compounds of Ricinus communis against DNA gyrase of Staphylococcus aureus by maestro 9.8 of Schrodinger show that the phytochemicals are effective against the target protein DNA gyrase. our studies suggest that the phytochemical from Ricinus communis such has INDICAN (G.Score 4.98) and SUPLOPIN-2(G.Score 5.74) can be used as lead molecule against Staphylococcus infections.

Keywords: euphorbiaceae, antimicrobial activity, Ricinus communis, Staphylococcus aureus

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718 Agrowastes to Edible Hydrogels through Bio Nanotechnology Interventions: Bioactive from Mandarin Peels

Authors: Niharika Kaushal, Minni Singh

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Citrus fruits contain an abundance of phytochemicals that can promote health. A substantial amount of agrowaste is produced from the juice processing industries, primarily peels and seeds. This leftover agrowaste is a reservoir of nutraceuticals, particularly bioflavonoids which render it antioxidant and potentially anticancerous. It is, therefore, favorable to utilize this biomass and contribute towards sustainability in a manner that value-added products may be derived from them, nutraceuticals, in this study. However, the pre-systemic metabolism of flavonoids in the gastric phase limits the effectiveness of these bioflavonoids derived from mandarin biomass. In this study, ‘kinnow’ mandarin (Citrus nobilis X Citrus deliciosa) biomass was explored for its flavonoid profile. This work entails supercritical fluid extraction and identification of bioflavonoids from mandarin biomass. Furthermore, to overcome the limitations of these flavonoids in the gastrointestinal tract, a double-layered vehicular mechanism comprising the fabrication of nanoconjugates and edible hydrogels was adopted. Total flavonoids in the mandarin peel extract were estimated by the aluminum chloride complexation method and were found to be 47.3±1.06 mg/ml rutin equivalents as total flavonoids. Mass spectral analysis revealed the abundance of polymethoxyflavones (PMFs), nobiletin and tangeretin as the major flavonoids in the extract, followed by hesperetin and naringenin. Furthermore, the antioxidant potential was analyzed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, which showed an IC50 of 0.55μg/ml. Nanoconjugates were fabricated via the solvent evaporation method, which was further impregnated into hydrogels. Additionally, the release characteristics of nanoconjugate-laden hydrogels in a simulated gastrointestinal environment were studied. The PLGA-PMFs nanoconjugates exhibited a particle size between 200-250nm having a smooth and spherical shape as revealed by FE-SEM. The impregnated alginate hydrogels offered a dense network that ensured the holding of PLGA-PMF nanoconjugates, as confirmed by Cryo-SEM images. Rheological studies revealed the shear-thinning behavior of hydrogels and their high resistance to deformation. Gastrointestinal studies showed a negligible 4.0% release of flavonoids in the gastric phase, followed by a sustained release over the next hours in the intestinal environment. Therefore, based on the enormous potential of recovering nutraceuticals from agro-processing wastes, further augmented by nanotechnological interventions for enhancing the bioefficacy of these compounds, lays the foundation for exploring the path towards the development of value-added products, thereby contributing towards the sustainable use of agrowaste.

Keywords: agrowaste, gastrointestinal, hydrogel, nutraceuticals

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