Assessment of the Effect on Periodontitis of Antibiotic Therapy and Bacterial Lysate Treatment.

Periodontitis is an inflammatory process that starts with soft tissue inflammation caused by the intervention of oral bacteria. By modulating local immunity, it is possible to supplement or replace current therapeutic methods. The aim of this study was to compare the effects of an immunostimulatory treatment with the antibiotherapy usually applied to periodontitis patients. On a model of periodontitis induced in 30 rats (divided into three equal groups) with bacterial strains selected from the human oral microbiome (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Streptococcus oralis), we administered antibiotics, bacterial lysates and saline for 10 days. Clinically, no significant lesions were observed between the groups, but hematologically, we detected a decrease in lymphocyte and neutrophil counts in both the antibiotic and lysate-treated groups. Immunologically, IL-6 remained elevated compared to the saline group, denoting the body's effort to compensate for bone loss due to bacterial action. Histopathologically, the results show more pronounced oral tissue regeneration in the antibiotic group and a reduced inflammatory reaction in the lysate group. We can conclude that the proposed bacterial lysate has similar effects to antibiotic therapy and can be considered an option in treating periodontitis, thus eliminating the unnecessary use of antibiotics.

Magnetic resonance imaging combined with histological evaluation of repair process using the microfracture technique in an association of osteocartilaginous and meniscal surgically induced lesions of the knee. In vivo experiment on a rabbit model.

The present research study aimed to assess magnetic resonance imaging (MRI) changes and histological findings in the therapeutic effects of microfractures in the treatment of complex animal knee lesions resulting from osteochondral and meniscal defects resulting from non-total meniscectomies. The anterior cruciate ligament lesions are also proven to facilitate the development of osteoarthritis in the knee and worsen the prognosis. Surgery was performed on the right knee joint of 22 male rabbits in order to partially remove the anterior horn of the internal meniscus and to induce an osteochondral defect at the level of the internal femoral condyle. The induced lesion complex was aimed to simulate a clinical situation that occurs frequently in orthopedic practice when young adults undergo partial meniscectomy and at the time of surgery, an osteochondral defect is diagnosed. Rabbits were separated into two study groups: the control (C1) group and the microfractures (MF2) group. After the induced cartilage defect and partial meniscectomy, both groups were followed-up for six months using detailed MRI. Also, anatomical specimens were histologically analyzed to show modifications and signs of healing process, along with complications, in the study group. The results showed that the microfracture group had better results concerning articular surface defect healing in comparison to the control group. Our results suggest that microfractures do improve results concerning surface contact healing and serial MRI studies can be useful in observing the remodeling process in dynamics.

Antimicrobial Activity of Sempervivum tectorum L. Extract on Pathogenic Bacteria Isolated from Otitis Externa of Dogs.

The present study investigated the antimicrobial activity, total phenolic content, and proanthocyanidin concentration of ethanolic extracts from fresh leaves of Sempervivum tectorum L. The extracts were phytochemically analyzed and evaluated for antimicrobial activity. The broth microdilution method was used to assess antimicrobial activity against pathogenic bacteria isolated from ear swabs taken from dogs with otitis externa. Many compounds were present in the ethanolic aqueous extracts, which exhibited a broad spectrum of antimicrobial activity. They showed strong antibacterial activity against standard clinical Gram-positive strains such as S. aureus and Gram-negative strains such as P. aeruginosa. In our study, the obtained quantity of total phenolic compounds in the ethanol:water extract of leaves was 126.17 mg GAE/g. The proanthocyanidin concentration in the tested Sempervivum tectorum L. extracts was 15.39 mg PAC/g material. The high contents of total phenolics and proanthocyanidin indicated that these compounds contribute to antimicrobial activity. The antimicrobial activity of the tested S. tectorum L. extracts ranged from 1.47 to 63.75 µg/mL, starting with 1.47 µg/mL and 1.75 µg/mL against S. aureus ATCC 25923 and P. aeruginosa ATCC 27853 strains, respectively. Likewise, S. tectorum L. ethanol extract demonstrated a bacteriostatic effect against S. aureus clinical isolate with a median MIC of 23.25 µg/mL and MBC of 37.23 µg/mL; and bactericidal against S. aureus ATCC 25923 with the median MIC of 20.33 µg/mL and MBC of 37.29 µg/mL. In the Gram-negative P. aeruginosa clinical and standard strains, the expressed MIC and MBC values were 24.234 and 20.53 µg/mL for MIC, and 37.30 and 37.02 µg/mL for MBC, respectively.

Effects of Aronia melanocarpa and Hypericum perforatum aqueous extracts on hexavalent chromium induced toxicity in rat's thyrocytes.

BACKGROUND: Hexavalent chromium known as oxidizing agent is able to form reactive oxygen species. Aronia melanocarpa and Hypericum perforatum are two plants known for their antioxidant effects. Our study aimed to establish if CrVI induces apoptosis and structural changes in thyrocytes and if its effect can be counteracted by the administration of both extracts. MATERIALS AND METHODS: Wistar rats divided in five groups: C - distilled water (DW), Cr - 75 mg/L CrVI in DW for 3 months, Cr 2 - 75 mg/L CrVI in DW for 3 months followed by 1 month DW, CrA - 3 months 75 mg/L CrVI in DW and 1 month Aronia 2.5% extract, CrH - 3 months 75 mg/L CrVI in DW and 1 month Hypericum 2.5% extract. Histological assessment and qRT-PCR for evaluation of BAX and Bcl2 protein levels performed on thyroid samples. RESULTS: The Cr and Cr2 groups were those with altered cytoarchitecture: increase in the diameter of many thyroid follicles, a decrease in their number, a decrease in the height of the follicular cells. The histological examination of the CrH group revealed almost recovery of structural architecture. The BAX gene levels were higher in the Cr and Cr2 groups indicating the apoptotic activity of chromium. In extract receiving groups the BAX gene expressions were significantly lower, but the lowest level presented the CrH group. Bcl2 gene expression levels indicate antiapoptotic activity being elevated in the Cr group, followed by CrA, Cr2, and CrH groups. The BAX/Bcl2 ratio which significantly increased in the case of the Cr and Cr2 group compared to the groups that were administered the two plant extracts. CONCLUSION: The results obtained in this study confirm that CrVI has toxic effects on thyroid endocrine cells and H. perforatum has stronger antioxidant properties against the action of hexavalent chromium in thyrocytes than A. melanocarpa.

Antibacterial Activity of Romanian Propolis against Staphylococcus aureus Isolated from Dogs with Superficial Pyoderma: In Vitro Test.

Staphylococcal infection treatment in dogs is frequently associated with adverse side effects, high costs, prolonged treatment, and resistant strain selection. Staphylococcus aureus is the most frequently isolated staphylococci in cases of canine superficial pyoderma. The number of Staphylococcus strains to exhibit primary resistance to various drugs in vitro is increasing. Propolis has a diverse chemical composition and well-known therapeutic properties against bacterial infections. The current investigation evaluated in vitro the antimicrobial activity of the commercial allopathic antimicrobials, Romanian propolis ethanolic extracts, against clinical Staphylococcus aureus strains isolated from superficial dermatitis clinical samples in dogs and two reference strains: Staphylococcus aureus ATCC 25923 and Staphylococcus aureus ATCC 43300, as the MSSA and MRSA positive controls, respectively, in western Romania. We used the microdilution broth technique to evaluate the susceptibility profile of the bacteria. The minimum inhibitory concentration (MIC) of the Romanian propolis ethanolic extract ranged from 6 to 10 µg/mL for all isolates, determined by the broth microdilution method. The MICs of ethanolic Romanian propolis extracts had a pronounced antibacterial activity. These results indicate that propolis can potentially be used and recommended for in vivo experiments as a promising therapeutic agent against Staphylococcus aureus infections in superficial dermatitis of dogs.

Occurrence, Pathogenic Potential and Antimicrobial Resistance of Escherichia coli Isolated from Raw Milk Cheese Commercialized in Banat Region, Romania.

The aim of the present study was to investigate the presence, pathogenic potential and antimicrobial susceptibility profile of Escherichia coli isolated from raw milk cheese, traditionally produced by farmers and marketed directly to the consumer in Banat region, Romania. A total of 81.1% (43/53) of the processed samples expressed positive results for E. coli, with a distribution of 83.8% (31/37), and 75.0% (12/16) in the cow- and sheep-milk-origin assortments, respectively. Overall, 69.8% (30/43) of the specimens had a contamination level ≤10 CFU/g. Molecular tests showed that, from the total number of E. coli isolates, 9.3% (4/43) harbored the stx2, and 2.3% (1/43), the stx1 virulence genes. The E. coli O157 (including H7) biovariety was identified in 7% (3/43) of the samples by the Vidas equipment. From the 27 antimicrobials tested with the Vitek2 automated system, the E. coli isolates displayed resistance to enrofloxacin (100%, 15 out of 15 tested isolates), ampicillin (39.5%, 17/43), norfloxacin (28.6%, 8/28), fosfomycin (25%, 7/28), amoxicillin/clavulanic acid (23.3%, 10/43), cefalexin (20%, 3/15), cefalotin (13.3%, 2/15), tetracycline (13.3%, 2/15), trimethoprim-sulfamethoxazole (9.3%, 4/43), piperacillin-tazobactam (7.1%, 2/28), cefotaxime (7.1%, 2/28), cefepime (7.1%, 2/28), ticarcillin/clavulanic acid (6.7%, 1/15), florfenicol (6.7%, 1/15), ceftazidime (3.6%, 1/28), and ertapenem (3.6%, 1/28). Ten (23.3%) strains were multidrug-resistant. The obtained preliminary results indicated hygienic-sanitary deficiencies throughout the cheese production process, and demonstrated that these products can harbor virulent and multidrug-resistant E. coli strains, which constitute a public health risk. However, future investigations, processing a higher number of samples, are still necessary to draw comprehensive conclusions.

Apiaceae Essential Oils: Boosters of Terbinafine Activity against Dermatophytes and Potent Anti-Inflammatory Effectors.

Dermatophyte infections represent an important public health concern, affecting up to 25% of the world's population. Trichophyton rubrum and T. mentagrophytes are the predominant dermatophytes in cutaneous infections, with a prevalence accounting for 70% of dermatophytoses. Although terbinafine represents the preferred treatment, its clinical use is hampered by side effects, drug-drug interactions, and the emergence of resistant clinical isolates. Combination therapy, associating terbinafine and essential oils (EOs), represents a promising strategy in the treatment of dermatophytosis. In this study, we screened the potential of selected Apiaceae EOs (ajowan, coriander, caraway, and anise) to improve the antifungal activity of terbinafine against T. rubrum ATCC 28188 and T. mentagrophytes ATCC 9533. The chemical profile of EOs was analyzed by gas chromatography. The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of EOs/main compounds were determined according to EUCAST-AFST guidelines, with minor modifications. The checkerboard microtiter method was used to identify putative synergistic combinations of EOs/main constituents with terbinafine. The influence of EOs on the viability and pro-inflammatory cytokine production (IL-1ß, IL-8 and TNF-α) was determined using an ex vivo human neutrophils model. The binary associations of tested EOs with terbinafine were found to be synergistic against T. rubrum, with FICI values of 0.26-0.31. At the tested concentrations (6.25-25 mg/L), EOs did not exert cytotoxic effects towards human neutrophils. Anise EO was the most potent inhibitor of IL-1ß release (46.49% inhibition at 25 mg/L), while coriander EO displayed the highest inhibition towards IL-8 and TNF-α production (54.15% and 54.91%, respectively). In conclusion, the synergistic combinations of terbinafine and investigated Apiaceae EOs could be a starting point in the development of novel topical therapies against T. rubrum-related dermatophytosis.

Antimicrobial Drug-Resistant Salmonella in Urban Cats: Is There an Actual Risk to Public Health?

The present study was undertaken to investigate the presence of Salmonella spp. in the faeces of client-owned cats in urban areas and to evaluate the risk that is posed to public health. Fresh faecal samples were collected directly from the rectums from 53 diarrhoeic and 32 non-diarrhoeic cats. The samples were individually screened for the presence of Salmonella spp. using standard methods and, in the case of positive findings, the resulting typical colonies were then biochemically confirmed using the VITEK®2 automated system. Subsequently, all of the Salmonella spp. isolates were molecularly tested for the presence of the invA gene. All of the isolates were serotyped using the slide agglutination technique according to the White-Kauffmann-Le Minor scheme. The phenotypic antimicrobial susceptibility profile of the isolated strains was obtained from the VITEK®2 system using specific cards from the Gram-negative bacteria. A total of 16 of the samples (18.82%) tested positive for Salmonella spp. according to conventional and molecular testing methods. Serotyping of the Salmonella isolates showed the presence of three serotypes, namely S. enteritidis (n = 9; 56.3%), S. typhimurium (n = 4; 25%), and S. kentucky (n = 3; 18.8%). All of the tested strains showed strong resistance towards cefazolin, cefepime, ceftazidime, and ceftriaxone. Additionally, resistance (listed in descending order of strength) was observed to trimethoprim/sulfamethoxazole (11/16; 68.8%), ampicillin (10/16; 62.5%), ampicillin/sulbactam (9/16; 56.3%), gentamicin (9/16; 56.3%), nitrofurantoin (8/16; 50.0%), and amikacin (5/16; 31.3%). No resistance was expressed against ciprofloxacin, ertapenem, imipenem, levofloxacin, piperacillin/tazobactam, and tobramycin. The results of this study highlight a substantial public health issue and medical concern, especially in vulnerable people, such as children, the elderly, and immunocompromised individuals.

Antibiotic Susceptibility Profile of Pseudomonas aeruginosa Canine Isolates from a Multicentric Study in Romania.

Treating infections caused by Pseudomonas aeruginosa is increasingly difficult due to high antibiotic resistance, materialized through the presence of multiple resistance strains, as well as due to rapid development of resistance throughout treatment. The present survey was conducted to investigate the antibiotic susceptibility profile of Pseudomonas aeruginosa pathogens in two University Veterinary hospitals from different geographical regions of Romania (i.e., Southwest Timișoara county and Northeast Iași county) involved in superficial canine infections. A total of 142 swab specimens were collected from dogs with superficial infections (superficial skin infections, otitis externa, and perianal abscess) to assess the presence of Pseudomonas aeruginosa, based on phenotypic and molecular characterization. According to their confirmed morphological and molecular features, 58 samples (40.84%; 58/142) were positive for Pseudomonas aeruginosa (according to their confirmed morphological and molecular features). Antibiotic susceptibility testing for 12 antibiotics was conducted using the Kirby-Bauer disc diffusion method. Drug resistance was observed in the case of all tested antibiotics. The susceptibility rate of P. aeruginosa strains that were tested in this study was in the following order: ceftazidime (53.44%; 31/58), followed by aztreonam (51.72%; 30/58), amikacin (44.82%; 26/58), azithromycin (41.37%; 24/58), gentamicin (37.93%; 22/58), cefepime (36.20%; 21/58), meropenem (25.86%; 13/58), piperacillin-tazobactam (25.86%; 13/58), imipenem (22.41%; 13/158), ciprofloxacin (17.24%; 10/58), tobramycin (8.62; 5/58), and polymyxin B (1.72; 1/58). The results highlight the importance of antibiotic susceptibility testing in Pseudomonas aeruginosa isolates from dogs with superficial infections to use an adequate treatment plan to manage the skin condition and other pathologies (otitis externa and perianal abscesses).

Aronia melanocarpa Ameliorates Adrenal Cytoarchitecture Against the Hexavalent Chromium-Induced Injury.

Hexavalent chromium is a toxin that penetrates the cell, triggering reactive oxygen species (ROS) production. Aronia melanocarpa, due to its proanthocyanidins, anthocyanins, and phenolic acid contents, is a valuable antioxidant. The aim was to observe the influence of hexavalent chromium Cr(VI) on the adrenal gland, and if this impact can be recovered by the administration of A. melanocarpa. Accordingly, 36 rats were divided into six groups: control; Aronia; Cr receiving Cr(VI) in distilled water for 3 months; CrA receiving a mix of Cr(VI) and A. melanocarpa at 2.5% aqueous extract for 3 months; Cr2 receiving, for 3 months, Cr(VI) in distilled water, and next, for 1 month, only distilled water; and respectively, CrA2 receiving, for 3 months, Cr(VI) in distilled water, followed by 1 month of Aronia at 2.5% extract administration. The adrenal gland samples were examined toward histological and molecular assessment, and results were statistically analyzed (ANOVA). Hexavalent chromium induced changes in the adrenal cortex expressed by focal or diffuse hypertrophies, cytoplasmic vacuolization (due to lipidic accumulation), and cells' shape and size alteration, including necrosis. These structural alterations were carried by Bax and Bcl2 gene expression: the Bax gene expression levels, increased significantly (p < 0.001) in all experimental groups, except the Aronia group, compared with control. In the Cr2, CrA, and CrA2 groups, notable reduction of Bax gene expression (p < 0.001) was reported compared with the Cr group. Regarding the Bcl2 gene expression (p < 0.001), a significant increase was observed in the experimental groups, compared with the control. The Bcl2 expression level had a similar pattern to Bax gene, consequently trying to compensate its overexpression. Aronia administered concomitantly, or after Cr(VI), diminished structural changes and expression of the studied genes, thus reducing the Bax/Bcl2 ratio and suggesting that the active ingredients from Aronia are capable of blocking apoptotic cascade induced by the pathway of Bax and Bcl2 proteins.

Is ECLIA Serum Cortisol Concentration Measurement, an Accurate Indicator of Pain Severity in Dogs with Locomotor Pain?

The purpose of determining serum cortisol level is to reflect the activity of stress axis, ethological alterations, acute and chronic pain, life quality, or psychogenic stress. Although it is stated that stress can produce a measurable influence on the cortisol level, a certified value of this pain biomarker in dogs was not generally accepted yet. This study aimed to investigate if serum cortisol measured follows allopathic treatments only, or it is associated with physiotherapy, point out pain level in dogs with orthopedic disease, which could reveal the healing progress. The diagnostic identified: hip dysplasia, cranial cruciate ligament rupture, or intervertebral disc disease. Ortolani and Barden tests, together with clinical examination, drawer sign, and tibia compression test, were done in dogs exhibiting postures, and motion alteration, and X-Ray confirmed. A total of 30 dogs were grouped in healthy (n = 10) and pain groups (n = 20), the blood sampling is done at the beginning of the investigation, and after ten days of the study. Dogs were handled in two ways: G1-treated with Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) only and respectively, G2-by therapy and physiotherapy. The analysis was performed on a Roche Cobas Analyzer (Roche, USA), serum cortisol being determined by Electrochemiluminescence immunoassay (ECLIA), and statistics using ANOVA, following Tukey's Multiple Comparison Test. The results revealed that, out of ten specimens in the Control group, nine were within the normal limits: 5-65 ng × mL-1 (24.76 ± 19.48678), and one sample under the set limit. In G1, it was observed that the plasmatic P1 values were below the levels of P2, in six situations. In G2, although the status of all subjects improved radically with the removal or evident reduction of pain, confirmed clinically and imagistically, the P2 values in five dogs were higher than the initial P1 values, and in contradiction with the observed clinical reality. Comparing results, the mean difference in G1 was 0.41, and in G2 = 2.54, with an SD for G1 = 13.38, and G2 = 16.66, registering moderate development. Standard deviation illustrated that the values of treated groups were highly spread throughout the interval, and the serum cortisol assay did not generate significant statistical differences between groups in our case. This inferred the doubt whether the used detection method or values registered correctly indicates the pain levels in dog species.

Gingival proliferative growth - stress and cytoarchitecture related with fixed and mobile orthodontic therapy.

The fixed orthodontic measures taken induce significant stress to the gingival growth process during arch wire maneuvers of aligning and leveling. We observed, for a period of one to four years, fixed orthodontic devices in 80 human subjects. From these, we selected 44 subjects (22 women and 22 men) where the inflammatory process exhibited following the orthodontic fixed treatment, and with vacuum-formed orthodontic retainers (VFR) succeeding to fixed treatment. Samples were collected from each patient and histological and immunohistochemical (IHC) methodology was made to analyze the cytoarchitecture. Statistics were made after one-way analysis of variance (ANOVA), with the Bonferroni's correction. The IHC examination performed in the early stage revealed the presence in the inflammatory infiltrate of CD8-type T-lymphocytes, and of dendritic cells in large numbers. The examination performed in the late stage revealed the presence in the inflammatory infiltrate of CD20-type B-lymphocytes, which are mature cells capable of immunoglobulin synthesis, their activation being an important step in the maturation of the antibody response. The stress generated by arch wires in both genders was significantly higher than in the case of VFR. This observation was pointed out also by the cytohistological investigation outcome but was also based on an original scale conceived by our research team, following gingival hyperplasia evaluation. Also, with statistical significance, the comparative obtained values for men (p=0.01) and for women (p=0.001) illustrate clinical observations, allowing to affirm that, in our case, men were more stressed in bearing arch wire devices (AWD) and VFR, in comparison with women.

The effect of cisplatin administration on certain trace elements homeostasis in rats and the protective effect of silver birch (Betula pendula) sap.

BACKGROUND: A clinically active structure with known antitumor activities is cisplatin (CDDP), but this it comes with toxicity characteristics which can be faded by the beneficial effects of Silver birch (Betula pendula) sap. OBJECTIVE: We aimed to assess the cisplatin activity on: Mn, Mg, Cu, Fe and Zn homeostasis in rats and to observe the effect of birch sap. METHODS: Healthy Wistar rats (n = 10/group) were divided in four groups: Control: receiving 1 mL saline I.P. way + water; E1: cisplatin 20 mg kgbw-1, I.P.; E2: cisplatin 20 mg kgbw-1, I.P. + birch sap and Control sap group: 1 mL saline I.P. + birch sap. Blood was collected: at the trial's start and after 48 h, and blood and organs (liver, kidney and spleen) for the cytoarchitecture investigation and readings were sampled after seven days. Samples were processed in nitric acid by microwave digestion and readings were completed by flame atomic absorption spectroscopy, the outcomes being statistically analyzed by ANOVA. RESULTS: Cisplatin produced a significant imbalance in the trace elements homeostasis, the birch sap administration recovering them usual homeostasis status. Comparatively with the Control, rats exposed to cisplatin presented a not significant (p > 0.05) decrease of Zn (-26.74%) and Mg (-10.25%), a significant (p < 0.05) decrease of Cu (-27.73%) at 48 h, a highly significant (p < 0.01) decrease of Cu (-56.08%) and Fe (-85.35%) at seven days after administration and a not significant (p > 0.05) increase of Mn (+28.16%). Birch sap administration after Cisplatin was followed by restoration or nevertheless significant increase (p < 0.05) of the investigated trace elements Zn (+56.88% to 48 h/+89.94% after seven days), Mg (+26.86%/+95.74%), Cu (+23.13%/+74.56%), Fe (+39.64%/+440.11%) and Mn (+4.30%/+15.87%), suggesting them defence against cisplatin. Histology revealed the order of main altered organs after the CDDP exposure: kidney, spleen and liver. CONCLUSIONS: The study recommended the important protective role of Betula pendula sap against diverse cisplatin deleterious side-effects.

PC2 Ovotransferrin: Characterization and Alternative Immunotherapeutic Activity.

Characterization and evaluation of immunotherapeutic potential of ovotransferrin PC2 (OTf PC2) were performed in this study. The ovoprotein was obtained from egg white from hens immunized with bacterial antigens, pathogenic for humans. For the negative control samples, OTf was extracted from eggs collected from Specific Pathogen-Free (SPF) hens and purified by affinity chromatography on Protein G-agarose column with two eluting peaks: I, representing ovalbumin, and II, ovotransferrin. The final apo-OTf form was reached by successive precipitation with ammonium sulfate and citric acid and the holo-OTf form by saturating the apo-form with FeCl3. Multiple OTf PC2 samples were analyzed through Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and, based on the molecular marker migration model, the ovotransferrin (76.5 kDa) and ovalbumin (45 kDa) were detected. The agglutination reaction exhibited statistically significant high specificity of the multiple OTf PC2, by reacting with the antigens used for hens' immunization. Following ELISA, it was established that OTf PC2 from hyperimmune eggs has specificity for all antigens; the antibody titer was high, indicating that OTf PC2 possesses immunological properties similar to immunoglobulin Y (Ig Y). This study suggests that OTf PC2 immunological activity may play a crucial role in the prevention and treatment of infections resistant to antibiotics and OTf PC2 can also act as a valuable nutraceutical.

The consequences of aluminium intake on reproductive function in male rats:a three-generation study.

BACKGROUND/AIM: The effects of aluminium exposure on reproductive biomarkers in male rats were followed in a three-generation study. MATERIALS AND METHODS: Forty Wistar male rats (F0) were divided into the following groups: control (C), receiving only tap water, and three experimental (E) groups, receiving aluminium sulphate (AS) (E1: 200 ppb, E2: 400 ppb, and E3: 1000 ppb) in drinking water for a 6-month exposure period. To obtain F1, three males from each group were mated with previously unexposed females (1:2 sex ratios) that during gestation and lactation were exposed to the same AS levels as males. The F1 generation male offspring were divided as described and exposed to the same AS levels. The protocol to obtain F2 was similar to that described for F1. RESULTS: Significantly lower testosterone levels in rats exposed to AS and in generations F1 and F2 compared to the parental one, luteinising hormone (LH) fluctuations in F0 and a significant LH decrease in F2 and F3 generations, testis weight decrease, increased immobile and abnormal sperm, and histoarchitecture alterations in the testes were observed. Moreover, interval between deliveries increased. CONCLUSION: Chronic exposure to AS was significantly deleterious, producing a pronounced decrease in the sperm count and testosterone levels in all experimental groups.

Germinated and Ungerminated Seeds Extract from Two Lupinus Species: Biological Compounds Characterization and In Vitro and In Vivo Evaluations.

In recent years, nutraceuticals attracted a great amount of attention in the biomedical research due to their significant contribution as natural agents for prevention of various health issues. Ethanolic extracts from the ungerminated and germinated seeds of Lupinus albus L. and Lupinus angustifolius L. were analyzed for the content in isoflavones (genistein) and cinnamic acid derivatives. Additionally, the extracts were evaluated for antimicrobial, antiproliferative, and anti-inflammatory properties, using in vitro and in vivo tests. Germination proved to be a method of choice in increasing the amount of genistein and cinnamic acid derivatives in both Lupinus albus L. and Lupinus angustifolius L. seeds. Biological evaluation of all vegetal extracts revealed a weak therapeutic potential for both ungerminated and germinated seeds.

The impact of exogenic testosterone and nortestosterone-decanoate toxicological evaluation using a rat model.

The impact of exogenic testosterone (T): 1.5 and 3.0 mg/kg.bw) and 19-nortestosterone 17-decanoate (ND): 1.5 and 7.5 mg/kg.bw) in castrated male rats was evaluated based on: (a) weight increase of the androgen target tissues, respecting the Hershberger methodology; (b) the 17α and ß-testosterone, 17 α and ß-estradiol and 17 α and ß-nortestosterone levels using the GC-MS/MS technique; and (c) observation of the serum free thyroxine levels (T4). Results revealed that T and ND significantly increased the weight of androgen target tissues as follows: ND was more influential on seminal vesicles, levator ani-bulbocavernosus muscle (LABC) and Cowper's glands and T (at a dose of 3.0 mg/kg.bw) influenced the weight of the ventral prostate and glans penis. Serum samples analyzed for steroid hormone levels showed the presence of 17ß-testosterone, 17ß-estradiol and 17ß-nor-testosterone, in castrated male rats injected with testosterone and nortestosterone, but no significant differences were found between thyroid responses and thyroid hormone levels. The results of this research proved the disrupting activity of T and ND when administered in high doses and the useful application of the Hershberger bioassay in the case of ND.

Cytoarchitecture of steroid dependent target tissues after testosterone administration compared to nandrolone decanoate in castrated rats in the aim of Hershberger bio test.

The objective was the cytoarchitecture evaluation of known steroid dependent target tissues after administering of testosterone, compared to action of its more active ester, nortestosterone (nandrolone decanoate) in castrated rat males in the aim of Hershberger bio test. Study was performed on 30 castrated male Wistar rats, aged between 35 and 39 days, in peripubertal period, divided into five groups. Androgen doses administration begun at the rats' age of 49 days. Animals were injected i.m., daily, for 10 consecutive days as follows: Aquatest (Balkan Pharmaceuticals Ltd., Moldova) testosterone aqueous solution: Testosterone I group (0.4 mg/animal); Testosterone II (0.8 mg/animal); (Deca-Durabolin, Balkan Pharmaceuticals); nandrolone decanoate oily solution: Nortestosterone I (1.5 mg/kg body weight); Nortestosterone II (7.5 mg/kg body weight) and Control (White sesame oil, Manicos, Romania, 0.1 mL/animal). Gonadectomy (GDX) induced modifications of target tissues wet weight accompanied by important modifications in cytoarchitecture. Changes following exogenous administration of testosterone and nortestosterone decanoate were found in: liver (granular dystrophy, mega-mitochondria, tubular intumescences), prostate (increasing of the structural elements), seminal vesicles (hyalinosis, thickening of cell walls and the hyaline presence), levator ani-bulbo-cavernosus muscle (muscle fibbers dilacerations), bulbourethral glands (muscular fibbers rarefaction by fluid accumulation) demonstrating the disruptor activity especially for overdosed nandrolone decanoate.