Group P3 Research Articles

Preventive effect of Nigella sativa on M1/M2 ratio, reducing risk of endothelial dysfunction in cigarette smoked Wistars

Background: Smoking is one of the top three causes of cardiovascular disease (CVD). Natural antioxidants including black cumin ( Nigella sativa) may inhibit the pathogenesis of initial process of atherosclerosis. The aim of this study was to determine the effect of black cumin (Nigella sativa) in preventing endothelial dysfunction mainly through macrophage M1/M2 inflammatory response in cigarette smoked male Wistars. Methods: In total, 50 Wistar rats were randomly allocated to five experimental groups: two control groups, namely no intervention (K-) and exposure to smoke of 40 cigarettes each day (K+); and three treatment groups: rats given a dose of 0.3 g (P1), 0.6 g (P2) or 1.2 g (P3) black cumin per kilograms bodyweight/ day, respectively, and exposed to smoke of 40 cigarettes each day. After 28 days of cigarette smoke exposure, macrophage M1/M2 ratio was evaluated by counting total M1 and M2 in ten microscope field of view. Data were analysed by Mann-Whitney test. Results: The M1 / ​​M2 ratio on K (-) was 0.9 7 ± 0.9 8 (<1) which means M2 was dominant, while the M1 / ​​M2 ratio on K (+) was of 4.97 ± 3.42 (> 1) which means M1 dominant. There was no significant difference in the number of M1 count in treatment groups P1, P2, P3 (p value = 0.996; 0.170; 0.884, respectively) when compared with K+. Additionally, P2 group has the lower M1 number with the highest significance value when compared to K+. The number of M1 counts on P1 did not differ significantly when compared to P2 with p = 0.121 and P3 with p = 0.936. Conclusions: In sum, ethanol extract of black cumin prevents endothelial dysfunction by inhibiting increase in macrophages M1 / M2 ratio in rats Wistar exposed to sub-chronic cigarette smoke.

Synthesis and characterization of a novel alkali mixed magnesium-aluminum phosphate with a layered structure–KMgAl(PO4)2

A novel phosphate compound - potassium magnesium-aluminum double phosphate with chemical formula KMgAl(PO4)2 - has been synthesized. Platelet-shaped single crystals have been grown from the flux using K2Mo2O7 as a solvent. The powder form has been obtained through a sol-gel reaction. It has been characterized by X-ray diffraction, differential thermal analysis, vibrational spectroscopy and electronic structure calculations. KMgAl(PO4)2 crystallizes in the trigonal centrosymmetric space group P3¯m1 (No. 164) with unit cell parameters a = b = 5.347(1) Å, c = 7.903(10) Å, γ = 120° (Z = 1). The main characteristic of the crystal is the two-dimensional (2D) lamellar framework formed by Mg|AlO4 and PO4 tetrahedrons sharing their vertices of oxygen atoms, with the K+ cations located within the interlayer space. The sample morphology and chemical composition were revealed by the scanning electron microscopy – energy-dispersive X-ray spectroscopy analysis. The IR spectroscopy of KMgAl(PO4)2 confirmed the existence of a single PO4 group in agreement with the structural study. A relatively large indirect band gap of 3.86 eV was determined.

Preventive effect of Nigella sativa on M1/M2 ratio, reducing risk of endothelial dysfunction in cigarette smoked Wistars.

Background: Smoking is one of the top three causes of cardiovascular disease (CVD). Natural antioxidants including black cumin ( Nigella sativa) may inhibit the pathogenesis of initial process of atherosclerosis. The aim of this study was to determine the effect of black cumin (Nigella sativa) in preventing endothelial dysfunction mainly through macrophage M1/M2 inflammatory response in cigarette smoked male Wistars. Methods: In total, 50 Wistar rats were randomly allocated to five experimental groups: two control groups, namely no intervention (K-) and exposure to smoke of 40 cigarettes each day (K+); and three treatment groups: rats given a dose of 0.3 g (P1), 0.6 g (P2) or 1.2 g (P3) black cumin per kilograms bodyweight/ day, respectively, and exposed to smoke of 40 cigarettes each day. After 28 days of cigarette smoke exposure, macrophage M1/M2 ratio was evaluated by counting total M1 and M2 in ten microscope field of view. Data were analysed by Mann-Whitney test. Results: The M1 / ​​M2 ratio on K (-) was0.97 ±0.98 (<1) which means M2 was dominant,while the M1 / ​​M2 ratio on K (+)was of4.97 ± 3.42(> 1) which means M1 dominant.There was no significant difference in the number of M1 count in treatment groups P1, P2, P3 (p value =0.996; 0.170;0.884, respectively) when compared with K+.Additionally, P2 group has the lower M1 number with the highest significance value when compared to K+.The number of M1 counts on P1 did not differ significantly when compared to P2 withp= 0.121 and P3 with p =0.936. Conclusions: In sum, ethanol extract of black cumin prevents endothelial dysfunction by inhibiting increase in macrophages M1 / M2 ratioinrats Wistar exposed to sub-chronic cigarette smoke.

The Effects of Green Betel Leaf (Piper betle) Extract Eye Drops on the Number of Staphylococcus aureus Colonies in Conjunctivitis Wistar Rats Model (Rattus novergicus)

Irrational use of antibiotics can cause resistance to some diseases such as bacterial conjunctivitis caused by Staphylococcus aureus. Previous research said that green betel leaf (Piper betle) contains antimicrobial compounds such as eugenol, cavalry, tannins, saponins, and flavonoids which also have antiseptic power as well as antibiotics. This study aims to prove the decrease in the number of Staphylococcus aureus colonies after giving green betel leaf eye drops (Piper betle) to conjunctivitis Wistar rats (Rattus norvegicus). This research was true experimental with a pretest-posttest control group design. 25 male white Wistar rats as objects were selected by simple random sampling and grouped into 5 groups randomly. The rats were given Staphylococcus aureus, control group K+ was given 0.5% levofloxacin eye drops, and K- was given aquadest as a comparison. Treatment groups P1, P2, and P3 were given different concentrations of betel leaf eye drops. Swab pretest was done 3 days after the rats were inoculated with Staphylococcus aureus and swab post-test was done 5 days after the rats were given green betel leaf eye drops, the calculation of Staphylococcus aureus bacteria colonies used the Total Plate Count (TPC) method. There was a significant decrease (p&lt;0.05) in the number of bacterial colonies for K-, K+, P1, P2, and P3 groups. In addition, there was a significant difference (p&lt;0.05) between K+ with P1 and K+ with P3. There was a decrease in the number of Staphylococcus aureus for all groups of conjunctivitis Wistar rats (Rattus norvegicus)

Effect Of Alpha-Tocopherol On Spermatozoa Death Of Apoptosis and Necrosis in Rats (Rattus Norvegicus) Exposed 2,3,7,8- Tetrachlorodibenzo-P-Dioxin

The aims of this research was to investigate effect of alpha-tocopherol on spermatozoa death in form of apoptosis and necrosis in rats (Rattus norvegicus) exposed 2,3,7,8- tetrachlorodibenzo-p-dioxin. Male rats were administered TCDD and alpha-tocopherol in experimental groups. Five experimental groups of a combination of TCDD and alpha- tocopherol were designed as follows; 0.5 ml of corn oil (control negative group, K-), 700 ng/kg/d of TCDD and 0.5 ml of corn oil (treatment control), 700 ng/kg/d of TCDD and 77 ng/kg/d of alpha-tocopherol (Group P1), 700 ng/kg/d of TCDD and 140 mg/kg/d of alpha-tocopherol (Group P2), 700 ng/kg/d of TCDD and 259 mg/kg/d of alpha- tocopherol (Group P3) respectively. Alpha-tocopherol and TCDD were given by oral gavage for 20 days. The result indicated that TCDD increased spermatozoa death in form of apoptosis and also necrosis. Alpha-tocopherol at 259 mg/kg/d most effective to decreased spermatozoa death number. The conclusion indicated that alpha-tocopherol at 259 mg/kg/d effective to decreased the spermatozoa death in form of apoptosis and necrosis in rats (Rattus norvegicus) exposed 2,3,7,8-tetrachlorodibenzo-p-dioxin.

The possibility of polymorphonuclear leukocyte activation in dental socket healing by freeze-dried Aloe vera induction

Background: Research has proved that Aloe vera has anti-inflammatory, anti-bacterial, and immunomodulator properties that can accelerate the healing process. These properties could be beneficial in the tooth extraction wound-healing process. Neutrophils are the first inflammatory cells to invade and are the predominant cell markers in the wound for 24 hours after injury. Polymorphonuclear leukocytes (PMN) provide initial protection against micro-organisms, as they engulf and digest foreign bodies. Purpose: The aim of this study was to prove the effect of Aloe vera 90% gel application on the PMN count in the healing process of Cavia cobaya following tooth extraction. Methods: This laboratory experimental research was done using Post-Test Only Control Group Design. The lower incisive of twelve male Cavia cobayas were extracted then divided into four groups. Group P1 and P3 had 90% Aloe vera gel applied into the socket and sutured; group K1 and K3, received no 90% Aloe vera gel application and acted as controls. Samples from group P1 and K1 were terminated and had their mandibula taken one day after tooth extraction, and those from group P3 and K3 were terminated after three days. Tissues were then processed into histology slides, and PMN cells were counted. Results: There were significant differences in the PMN count between groups K1, P1, and K3 compared with P3 (p &lt; 0.05). Higher PMN count was shown on the group containing Aloe vera 90% gel compared with control group, both on day one and three after tooth extraction. Conclusion: Aloe vera 90% gel can increase the PMN count in the inflammation phase of the tooth extraction wound-healing process.

Skin barrier before and after topical Adipose Stem Cell-Conditioned Medium (ASC-CM) treatment in photoaging

Background: Photoaging is a type of aging mainly caused by ultraviolet irradiation. The skin barrier is affected in photoaging and it will result in coarseness, roughness and dry skin. It is probably because Ultraviolet (UV) affects the extracellular matrix in the skin. Adipose Stem Cell-Conditioned Medium (ASC-CM), which contains many growth factors, becomes a potential agent as a novel anti-photoaging treatment by repairing the skin barrier. This study aims to evaluate the skin barrier function before and after treatment with ASC-CM in photoaging skin by evaluating changes in Transepidermal Water Loss (TEWL) and skinfold thickness.Methods: A true experimental study was conducted among 28 male Wistar rats aged 10-12 weeks. They were acclimated for 1 week and randomly divided into 4 groups: UV irradiated group with topical ASC-CM (Group P1); UV irradiated group with topical vehicle only (Group P2); only topical ASC-CM, non-UV irradiated (Group P3), and non-UV irradiated and no topical treatment (Group P0). The ASC-CM was given twice a day for four weeks and the total irradiance dose for 6 weeks was 4,2 J/cm2. Data were analyzed using SPSS version 20 for Windows.Results: There was a statistically significant difference of TEWL between pre-and post-treatment in group P3 (p=0.043) and P0 (p=0.018). In addition, a statistically significant difference in skin thickness was also found between pre-and post-treatment in group P2 (p=0.005) and P3 (p=0.003). There was a statistically significant difference in skin thickness post-treatment (p=0.000); however, there was no statistically significant difference of TEWL post-treatment by One-Way ANOVA test (p=0.138).Conclusion: ASC-CM could be a potential option for photoaging skin based on the TEWL and skin thickness evaluation. However, a longer study period was needed to know the further effect of ASC-CM.

Pengaruh Aflatoksin 9,58 ppb dalam Pakan Terhadap Biometrik Saluran Cerna dan Berat Badan Ayam Petelur Periode Awal Pertumbuhan

The purpose of this study was to determine the effect of aflatoxin content of 9.58 ppb in feed on gastrointestinal biometrics and weight gain of laying hens in the early growth period. Thirty-six day old chicks (DOC) were used in this study and were divided into two groups P0 and P1. P0 (control group) was fed with 100% basal diet and P1 (treatment group) was fed a diet with a composition of 80% basal feed + 20% aflatoxin-containing feed. After the DOC was adapted, the first feeding containing aflatoxin was given from the 20th day to the 60th day. Data collection was carried out three times, namely the 20th, 40th, and 60th days. At the end of each period, the chickens would be euthanized to collect data on the weight and length of the digestive tract and body weight. Weight gain with time increased, but had no significant effect (p&gt;0.05) between the control and treatment groups. Body weight continues to increase because the body's homeresis function is still running well. Observations on gastrointestinal biometrics showed significant differences (p&lt;0.05) in oesophagus weight, proventriculus weight, ventriculus weight, jejunum weight, ileum weight and length, and cecum weight because aflatoxins have a direct effect on the digestive tract they pass. The conclusion of this study was that the provision of 20% aflatoxin-containing feed did not affect weight gain but had shown an effect on the biometrics of several gastrointestinal tracts.

Beetroot Powder (Beta vulgaris L.) Decrease Oxidative Stress by Reducing of Malondialdehyde (MDA) Levels in Hyperuricemia

Introduction: Hyperuricemia is a metabolic disease associated with lifestyle habits, age, sex, and inherited factors. High uric acid levels in a hyperuricemia state cause oxidative stress that can lead to increased malondialdehyde (MDA) levels. Beetroot powder has the potential as a functional food rich in antioxidants that can reduce MDA levels. This study aims to determine the beetroot powder effect on MDA level reduction in hyperuricemia rat models. Methods: This research was an experimental study with fifteen hyperuricemia rats were divided into three groups by randomized pre-post control group design. They were divided into three groups: P1 were healthy rats + standard feed (comfeed), P2 were hyperuricemia rat models + standard feed (comfeed) and P3 were hyperuricemia rat models + beetroot powder 3,12 g/kg/bw for 14 days. The MDA level examinations were conducted on day 0 and day 14. Results: After 14 days of treatment, the P3 group has a significant decrease in MDA levels compared to day 0 (p = 0.042). Groups P1 and P2 have a significant increase in MDA levels (p = 0.043). MDA levels between groups were significantly different (p = 0.002) on day 14. Conclusion: Giving beetroot powder for 14 days with dose 3,12 g/kg/bw for 14 days significantly reduced MDA levels in hyperuricemia.

EFFECT OF AFRICAN LEAF EXTRACT (Vernonia amygdalina) ON ALLOXANINDUCED DIABETES MELLITUS LEYDIG CELL MYTOSIS OF WISTAR RATS

Introduction: Diabetes mellitus can reduce the quality of human life when accompanied by complications, one of which is sexual dysfunction. In diabetics, there is an increase in ROSproduction which can cause damage to local Leydig cell proliferation so that Leydig cell activity and testosterone secretion decrease. African leaves can be used as a control of blood sugar levels and antioxidants to overcome oxidative stress. Purpose: To determine the effect of administration of African leaf extract (Vernonia amygdalina) on alloxan-induced Leydig cell mitosis in male rats with diabetes mellitus. Method: This experimental study used 31 male Rattus norvegicus white rats divided into K, P0, P1, P2, and P3 groups. Each group consisting of six rats and one rat outside the study group was used as a reference. Experimental animals will be induced alloxan with a single dose of 150 mg/kgBW for four days. African leaf extract was administered using a probe to groups P1, P2, P3 at a dose of 100, 200, and 400 mg/kgBW for 14 days. Rats were anaesthetized and euthanized so that the testes could be used to make preparations with HE staining and analysis of preparations using a 400x magnification light microscope. The research data were analyzed using One-Way Annova with P&lt;0.05. Results: The administration of African leaf extract affected increasing the number of mitotic Leydig cells (P=0.026). There was a significant increase in the number of mitotic Leydig cells at a dose of 100 mg/kgBW (P=0.021), but not effective at a dose of 200 mg/kgBW (P=0.188) and 400 mg/kgBW (P=0.823). Conclusion: The administration of African leaf extract at a dose of 100 mg/kgBW has been shown to increase the number of mitotic Leydig cells so that it can be used to improve sexual function in diabetics.

Kebar Grass Extract (Biophitum Petersianum) Increases Diameter Of The Seminiferous Tubules Of Diabetic Mice

Diabetes is a metabolic disease that can affect the male reproductive system. The condition of hyperglycemia will increase the production of ROS, causing oxidative stress. Oxidative stress causes microangiopathy, triggers cell apoptosis, interferes with spermatogenesis, thereby affecting the diameter of the seminiferous tubules. Kebar grass contains of flavonoid and vitamin E which can be used to reduce ROS reactions and inhibit oxidative stress. This research was to prove that kebar grass extract can increase diameter of the seminiferous tubules of diabetic mice. Total of 30 samples were divided into 5 groups: negative control group (K-) was given CMC-Na, positive control group (K+) was given metformin 2mg/kg, groups P1, P2 and P3 received 2mg/kg metformin and kebar grass extract with different dosages (67.5mg/kg, 135mg/kg and 270mg/kg). The treatment lasted for 35 days. On the 36th day the mice were sacrificed and the testes were taken, then histological observations were made with HE staining to measure the diameter of tubules. The result showed that there was a significant difference in the diameter of seminiferous tubules with p value=0,000. Kebar grass extract can increase the diameter of seminiferous tubules of diabetic mice.

Study of the magnetic properties of YbMn2Sb2 single crystals by [formula omitted]SR

The crystal growth and the structural, transport and magnetic properties of YbMn2Sb2 single crystals are reported. The crystals show a trigonal symmetry of La2O2S-type structure (space group P3¯m1, N. 164, hP5), where corrugated honeycomb layers of MnSb are separated by Yb atoms. No structural phase transition was observed down to 22 K. The resistivity measurements show a predominantly insulating behavior. The combined resistivity, dc susceptibility and specific heat measurements confirm successive features at 40 K, 116 K and 225 K, being the phase transition at TN=116 K due to the Mn+2 lattice antiferromagnetic ordering. Muon spin rotation experiments (μSR) reveal a more complicated scenario, with temperature dependence of the magnetic volume fraction reflecting a static and strongly disordered magnetic ground state. The role of spin–lattice coupling and its relationship with exchange interactions between Mn moments is discussed as a possible cause of the magnetic behavior observed.

Current approaches to the use of antiplatelet and anticoagulant drugs in pregnant women with thrombocytopenia caused by hemostasis activation

Introduction. At present, hemostasis disorders still hold one of the lead places in the pathogenesis of reproductive losses associated with high risk of miscarriage and remain one of the major causes for developing serious obstetric complications. Relevance, efficacy and safety of using drugs containing acetylsalicylic acid (ASA) and low molecular weight heparins (LMWH) in pregnant women with thrombocytopenia comorbid with intravascular coagulation activation remain pressing issues. The study of thrombocytopenia pathogenesis during pregnancy and search for new methods of hemostasis correction are accounted for by high risk of developing diverse perinatal complications and necessity to conduct prophylactic measures for their prevention.Aim: to study hemostasis changes and clinical outcomes in pregnant women with thrombocytopenia, to substantiate the use of antiplatelet agents and anticoagulants in pregnant women with activated intravascular blood coagulation for preventing development of thromboembolic and placenta-associated complications of pregnancy.Materials and Methods. A multicenter prospective study was conducted by enrolling 299 pregnant women at a gestational age of22.85 [22.00; 25.00] weeks, whose average age was 31.06 [27.75; 35.00] years. There were distinguished three groups: main group (n = 124) consisted of pregnant women with lowered platelet count and activated intravascular blood coagulation, who received ASA preparations and LMWH in prophylactic doses for 4 weeks; pregnant comparison groups (n = 125) received no such drugs; control group (n = 50) consisted of women with normal platelet counts during physiological pregnancy. All patients underwent clinical, anamnestic and laboratory examination, and the clinical outcomes of pregnancy were assessed.Results. It was found that use of ASA (a combined drug: ASA 75 mg + magnesium hydroxide 15.2 mg/day) and LMWH (enoxaparin sodium) in pregnant women from the main group were noted to improve hemostasis parameters: via markedly increased platelet count, decreased fibrinogen level, prothrombin index as well as normalized antithrombin III level (p &lt; 0.01). It was also found during treatment that the D-dimer level and the rate (83.6 ± 2.3 % and 72.4 ± 2.7 %) and the degree (86.4 ± 2.7 % and 74.4 ±2.8 %) of platelet aggregation in the main group (in contrast with comparison group) were significantly decreased (p &lt; 0.01). The frequency of detected chronic placental insufficiency was significantly higher (p &lt; 0.01) in the comparison group (n = 50; 41.32 %) vs. the main group after receiving ASA and LMWH (n = 20; 16.52 %). The incidence of moderate (p = 0.016) and severe (p = 0.018) preeclampsia was significantly higher in the comparison group vs. the main group. While comparing the volume of blood loss during delivery, there were no differences between the main group and the comparison group (p = 0.46); it was noted only a statistically significant difference compared to the control group (p = 0.04), in which the lowest blood loss was found (337.9 [200.0; 600.0] ml) in comparison with the other two groups (p1,3 = 0.05; p2,3 = 0.04). It should be noted that the volume of blood loss in all groups remained within the physiological permissible range.Conclusion. Our clinical and laboratory study allowed to substantiate relevance of using ASA and LMWH drugs in pregnant women with thrombocytopenia comorbid with confirmed activation of intravascular blood coagulation, for prevention of thromboembolic and placenta-associated complications of pregnancy as well as proved safety of such drugs during pregnancy.

Growth response and vital statistics of fat and thin tailed sheep with soybean husk supplements in Malang District

This study aimed to determine the pattern of body weight gain and vital statistical measures of fat sheep and thin tails and to determine the response of local sheep production to the provision of soybean meal/skin. The method used in this research is a case study and experimental. The sampling technique is done by a simple random method on vital statistical measures performed by measuring the chest (using a measuring tape) and body length (using a measuring stick). The tabulated data were first analyzed for homogeneity and normality, which were then tested by an independent sample t-test using SPSS. As for the growth response, the material used was 16 male FTS and TTS aged under one year. Daily body weight growth between Fat tail Sheep (FTS) and Thin Tail Sheep (TTS) showed very significant differences (P <0.01). Statistical analysis showed that body length between FTS and TTS had no difference (P> 0.05), chest circumference between FTS, and TTS; there was no difference (P> 0.05). Adding bodyweight FTS and TTS were respectively 93.29 ± 26.73 g / head / day and 78.18 ± 27.01 g / head / day. The FTS and TTS bodies’ length was 49.81 ± 4.06 cm and 49.34 ± 4.80 cm, respectively, while the chest circumference between FTS and TTS was 63.61 ± 3.98 cm and 62, respectively. 17 ±4.10 cm. The daily body weight gain of rams fed with additional soybean husk feed statistically results obtained significant differences (P <0.05), the results of the study showed that the Daily Weight Gain (DWG) FTS male respectively P0, P1, P2, and P3 groups respectively namely: 105.07 ± 3.58; 118.08 ± 2.65; 140.38 ± 4.40; 155.01 ± 4.01 g / head/day. The results showed that the feed efficiency of male rams in each group P0, P1, P2, and P3 were: 7.84 ± 0.27; 8.25 ±0.18; 9.27 ± 0.29; and 9.75 ± 0.32%. While for TTS also obtained a significant difference (P <0.05) DWG at P0, P1, P2, P3 is 101.02 ± 2.18; 116.9 ± 2.88; 127.82 ± 3.20; 140.31 ± 2.41 g / head / day, so that the efficiency of feed obtained for TTS is P0, P1, P2, P3 respectively 6 ± 0.19; 7.14 ± 0.11; 8.87 ± 0.22; 9.85 ± 0.12. It was concluded that the provision of soybean meal had a significant positive effect on growth and vital statistics, both for sheep with fat tails and thin tails.

Structure of Enterohemorrhagic Escherichia coli O157:H7 Intimin Virulence Factor Bound to Nanobodies

Enterohemorrhagic E. coli O157:H7 (EHEC) is a food and water-born pathogen that presents a significant risk to human health worldwide. EHEC naturally resides in the intestinal tract of cattle as normal flora but can be transmitted to humans when contaminated food is ingested. Once EHEC invades and colonizes the human intestinal tract, it secretes Shiga-like toxins causing serious and potentially fatal gastrointestinal diseases. Unlike most foodborne bacterial diseases, antibiotics are not recommended for treatment of EHEC as antibiotics can induce enterotoxin release, increasing the risk of hemolytic uremic syndrome. There is currently no effective treatment or vaccine against enterohemorrhagic E. coli. EHEC pathogenicity involves intimin, a virulence factor which mediates bacterial colonization of the host GI tract. Due to its critical role in EHEC pathogenesis, intimin has attracted attention as an antibacterial drug target. Inhibiting the interaction of intimin with its cognate bacterial secreted receptor Tir could neutralize bacterial colonization of the gastrointestinal tract. One novel solution for treating EHEC infection comes from a unique class of antibodies known as nanobodies. Nanobodies are the antigen binding domain of heavy chain antibodies produced by the Camelid family. Intimin specific nanobodies were generated by immunizing a llama followed by phage display and selection for high affinity intimin binding nanobodies. The resultant intimin-specific nanobodies neutralized EHEC in vitro. We hypothesize that nanobodies specifically bind the Tir-binding domain of intimin in order to neutralizing EHEC infection. The goal of this research project is to determine the complex crystal structure of the nanobodies bound to intimin. The structure of nanobodies-intimin would show their protein-protein interaction binding site revealing key amino acids involved in binding. These residues could also be involved in the binding of intimin to its natural ligand, Tir. Thus, revealing such residues could form the bases for identifying and developing novel therapeutics and biosensors for treating and preventing EHEC pathogenesis.We show through size exclusion chromatography that nanobodies bind intimin and form stable complexes. We have set up crystal trails of five nanobodies (Int1, Int2, Int3, Int4, and IntN1) complexed to intimin. Two of the five complexed proteins, Int2-intimin and Int3-intimn formed crystals. The crystal condition for the two protein complexes Int2 and Int3 bound to intimin were optimized by varying the concentration of precipitants and pH range. Crystals of the protein complexes were sent to the Canadian Light Source for X-ray diffraction data collection. Diffraction measurements of Int2-intimin and Int3-intimin complexes were collected and processed in space groups P1 and P222, respectively. Currently, we are analyzing the structure by molecular replacement.

The effectiveness of the combination of Moringa oleifera extract and propolis on Porphyromonas gingivalis biofilms compared to 0.7% tetracycline

Background: Periodontitis is an inflammatory disease that occurs in periodontal tissues. Porphyromonas gingivalis is also known as a bacterium commonly associated with the pathogenesis of periodontitis. Tetracycline is one of the antibiotics often used in periodontal tissue treatment. Propolis and Moringa oleifera are also known to have certain compounds assumed to be able to inhibit biofilm growth. Purpose: This study aims to understand the effectiveness of the combination of Moringa oleifera and propolis on porphyomonas gingivalis biofilms compared to 0.7% tetracycline. Methods: A biofilm inhibition activity test was performed using the broth micro dilution method. First, bacteria were prepared by making a suspension in brain heart infusion media and adjusting it to 0.5 McFarland I standard. Second, fifteen samples were divided into five groups; group K as control group (0.1% sodium carboxymethyl cellulose), T (0.7% tetracycline), and treatment groups with the combination of propolis and Moringa oleifera in various concentrations, such as P1(10%+20%), P2(10%+40%), and P3(10%+80%). Third, the result data obtained in the form of optical density (OD) was read by using an ELISA reader. Next, statistical analysis using analysis of the variance test was conducted (p&lt;0.05. Results: There was no significant difference between group T and group P1 (0.075). Nevertheless, there were significant differences between group T and group P2 as well as between group T and group P3 (0.00) (p=&lt; 0.05). Conclusion: The combination of 10% propolis and 40% Moringa oleifera as well as the combination of 10% propolis and 80% Moringa oleifera have better antibacterial effectiveness against Porphyromonas gingivalis biofilm than 0.7% tetracycline.

The Effect of Sambiloto Extract on the Expression of β Estrogen Receptors in Ovaries and Lust Cycle in Mice Model PCOS - Insulin Resistance

Infertility was a serious problem in the reproductive period, one of the caused of infertility were common in women of childbearing age group is Polycystic Ovary Syndrome (PCOS).Sambiloto (Andrographis paniculata Nees), had been already known contain flavonoid and lactones. The purpose of this research had to determine the effect of various doses of sambiloto extract with the expression of β estrogen receptor and estrus cycle in the mice model of PCOS - insulin resistance. This research was an animal model experimental laboratory research with a completely randomized design (CRD).The results showed that sambiloto extract therapy in treatment group had an improvement estrus cycle compared with control group. Positive control group, almost 90% had persistent anestrus condition. In treatment group 1 and 2 in the last vagina smear obtained anestrus condition as much as 60% in diestrus and medestrus phase. Whereas treatment groups 3 all experimental animals in a estrus state by proestrus and estrus phase. β Estrogen receptor expression had tested by Kruskal Wallis test for overall treatment, obtained significantly different results (p ≤ 0.002) followed by Mann - Whitney, showed that immunoreactive cells score from highest to lowest occurred in each group Negative control (KN), P3, P2, P1 and Positive Control (KP).It could be concluded that sambiloto extract at dose 18mg/kgbb, 36mg/kgbb, and 72mg/kgbb had been given an overview of the differences expression of β estrogen receptor, could change the estrus cycle of female mice models of insulin resistance PCOS.

Antihyperglycemic Test of Ethanol Extract of Soursop Leaf (Annona muricata L.) on Blood Sugar Levels of Male Mouse (Mus musculus) Induced by Glucose

Soursop leaf (Annona muricata L.) is an herbal plant that is widely used by the public empirically because it has many properties. The sling leaf contains tannin compounds, alkaloids, as well as several other chemical compounds including annonaceous acetogenin, which can be used as a treatment. This study aims to find out if soursop leaf extract has the effect of deducting blood glucose levels in mice test animals (Mus musculus) which induced a 50% glucose solution. The study used the RAL method (complete random design) and 20 male mice were divided into 5 groups with 4 repetitions. Group P1, P2, and P3 (treatment group) administered ethanol extract of sine leaves at doses of 250, 500, and 750 mg/kg bb respectively, while k- and k+ (control group) administered CMC 0.5% and glibenclamide with oral administration. The data obtained were statistically analyzed with ANOVA (Variance analysis) one-way to find out the differences between treatment groups and continued with the BNT test (The smallest real difference) to find out meaningful differences between groups. The results showed that there were differences between the treatment group and the control group. On the 7th day of the BNT test, the treatment group with a dose of 750 mg/kg bb showed significant differences between the negative Control group (CMC 0'5%), Positive control (Glibenclamide), P1 (250 mg/kg bb), and P2 (500 mg/kg bb), while on the 14th day showed no significant difference with the treatment group of 500 mg/kg bb. concluded from this study that all concentrations of ethanol extract of sling leaves could lower blood glucose levels in test animals, with the most optimal decrease being the group at a dose of 750 mg/kg bb.

Pengaruh antioksidan Probucol terhadap kadar Malondialdehide (MDA) dan jumlah “Circulating Endothel Cells” pada tikus putih yang menerima stressor

The obyective of the study was to find out the effect of Probucol as an antioxidant on Malondialdehyde (MDA) level and total circulating endothelial cells of rats exposed to stressor.This study was a post test only control group design using 35 three- months old male wistar rats devided into 5 groups consisting of 7 rats each namely the control groups (Po1, Po2) and the experiment groups (P1,P2, P3). Details of the treatment to each groups ere as follows:Group Po1 were given per oral Probucol dilution solution for 3 weeksGroup Po2 were given per oral Probucol dilution solution and stressor for 3 weeks Group P1 were given per oral Probucol dose I (5 mg/rat) and stressor for 3 weeksGroup P2 were given per oral Probucol dose II (10 mg/rat) and stressor for 3 weeks, andGroup P3 were given per oral Probucol dose III (20 mg/rat) and stressor for 3 weeksMalondialdehyde (MDA) level and total circulating endothelial cells were measured using “Buege and Aust (1978) and Hladovic and Rossman (1973)“ method modified by Wijayanto (1996). Data on MDA level and total circulating endothelial cells were analyzed by using analysis of variant (Anova). The presence of significant difference was confirmed by using LSD test. Correlation between MDA level and total circulating endothelial cells was analysed by using “scarter plot” and r (rho) tests. The results showed a significant increase of MDA level of Po1 compared to Po2 (P less than 0.05). Significant decrease of MDA level (P less than 0,05) was observed in group P1, P2 and P3 compared to that of the Po1 and Po2 control groups. The results also showed a significant increase of total circulating endothelial cells of the control group Po1 compared to that in the control group Po2 (P less than 0,05), while significant decrease of total circulating endothelial cells (P less than 0,05) was observed in group P1, P2 and P3 compared to that of the control group Po2. It can be concluded that stressor may increase MDA level and total circulating endothelial cells

Pengaruh Pemberian Minuman Energi Terhadap Kadar Serum Kreatinin Rattus Norvegicus

The energy drink is a beverage that is increasingly being used by the community with the aim to improve work performance, vitality, and concentration. The energy drink is also suspected to affect kidney function. This study aims to determine whether there is influence of various brands of energy drinks (M1, M2, M3, M4) of the serum creatinine levels in male rats Rattus norvegicus.This is a study with randomized post test only control group design. The study used male rats Rattus norvegicus as many as 25 animals that were divided into 5 groups. The control group, the group P1 = M1 energy drinks, group P2 = M2 energy drinks, group P3 = M3 energy drinks, group P4 = M4 energy drinks. The energy drinks were given as many as 2.5 ml / 200 g weight / day for 1 month. Cardiac blood sampling performed to measure the levels of serum creatinine and then the results were analyzed using One-Way Anova to compare the levels of serum creatinine in each group of this study. Results of One-Way Anova test obtained by sig α = 0.000 which means there is significant difference between the treatment group and the control group. It can be concluded that the administration of energy drinks to the Rattus norvegicus would be altered the kidney function. It is caused by various substances contained in energy drinks such as taurine, sugar, vitamin B, caffeine, and other supplementary materials.