Additive potential of ginger starch on antifungal potency of honey against Candida albicans

Objective:To evaluate the additive action of ginger starch on the antifungal activity of honey against Candida albicans(C.albicans).Methods:C.albicans was used to determine the minimum inhibitory concentration(MIC)of four varieties of Algerian honey.Lower concentrations of honey than the MIC were incubated with a set of concentrations of starch and then added to media to detennine the minimum additive inhibitory concentration(MAIC).Results:The MIC for the four varieties of honey without starch against C.albicans ranged between 38%and 42%(v/v).When starch was incubated with honey and then added to media,a MIC drop was noticed with each variety.MAIC of the four varieties ranged between 32%honey(v/v)with 4%starch and 36%honey(v/v) with 2%starch.Conclusions:The use of ginger starch allows honey benefit and will constitute an alternative way against the resistance to antifungal agents.

Surrogate potency assays:Comparison of binding profiles complements dose response curves for unambiguous assessment of relative potencies

Surface plasmon resonance(SPR) systems are widely used for detailed characterization of antibody activities including antigen and Fc-receptor binding. During the later stages of development, where the focus is to ensure that established critical quality attributes(CQAs) are maintained during cell culture,purification and formulation processes, analysis is simplified, and relative potencies are often determined. Here, simulation of binding data revealed that relative potency values, determined via parallel line analysis(PLA) and half maximal effective concentration(EC50) analysis accurately reflect changes in active concentration only if binding kinetics remain unchanged. Changes in the association rate constant shifted dose response curves, and therefore relative potencies, in the same way as changes in analyte concentration do. However, for interactions characterized by stable binding, changes in the dissociation rate constant did not result in any shift, suggesting that this type of change may go unnoticed in the dose response curve. Thus, EC50 and PLA analyses of dose response curves obtained with an anti-TNF-α antibody were complemented with the Biacore functionality for sensorgram comparison analysis, whereby changes in antigen and Fc-receptor binding profiles could be detected. Next, analysis of temperature stressed TNF-α antibody revealed that calibration free concentration analysis(CFCA) data correlated perfectly with relative potency values. Together, these results demonstrate that combinations of SPR based dose response curves, sensorgram comparison and CFCA can be used to strengthen the confidence in relative potency assessments, and suggest that SPR can potentially be used as a surrogate potency assay in the quality control of biotherapeutic medicines.

Immunohistochemical Analysis of the Acid Secretion Potency in Gastric Parietal Cells

Gastric parietal cells are important in acid secretion, but it is unclear which cells throughout the gastric gland have the highest secretion potency. Here, we used immunohistochemical methods with anti-H+, K+-ATPase, phosphoryl ezrin and CD44 antibodies to study the distribution of gastric acid secretion activity. Stomach tissues from freely fed and starved rats were cryofixed for light microscopy or fixed by high-pressure freezing for electron microscopy. Parietal cells from freely fed animals corresponded to the active secretion phase and to the inactive resting phase from starved rats. Anti-H+, K+-ATPase and anti-phosphoryl ezrin labeling were observed on the membrane of the intracellular canaliculi and the tubulovesicle from freely fed rats, while cells from starved animals showed weak labeling with anti-phosphoryl ezrin antibody staining. Morphometrical analysis at the electron microscopic level was performed on active and inactive acid secretory phases between the upper and base regions of the gland. H+, K+-ATPase and CD44 were distributed on both sites of the microvillous and tubulovesicle membrane in the same cells, but phosphoryl ezrin localized predominantly on the microvillous membrane in active cells of the glandular neck and upper base. Therefore, the highest secreting potency appeared to be in cells of the glandular neck and upper base.

The biological potency of benzapyrene in the humates composition

The aim of the paper was to determine benz(a)-pyrene in the preparations containing humates and study the benz(a)pyrene biological potency for the agricultural plants. The research methodology included the determination of the dependencies in the system “substance concentration (dose)—effect on the plant”. Concentrations of benz(a)pyrene in 12 samples of the humates preparations and fertilizers based on their trademarks “Irkutsk humates”, obtained from brown coal, varied in the range from 0.3 to 50 mcg/kg, which creates no soil contamination in conditions of the use of preparations. Between contents of benz(a)pyrene and humates there is a correlation (rxy = 0.95;α = 0.05). It is ascertained that the effects of stimulation and/or inhibition of the growth and yield of agricultural plants depend on the concentration of benz(a)pyrene and the method of plant processing. Optimal concentrations of benz(a)pyrene were 150-200 ng/dm3 for preplant way of processing of potato tubers, 3-10 ng/dm3—for top dressing (spraying) and 0.1-0.3 ng/dm3—for dressing under the roots (hydroponic). The obtained results allowed us to offer one of the possible mechanisms of biological potency of humates as the plant growth stimulants, and also a way testing of the preparations by screening of their benz(a)pyrene content.

Determination of the Potency of Extracted, Purified and Formulated Insulin from the Pancreatic Organs of the Sudanese Beef Cattle

The treatment of type 1 diabetes is mainly dependent on insulin therapy and current formulated insulin formulations are used for its control all over the world. The presented study was designed to evaluate the potency of extracted, purified and formulated insulin from the pancreatic organs of the Sudanese beef cattle. Twenty healthy rabbits were used to conduct the study following subcutaneous administration of the sample insulin, to determine the hypoglycemic effect and to analyze the potency of the testing insulin by the hypoglycemic seizure method, blood sugar method and glucose enzymatic colorimetric test (GOD-PAP) respectively. The potency of the injected insulin samples was estimated by comparing the variation in blood glucose levels produced in the treated animals with that produced by a standard insulin preparation under the suitable conditions of the blood sugar method. The results revealed that the potency of the testing beef insulin samples was slightly higher (i.e., 2.2 USP units/ml, 9%) compared to the standard and assumed potency of the prepared insulin preparations (i.e., 1-2 USP units/ml) which indicated that the solvents and diluents used to prepare the assay dilution might be of higher potency and must be diluted to such an extent that the testing insulin potency must be compatible with the standard dilutions. Furthermore, to determine the choice of an assay to analyze the potency of insulin preparations, not only the accuracy of the result but also the purpose for which the test is to be used and the time limit must be taken into consideration.

Photosynthetic Performance and Potency of <i>Cannabis sativa</i>L. Grown under LED and HPS Illumination

Light-emitting diodes (LEDs) can be used as an energy efficient alternative to high-pressure sodium (HPS), which have historically been the standard for supplemental lighting in cannabis cultivation. However, there is a lack of scientific understanding in the cannabis industry regarding plant physiology, which has resulted in the adoption of cannabis cultivation methods based on hearsay rather than scientific research. The goals of this study were to 1) compare LED lighting options that are commonly used in the cannabis industry and 2) compare the top performing LED light with an industry standard HPS light. Specifically, three LED lights were compared (California Light Works ((SolarSystem 1100), BIOS Lighting (Icarus Gi2), and Fluence Bioengineering (now Fluence by Osram) (SPYDR xPLUS)), based on light distribution, leaf temperature, and photosynthetic performance indices. The LED versus HPS comparison was based on light response curves measured at photosynthetic photon flux densities (PPFD) of (0, 100, 200, 300, 400, 500, 750, 1000, 1250, 1500, 1750 and 2000 μmol∙m−2∙s−1), carbon assimilation rates (A) μmol CO2 m−2∙s−1 using a LiCor-6800 and resulting cannabinoid potency (THCA). The SPYDR xPLUS-Fluence by Osram had the highest performing LED light used in the LED comparison. At the suggested distance from bulb to canopy in the HPS versus LED comparison (6 inches for LEDs and 4 ft for HPS), carbon assimilation rates displayed a 142% percent increase in plants grown under LED vs. HPS with average photon flux densities of 795 and 298 μmol∙m−2∙s−1 for LED and HPS, respectively. All cultivars of Cannabis sativa L. showed increased cannabinoid potency when grown under LED illumination. The results of this study provide further insight regarding the selection of supplemental light to achieve maximum productivity of Cannabis sativa L.

Confidence limit calculation for antidotal potency ratio derived from lethal dose 50

AIM:To describe confidence interval calculation for antidotal potency ratios using bootstrap method.METHODS:We can easily adapt the nonparametric bootstrap method which was invented by Efron to construct confidence intervals in such situations like this.The bootstrap method is a resampling method in which the bootstrap samples are obtained by resampling from the original sample.RESULTS:The described confidence interval calculation using bootstrap method does not require the sampling distribution antidotal potency ratio.This can serve as a substantial help for toxicologists,who are directed to employ the Dixon up-and-down method with the application of lower number of animals to determine lethal dose 50 values for characterizing the investigated toxic molecules and eventually for characterizing the antidotal protections by the test antidotal systems.CONCLUSION:The described method can serve as a useful tool in various other applications.Simplicity ofthe method makes it easier to do the calculation using most of the programming software packages.

Dual role of lipids for genome stability and pluripotency facilitates full potency of mouse embryonic stem cells

While Mek1/2 and Gsk3βinhibition("2i")supports the maintenance of murine embryonic stem cells(EsCs)in a homogenous naive state,prolonged culture in 2i results in aneuploidy and DNA hypomethylation that impairs developmental potential.Additionally,2i fails to support derivation and culture of fully potent female ESCs.Here we find that mouse ESCs cultured in 2i/LIF supplemented with lipid-rich albumin(AlbuMAx)undergo pluripotency transition yet maintain genomic stability and full potency over long-term culture.Mechanisticaily,lipids in AlbuMAx impact intracellular metabolism including nucleotide biosynthesis,lipid biogenesis,and TCA cycle intermediates,with enhanced expression of DNMT3s that prevent DNA hypomethylation.Lipids induce a formative-like pluripotent state through direct stimulation of Erk2 phosphorylation,which also alleviates X chromosome loss in female ESCs.Importantly,both male and female"all-ESc"mice can be generated from de novo derived ESCs using AlbuMAXbased media.Our findings underscore the importance of lipids to pluripotency and link nutrient cues to genome integrity in early development.

Effects of linker amino acids on the potency and selectivity of dimeric antimicrobial peptides

Dimerization is an effective strategy for designing antimicrobial peptides that combine the advantages of different native peptides. In this study, we explored the effects of different linker amino acids, including leucine, proline and aminocaproic acid, on the anticancer, antimicrobial and hemolytic activities of the heteromeric antimicrobial peptides AM-1, AM-2, and AM-3. Proline and aminocaproic acid are ideal linkers for increasing the potency and selectivity of heteromeric antimicrobial peptides. The results of MD simulations provided a rationalization for this observation. Both AM-2, which had a proline linker,and AM-3, which had an aminocaproic acid linker, adopted a compact conformation in water and a bent conformation in membranes. This change in the flexible structures of AM-2 and AM-3 could have resulted in decreased binding of these peptides to zwitterionic lipid bilayers and increased damage to mixed lipid bilayers containing acidic phospholipids. In short, these findings obtained via assessing the effects of linker amino acids will contribute to the design of ideal heteromeric antimicrobial peptides with high selectivity and potency.

重组全人源抗表皮生长因子受体单克隆抗体生物学活性检测方法的Potency验证

目的对已建立的重组全人源抗表皮生长因子受体(epidermal growth factor receptor,EGFR)单克隆抗体生物学活性检测方法进行Potency验证。方法共4个验证批次,每个批次包括1个对照组和3个考察组,对照组抗体浓度设定为100%,8个浓度梯度,考察组抗体浓度为对照组的60%、70%、80%、100%、120%、130%和140%。用已建立的生物学活性检测方法测定对照组和各考察组的抗体活性,得到的结果用PLA 3.0软件进行分析,判断回归、线性、平行性、等效性是否合格,并得到实测效能。准确度以回收率表示。并以标称效能为横坐标,实测效能为纵坐标作线性回归,验证其线性和范围。结果各考察组的回归、线性、平行性、等效性均合格;各考察组的回收率均值在80%~125%范围内;标称效能与实测效能作线性回归,y轴截距为-1.716 5,斜率为0.984 6,相关系数为0.867 9,均符合标准。结论该方法用于评价目标活性的样品浓度区间在上述考察范围内(即60%~140%)时,可以真实地反映抗EGFR单抗的生物学活性。

Antibacterial potency screening of Capparis zeylanica Linn.

Objective:To conduct the antibacterial potency and minimum inhibitory concentration of extracts(n-hexane,acetone,chloroform and methanol)obtained from the root,leaf and stem of Capparis zeylanica.Methods:The powdered leaf,root and stem samples were Soxhlet extracted sequentially in n-hexane,acetone,chloroform and methanol.Antibacterial potency was evaluated by following the agar diffusion method and amoxicillin disc was used as a control.Results:In vitro antibacterial activity against 12 bacteria was performed with crude extracts.Among them,all the bacteria showed the moderate activity but chloroform and methanolic extracts showed promising antibacterial potency against Staphylococcus aureus,Sarcina lutea,Bacillus megaterium,Bacillus subtilis,Salmonella typhi and Shigella dysenteriae(leaf>root>stem).This activity was evaluated using disc diffusion method with a standard antibiotic,30μg/disc of amoxicillin.Conclusions:Strong antibacterial potency of chloroform and methanolic extracts provides new antibacterial compounds.

“Too Soon on Earth”: A Biophilosophical Model of Schizophrenia. Some Implications for Humanoid Robots

This paper presents a new explanatory model for schizophrenia based upon philosophical, molecular and neurobiological hypotheses as well as on years of experience in observing and treating these patients. To start with, a novel interpretation of the Hegelian concept of mediation is presented. Mediation is defined as the rejection of non-realizable programs, such as thoughts and ideas, at a certain point in time in the evolution of a living system. Whenever a system treats non-realizable programs as if they were realizable, its ability to “test the reality” is lost, and consequently a loss of ego-boundaries may occur. On the molecular level, I will try to show how “non-splicing” of introns during the mRNA splicing process is equivalent to a loss of the rejection function corresponding to mediation. At the cellular level in the brain, mediation can be explained in terms of glial-neuronal interactions. Glia exert a spatio-temporal boundary setting function determining the grouping of neurons into functional units. Mutations in genes that result in non-splicing of introns can produce truncated (“chimeric”) neurotransmitter receptors. I propose that such dysfunctional receptors are generated in glial cells and that they cannot interact properly with their cognate neurotransmitters. The glia will then lose their inhibitory-rejecting function with respect to the information processing within neuronal networks. This loss of glial boundary setting could be an explanation for the loss of ego or body boundaries in schizophrenia. Pertinent examples of case studies are given attempting to deduce the main symptoms of schizophrenia from the proposed hypothesis. Some implications for the design of delusional robots are also discussed. Finally, the evolutionary potency of non-coding introns is philosophically interpreted that schizophrenics may be “too soon on earth”.

In-silico Prediction of the Sweetness of Aspartame Analogues from QSPR Analysis

The extensive utilization of the low-energy dipeptide sweetener aspartame in foods leads to various studies on searching for new sweeteners in series. However, the real mechanistic cause of their sweetness power is still not completely known owing to their complex interactions with human sweet receptor, which may be different from that of other sweeteners to some extent. In this contribution, predictive quantitative structure-property relationship(QSPR) models have been developed for diverse aspartame analogues using Materials Studio 5.0 software. The optimal QSPR model(r2 = 0.913, r2 CV = 0.881 and r2 pred = 0.730) constructed by the genetic function approximation method has been validated by the tests of cross validation, randomization, external prediction and other statistical criteria, which shows that their sweetness power is mainly governed by their electrotopological-state indices(S_sssCH and S_ssNH), spatial descriptors(Shadow length: LX, ellipsoidal volume and Connolly surface occupied volume) and topological descriptors(Chi(3): cluster and Chi(0)(valence modified)), which partially supports both multipoint attachment theory proposed by Nofre and Tinti et al. and B-X theory proposed by Kier et al.. Present exploited results provide the key structural features for the sweetness power of aspartame analogues, supplement the mechanistic understanding of the sweet perception, and would be also helpful for the design of potent sweetener analogs prior to their synthesis.

Quality of compounded topical 2% diltiazem hydrochloride formulations for anal fissure

AIM:To investigate the quality of topical 2%diltiazem formulations extemporaneously compounded by retail pharmacies openly offering drug-compounding services.METHODS:A participating healthcare professional wrote 12 prescriptions for compounded 2%diltiazem cream,with 2 refills allowed per prescription.The 12sets of prescriptions were filled,at intervals of 1-2 wk between refills,at 12 different independent retail pharmacies that openly offer drug-compounding services in a major metropolitan region.The 36 resultant preparations,provided as jars or tubes,were shipped,as soon as each was filled,at ambient temperature to the study core laboratory for high-performance liquid chromatography(HPLC)analysis,within 10 d of receipt.For the HPLC analysis,8 different samples of the topical diltiazem,each approximately 1 g in weight,were taken from prespecified locations within each container.To initiate the HPLC analysis,each sample was transferredto a 100 mL volumetric flask,to which methanol was added.The HPLC analysis was conducted in accordance with the laboratory-validated method for diltiazem in cream,ointment,and gel formulations.The main outcome measures were potency(percentage of label claim)and content uniformity of the compounded topical 2%diltiazem formulations.RESULTS:Of the 36 prescriptions filled,30 were packaged in jars and 6 were packaged as tubes.The prescriptions were specifically for cream formulations,but6 of the 12 pharmacies compounded 2%diltiazem as an ointment;for another pharmacy,which had inadequate labeling,the dosage form was unknown.The United States Pharmacopoeia(USP)standard for potency is 90%-115%of label claim.Of the 36 preparations,5(13.89%)were suprapotent and 13(36.11%)were subpotent.The suprapotent prescriptions ranged in potency from 117.2%to 128.5%of label claim,and the subpotent prescriptions ranged in potency from34.8%to 89.8%of label claim.Fourteen(38.9%)preparations lacked content uniformity according to the USP standard of 90%-110%potency and<6%relative standard deviation.Of the 30 formulations packaged in jars,12(40%)lacked content uniformity,while of the6 formulations packaged in tubes,2(33.3%)lacked content uniformity.Nine of the 12 pharmacies(75%)failed USP potency or content-uniformity specifications for at least 1 of the 3 prescription fills.For 5 of the 12pharmacies(41.7%),the mean potency across all three prescription fills was<90%of label claim.CONCLUSION:Patients prescribed topical 2%diltiazem for treatment of anal fissure frequently receive compounded formulations that are misbranded with respect to potency and that lack content uniformity.

Comparison of functional outcomes after retropubic, laparoscopic and robot-assisted radical prostatectomy: A meta-analysis

AIM: To assess the 6-mo and 12-mo functional outcomes after retropubic, laparoscopic and robot-assisted laparoscopic radical prostatectomy retropubic radical prostatectomy(RRP) laparoscopic radical prostatectomy(LRP); robot-assisted laparoscopic prostatectomy(RARP). METHODS: A literature search was conducted using the Pub Med, EMBASE, The Cochrane Library and the Web of Knowledge databases updated to March, 2014 for relevant published studies. After data extraction and quality assessment via the Newcastle-Ottawa Scale or the Cochrane collaboration's tool for assessing risk ofbias, meta-analysis was performed using Rev Man 5.1. Either a random-effects model or a fixed-effects model was used. Potential publication bias was assessed using visual inspection of the funnel plots, and verified by the Egger linear regression test. RESULTS: Thirty-seven studies were identified in total: 14 articles comparing LRP with RRP, 12 articles comparing RARP with RRP, and 11 articles comparing RARP with LRP. For urinary continence, a statistically significant advantage was observed in RARP compared with LRP or RRP both at 6 mo [odds ratio(OR) = 1.93; P < 0.01, OR = 2.23; P < 0.05, respectively] and 12 mo(OR = 1.47; P < 0.01, OR = 2.93; P < 0.01, respectively) postoperatively. The continence recovery rates after LRP and RRP, with obvious heterogeneity(6-mo: I2 = 74%; 12-mo: I2 = 75%), were equivalent(6-mo: P = 0.52; 12-mo: P = 0.75). In terms of potency recovery, for the first time, we ranked the three surgical approaches into a superiority level: RARP > LRP > RRP, with a statistically significant difference at 12 mo [RARP vs LRP(OR = 1.99; P < 0.01); RARP vs RRP(OR = 2.66; P < 0.01); LRP vs RRP(OR = 1.34; P < 0.05)], respectively. Meta-regression and subgroup analyses according to adjustment of the age, body mass index, prostate volume, Gleason score or prostate-specific antigen did not vary significantly. CONCLUSION: Current evidence suggests that minimally invasive approaches(RARP or LRP) are effective procedures for functional recovery. However, more high-quality randomized control trials investigating the long-term functional outcomes are needed.

Testing for Normality from the Parametric Seven-Number Summary

The objective of this study is to propose the Parametric Seven-Number Summary (PSNS) as a significance test for normality and to verify its accuracy and power in comparison with two well-known tests, such as Royston’s W test and D’Agostino-Belanger-D’Agostino K-squared test. An experiment with 384 conditions was simulated. The conditions were generated by crossing 24 sample sizes and 16 types of continuous distributions: one normal and 15 non-normal. The percentage of success in maintaining the null hypothesis of normality against normal samples and in rejecting the null hypothesis against non-normal samples (accuracy) was calculated. In addition, the type II error against normal samples and the statistical power against normal samples were computed. Comparisons of percentage and means were performed using Cochran’s Q-test, Friedman’s test, and repeated measures analysis of variance. With sample sizes of 150 or greater, high accuracy and mean power or type II error (≥0.70 and ≥0.80, respectively) were achieved. All three normality tests were similarly accurate;however, the PSNS-based test showed lower mean power than K-squared and W tests, especially against non-normal samples of symmetrical-platykurtic distributions, such as the uniform, semicircle, and arcsine distributions. It is concluded that the PSNS-based omnibus test is accurate and powerful for testing normality with samples of at least 150 observations.

Equi-Quantity, Equi-Calorie and Dose of Rice on Relative Glycemic and Insulinemic Response in Diabetic Patients

Background and Aim: Diabetes mellitus is a chronic metabolic disorder with high blood sugar level. The postprandial glycemic impact of foods depends on the insulin status, which is deranged completely in a type 2 diabetic person. Dietary management of this group largely focuses on the low glycemic index (GI) food, based on equi-carbohydrate comparison, to keep the blood sugar level close to normal. But we consume whole food, along with other co-nutrients, moisture, fibre etc. The present study is aiming to assess the impact of main staple food rice with regards to Equi-Quantity, Equi-Calorie and Dose on relative glycemic and insulinemic response in diabetic patients as compared to normal group. Method: Blood samples of diabetic patients with stable blood sugar under medicinal treatment and paired normal patients (n = 6 + 6) were collected after an overnight fast and up to 2 hours post consumption of test and standard food on different occasions. Glucose and insulin levels were measured using glucometer (Abbott pharmaceuticals) and ECLIA method. Result: Equi-quantity of rice exerts a much lower glycemic and insulinemic response in comparison with bread in both normal and diabetic individuals and the response to rice does not show a proportional increase even when the quantities are doubled. Rice has higher moisture content which acts as energy diluent, decreasing the total starch in equivalent quantities. Equi-calorie (132 kcal) quantity comparison of rice (100 g) and bread (50 g) showed a much lower glycemic and insulinemic impact on rice in diabetic individuals, even though quantity is double and satiety level reaches earlier than low moisture food bread in equi-calorie quantity. The normal individuals, with normal insulin response can control the glycemic response to lower levels than those of diabetic subject. Conclusion: Rice having lower glycemic and insulinemic impact is a better suited food for diabetic individuals who already have a compromised insulin status.

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 107 FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.

Synthesis of Gemcitabine-(C₄-<i>amide</i>)-[anti-HER2/<i>neu</i>] Utilizing a UV-Photoactivated Gemcitabine Intermediate: Cytotoxic Anti-Neoplastic Activity against Chemotherapeutic-Resistant Mammary Adenocarcinoma SKBr-3

Gemcitabine is a pyrimidine nucleoside analog that becomes triphosphorylated intracellularly where it competitively inhibits cytidine incorporation into DNA strands. Another mechanism-of-action of gemcitabine (diphosphorylated form) involves irreversible inhibition of the enzyme ribonucleotide reductase thereby preventing deoxyribonucleotide synthesis. Functioning as a potent chemotherapeutic gemcitabine promote decreases in neoplastic cell proliferation and apoptosis which is frequently found to be effective for the treatment of several leukemias and a wide spectrum of carcinomas. A brief plasma half-life in part due to rapid deamination and chemotherapeutic-resistance restricts the utility of gemcitabine in clinical oncology. Selective “targeted” delivery of gemcitabine represents a potential molecular strategy for simultaneously prolonging its plasma half-life and minimizing innocient tissues and organ systems exposure to chemotherapy. The molecular design and an organic chemistry based synthesis reaction is described that initially generates a UV-photoactivated gemcitabine intermediate. In a subsequent phase of the synthesis method the UV-photoactivated gemcitabine intermediate is covalently bonded to a monoclonal immunoglobulin yielding an end-product in the form of gemcitabine-(C4-amide)-[anti-HER2/neu]. Analysis by SDS-PAGE/chemiluminescent auto-radiography did not detect evidence of gemcitabine-(C4-amide)-[anti-HER2/neu] polymerization or degradative fragmentation while cell-ELISA demonstrated retained binding-avidity for HER2/neu trophic membrane receptor complexes highly over-expressed by chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3). Compared to chemotherapeutic-resistant mammary adenocarcinoma (SKBr-3), the covalent immunochemotherapeutic, gemcitabine-(C4-amide)-[anti-HER2/neu] is anticipated to exert greater levels of cytotoxic anti-neoplastic potency against other neoplastic cell types like pancreatic carcinoma, small-cell lung carcinoma, neuroblastoma, glioblastoma, oral squamous cell carcinoma, cervical epitheliod carcinoma, or leukemia/lymphoid neoplastic cell types based on their reported sensitivity to gemcitabine and gemcitabine covalent conjugates.

Anti-trypanosomal Activity of Bufonidae(Toad)Venom Crude Extract on Trypanosoma brucei brucei in Swiss Mice

Trypanosomiasis afflicts about 6~7 million people globally and to a large extent impedes livestock production in Africa.Naturally,trypanosomal parasites undergo genetic mutation and have developed resistance over a wide range of therapies.The utilization of animals and plants products has presented therapeutic potential for identifying novel anti-trypanosomal drugs.This study evaluated toad venom for anti-trypanosomal potency in-vivo in Swiss mice.Toads were collected from July to August 2019.The acute oral toxicity and biochemical characterization of the toad venom were determined.The experimental mice were administered various doses(130 mg/kg,173 mg/kg and 217 mg/kg)of the toad venom crude extract and 0.75 mg/mL of Diamizan Plus standard drug for the treatment of trypanosomiasis,once daily for 3 days.The in-vivo anti-trypanosomal activity was evaluated by a curative test,after infecting the mice with Trypanosoma brucei brucei.The pre-patent period was 72 hours before treatment commenced.The overall results showed that trypanosomal load was highest in the control group while the group treated with Diamizan drug had the least trypanosomal load.As such,the mean trypanosomal load in relation to treatments showed a very high significant difference(P<0.05).Also,the mean trypanosomal load in Swiss mice in relation to the highest dosage of toad venom versus Diamizan drug showed a very high significant difference(P<0.05).The mean change in relation to the haematological parameters across treatments groups varied significantly(P<0.05)with the exception of Hb which showed no significant difference(P>0.05)across treatment groups.The over 50%reduction in the trypanosomal load in the 130 mg/kg group in comparison with the control group brings to bare the anti-trypanosomal potency of the toad venom.The anti-trypanosomal activity demonstrated by the toad venom has provided basis for development of new therapeutic agents from different toad species.The study recommends further studies(both in-vivo and in-vitro)followed by the characterization of the active compounds present in the toad venom responsible for the anti-tyrpanosomal activity observed alongside the management and conservation of these species.