Category Archives: Original Research

Surgical procedure for acellular vascular xenografts testing in sheep carotid artery

Marius Mihai Harpa1, Alexandra Iulia Puscas2, Emanuel David Anitei3, Dan Teodor Simionescu4, Raluca Ionela Truta1, Hamida Al-Hussein2, Ionela Cotfas1, Claudiu Ghiragosian2, Cosmin Marian Banceu2, Tudor Capilna2, Horatiu Suciu2, Hussam Al-Hussein2

1. Regenerative Medicine Laboratory, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Emergency Institute for Cardiovascular Diseases and Transplantation Targu Mures, Romania
4. Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University, Clemson, South Carolina, USA

DOI: 10.2478/amma-2024-0024

Objective: The objective of this study is to demonstrate the safety and reproducibility of our surgical technique for implanting decellularized vascular xenografts in the carotid artery of sheep.
Methods: Acellular porcine carotid arteries were implanted as interposition xenografts in seven sheep. An intravascular shunt was used for cerebral protection, and a flowmeter was utilized to assess graft performance.
Results: There were no intraoperative deaths or postoperative neurological complications observed. Acute graft thrombosis occurred in one sheep during surgery, but was successfully managed with thrombectomy to restore blood flow. Post-implantation flowmetry and Doppler ultrasound confirmed graft functionality.
Conclusions: Our study demonstrates the successful application of our surgical method for implanting decellularized vascular xenografts in the carotid artery of sheep. The implanted grafts maintained patency, normal blood flow, and favorable wound healing and neurological outcomes post-surgery.

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Qualitative phytochemical screening and in vitro
antioxidant activities of aqueous extracts of
Rhaphiolepis bibas (Lour.) Galasso & Banfi

Ibrahim Waweru Kariuki, Mathew Ngugi, John Maingi

Kenyatta University, Nairobi, Kenya

DOI: 10.2478/amma-2024-0022

Reactive oxygen species causes harm to cell membranes and biomolecules, wherefore chronic diseases develop. Antioxidants scavenge such free radicals combating oxidative stress. This research aimed to determine the antioxidant potential of the aqueous stem bark, root and leaf extracts of Rhaphiolepis bibas against standards. DPPH radical scavenging activity was high from th stem bark extract at 72.33% with root extract at 65.85% and leaf extract at 55.91%, while ascorbic acid scavenged 89.53% of DPPH radicals. The leaf extract had the highest H2O2 scavenging activity of 91.92% with stem bark at 91.17% and the root extracts at 89.12%. The aqueous root extract of R. bibas had a significantly higher FRAP capacity in comparison to the leaf extracts and the stem bark. The abilities to chelate iron by the leaf extract were statistically higher compared to the root and stem bark extracts. Stem bark extracts had the highest phenol content of about 149.44 followed by the root extract at 141.14 and the least amount of phenol was found in the aqueous leaf extract having 73.012 Gallic acid equivalent/g. The root extracts had the highest total flavonoid 377.66-milligram quercetin equivalent/gram dry weight followed by stem bark extract at 255.72 and the least amount was found in aqueous leaf extract having 164.52 mgQE/g of sample dry weight. The existence of secondary metabolites linked to antioxidant action was shown by the qualitative phytochemical screening.

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Considerations regarding the attitude of medical staff towards administrative risks within the Pathology Department

Aurora Maria Popa1, Bianca Andreea Lazar2, Liviu Onoriu Marian1, Ovidiu Simion Cotoi2,3, Septimiu Voidăzan4

1. Faculty of Mechanical Engineering, Technical University of Cluj-Napoca, Romania
2. Pathology Department, Mures Clinical County Hospital, Targu Mures, Romania
3. Department of Pathophysiology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
4. Department of Epidemiology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2024-0023

Objective: In the context of public health, there is a common misconception that only Romania’s system faces issues and generates societal dissatisfaction. However, globally, there is significant room for improvement, as the performance of the sector is often unsatisfactory. This paper presents findings from an exploratory study on the attitudes of medical personnel in pathology services towards managerial and administrative risks, supported by a survey that provides a wealth of useful information.
Methods: The survey encompassed a geographic area including the counties of Mureș, Harghita, Sibiu, and Alba, involving 12 healthcare units: 2 university hospitals, 2 county hospitals, and several municipal or town hospitals. These institutions housed anatomical pathology service structures staffed by approximately 240 physicians and assistants, with a ratio of 1 physician to 2.5 assistants. The questionnaire aimed to test four hypotheses through questions that invited respondents to select from formatted answers.
Results: Of the four hypotheses formulated and their corresponding items, only two were confirmed. This outcome suggests that the Romanian healthcare system is not unequivocally adaptable to the needs of medical product consumers. Specifically, the data indicate that hospitals face dire financial conditions amidst various challenges and vulnerabilities.
Conclusion: Differences of opinion between the two categories of respondents are evident in certain risk categories. This discrepancy arises from the specific nature of their activities and their respective contributions to ensuring the quality and efficiency of the service.

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The influence of the COVID-19 pandemic on pediatric hospitalizations for type 1 diabetes mellitus

Măriuca Mănescu1, Ion-Bogdan Mănescu2, Alina Grama1,3

1. Department of Pediatrics, Emergency County Clinical Hospital of Targu Mures, Targu Mures, Romania
2. Department of Laboratory Medicine, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Targu Mures, Romania
3. Department of Pediatrics, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Targu Mures, Romania

DOI: 10.2478/amma-2024-0021

Objective: In this retrospective cohort study, we aimed to provide a snapshot of how the pandemic has affected pediatric type 1 diabetes mellitus (T1D) admissions in our hospital.
Methods: This study included 117 patients aged 0-18 classified based on period (pre-pandemic vs. pandemic period 2020-2022) and type of diagnosis at admission: new-onset T1D (nT1D) or diabetic ketoacidosis (DKA)-decompensated T1D. We investigated the effect of the COVID-19 pandemic on the demographic, clinical, and laboratory characteristics of these patients.
Results: Out of all T1D-related admissions, the proportion of admissions for nT1D increased compared to the pre-pandemic period: 71.6% vs 53.4%, p=0.048. Unrelated to the pandemic, the type of diagnosis at admission was associated with 1) the sex distribution (males – more nT1D admissions, females – more frequent DKA admissions, p=0.01), and 2) hospitalization duration (longer for nT1D admissions than for DKA-decompensated T1D admissions, p=0.001). Blood glucose and HbA1c levels were influenced neither by the pandemic period nor by the type of diagnosis. During the pandemic, a change in the T1D seasonality became apparent. A potential association pattern between new COVID-19 cases, number of T1D admissions, and stringency of restrictions was observed.
Conclusions: During the COVID-19 pandemic, the proportion of nT1D admissions increased, as well as the severity of DKA-decompensated T1D cases. In addition, the pandemic period brought about notable shifts in the seasonality of pediatric T1D.

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Parameters of dynamic spirometry before and after administration of salbutamol in COPD patients

Medžida Rustempašić1, Muamer Dervišević2

1. Private Health Institution Eurofarm, Sarajevo, Bosnia and Herzegovina
2. University Clinical Center, Clinic for Lung Diseases and Tuberculosis “Podhrastovi”, Sarajevo, Bosnia and Herzegovina

DOI: 10.2478/amma-2024-0019

Introduction: The aim of this research was to examine the existence of broncho-obstruction of the airways in patients suffering from chronic obstructive pulmonary disease, as well as changes in resistance values in this part of the bronchial tree after inhalation of salbutamol.
Methods: This study was designed as a prospective, interventional clinical trial that included a sample of 147 patients suffering from COPD. Patients were stratified into four groups of thirty patients each based on the severity of airflow limitation (based on the post-bronchodilator FEV1 value), according to the GOLD grade. The test was conducted at the University Clinical Center Sarajevo, Clinic for Lung Diseases and Tuberculosis “Podhrastovi”
Results: The results of our research showed that the average values of FEV1 compared to the predicted values of this parameter in subjects in the GOLD 4 group before the administration of salbutamol were statistically significantly lower than the average values of FEV1 in other subjects of the GOLD group. After the administration of salbutamol, a statistically significant increase in the value of FEV1 was registered in all tested groups. When the response to salbutamol was compared among the GOLD groups, it was assessed that the difference in the percentage increase in predicted FEV1 values after the administration of salbutamol among the tested groups was not statistically significant.
Conclusion: Bronchodilation with salbutamol and additional ipratropium had a significant effect on both mentioned parameters, especially in the COPD group, which speak in favor of the presence of increased peripheral resistance in all groups of patients. A statistically significant bronchodilator response was obtained in GOLD 1 and GOLD 2 groups, i.e. in groups of patients with milder forms of the disease.

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In vitro modulation of seizure-like activity with beta-cyclodextrin-complexed rufinamide

Rita-Judit Kiss1, Zsolt András Nagy2, Ádám Szentes2, Ágnes Csüdör2, Andrea Máthé2, Henrietta Makó2, Előd Bomher2, Ádám József Berki3, Zsolt Gáll4, Tibor Szilágyi1, Károly Orbán-Kis1

1. Physiology Department, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Department of Neurology, Semmelweis University, Budapest, Hungary
4. Pharmacology and Clinical Pharmacy Department, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2024-0020

Objective: The global health concern of pharmacoresistant epilepsy necessitates innovative therapeutic strategies. Drug resistance often arises due to complex pharmacokinetic challenges. Beta-cyclodextrin, known for enhancing drug solubility and stability, offers a potential solution for improving the efficacy of antiseizure medications. This study aims to investigate the impact of beta-cyclodextrin-complexed rufinamide on seizure-like activity using an in vitro model of temporal lobe epilepsy.
Methods: Seizure-like neuronal activity was induced using a low-magnesium model. Local field potentials were recorded from transverse rat hippocampal slices. Rufinamide was solubilized using beta-cyclodextrin and administered at 100 micromolar concentration. The impact on various seizure-like parameters and time-resolved phase-amplitude coupling was assessed.
Results: Rufinamide increased the duration of the preictal phase while reducing the duration of ictal and postictal phases. The frequency of seizure-like events was higher in rufinamide. No significant change was observed in the firing rate of the first 10 ictal spikes, but the firing frequency of the second set of 10 ictal spikes was higher during rufinamide perfusion. Time-resolved phase-amplitude coupling maximum analysis did not reveal significant differences between the control and rufinamide treatment.
Conclusions: Beta-cyclodextrin-solubilized rufinamide significantly modulates seizure-like event dynamics, exhibiting both anticonvulsant and proconvulsant effects. While the compound shortened seizure-like activity, it increased the frequency of seizure-like events. Our observations suggest a need for further investigation into the solubilization method and its impact on rufinamide’s bioavailability. Dose-dependent effects and underlying molecular mechanisms should also be explored to enhance the pharmacological properties of antiseizure medications.

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Purple sweet potato (Ipomoea batatas L.) reduces the wound temperature and erythema in STZ-induced diabetic rats

Ratih Paramita Suprapto1, Heri Kristanto2

1. Department of Physiology, Faculty of Medicine, Universitas Brawijaya, Jl Veteran, East Java, Indonesia
2. School of Nursing, Faculty of Health Science, Universitas Brawijaya, Puncak Dieng Eksklusif, East Java, Indonesia.

DOI: 10.2478/amma-2024-0018

Objective: Wound temperature is one of the wound biomarkers representing the wound healing progress. The diabetic wound is associated with a prolonged inflammation phase marked by an increasing wound temperature and erythematous wound. Purple sweet potato extract (PSP), an anthocyanin-rich plant, improved wound healing in both diabetic and non-diabetic wounds in animal studies. This study aims to know the effect of purple sweet potato ethanol extract on wound temperature and erythema in streptozotocin-induced diabetic rats.
Methods: Rats were divided into four groups: normal rats + gel placebo; diabetic rats + gel placebo; diabetic rats + PSP 10%; and diabetic rats + PSP 15%. Diabetes mellitus was induced by streptozotocin injection. After diabetic confirmation, the back of the rats was excised and the gel was administered daily for 14 days. The wound temperature was measured at the wound surface using infrared thermography on days 0, 3, 7, and 14. The wounds were photographed and erythema analysis was conducted using Corel Photo paint®.
Results: Diabetic wounds exhibited higher surface temperature compared with the normal wound (37.08°C ± 0.29 vs 35.13°C ± 0.46) on day 14 of examination and topical application the purple sweet potato ethanol extract gel 10% and 15% markedly decreased the wound temperature at day 7 and 14 compared with the diabetes wound (p<0.0001). Wound erythema was significantly reduced in the PSP 10% and 15% diabetic wound treatment groups on day 14 (p<0.05).
Conclusion: Purple sweet potato extract gel treatment was found to have the potential to reduce inflammation in diabetic wounds.

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The effects of pearl oyster shell-derived bone grafts on TNF-alfa levels: An in vivo study in Cavia porcellus

Sri Oktawati1, Nurlindah Hamrun2, Muthmainnah Muthmainnah1, Dian Setiawati1, Andi Sitti Hajrah Yusuf3

1. Department of Periodontology, Faculty of Dentistry Hasanuddin University, Makassar, Indonesia
2. Department of Oral Biology, Faculty of Dentistry Hasanuddin University, Makassar, Indonesia
3. Department of Oral and Maxillofacial Surgery, Faculty of Dentistry Hasanuddin University, Makassar, Indonesia

DOI: 10.2478/amma-2024-0015

Objective: This study aims to assess the effect of bone graft materials derived from pearl oyster shells on the tumor necrosis factor-alpha (TNF-α) level.
Methods: This research utilized a bone graft containing hydroxyapatite Pinctada maxima (HPM) powder obtained from pearl oyster shells. Material testing was conducted on 30 male guinea pigs aged 8-10 weeks, divided into three groups: the negative control group, the group treated solely with HPM, and the group treated with HPM combined with platelet-rich fibrin (PRF). Each treatment group was observed for 7 and 14 days. The results were analyzed using One-way ANOVA with a significance level of p<0.05.
Results: The findings demonstrated a decrease in the average TNF-α levels across all groups. On observation days 7 and 14, the HPM group exhibited a significant difference compared to the negative control group, indicating reduced TNF-α levels. However, no significant difference was observed when comparing the HPM group with the HPM and PRF groups.
Conclusions: Bone grafts derived from pearl oyster shells possess high levels of hydroxyapatite, and the addition of PRF into the bone graft effectively reduces TNF-α.

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Exploring the impact of high shear mixing process parameters on the physical characteristics of excipient powder blend by design of experiments

Béla Kovács1,2, Erzsébet Orsolya Tőkés2,3, Éva Katalin Kelemen2, Katalin Zöldi2, Edit Suba2, Boglárka Kovács-Deák3, István Székely-Szentmiklósi2,3, Francisc Boda4, Tibor Casian5

1. Department F1, Biochemistry and Environmental Chemistry, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. Gedeon Richter Romania, Targu Mures, Romania
3. Doctoral School, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
4. Department F1, General and Inorganic Chemistry, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
5. Department of Pharmaceutical Technology and Biopharmacy, “Iuliu Hațieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania

DOI: 10.2478/amma-2024-0016

Objective: Dry-route manufacturing technology development poses challenges to pharmaceutical technology research specialists, relying on active substance characteristics, excipient selection, and parameter optimization. Amongst various technological possibilities high shear mixing generally ensures dosage uniformity and tablet dissolution through influential shear forces, potentially enhancing dry powder blend processability. This study explores the processability of a placebo formulation within the quality by design framework to address some of the aforementioned challenges.
Methods: A 2^4 full-factorial experimental design was used to assess the manufacturability of a placebo formulation via high shear mixing. The effect of impeller and chopper speed, high shear mixing time, and homogenization/lubrication times on powder blend rheology and compression properties was investigated.
Results: The findings of the present study showed that product critical quality attributes like resistance to crushing or disintegration time are mainly dependent on the mixing efficiency translated by the impeller speed and high shear mixing time. Software augmented process development enabled the attainment of the design space, ensuring the fulfilment of desired product performance criteria. Furthermore, the study has also shown that the careful selection of excipients is crucial in the case of dry-route manufacturing technologies, as sodium lauryl sulphate can noticeably influence the processability of powder blends due to its lubricant properties.
Conclusions: Considering the advantages and challenges raised by high-shear mixing, software aided data analysis can further augment the formulation, scale-up and lifecycle management of products developed using this technological process.

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Beyond the gut – Atypical presentation of Salmonella spp. infection

Anca Cighir1,2, Corina Eugenia Budin2,3, Teodora Cighir2, Anca Delia Mare1,2, Adrian Man1,2

1. Department of Microbiology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Targu Mures, Romania
2. Mureș Clinical County Hospital, Targu Mures, Romania
3. Department of Pathophysiology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Targu Mures, Romania

DOI: 10.2478/amma-2024-0014

Salmonella is a Gram-negative, non-spore-forming, motile, facultative anaerobic rod. The most studied species are Salmonella typhi and paratyphi (causing typhoid fever) and non-typhi Salmonella species (which can cause different clinical syndromes – gastroenteritis, disseminated infections, etc.). A 61-year-old male patient with multiple comorbidities (including myelofibrosis) presented to the Pulmonology outpatient department, Clinical County Hospital of Târgu Mureș, with a mucopurulent nocturnal cough. Paraclinical examinations showed the presence of a pleural empyema, which was evacuated in local anesthesia. The pleural fluid was sent to the Microbiology Department for bacteriological testing, where a fluoroquinolone-resistant strain of Salmonella spp. was detected. The patient received antibiotic treatment according to the antibiotic susceptibility testing. Due to the persistence of the symptoms, the patient returned two weeks later, when another puncture was performed. Salmonella was isolated again, but the strain showed a higher resistance to antibiotics. The two strains were compared using molecular methods of diagnosis (Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction – ERIC-PCR), the results showing a similarity of 92%. The occurrence of an extra amplicon band in ERIC-PCR suggests an important change in the bacterial genetic material, potentially related to acquisition of antibiotic resistance factors.

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