Category Archives: Original Research

Ex vivo permeability study and in vitro solubility characterization of oral Canagliflozin self-nanomicellizing solid dispersion using Soluplus® as a nanocarrier

DOI: 10.2478/amma-2024-0011

Background and objective: Self-nanomicellizing solid dispersion SNMSD is a new formulation that combines solid dispersion and nanomicelle strategies; the strategy involves utilizing a suitable carrier that self-assembles into nanomicelles when interacting with gastrointestinal fluids. Canagliflozin, a sodium-glucose cotransporter-2 inhibitor for treating type 2 diabetes, has been linked to poor absorption due to its insolubility in aqueous media. The study aimed to create self-nanomicellizing solid dispersion systems for canagliflozin to overcome its pharmaceutical limitations and improve oral bioavailability.
Materials and Methods: Soluplus® was chosen as a nanocarrier to improve canagliflozin solubility after screening several polymers using a phase solubility study. The solvent evaporation method was selected for preparing the solid dispersion. The optimal formula was characterized through ex vivo permeability and in vitro studies.
Results: The CFZ-SNMSD formula, with a particle size PS of 60.77±1.00 nm and polydispersity index PDI of 0.06±0.02, has a stable distribution upon dilution to 20-fold with water. The apparent solubility of canagliflozin in the optimized CFZ-SNMSD formula was enhanced by 904.40±4 folds due to amorphization and nanomicellization, as demonstrated by transmission electron microscopy. CFZ-SNMSD formula showed a significant enhancement in dissolution rate compared to the physical mixture and pure drugs. The dissolution efficiency parameter confirms these findings (DE30, CFZ-SNMSD = 77.20% compared to DE30, pure drug = 18.28%). Studies show that canagliflozin’s permeability increases exponentially over time due to Soluplus® dispersibility, solubilization, and glycoprotein inhibitory effect, enhancing bioavailability and overcoming GIT membrane barriers.
Conclusions: The study indicates that canagliflozin self-nanomicellizing solid dispersion systems are promising methods for improving the oral bioavailability of canagliflozin medication.

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Beneficial effects of Anemone palmata extracts on male rabbit fertility and reproduction

DOI: 10.2478/amma-2024-0010

Objective: The current study is intended to offer an insight on positive benefits of the herb Anemone palmate belonging to Ranunculaceae family, as an enhancing agent of male rabbit’s fertility.
Methods: Twelve male rabbits, in the growth stage, were divided randomly into four equal groups: one serving as the control and three subjected to different treatments. Anemone palmata extracts (50, 100, and 200 mg/kg) were administered orally to the groups. Blood samples were collected to measure the serum levels of testosterone four weeks later. The weight of the testis and epididymis, sperm count, and sperm motility were determined histologically.
Results: The findings revealed a significant increase in the relative weights of the reproductive organs (testis and epididymis) and plasma testosterone levels in the groups that received the 100 and 200 mg/kg doses. Moreover a notable enhancement in the spermatozoa biology, including concentration, motility, and speed, was observed in the groups treated with 100 and 200 mg/kg doses compared to the control and the group treated with 50mg/kg. The histological study revealed some changes in the spermatogenesis and the structure of the organs, involving the presence of the spermatid phase. A significant increase in the thickness of seminiferous walls, a decrease in the interstitial space, and a reduction in the lumen and intra-tubular spaces were observed as well.
Conclusions: it is well indicated that Anemone palmata could improve fertility factors, and exhibit good effects on gonad and sperm parameters in rabbits.

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Assessing the control of the disease on current treatments available in Romania for hereditary angioedema patients

DOI: 10.2478/amma-2024-0009

Background: Acute treatment must be permanently accessible for every patient diagnosed with hereditary angioedema (HAE). In many cases this type of therapy does not provide/offer sufficient control of the disease, so long-term prophylaxis (LTP) is recommended. In the case of regular and prolonged/extended administration of drugs, the route of administration is essential. The aim of the investigation was to assess the control of HAE among patients in Romania receiving the available medications, while also examining potential correlations within the outcomes.
Material and methods: A phone call was made to all adult patients registered in the Romanian HAE Registry. Patients with confirmed diagnosis of HAE who had at least one angioedema attack in the last three months were asked to complete, online, the angioedema control test (AECT) for one- and three months respectively. AECT scores were calculated according to the authors’ instructions.
Results: A total of 121 patients were contacted. Of these, 83 complies with the eligibility criteria and 56 completed the questionnaires (response rate 67.4%), 18 (32.1%) men and 38 (67.9%) women. Acute, home administered treatment with Icatibant or pdC1-INH was available for every patient during the study time. Nine (14.5%) participants used LTP too, with pdC1-INH. These treatments ensured an adequate control of the disease in only 13 patients (21%) in case of the three-month AECT, of whom 2 used LTP. The one-month questionnaire showed a well-controlled disease in 14 patients (23%), from which only 1 was on prophylactic therapy.
Conclusion: In most Romanian HAE patients, the available drugs do not offer a proper control of the disease. Even though a first-line drug for LTP is available, its administration route by intravenous injections makes it inconvenient for many patients, highlighting the necessity for new, easy-to administer drugs for HAE patients from our country.

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Effect of carbamazepine-beta-cyclodextrin inclusion complex on seizure-like events in an in vitro model of temporal lobe epilepsy

DOI: 10.2478/amma-2024-0007

Objective: Pharmacoresistant epilepsy represents a significant global health challenge, necessitating novel therapeutic approaches. Despite advances in antiseizure medications, many patients remain treatment-resistant partially due to complex pharmacokinetic issues. Beta-cyclodextrin, known for enhancing drug solubility and stability, offers potential solutions by forming inclusion complexes, thereby improving anti-seizure medication’s efficacy. This study aimed to investigate the effect of beta-cyclodextrin and beta-cyclodextrin-complexed carbamazepine on epileptiform activities, using an in vitro model of temporal lobe epilepsy.
Methods: Seizure-like neuronal activity was induced using the low-magnesium model. Local field potentials were recorded from transverse rat hippocampal slices immersed in epileptogenic artificial cerebrospinal fluid, followed by the administration of either beta-cyclodextrin or carbamazepine, the latter in 100 micromolar concentration.
Results: Beta cyclodextrin, applied alone, significantly reduced the duration of interictal and ictal phases while increasing the frequency of seizure-like events. Carbamazepine exhibited an important anticonvulsant effect, significantly reducing ictal and postictal phase durations. However, the frequency of seizure-like events was increased. Notably, in some of the slices, carbamazepine completely suppressed epileptiform activity.
Conclusions: Beta cyclodextrin had an effect on its own; it shortened seizure durations and increased their frequency. Carbamazepine in complexed form, as used in our study, exhibited anticonvulsant efficacy, emphasizing the feasibility of solubility enhancement by this method. This study provides insights into potential therapeutic strategies for pharmacoresistant temporal lobe epilepsy, improving the pharmacological properties of the drugs. As cyclodextrins emerge as promising excipients for antiepileptic drugs with poor solubility, more effort is needed in order to elucidate the underlying mechanisms of their effects.

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Anti-obesity and Antioxidant Effects of Ethanol Extract of Fingerroot Rhizomes (Boesenbergia pandurata Roxb.) on High-Carbohydrate Diet-induced Mice

DOI: 10.2478/amma-2024-0005

Objective: Obesity is defined as the abnormal and excessive accumulation of fat. Enlargement of white adipose tissue due to obesity activates the sympathetic nervous system to stimulate lipolysis to break down fat extensively. This causes a lot of free fatty acids to circulate in the body. Excessive circulating free fatty acids affect many cells and produce oxidative stress, which spreads throughout the body. This research purpose is to determine the anti-obesity and antioxidant activities of ethanol extract of B. pandurata rhizome (EEBP).
Methods: Fingerroot rhizomes were extracted using maceration and the extract was used for in vivo, antioxidant, and total flavonoid concentration tests. Anti-obesity test was carried out by dividing mice into 6 groups such as normal controls, obese controls, standard groups, and extract-treated groups (100, 200, 400 mg/kg b.w). The in vivo test parameters observed included measurement of body weight and also liver, kidney, spleen, and retroperitoneal fat index measurement. Furthermore, in this research antioxidant test was performed using the DPPH method.
Results: In vivo test showed that the giving of EEBP at a dose of 400 mg/kg b.w. effectively decreased body weight and retroperitoneal fat, but slightly affected the organ index of mice such as the liver, renal, and spleen. Furthermore, the antioxidant test showed that the IC50 EEBP results obtained were 37,05 µg/ml. In addition, the total flavonoid content found in the EEBP is 15,775 mgQE/g.
Conclusion: The present study showed that EEBP may have a considerable potential anti-obesity agent and also has a very strong antioxidant effect.

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Biological profiles of Q. cerris, Q. dalechampii, and Q. robur bark extracts: A characterization study

DOI: 10.2478/amma-2024-0003

Objective: The main objective of the present study was to characterize the extracts obtained from the bark of three oak species in order to assess their use in potential cosmetic products.
Methods: The extracts were obtained from the oak barks (periderm and rhytidome) using ultrasound-assisted extraction. The total polyphenolic content was assessed afterward, using the Folin–Ciocâlteu method, while the antioxidant capacity was determined using methods based on the neutralization of the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radicals. To assess the tyrosinase inhibitory effect, a protocol using L–DOPA as the substrate of the enzyme was employed.
Results: The extracts presented high levels of polyphenolic compounds, with Q. cerris having the highest content. Because of the high concentration of the extracts in polyphenolic compounds, they revealed a great reducing capacity against both DPPH and ABTS radicals, but unfortunately the tyrosinase inhibitory activity of the tested extracts was very weak compared to the positive control.
Conclusions: The extracts may have beneficial effects when used in cosmetic products because of the antioxidant effects, but more studies must be conducted for the determination of the main phytochemical compounds comprised in the extracts and their correlation to the biological effects.

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Non-invasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) in Mureș County, Romania: Incidence and impact of reclassification over a six-year period (2016-2021)

DOI: 10.2478/amma-2023-0048

Objective: In 2016 a group of expert endocrine pathologists proposed a new terminology, NIFTP (Non-invasive follicular thyroid neoplasm with papillary-like nuclear features) for a histological subtype of thyroid carcinoma (encapsulated follicular variant of papillary thyroid carcinoma, non-invasive type). In this study, we aimed to assess on a retrospective basis the epidemiological trend and overall incidence of NIFTP in Mureș county, Romania, over a six-year period (2016-2021).
Methods: All NIFTPs registered between 2016-2021 in Târgu-Mureş Pathology Departments were reevaluated. NIFTP’s incidence was calculated by dividing the number of NIFTPs with the number of papillary thyroid carcinomas (PTCs). Further on, we compared NIFTP’s incidence between 2016-2018 with NIFTP’s incidence between 2019-2021, as the diagnostic criteria of NIFTP were revised in 2018.
Results: Forty-six cases of NIFTP were registered in our departments between 2016-2021, resulting in an overall NIFTP incidence of 17.9 % [CI (95%): 7.6-32.5] among the 257 PTCs. When we compared the NIFTP’ incidence between the two study periods (2016-2018 versus 2019-2021), our data revealed that the NIFTP’s overall incidence has dropped from 21.4 % [CI (95%): -14.5-60.1] to 15% [CI (95%): -8.2-43] in the second evaluated period (p=0.034).
Conclusion: The overall NIFTP incidence was low in our departments between 2016-2021. Moreover, NIFTP’s incidence decreased after 2018, following revision of the diagnostic criteria. Our results highlight that very stringent morphologic criteria need to be apply when making a diagnosis of NIFTP, in order to avoid a misdiagnosis and/or an overtreatment of a tumor with indolent behavior.

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Determination of isohydrafural and methyluracil in ear drops by high performance liquid chromatography

DOI: 10.2478/amma-2023-0049

Objective: The aim of current study was the development and validation of a reversed-phase high-pressure liquid chromatography (RP-HPLC) method for the quantitative determination of two active ingredients, isohydrafural (IHF) and methyluracil (MU) in fixed dose combination ear drops.
Methods: An efficient separation of the two compounds was achieved on a Teknokroma C1 5μm (150*4,6mm) column, with a methanol: water 60:40 (V: V) mobile phase, at 300C temperature and 0.6 mL/min flow-rate. The total analysis time was 5.5 min.
Result: The verified validation parameters were: linearity, selectivity, specificity, precision (repeatability and reproducibility), robustness, limit of detection and quantification.
Conclusions: Good separation and lack of interference from other chromatographic peaks, rapidanalysis times were obtained. The newly developed chromatographic method can be applied for the analysis of fixed dose combination ear drops with isohydrafural and methyluracil.

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Packed cell volume and its relation to obesity, gender and smoking status

DOI: 10.2478/amma-2023-0047

Objective: Our aim was to evaluate the packed cell volume (PCV) correlation with body mass index, body fat percentage, also differences between genders and smokers & non-smokers, among Iraqi students.
Methods: A cross-sectional study was done on 112 healthy individuals (from which 52.7% were males), aged between 18-23 years old, in December 2022. A short informative history was taken through a questionnaire, anthropometric measures were taken to calculate body mass index and body fat percentage, and the determination of packed cell volume was done by the Microhematocrit method.
Results: Packed cell volume among males was higher 47.45±3.409% than for females 39.90±3.169%, with a difference statistically significant (p=0.000), also the correlation of PCV revealed direct significance with body mass index (p=0.011) and indirect statistically significant with body fat percentage (p=0.000). The prevalence rate of smoking was 13.4% and the level of PCV among smokers was 46.80±6.085%, significantly higher (p=0.015) than among non-smokers 43.43±4.702%.
Conclusion: Packed cell volume showed a direct correlation with body mass index, an inverse one with the body fat percentage, and was significantly higher among young smokers, emphasising the idea that this parameter can help to evaluate the health risk and to be included in preventive programs and assessment protocols.

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The outcome in critically ill patients admitted for thoracic trauma – A single center analysis over one year

DOI: 10.2478/amma-2023-0046

Objective: The main objective of the current study was to examine the outcome of critically ill patients in relation to clinical and thoracic injuries. The secondary objectives were to assess the role of CT in the evaluation of the diaphragm and to provide an analysis of prognostic abilities with respect to diaphragm changes.
Methods: This single-center retrospective observational cohort study was conducted in the ICU of Târgu Mureș County Emergency Clinical Hospital, Romania. This study identified 52 critically ill blunted or penetrating chest trauma patients admitted to the ICU from 01 January 2021 to 31 December 2021. CT scan was used to identify thorax injuries and diaphragm thickness. The outcome of all patients was analyzed.
Results: Most of the patients experienced traffic accidents (44.23%) or falls (26.93%). The predominant characteristics associated with chest trauma were rib fractures (92.30%), lung contusions (63.50%), and pneumothorax (53.80%). The most common injury seen in the study was rib fractures, accounting for 92.30% of cases. This was followed by lung contusions, which were present in 63.50% of patients, and pneumothorax, which occurred in 53.80% of cases. It was examined ROC AUC for thickness of the right and the left diaphragm and severity scores. When assessing the thickness of the diaphragm in deceased and survivors, no statistically significant differences were found.
Conclusion: Although no significant differences were found regarding the prognosis between the survivors and the deceased, diaphragm thickness might potentially serve as a predictor for the severity of the injury.

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