Pharmacology Laboratory Research Articles

Effectiveness of Short-Term Use Denosumab and Risedronate Using β-Crosslaps and Histopathology as a Parameter in Osteoporotic Rat Model

Osteoporosis is a condition of decreased bone mass density. Pharmacological management uses drugs that decrease bone resorption or increase bone formation. The most commonly used drugs are bisphosphonates. Risedronate is one of these bisphosphonates. It inhibits osteoclasts, resulting in osteoclast apoptosis. Denosumab is a human monoclonal antibody that inhibits receptor activator kappa-B ligand, inhibiting osteoclast activation. Both drugs are widely used. Research on the efficacy of these two mechanisms has yet to obtain conclusive results. β-cross-laps is a parameter that can be used to evaluate the effectiveness of therapy through monitoring the bone resorption process. This experimental study used female rats >9 weeks old and was conducted at the Pharmacology and Therapeutic Laboratory, Universitas Padjadjaran Bandung, Indonesia, from June to September 2021. This study utilized a simple random sampling to allocate 24 experimental animals into three groups: control, risedronate, and denosumab. β-Crosslaps expression values before ovariectomy, post ovariectomy before receiving medication, and post ovariectomy and receiving medication was recorded and statistically analyzed using the SPSS version 24.0. The analysis of 24 samples revealed a statistically significant decrease in the median value of β-Crosslaps after ovariectomy in the denosumab group (p=0.036) when compared to the control group, whereas the decrease in the risedronate group was not significant (p=0.687). Administration of denosumab in rat models is more effective in reducing bone resorption compared to risedronate.

Potential of 6′‑hydroxy justicidin B from Justicia procumbens as a therapeutic agent against coronavirus disease 2019

BackgroundSince the onset of the coronavirus disease 2019 (COVID-19) pandemic, remarkable advances have been made in vaccine development to reduce mortality. However, therapeutic interventions for COVID-19 are comparatively limited despite these intensive efforts. Furthermore, the rapid mutation capability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a characteristic of its RNA structure, has led to the emergence of multiple variants, necessitating a shift from a predominantly vaccine-centric approach to one that encompasses therapeutic strategies. 6′-Hydroxy justicidin B (6′-HJB), an arylnaphthalene lignan isolated from Justicia procumbens, a traditional Chinese medicine, is known for its antiviral properties. Hypothesis/PurposeThe aim of the present study was to assess the effectiveness and safety of 6′-HJB against SARS-CoV-2 in order to determine its potential as a therapeutic agent against COVID-19. MethodsThe efficacy of 6′-HJB was evaluated both in vitro using Vero and Calu-3 cell lines and in vivo using ferrets. The safety assessment included toxicokinetics, safety pharmacology, and Good Laboratory Practice (GLP)-compliant toxicity evaluations following single- and repeated-dose toxicity studies in dogs. ResultsThe anti-SARS-CoV-2 efficacy of 6′-HJB was evaluated through dose-response curve (DRC) analysis using immunofluorescence; 6′-HJB demonstrated superior inhibition of SARS-CoV-2 growth and lower cytotoxicity than remdesivir. In SARS-CoV-2-infected ferret, 6′-HJB showed efficacy comparable to that of the positive control, Truvada. Further GLP toxicity studies corroborated the safety profile of 6′-HJB. Single-dose and 4-week repeated oral toxicity studies in Beagle dogs demonstrated minimal harmful effects at the highest dosages. The lethal dose of 6′-HJB exceeded 2,000 mg kg-1 in Beagle dogs. Toxicokinetic and GLP safety pharmacology studies demonstrated no adverse effects of 6′-HJB on metabolic processes, respiratory or central nervous systems, or cardiac functions. ConclusionThis research highlights both the antiviral efficacy and safety profile of 6′-HJB, underscoring its potential as a novel COVID-19 treatment option. The potential of 6′-HJB was demonstrated using modern scientific methodologies and standards.

Anesthesia and Analgesia Recommended for Painful Procedures in Small Animals

The non-invasive but painful and invasive procedures are frequently performed in experimental animal studies, such as blood withdrawal, route of administration, and surgical procedures. Most of these procedures commonly conducted under anesthesia and/or analgesia and should be complied with the ethical requirements. However, pain is difficult to assess in animals, therefore, it is advisable to provide the analgesics pre-emptively or if painful condition is expected. Preanesthetic like tranquilizer is generally used to reduce the stress in animal and further it also aids in the smooth induction of anesthesia and recovery. In view of this, the objective is to demonstrate and discuss the guidelines of anesthesia and analgesia in small animals, especially mouse and rat with principal focus to ethical purview. These guidelines can be applicable to the animal house facility, pharmacology laboratory, and to a large extent for veterinary care.

In vitro and In vivo Studies for Screening of Anti-inflammatory, Thrombolytic and Acute Toxicological Activity in Camellia Chrysantha (Hu) Tuyama

Purpose: The objective of this study was to investigate the impact of methanol-prepared leaves extract of Camellia Chrysantha (Hu) Tuyama. on several in vitro activities, including anti-inflammatory and thrombolytic effects. Additionally, the study aimed to assess the acute toxicological activity of the extracts in an animal model. Place and Duration of Study: The research study was carried out from Nov 2023 to Feb 2024 at the Bangladesh Council of Scientific and Industrial Research (BCSIR), Laboratory of Pharmacology, University of Asia Pacific and the Laboratory of Phytochemistry and Pharmacology, Department of Pharmacy, as well as the Laboratory of Microbiology at Stamford University Bangladesh, Dhaka. Methodology: The methodology involved various doses of methanolic leaves extract of Camellia Chrysantha (Hu) Tuyama, and employed techniques such as the Inhibition of Protein Denaturation Assay for anti-inflammatory test and the clot lysis test for thrombolytic assay. The acute toxicological activity was assessed by Cinnamon oil induced toxicological and administration of acetic acid induced writhing responses in Swiss albino mice. Result: The Inhibition of Protein Denaturation Assay test yielded findings indicating its anti-inflammatory properties against all strains used in this research. The findings from the study examining the anti-inflammatory properties indicate that Chrysantha (Hu) Tuyama exhibits a significant efficacy of 86.54% in inhibiting inflammation at a concentration of 1000 µg/mL. This level of efficacy is comparable to that of acetylsalicylic acid, which also has a similar efficacy of 98.54 % at the same concentration. Thrombolysis test is a controlled laboratory procedure that assesses the ability of plant extracts to dissolve blood clots. The study found that MECCL can reduce blood clots. Table 2 shows that MECCL's thrombolytic potential was 95.69%, which is much higher than the usual value of 91.304%. Our research found that MECCL did not induce any harmful effects in mice during the acute toxicity test, as determined by the toxicological assessment. No signs of illness or death were reported throughout the acute toxicity trial. Conclusion: The Camellia chrysantha (Hu) Tuyama plant exhibits privileged qualities that could lead to the discovery of novel therapeutic substances. It has anti-inflammatory, thrombolytic, and mild acute toxicity properties. Further exploration of this plant could be beneficial. Our study results contribute to the growing body of evidence demonstrating that natural products may potentially have favourable effects on human health.

CHEMICAL ADHERENCE IN RESISTANT HYPERTENSION: PILOT STUDY IN AN ESH EXCELLENCE CENTER

Objective: Arterial hypertension (HTN) is responsible for the largest proportion of preventable mortality in the world. The primary cause of resistant hypertension (rHTN) is poor medication adherence. Blood or urine drug assays enable us to assess chemical adherence more accurately than self-reported questionnaires. We assayed anti-hypertensive drugs in patients with rHTN at our Center of Excellence in Hypertension, in collaboration with the Pharmacology and Toxicology Laboratory. Design and method: Patients with rHTN (office SBP/DBP >140/90 mmHg or out of office daytime >135/85 mmHg despite being prescribed 3 drugs at optimal dosage, including a diuretic, a renin angiotensin system inhibitor (RASI) and a calcium channel blocker (CCB), were informed of the study and agreed to have blood drugs concentrations measured. We describe the results for chemical adherence in the first 54 patients included between January 1, 2021 and September 1, 2023 in our center. Results: Of the 54 patients included, 38 (70%) were men, with a mean age of 65 +/- 11 years. The mean number of antihypertensive treatments prescribed was 5 +/- 1 (min 3, max 8). The proportions of treatments were RASI 87%, CCB (mainly dihydropyridines) 81%, diuretics 81%, beta-blockers 76%, alpha-blockers 57%, mineralocorticoid receptor antagonists (MRA) 37%, and central antihypertensives 26%. The best compliance was observed with alpha-blockers 81%, followed by CCB 50%, RASI 45%, diuretics 37% and beta-blockers 34%. The least compliant were central antihypertensives 0% and MRA 5%. All prescribed drugs were completely absent in the blood in 26% of patients, and 63% of patients had incomplete adherence (1 or more but not all drugs present in the blood), of whom 1/3 were non-compliant to 3 or more medications. Only for 9% of patients chemical adherence was excellent. Conclusions: Chemical adherence when measured at clinical visit was poor in this cohort of patients followed in a center of excellence. Limitations of this study are lack of follow-up for the same patient to assess variation in adherence over time, and small sample size. These results suggest the need to develop therapeutic education to improve patient involvement in the management of HTN and compliance.

The Amentoflavone-Rich Extract of Ouratea fieldingiana Leaves Presents Antioxidant and Anti-inflammatory Activity in Mice

Aims: To evaluate the antioxidant and anti-inflammatory effects of the Ouratea fieldingiana leaf extract (OFE) and its main constituent amentoflavone (AMT). Study Design: The phenolic compounds of Ouratea fieldingiana leaves (OFE) were quantified, the chemical structure characterized and the anti-inflammatory effect of OFE and amentoflavone was evaluated in mice paw edema and peritonitis. Place and Duration of Study: The work involved the partnership of the Natural Products Chemistry Laboratory and Inflammation Physio pharmacology Laboratory, between July 2020 and June 2021. Methodology: The OFE was obtained from 70% ethanol extract of O. fieldingiana. OFE was subjected to classic chromatographic column furnishing AMT. The phenolic compounds were characterized by HPLC and quantified by Follin-Ciocalteu methodology. The antiradical potential was evaluated by DPPH and ABTS methods. Anti-inflammatory activity was determined in female Swiss mice received per oral OFE (0.1-10 mg/kg) or sterile saline 60 min before stimulation with carrageenan (300 mg) for quantification of the parameters: edema, abdominal hypernociception, neutrophil migration and oxidative stress markers (reduced glutathione, malondialdehyde, myeloperoxidase and catalase activities). Results: AMT was identified as the main compound in the OFE (292.64 ± 3.87 mg/g extract). OFE has an average inhibitory concentration of 9.81 ± 0.17 µg/mL extract to inhibit the DPPH radical. OFE and AMT inhibited paw edema [OFE: 32% (0-2 h), 28% (2-4 h); AMT: 55% (0-2h); 51% (2-4 h)], neutrophil migration (OFE: 65%; AMT: 67%); myeloperoxidase activity (OFE: 37%; AMT: 45%) and abdominal hypernociception (OFE: 31%; AMT: 35%). OFE, but not AMT, increased catalase activity (57%) and reduced glutathione (62%), but decreased malondialdehyde (69%). Conclusion: The amentoflavone-rich extract from O. fieldingiana leaves presents antioxidant activity in vitro and in vivo, and anti-inflammatory activity in vivo, corroborating the popular use of the plant.

Exploration of “Bilingual + Ideological and Political Theory” Integration in Pharmacology Laboratory Teaching of Pharmaceutical Engineering Majors

Exploration of “Bilingual + Ideological and Political Theory” Integration in Pharmacology Laboratory Teaching of Pharmaceutical Engineering Majors

Assessing drug safety by identifying the axis of arrhythmia in cardiomyocyte electrophysiology

Many classes of drugs can induce fatal cardiac arrhythmias by disrupting the electrophysiology of cardiomyocytes. Safety guidelines thus require all new drugs to be assessed for pro-arrhythmic risk prior to conducting human trials. The standard safety protocols primarily focus on drug blockade of the delayed-rectifier potassium current (IKr). Yet the risk is better assessed using four key ion currents (IKr, ICaL, INaL, IKs). We simulated 100,000 phenotypically diverse cardiomyocytes to identify the underlying relationship between the blockade of those currents and the emergence of ectopic beats in the action potential. We call that relationship the axis of arrhythmia. It serves as a yardstick for quantifying the arrhythmogenic risk of any drug from its profile of multi-channel block alone. We tested it on 109 drugs and found that it predicted the clinical risk labels with an accuracy of 88.1–90.8%. Pharmacologists can use our method to assess the safety of novel drugs without resorting to animal testing or unwieldy computer simulations.

The effectiveness of Jamblang (Syzygium cumini (L) Skeels) extract on the atherogenic index in a diabetic rat model

Jamblang fruits (Syzygium cumini (L.) Skeels) have the potential as an antioxidant by facilitating glucose and lipid metabolism for reducing the atherogenic index (AI). This study aimed to explore the effect of the Jamblang extract on the atherogenic index in a diabetic rat model. This pre- and post-control group design study was conducted in the Pharmacology Laboratory of Universitas Sumatera Utara. Twenty-five rats were divided into five groups. Alloxan 120 mg/kg BW was administrated intraperitoneally, and the rats' fasting blood glucose levels were measured four days later. Group I (negative control) were treated with aqua dest, group II (positive control) was treated with glibenclamide 1,25 mg /kg BW, and group III, IV, and V were given an extract of Jamblang at a dose of 125, 250, and 500 mg/kg BW for seven days. Trigliserida and HDL-C measured using spektrofotometer microlab. The data were subjected to the Kruskal-Wallis test to examine the effect of Jamblang extract in different doses on the atherogenic index, followed by a Mann- Whitney Post hoc test. The result revealed that the extract Jamblang dose of 125, 250, and 500 mg/kg BW significantly decreased the atherogenic index (p value= 0,012<0,05). In conclusion, the Jamblang (Syzygium cumini (L.) Skeels) with the best-tested dose was 500 mg/kgBW to decrease the atherogenic index until 0,42 in vivo study.

Linta meliala,roito yuliana UJI EFEKTIVITAS ANTIDIARE EKSTRAK ETANOL DAUN GAGATAN HARIMAU (Vitisgracilis BL) PADA TIKUS JANTAN PUTIH

Abstract Diarrhea is the abnormal passage of loose or watery stools, with a frequency of more than three times a day. Diarrhea can be caused by bacteria. Gagatan Harimau leaves are known to have antibacterial properties. This research was conducted using an experimental method in the field of pharmacology at the Pharmacology Laboratory, Faculty of Pharmacy, Deli Husada Health Institute Delitua, using 25 rats induced with castor oil and divided into 5 treatment groups: negative control (1% CMC Na), positive control (Loperamide HCl), and treatment groups with ethanol extract of Gagatan Harimau leaves at doses of 50 mg/kg body weight (BW), 100 mg/kg BW, and 200 mg/kg BW. The sample used was Gagatan Harimau leaves (Vitis graillis BL), which were extracted by maceration using 96% ethanol as the solvent and concentrated using a rotary evaporator. The phytochemical screening results of Gagatan Harimau leaves showed the presence of flavonoids, saponins, alkaloids, and tannins. The results of the antidiarrheal effect test showed that the ethanol extract of Gagatan Harimau leaves at doses of 50, 100, and 200 mg/kg BW had antidiarrheal effects on male rats. The 200 mg/kg BW dose of ethanol extract showed the most effective antidiarrheal effect compared to the 50 and 100 mg/kg BW doses. The extract from Gagatan Harimau leaves (Vitis gracillis BL) exhibited antidiarrheal effects on male white rats induced with castor oil.

BINAHONG [Anredera cordifolia (Tenore) Steenis] LEAF INFUSA FOR SUTURE WOUND INFECTION CONVALESCENCE

Background: Postpartum infections are the top 4 main causes of the high maternal mortality rate in Indonesia at 2022 of 183 per 100,000 live births. Efforts to prevent complications due to infection of puerperal suture wounds with pharmacological therapy using antibiotics and antiseptics, as well as non-pharmacological ones using plant extracts. Binahong is believed to contain flavonoids, terpenoids, saponins, tannins, essential oils, and alkaloids, which are especially useful in healing wound infections. The aim of the study was to determine the effectiveness of infusion of binahong leaves (Anredera cordifolia (Tenore) Steenis) on the healing of suture wound infections in rats (Rattus norvegicus) experimental animals. Method: The research method was a true-experimental posttest only control group design, the study was conducted from June to August 2023 at the Microbiology and Pharmacology Laboratory of Muhammadiyah Sidoarjo University, the research variables were binahong leaf infusion and healing of suture infection, assessment and data collection using the REEDA scale instrument, statistical analysis with the one-way ANOVA, and post-hoc follow-up tests. Result: The results showed p <0.05, so there was a significant effect of giving binahong leaf infusion on the healing of suture wound infections. In terms of the difference in the mean value and the results of the post-hoc significant difference test, it appears that the 5% binahong leaf infusion group showed the most effective results in healing suture wound infections and was significantly different from the negative control group with less effective healing of suture infections. Conclusion : This is in line with research that states that the group given a 5% concentration of binahong extract was more effective than the group with a higher concentration. Thus, administration of a 5% concentration of binahong leaf infusion can be an alternative for healing suture wound infections due to the important content in it in the form of flavonoids, terpenoids, saponins, tannins, essential oils, and alkaloids. Further research is needed with lower concentrations of binahong leaf infusion to find out the minimum concentration that can have a good effect on the convalescence of suture wound infections.

The history of the Department of Pharmacology, Institute of Experimental Medicine (to the 100th anniversary of the department)

The Department of Pharmacology of the Institute of Experimental Medicine was established in the autumn of 1923 on the initiative of the institutes leadership. Professor Nikolai Pavlovich Kravkov (18651924), the most respected domestic pharmacologist at that time and the head of the Department of Pharmacology of the Military Medical Academy, was invited as the alleged head of the department. However, N.P. Kravkovs untimely death in April 1924 precluded the department from developing extensive research in Pharmacology. Professor Vladimir Vasilyevich Savich (18741936), a student of I.P. Pavlov who focused his main research on the effect of medicinal substances on the nervous regulation of the cardiovascular system, gastrointestinal tract, endocrine system, and water metabolism, led the department from 1924 to 1936. The mechanism of the direct impact of drugs and poisons on organs and tissues was studied using both isolated organs, notably endocrine glands, and classical conditioned reflex techniques. In 1936, the department was disbanded in connection with the death of the head (V.V. Savich) and the reorganization of the institutes structure. Professor S.V. Anichkov (18921981), who later became the Hero of Socialist Labor, laureate of the Lenin and State Prizes of the USSR, and academician of the USSR Academy of Medical Sciences, revived the department in 1948 and remained its permanent head until his death (in July 1981). Along with him were well-known researchers Professors V.M. Karasik, N.V. Khromov-Borisov, I.S. Zavodskaya (who led the department from 1981 to 1984), Y.S. Borodkin (who led the department from 1984 to 1992), N.A. Kharauzov, V.E. Ryzhenkov, N.S. Sapronov (who led the department from 1992 to 2011), P.P. Denisenko, and N.A. Losev. The main focus of the department from 1948 to 1984 was the integration of fundamental pharmacological advancements with the introduction of drugs into healthcare practice. Currently, the Department of Neuropharmacology. S.V. Anichkova is one of the largest research centers in Russia, focusing on fundamental research in Pharmacology. Professor P.D. Shabanov has been the head of the department since 2011. The department divided into four laboratories: the Laboratory of Chemistry and Pharmacology of Medicinal Products (led by Doctor of Medical Sciences, E.R. Bychkov), the Laboratory of General Pharmacology (led by Doctor of Biological Sciences, Prof. A.A. Lebedev), the Laboratory of Biochemical Pharmacology (led by Doctor of Medical Sciences, Prof. P.D. Shabanov), and the Laboratory of Synthesis and Nanotechnology of Medicinal Substances (led by Doctor of Biological Sciences, Prof. L.B. Piotrovsky).

Nephroprotective effect of ethanol extract Abelmoschus manihot l. Leaves in gentamicin-induced mice

Nephrotoxicity is the most common side effect of gentamicin, an aminoglycoside antibiotic group. It was mediated by oxidative stress mechanisms. Abelmoschus manihot L. leaves (AML) have antioxidant activity flavonoids. This study aims to determine the nephroprotective effect of AML in gentamicin-induced nephrotoxicity using mice models. It conducted an experimental design study in a pharmacology laboratory using mice models. It was conducted experimentally in 7 groups. There are normal, solvent, negative control, and four test groups (given an ethanolic extract of AML at doses of 50, 100, 200, and 400 mg/kgBW) which were administered orally for 7 days. Gentamicin 112 mg/kgBW was induced intraperitoneally on the 8th day. All mice were anesthetized on the 11th day to collect blood serum and renal organs for further examination. The data were analyzed using SPSS version 26 for One Way ANOVA test and followed by Duncan's post-hoc analysis test. The difference is considered significant if the p-value <0.05. Gentamicin successfully induced renal damage based on increasing creatinine and blood urea nitrogen (BUN) levels. This was by histopathology analysis which showed renal necrosis in the negative control group. Ethanolic extract of AML prevents nephrotoxicity due to gentamicin in a dose-dependent manner in creatinine level but not in BUN and histopathology. However, taken together all results showed that ethanol extract of AML has a nephroprotective effect in gentamicin-induced nephrotoxicity.
 Keywords: Abelmoschus manihot L., gentamicin, nephroprotection, renal histopathology

Evaluation of In-vitro Antimicrobial, Anti-arthritis and In-vivo Analgesic and Neuropharmacological Investigation of the Bark Extract of Solanum americanum Milli

Purpose: The objective of this study was to investigate the impact of methanol-prepared bark extract of Solanum americanum Milli. Bark on several in vitro activities, including antimicrobial and anti-arthritis effects. Additionally, the study aimed to assess the analgesic and neuropharmacological properties of the extracts in an animal model.
 Place and Duration of Study: The research study was carried out from March 2023 to July 2023 at the Bangladesh Council of Scientific and Industrial Research (BCSIR) and the Laboratory of Phytochemistry and Pharmacology, Department of Pharmacy, as well as the Laboratory of Microbiology at Stamford University Bangladesh, Dhaka.
 Methodology: The methodology involved various doses of methanolic bark extract of Solanum americanum Milli (MESAB), and employed techniques such as the Disk Diffusion Method for antimicrobial test and the Inhibition of Protein Denaturation Assay for anti-arthritic test. The analgesic efficacy was assessed by the hot plate test and administration of acetic acid induced writhing responses in Swiss albino mice. The mechanism test is conducted through the cyclic guanosine monophosphate (cGMP) test. The neuropharmacological inquiry included the measurement of locomotor activity through the utilization of open field and hole cross tests.
 Results: The Disk Diffusion Test of MESAB yielded findings indicating its antimicrobial properties against all strains used in this research. The findings from the study examining the anti-arthritis properties indicate that MESAB exhibits a significant efficacy of 94.59% in inhibiting arthritis at a concentration of 1000 µg/mL. This level of efficacy is comparable to that of diclofenac sodium, which also has a similar efficacy of 98.19% at the same concentration. The findings of the present study demonstrated that MESAB displayed analgesic properties in both analgesic models, suggesting the involvement of both central and peripheral pathways. Based on the results of the study, it was observed that the administration of MESAB at a dosage of 400 mg/kg had a significant analgesic effect. The application of MB before to treatment resulted in a further enhancement of this effect, leading to an increase in the percentage of inhibition from 66.75% to 79.70%, respectively. The consideration of the involvement of the GABA-benzodiazepine receptor in neuropharmacological activity holds significant importance. In the Open Field and Hole Cross Tests, it was shown that the extract exhibited significant result compared to Diazepam in terms of increasing motor coordination.
 Conclusion: In the future, plant-derived pharmacological compounds may offer potential benefits for various clinical disorders, such as neurodegenerative illnesses. These substances possess neuropharmacological properties, antibacterial effects, and anti-arthritic activity, which could potentially serve as alternatives to non-steroidal anti-inflammatory drugs (NSAIDs).

Control of Chemical Contaminants and Evaluation of Oral Subacute Toxicity of an Alcoholic Polyherbal Product “Plaie de ventre” Marketed for Health Claims in Yopougon (Côte d’Ivoire)

Objective: Alcoholic mixtures are highly appreciated by the Ivorian population to their low cost and for their health claims. Contaminants control and toxicity tests were carried out in order to check the harmlessness of extractable from polyherbal product “Plaie de ventre”.
 Place and Duration of Study: The study was conducted at the Laboratory of Pharmacology and Clinical Pharmacy of Félix Houphouët-Boigny University for a period of 4 months.
 Methods: The search for pesticide residues and metallic trace elements was carried out respectively by high-performance liquid phase chromatography and by atomic absorption with air-arc flame ethylene AAS 20, respectively. Subacute oral toxicity study involved daily administration of the extractable from the polyherbal product to three groups of rats for 28 days at doses of 21.06, 42.12, and 84.24 mg/kg bw.
 Results: The detected residues of pesticides and trace metals in product “Plaie de ventre” mixture were consistent with the standards for their use. Moreover, the administration of extractable from this mixture did not result in any alteration of the body or relative weights of the rats, the structure of the kidney and liver at different doses. Finally, the analysis of the hematological and biochemical parameters indicated a noteworthy escalation (p < 0.05) in levels of white blood cells, blood platelets, and serum triglycerides.
 Conclusion: The extract from the polyherbal “Plaie de ventre” caused changes in hematological and biochemical parameters compared to control group in treated rats. Consumers may face a long-term risk of developing hyperlipidemia.

Influence of the probiotic “Bioseven” on the intestinal biocenose of white rats

The use of probiotics in animal husbandry ensures the maintenance of homeostasis of the digestive tract and prevents the development of factor infections in young animals – colibacteriosis, dyspepsia, and others, promotes the growth of animals with high survival rates. The main advantages of probiotics over chemotherapeutic drugs and antibiotics are that they are harmless to animal organism and are ecologically clean. In connection with the great attention paid to probiotics as ecologically safe drugs, the study of biological properties and the selection of bacterial strains, the most promising in the probiotic sense, are currently being intensified. This is the direction of the selection of species-specific strains for the intestinal biocenosis of a specific animal species, which have high colonization and antagonistic properties. Therefore, the development of the scientific basis for the creation of new probiotic preparations gave an impetus to their improvement and continued research in this direction. The aim of the work was to study the effect of the probiotic “Bioseven” on the intestinal biocenosis of white rats depending on the dose. Objectives of the study: to establish the toxicity of the drug “Bioseven” on laboratory animals with a single dose (“acute toxicity”) according to microbiological indicators; to investigate the toxicity of “Bioseven” on laboratory animals with long-term administration (“subacute toxicity”) according to microbiological indicators. Research work was carried out during 2022 at the department of veterinary and sanitary examination, hygiene of animal husbandry products and pathanatomy named after J. S. Zagaevskii. The toxicological characterization of the probiotic drug “Bioseven” (study of subacute and acute toxicity) was carried out under the conditions of the State Research Control Institute of Veterinary Medicines and Feed Additives (Laboratory of Pharmacology and Toxicology), (Lviv), the manufacturer of the drug was PP “BTU-CENTER” Ladyzhyn, Vinnytsia region. On the 31st day of use of the drug “Bioseven” (a feed additive with probiotic action) in a therapeutic dose, microorganisms of the genus Lactobacillus were sown 0.3 lg CFU/g more from the large intestine of experimental rats, microorganisms of the genus Bifidobacterium were sown at 0.5 lg CFU/g more than in the control group. The number of Candida fungi decreased by 0.6 lg CFU/g, and the total number of Escherichia coli – 0.2 lg CFU/g. When using the “Bioseven” probiotic in a 5-fold therapeutic dose, 0.5 lg CFU/g more microorganisms of the Lactobacillus genus and 0.6 lg CFU/g more Bifidobacterium microorganisms were sown from the large intestine of experimental rats than in the control group. At the same time, the number of Candida fungi decreased by 0.7 lg CFU/g, and the total number of Escherichia coli to 0.4 lg CFU/g. Under the conditions of use of the drug “Bioseven” in a 10-fold therapeutic dose, microorganisms of the genus Lactobacillus were sown from the large intestine of experimental rats, 0.9 lg CFU/g more, microorganisms of the genus Bifidobacterium 1.2 lg CFU/g more than in the control the group At the same time, the number of Candida fungi decreased by 1.1 lg CFU/g, and the total number of Escherichia coli by 0.8 lg CFU/g. Prospects for further research consist in the study of the impact of the biotic drug “Bioseven” on indicators of metabolic processes and productive qualities of farm animals.

Pharmacological Investigation of In-vitro Anti-inflammatory, Antimicrobial, Thrombolytic, Cytotoxic and In vivo Analgesic Activities of Ethanolic Leaf Extract of Diospyros malabarica

Aims: This study was designed to investigate the effects of leaf extracts of Diospyros malabarica plant (EEDML) on a number of in vitro activities, including antibacterial, anti-inflammatory, cytotoxic, and thrombolytic activities, as well as in vivo analgesic activities.
 Place and Duration of Study: The Laboratory of Phytochemistry and Pharmacology at the Department of Pharmacy and the Laboratory of Microbiology at Stamford University Bangladesh and the Bangladesh Council of Scientific and Industrial Research, Dhaka, conducted this investigation from January 2022 to June 2023.
 Methodology: The study employed the Egg Albumin Protein Denaturation Assay to evaluate the anti-inflammatory effects of various doses. The Disk Diffusion Method was employed for conducting an antimicrobial assay. Different tests were conducted to evaluate the effects of thrombolytic and cytotoxic substances. These tests included the Clot Lysis assay and the Brine Shrimp Lethality Assay. In-vivo analgesic testing was conducted using the hot plate method and the acetic acid-induced writhing test.
 Results: The experimental tests showed significant pain-relieving effects, with inhibition percentages of 46.2% and 66.61% observed at doses of 200 and 400 mg/kg, respectively. The anti-inflammatory test showed a significant inhibition of EEDML at a concentration of 1000μg/mL, with an inhibition rate of 85.45%. The antimicrobial test showed a moderate antimicrobial effect, with a zone of inhibition ranging from 07-15, depending on the microorganism and dosage. The thrombolytic test showed a clot lysis rate of 85.37%, while the cytotoxic test indicated an LC50 value of 1.47.
 Conclusion: EEDML's phytochemical displays strong pharmacological properties, suggesting its potential for drug discovery in diverse areas.

PELATIHAN PENANGANAN DASAR HEWAN COBA BAGI LABORAN BERBAGAI PERGURUAN TINGGI FARMASI DI PROVINSI NTB

The use of experimental animals in drug research needs to be done before testing on humans. The increase in the use of experimental animals and ethical demands in pharmaceutical research have encouraged an increase in the capacity of laboratory staff in a sustainable manner, including in the Province of West Nusa Tenggara (NTB). Animal welfare greatly influences research results. The concept of welfare in question is "Five Freedom". In addition, the principles of replacement, reduction, refinement must also be fulfilled. This needs to be considered in order to get valid research results. The purpose of this activity is to train and support the basic knowledge and skills of laboratory assistants in handling experimental animals at Pharmacy colleges in West Nusa Tenggara. The training was conducted at the pharmacology laboratory of the Pharmacy Study Program, Faculty of Medicine, University of Mataram. The training begins with giving the participants a pretest, followed by delivery of materials and workshops to increase knowledge regarding the process of handling experimental animals, then ends with the participants filling out the posttest. After delivering the material and workshops, an increase in evaluation results was obtained in the form of an increase in the posttest scores of the participants related to knowledge about the material handling of the experimental animals presented. The average pretest score was 63.00 ± 21.11 with a score of 20.00 (lowest) and 90.00 (highest), experiencing an increase in knowledge based on the posttest average score of 74.00 ± 14.97 with a score of 60.00 (lowest) and 100.00 (highest). Conclusion: This training activity can increase the knowledge and skills of laboratory assistants at various pharmaceutical colleges in NTB regarding the basic handling of experimental animals.

In Memoriam: Prof. Olena Petrivna Sotnikova

Professor Olena Petrivna Sotnikova, Head of Pharmacology and Tissue Therapy Laboratory at the Filatov Institute of Eye Diseases and Tissue Therapy, passed away on June 9, 2023. She was born in the town of Makiivka, Donetsk region, on June 26, 1939. After graduating cum laude from the Faculty of General Medicine, Odesa Medical Institute, in 1969, Olena Petrivna began her career as a junior research associate at the Pharmacology and Tissue Therapy Laboratory at the Filatov Institute of Eye Diseases and Tissue Therapy, NAMS of Ukraine. Completing her Cand Sc (Med) thesis, Cytochemical Changes in the Corneal Epithelium and Nasal Mucosa in Impaired Sensory Innervation, under the co-supervision of Drs Sc (Med) V.D. Dragomiretskyi and R.K. Marmur, was her first serious achievement in science. In 1981, O.P. Sotnikova was awarded with a silver medal and the Certificate of Participation in the Exhibition of Economic Achievements of the USSR as a co-developer of a preparation named “aloe tablets covered with a film shell”. She defended her Dr Sc (Med) dissertation and became head of the Pharmacology and Tissue Therapy Laboratory in 1990 and was promoted to professorship in 1999. Professor Sotnikova’s study on molecular and cellular bioregulative and protective mechanisms of tissue preparations in the presence of the impact of adverse factors on the body has become an important contribution to the Acad. Filatov’s fundamental theory of “biogenic stimulators”. Two components had been incorporated into the science and practice activities of the laboratory under her guidance: first, the ocular pharmacological research aimed at the development of single and multicomponent ophthalmic dosage forms from national plant, animal and geo-organic raw materials, and, second, the preclinical evaluation of the curative and preventive efficacy of these forms in ophthalmopathy models. Twelve individuals completed their Cand Sc (Med) degrees under the supervision of Prof. Sotnikova. She produced about 400 publications and authored or co-authored 20 patents. Olena Petrivna was a person with a capital P. She was a remarkably talented scientist and manager, and her job was her passion. She also was a benevolent and empathetic person who had never remained indifferent to the problems of others and was always ready to assist in any way she could. Olena Petrivna’s memory will leave on forever through her numerous published works, scientific ideas and research results, and the apprentices that she mentored.

The effect of giving moringa tempeh flour on hemoglobin levels and body weight of Wistar Rats

Background: Tempeh is a fermented product that has earned the nickname "the miracle food" because of its nutritional content, especially amino acids and fatty acids, which are good for health. Moringa leaves, which are also called "the miracle leaves," have complete nutritional content, especially iron, and vitamin C. Vitamin C plays a significant role in the absorption and metabolism of ironObjectives: Was to increase absorption of iron, which can increase hemoglobin blood and body weight of ratsMethods: There were 4 groups, each consisting of 6 rats. The rats were kept in animal laboratories at room temperature (25–27°C) for 21 days. This research was conducted from 7 to 27 October 2020 in the Pharmacology Laboratory of Medicine, Faculty of Medicine Universitas Muhammadiyah Malang. This study used a Completely Randomized Design, used 24 Rattus norvegicus with treatment conditioned with anemia. The four groups were divided into control anemia rats, control non-anemia rats, anemia rats that consumed tempeh, and anemia rats that consumed tempeh with moringa leaves of 4%. The rats body weight was weighed daily and the blood Hb was taken at the end of the treatment. Statistical test used One-way Analysis of Variance (α = 0,01).Results: The results of statistical tests with ANOVA show not significant influence. The results of the hemoglobin and body weight rats had no significant effect.Conclusion: The value was not significant between groups in the hemoglobin assessment, but the treatment group that was given moringa leaves tempeh was higher than the anemia control group and the pure tempeh group.