The Effect of Rubus idaeus Polyphenols Extract in Induced Endometriosis in Rats.

Endometriosis is a common gynecological condition with a complex physio-pathological background. This study aimed to assess the role of Rubus idaeus leaf extract (RiDE) as a potential therapeutic agent in reducing the size of the endometriotic lesions and modulate the plasma expression of MMP-2, MMP-9, and TGF-ß1. The endometriotic lesions were induced in a rat model by the autologous transplant of endometrium. Thirty-six female rats, Wistar breed, with induced endometriosis, were divided into four groups and underwent treatment for 28 days. The CTRL group received 0.5 mL/day of the vehicle; the DG group received 1 mg/kg b.w./day dienogest; the RiDG group received 0.25 mL/kg b.w./day RiDE and the D+RiDG group received 1 mg/kg b.w./day dienogest and 0.25 mL/kg b.w./day RiDE, respectively. Rats' weight, endometriotic lesion diameter and grade, and plasma levels of MMP-2, MMP-9, and TGF-ß1 were assessed before and after treatment. The administration of RiDE in association with dienogest vs. dienogest determined a lower weight gain and a reduction in diameter of the endometriotic lesions. RiDE administration restored MMP2 and MMP9 plasma levels to initial conditions. Rubus idaeus extract may help in reducing dienogest-associated weight gain, lower the size of endometriotic lesions, and have anti-inflammatory effects through MMP2 and MMP9 reduction.

Anti-inflammatory and antioxidant efficacy of lavender oil in experimentally induced thrombosis.

BACKGROUND: Lavender oil (LO) possesses anti-inflammatory, antioxidant, antifungal, antibacterial, sedative, cardio-protective, and antinociceptive properties. Thrombosis and inflammation are interplayed processes that interact and influence one another. Our research compared three routes of administration to assess the efficacy of pretreatment with LO on carrageenan-induced thrombosis in rat tail. MATERIALS AND METHODS: Wistar-Bratislava white rats were randomly divided into five groups of ten rats each and pretreated 3 consecutive days prior the inducement of thrombosis to with one dose of LO (150 mg/kg body weight (b.w.)): per os by gavage (TLOPO group), intraperitoneal (TIPLO group) and subcutaneous (TSCLO group). We also have a control (C, received saline solution 0.9% and DMSO (vehicle) 1 ml intraperitoneal (i.p.)) group and a group with thrombosis (T group, received saline solution 0.9% plus vehicle 1 ml i.p.). Histopathological examinations were conducted together with measurements of the circulating levels of three oxidative stress markers, antioxidant effect (TAC and THIOL), and three proinflammatory cytokines (TNF- α, RANTES, and MCP-1). RESULTS: When administered intraperitoneally, lavender oil has the best efficacy on circulating levels of oxidative stress parameters (MDA, NOx, TOS), one oxidative stress marker (THIOL), and all studied proinflammatory cytokines (p-values < 0.02). Moreover, TIPLO displayed the closest values for bleeding and clotting time to the C group, as well as the lowest length of the thrombus than the T, TPOLO, and TSCLO groups (p-values < 0.001). The TIPLO group has histological appearance comparable to the C group, with the exception of the presence of oedema. CONCLUSIONS: Lavender oil pretreatment with intraperitoneal administration as three days, one-dose per day, showed anti-inflammatory and antioxidant efficacy in experimentally induced thrombosis.

Intestinal Dysbiosis in Rats: Interaction between Amoxicillin and Probiotics, a Histological and Immunohistochemical Evaluation.

Gastrointestinal microbiota can be easily altered by common treatments, such as antibiotic therapy. However, the dysmicrobism induced by such a treatment may be counteracted by the administration of different beneficial microbes, such as probiotics. Therefore, this study aimed to establish the interaction between intestinal microbiota, antibiotic therapy, and sporulated bacteria, correlated with the evolution of growth indices. Twenty-five Wistar rats, females, were divided into five groups. Amoxicillin and a probiotic combination of Bacillus subtilis, Bacillus licheniformis, and Pediococcus acidilactici were administered according to each group's purpose. The conventional growth indices were calculated and histological and immunohistochemical assessments were realized from intestinal samples. The results of the conventional growth indices suggested a beneficial effect when the antibiotic therapy was accompanied by probiotics, while for the groups where the dysmicrobism was present, the values for feed conversion ratio were negative. These findings were supported by the microscopic aspects of the intestinal mucosa, which suggested a decreased absorption capacity due to significant morphological changes. Moreover, the immunohistochemical reaction of the inflammatory cells from intestinal lamina propria was intensely positive for the same affected groups. However, for the control group and the group with antibiotic and probiotic treatment, the immunopositivity was significantly decreased. Probiotics based on bacillus spores administered simultaneously with the antibiotic offered the best restoration of the gut microbiota, a fact suggested by the absence of intestinal lesions, a normal food conversion ratio, and low expression of TLR4 and LBP immunomarkers.

In Vitro and In Vivo Characterisation of a Mucoadhesive Buccal Film Loaded with Doxycycline Hyclate for Topical Application in Periodontitis.

Mucoadhesive films loaded with doxycycline hyclate (Doxy Hyc), consisting of mixtures of hydroxypropylmethyl cellulose (HPMC) E3, K4 and polyacrylic acid (Carbopol 940), were prepared by casting method, aiming to design a formulation intended for application in the oral cavity. The obtained film formulations exhibited a Doxy Hyc content between 7.52 ± 0.42 and 7.83 ± 0.41%, which had adequate mechanical properties for application in the oral cavity and pH values in the tolerance range. The x-ray diffraction studies highlighted the amorphisation of Doxy Hyc in the preparation process and the antibiotic particles present on the surface of the films, identified in the TEM images, which ensured a burst release effect in the first 15 min of the in vitro dissolution studies, after which Doxy Hyc was released by diffusion, the data presenting a good correlation with the Peppas model, n < 0.5. The formulation F1, consisting of HPMC K4 combined with C940 in a ratio of 5:3, the most performing in vitro, was tested in vivo in experimentally-induced periodontitis and demonstrated its effectiveness in improving the clinical parameters and reducing the salivary levels of matrix metalloproteinase-8 (MMP-8). The prepared Doxy Hyc loaded mucoadhesive buccal film could be used as an adjuvant for the local treatment of periodontitis, ensuring prolonged release of the antibiotic after topical application.

Immunomodulatory Effect of Novel Electrospun Nanofibers Loaded with Doxycycline as an Adjuvant Treatment in Periodontitis.

The immunomodulatory effect of a novel biomaterial obtained through electrospinning, based on polylactic acid (PLA) and nano-hydroxyapatite (nano-HAP), loaded with doxycycline (doxy) was evaluated in an animal model. The treatment capabilities as a local non-surgical treatment of periodontitis was investigated on the lower incisors of Wistar rats, after the induction of localized periodontitis using the ligature technique. Following the induction of the disease, the non-surgical treatment of scaling and root planing was applied, in conjunction with the application of the new material. The results of the treatment were evaluated clinically, using the tooth mobility and gingival index scores, as well as histologically. The salivary concentrations of matrix metalloproteinase 8 (MMP-8) and plasmatic concentrations of interleukin 1 (IL-1), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) were also monitored. Two weeks after the ligature application, the periodontal disease was successfully induced in rats. The application of the novel biomaterial obtained through electrospinning was proven to be more effective in improving the clinical parameters, while decreasing the salivary MMP-8 and plasmatic IL-1 and TNF-α concentrations, compared to the simple scaling and root planing. Thus, the novel electrospun biomaterial could be a strong candidate as an adjuvant to the non-surgical periodontal therapy.

Inhibition of Tumor Growth and Modulation of Antioxidant Activity of Rhodoxanthin Isolated from Taxus baccata Aril against B16F10 Murine Malignant Melanoma.

Malignant melanoma is the most aggressive type of skin cancer, and due to the numerous limitations of current treatment methods, there is an urgent need to develop novel approaches for both the prevention and treatment of malignant melanoma, with research-oriented bioactive substances representing a notable first step. The current study decided to expand on previous rhodoxanthin research by investigating the possible anti-tumor effect as well as the effect on the antioxidant status in the case of murine melanoma in an experimental model. The 21-day study was carried out on female C57BL/6J mice. On the first day of the experiment, they were subcutaneously inoculated with 106 B16F10 cells and were given rhodoxanthin orally until the end of the study. Rhodoxanthin supplementation significantly reduced tumor growth (42.18%) and weight (15.74%). Furthermore, the epidermal growth factor (EGF) activity was reduced and the concentration of 8-OHdG dropped in the treated melanoma-bearing mice compared to the untreated ones, demonstrating the role of rhodoxanthin in slowing tumor growth, one of the mechanisms being the reduction of EGF level and the decrease of DNA oxidation. The administration of rhodoxanthin determined variations in antioxidant enzymes, both at the plasma level and at the tissue level.

Morphometric Features and Microanatomy of the Lingual Filiform Papillae in the Wistar Rat.

The mammalian tongue plays a fundamental role in various physiological and behavioral activities. Significant morphological variations have been recorded in the tongue of several species. This study aims to obtain detailed histological and morphometric information about the filiform papillae on the surface of rat tongue. The tongues of five 10-month-old Wistar rats were utilized, which were later examined with a stereo-microscope. Fragments from the three regions of the tongue were collected for histological investigations. The tongue of the Wistar rat has an intensely keratinized stratified squamous epithelium, with the highest degree of keratinized epithelium covering the filiform papillae. The filiform papillae differ in density, with the highest density recorded on the posterior part of the lingual body and the lowest density on the protuberance. The shortest filiform papillae were observed on the apex of the tongue and the tallest on the anterior part of the lingual body. Interestingly, the orientation of the filiform papillae on the lingual protuberance was inclined posteroanteriorly, in the opposite way as compared to the papillae from all the other regions of the tongue. Histologically, a difference was recorded in the structure of the covering epithelium of the anterior vs. the posterior face of the filiform papillae.

Evaluation of Oxidative Stress Biomarkers, Pro-Inflammatory Cytokines, and Histological Changes in Experimental Hypertension, Dyslipidemia, and Type 1 Diabetes Mellitus.

The present study aims to compare the oxidative stress biomarkers, pro-inflammatory cytokines, and histological changes induced by three cardiovascular risk factors, namely, hypertension, dyslipidemia, and type 1 diabetes mellitus. Hypertension was induced with 40 mg/kg body weight (b.w.) of N omega-nitro-L-arginine-methyl (L-NAME) administered orally. Dyslipidemia was induced by the administration of a diet with a high cholesterol (2%) content. Diabetes mellitus was induced by intraperitoneal administration of a single dose of streptozocin (65 mg/kg). Malondialdehyde (MDA) and total oxidative status (TOS) are increased by all three cardiovascular risk factors (up to 207%). The indirect assessment of NO synthesis (NOx) is observed to be reduced after L-NAME administration (43%), and dyslipidemia induction (16%), while type 1 diabetes mellitus is associated with the highest levels of NOx (increased 112%). Hypertension, dyslipidemia, and type 1 diabetes reduced the total antioxidative capacity (TAC) and total thiol (SH) levels (up to 57%). The values of evaluated pro-inflammatory cytokines, tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß), assessed from the ascending aorta were elevated by all three cardiovascular risk factors, with the highest levels induced by type 1 diabetes mellitus (up to 259%). The histopathological examination of the ascending and descending aorta revealed reversible pro-atherogenic changes consisting of the accumulation of lipid droplets in the subendothelial connective tissue on rats with hypertension and dyslipidemia. Irreversible pro-atherogenic changes consisting of a reduction of the specific elasticity of the arteries were observed in rats with type 1 diabetes mellitus. Type 1 diabetes mellitus demonstrates an alteration of the oxidative stress parameters, the elevation of tissue levels of the pro-inflammatory cytokines and causing irreversible pro-atherogenic changes on the aortic wall.

Protective Effect of Liposomal Epigallocatechin-Gallate in Experimental Gentamicin-Induced Hepatotoxicity.

Our study aimed to assess the effect of liposomal epigallocatechin-gallate (LEGCG) compared with epigallocatechin-gallate (EGCG) solution on hepatic toxicity induced by gentamicin (G) administration in rats. Five groups were evaluated, a control group (no G administration) and four groups that received G (1 mL, i.p, 80 mg/kg b.w. (body weight/day), for 7 days) to which we associated daily administration 30 min before G of EGCG (G-EGCG, 2.5 mg/0.1 kg b.w.), LEGCG (G-LEGCG, 2.5 mg/0.1 kg b.w.) or silymarin (100 mg/kg b.w./day). The nitro-oxidative stress (NOx), catalase (CAT), TNF-α, transaminases, creatinine, urea, metalloproteinase (MMP) 2 and 9, and liver histopathological changes were evaluated. LEGCG exhibited better efficacy than EGCG, improving the oxidant/antioxidant balance (p = 0.0125 for NOx and 0.0032 for CAT), TNF-α (p < 0.0001), MMP-2 (p < 0.0001), aminotransferases (p = 0.0001 for AST and 0.0136 for ALT), creatinine (p < 0.0001), urea (p = 0.0006) and histopathologic liver changes induced by gentamicin. Our study demonstrated the beneficial effect of EGCG with superior results of the liposomal formulation for hepatoprotection in experimental hepatic toxicity induced by gentamicin.

The influence of haematogenous bone marrow on the early osseointegration of a titanium implant which penetrates the endosteum / La influencia de la médula ósea hematógena en la osteointegración temprana de un implante de titanio que penetra en el endostio

SUMMARY: The utility of metallic bio-medical implants in osseous or dental affections is irrefutable. The paper aims to test the tolerance of the bone marrow to titanium implants. Titanium implants were inserted in the femur of 11-months old rabbits. The implants penetrated the endosteum, half of their length getting into the haematogenous bone marrow. Seven days after the insertion we collected bone fragments containing the implant. The CT exam revealed a significant decrease in the density of the bone at the interface with the implant and a more discrete one aloof from the insertion area. The histologic exam after 7 days revealed osseous reparatory processes only in the endosteal area from where it expanded on the surface of the implant which was inside the marrow. The presence and intensity of the osseous reparatory processes after only seven days post-implant demonstrates that the marrow actively participates in bone regeneration and implants osseointegration.

RESUMEN: La utilidad de los implantes biomédicos metálicos en afecciones óseas o dentales es irrefutable. El documento tiene como objetivo probar la tolerancia de la médula ósea a los implantes de titanio. Se insertaron implantes de titanio en el fémur de conejos de 11 meses. Los implantes penetraron en el endostio y la mitad de su longitud penetró en la médula ósea hematógena. Siete días después de la inserción, recolectamos fragmentos de hueso que contenían el implante. El examen de TC reveló una disminución significativa en la densidad del hueso en la interfaz con el implante y una más discreta alejada del área de inserción. El examen histológico a los 7 días reveló procesos de reparación ósea solo en el área endóstica desde donde se expandió en la superficie del implante que estaba dentro de la médula. La presencia e intensidad de los procesos de reparación ósea después de solo siete días del implante demuestra que la médula ósea participa activamente en la regeneración ósea y en la osteointegración de los implantes.

Histochemical assessment of mucin-secreting cells in the stomach of domestic rabbit / Evaluación histoquímica de células secretoras de mucina en el estómago de conejo doméstico

SUMMARY: The mucous substances of the stomach in mammals are important not only for the protection of the gastric epithelium from the acid environment and grinding actions, but it facilitates some other functions of the stomach such as antibacterial, antimetastatic, and immunological roles. The goal of the study is to highlight the distribution of mucin-secreting cells in the gastric mucosa in domestic rabbits, including the type of mucus synthesized. The gastric samples collected from ten individual rabbits were fixed in 10 % buffered formalin and underwent later standard paraffin tissue sample processing, which included dehydration, clarification, and embedding in paraffin. The tissue sections were eventually stained histochemically by PAS reaction and by Alcian blue method (pH 2.5) for neutral and acidic mucins detection, respectively. The quantification of mucins in the cytoplasm of mucus-secreting cells was performed by grading the gastric tissue samples from negative (-) to intensely positive (++). The mucus elaboration was observed in all the regions of the stomach (i.e., cardial, fundic, and pyloric regions), but only for the neutral mucin. The acidic mucin synthesis occurred only in the secretory units of the gastric glands from the cardial region in the stomach. Pyloric glands synthesized the largest amounts of neutral mucins, followed by moderate amounts elaborated by cardial glands, while the fundic region does not synthesize it at all. The description of new microscopic features of the stomach in rabbits is fundamental not only for comprehending species-related physiological features but gastric pathological processes.

RESUMEN: Las sustancias mucosas del estómago en los mamíferos son importantes no solo para la protección del epitelio gástrico del ambiente ácido y las acciones de trituración, sino que facilitan además otras funciones del estómago, como son las funciones antibacterianas, antimetastásicas e inmunológicas. El objetivo del estudio fue resaltar la distribución de las células secretoras de mucina en la mucosa gástrica de conejos domésticos, incluido el tipo de moco sintetizado. Las muestras gástricas recolectadas de diez conejos se fijaron en formalina tamponada al 10 % y se sometieron a un procesamiento que incluyó deshidratación, clarificación e inclusión en parafina. Las secciones de tejido finalmente se tiñeron histoquímicamente mediante la reacción de PAS y el método del azul de Alcian (pH 2,5) para la detección de mucinas neutras y ácidas, respectivamente. La cuantificación de mucinas en el citoplasma de las células secretoras de moco se realizó clasificando las muestras de tejido gástrico desde negativas (-) hasta intensamente positivas (++). La elaboración de moco se observó en todas las regiones del estómago (es decir, cardias, fúndica y pilórica), pero solo para la mucina neutra. La síntesis de mucina ácida ocurrió solo en las unidades secretoras de las glándulas gástricas de la región correspondiente al cardias del estómago. Las glándulas pilóricas sintetizaron la mayor cantidad de mucinas neutras, seguidas de cantidades moderadas elaboradas por las glándulas cardiales, mientras que la región fúndica no las sintetizó en abso- luto. La descripción de nuevas características microscópicas del estómago en conejos es fundamental no solo para comprender las características fisiológicas relacionadas con las especies sino también para entender los procesos patológicos gástricos.

First report of deep crypt secretory cells in ascending colon in chinchilla (Chinchilla lanigera): a histological and histochemical study / Primer reporte de células secretoras de criptas profundas en colon ascendente en chinchilla (Chinchilla lanigera): un estudio histológico e histoquímico

SUMMARY: The normal morphology of the colon differs among mammal species.The ascending colon presents several types of cells, responsible for carrying different functions for this organ. Among them, the mucus-secreting cells ensure the integrity of the mucosa, local defense, protection against different external factors, inflammatory diseases, cancer, etc. The ascending colon from 5 adult male chinchillas were processed for paraffin embedding and stained with three methods: Goldner's trichrome, PAS reaction, and Alcian blue staining procedure. The results showed that the structure of the ascending colon is similar to the one described in other species, i.e. mucosa, submucosa, muscularis externa, and serosa. Regarding the mucus-secreting cells present in the deeper part of the mucosal crypts (deep crypt secretory or DCS cells) turned out to be different not only morphologically from the surface goblet cells but also regarding the type of mucus synthesized. DCS cells have a multivacuolated, faintly stained cytoplasm with moderately PAS-positive reaction and intensely positive reaction to Alcian blue stain. The mean surface of DCS cells was 521.6 μm2 as compared to 437.9 μm2 for goblet cells (p<0.05). In conclusion, our study describes for the first time in chinchilla (Chinchilla lanigera) the presence of formerly known non-goblet or vacuolated cells, and recently entitled DCS cells in the glandular epithelium of the colon. The understanding of morphological peculiarities in chinchilla may serve as a good basis to understand the pathophysiology of various conditions that may arise.

RESUMEN: La morfología normal del colon es diferente entre las especies de mamíferos. El colon ascendente presenta varios tipos de células, encargadas de llevar varias funciones a este órgano. Entre ellos, las células secretoras aseguran la integridad de la mucosa, defensa local, protección frente a diferentes factores externos, enfermedades inflamatorias, cáncer, etc. Se procesaron para su inclusión en parafina el colon ascendente de 5 chinchillas machos adultos y se tiñeron con tres métodos: tricrómico de Goldner, reacción PAS y Azul de Alcian. Los resultados mostraron que la estructura de del colon ascendente es similar a la descrita en otras especies, es decir, mucosa, submucosa, muscular externa y serosa. Las células secretoras de la mucosa presente en la parte más profunda de las criptas mucosas (células secretoras de la cripta profunda o células DCS) resultaron ser diferentes morfológicamente de las células caliciformes superficiales, con citoplasma levemente teñido con reacción PAS positiva moderada y reacción intensamente positiva a Azul de Alcian. La superficie media de las células DCS fue de 521,6 μm2 en comparación con 437,9 μm2 de las células caliciformes (p <0,05). En conclusión, nuestro estudio describe por primera vez en chinchilla (Chinchilla lanigera) la presencia de células no caliciformes o vacuoladas anteriormente conocidas, y recientemente denominadas células DCS en el epitelio glandular del colon. La comprensión de las peculiaridades morfológicas de la chinchilla puede servir como una buena base para comprender la fisiopatología de las diversas afecciones.

Protective Effects of Liposomal Curcumin on Oxidative Stress/Antioxidant Imbalance, Metalloproteinases 2 and -9, Histological Changes and Renal Function in Experimental Nephrotoxicity Induced by Gentamicin.

BACKGROUND: Our study aimed to assess the efficiency of Curcumin nanoformulation (LCC) on experimental nephrotoxicity induced by Gentamicin in rats. METHODS: Six groups of seven rats were used: C-(control group) received saline solution i.p. (i.p. = intraperitoneal), G-gentamicin (G, 80 mg/kg body weight (b.w.)), GCC1 and GCC2-with G and CC solution (single dose of 10 mg/kg b.w.-CC1, or 20 mg/kg b.w.-CC2), GLCC1 (10 mg/kg b.w.) and GLCC2 (20 mg/kg b.w.) with G and LCC administration. Oxidative stress parameters (NOx = nitric oxide, MDA = malondialdehyde, TOS = total oxidative stress), antioxidant parameters (CAT = catalase, TAC = total antioxidant capacity), matrix metalloproteinases (MMP-2 and MMP-9), and renal function parameters (creatinine, blood urea nitrogen, and urea) were measured. Kidneys histopathologic examination was made for each group. RESULTS: Pretreatment with CC and LCC in both doses had significantly alleviating effects on assessed parameters (NOx, MDA, TOS, CAT, TAC, MMP-2, and -9) as compared with the untreated group (p < 0.006). Histopathological aspect and renal function were significantly improved in CC and LCC groups. Liposomal formulation (LCC) showed higher efficiency on all examined parameters compared to CC (p < 0.006). CONCLUSIONS: Our results demonstrated improving renal function and kidney cytoarchitecture, oxidative stress/antioxidant/balance, and MMPs plasma concentrations with better dose-related efficacity of LCC than CC.

Glandular structures in the major interlobular and extrapancreatic ducts in the guinea pig.

The morphology of the pancreatic duct system did not receive much attention as compared to the microanatomy of the exocrine and endocrine pancreas. The histological peculiarities of the excretory duct system are of major importance especially in laboratory animals like guinea pigs. The paper describes the histological peculiarities of the major interlobular and extrapancreatic ducts in guinea pigs. The pancreatic tissue samples were collected from five guinea pigs. For histological investigation, several pancreatic fragments underwent fixation in 10% buffered formalin and were later processed by the standard paraffin technique. Subsequentially, tissue sections were stained by Goldner's trichrome staining. The mucous substances were assessed by Alcian blue and Periodic acid-Schiff staining methods. The interlobular and main pancreatic ducts in the guinea pig present a simple columnar epithelium surrounded by a thick layer of dense connective tissue. The aforementioned epithelium of the main pancreatic ducts includes principal cells, goblet cells and basal cells. Additionally, the ductal epithelium presents occasional unicellular multiloculated intraepithelial mucous glands and prominent extraepithelial glands. The latter adopts a simple or compound tubular feature. The mucus elaborated by the three glandular types is mostly neutral in goblet cells, predominantly acidic in extraepithelial ductal glands, and a similar amount of acidic and neutral mucin in intraepithelial glands. In conclusion, the epithelium-associated mucous glands in the interlobular and main pancreatic ducts in the guinea pig are restricted not only to goblet cells. A substantial mucous discharge probably with a protecting role against irritative pancreatic juice derives from the main ductal intraepithelial and extraepithelial glands.

Effect of Liposomal Curcumin on Acetaminophen Hepatotoxicity by Down-regulation of Oxidative Stress and Matrix Metalloproteinases.

BACKGROUND/AIM: The hepatoprotective role of various molecules in drug-induced hepatotoxicity arouses great interest. We investigated the effect of liposomal curcumin (LCC) on experimental acetaminophen (APAP)-induced hepatotoxicity. MATERIALS AND METHODS: Rats were randomly allocated into 5 groups, and the effect of two LCC concentrations was studied: group 1 - 1 ml intraperitoneal (i.p.) saline, group 2 - APAP pretreatment, group 3 - APAP+silymarin (extract of the silybum marianum with anti-inflammatory, anti-oxidant, and anti-fibrotic properties), group 4 - APAP+LCC1, group 5 - APAP+LCC2. The biomarkers of oxidative stress (nitric oxide and malondialdehyde) and antioxidant status of plasma (thiols and catalase), TNF-α, MMP-2 and MMP-9 serum levels were evaluated. RESULTS: An improvement in oxidative stress, antioxidant status, and TNF-α, MMP-2 and MMP-9 levels was obtained in groups pretreated with LCC compared to silymarin treatment, in a dose-dependent manner. Histopathological examination reinforced the results. CONCLUSION: Liposomal curcumin improves the oxidative stress/antioxidant balance and alleviates inflammation in experimental APAP-induced hepatotoxicity.

A novel mitochondria-targeting method using special staining for the detection of apoptotic hepatocytes.

Early detection of apoptotic cells on histological slides is of major importance for both diagnostic and research areas. In the current study, the aim was to propose a convenient method to stain the mitochondria and establish whether hepatocytes undergoing apoptosis can be identified in tissue sections using the proposed method. Liver tissue from five adult chinchillas was fixed with 10% neutral buffered formalin for Goldner's trichrome (GT) and Groat's iron hematoxylin and eosin (HE) stains and with Kolster's fixative for the Heidenhain's iron hematoxylin procedure. The HE and GT-stained sections showed the morphological features consistent with apoptosis i.e., homogenous intensely acidophilic cytoplasm, cell shrinkage with an irregular outline, nuclear shrinkage with cloudy karyoplasm, and karyopyknosis in the late stage. Sections stained with Heidenhain's iron hematoxylin method was used to pinpoint mitochondria and revealed cells which were undergoing the first stages of the apoptosis process i.e., disappearance of mitochondria from the cell, chromatin condensation and margination, paracentral localization of nucleoli, and vacuolated nuclei. In more advanced stages of apoptosis, cells presented significant nuclear and cytoplasmic changes. It was concluded that this is the first report targeting the mitochondria, by performing inexpensive histological staining techniques, in order to assess dead cells in situ.

The Effect of Mofettes on Oxidative Stress/Antioxidant Balance in Experimental Myocardial Ischemia.

BACKGROUND/AIM: Natural mofettes are gases resulting from post-volcanic emanations. This study aimed to examine the effect of mofette therapy on plasma oxidative stress and antioxidant parameters in rats after experimental induction of myocardial ischemia, as well as on structural changes in myocardial tissue. MATERIALS AND METHODS: White Wistar-Bratislava rats were divided into three groups. In groups 2 and 3, myocardial ischemia was induced by isoproterenol. Rats in group 3 were additionally exposed to high levels mofettes. Oxidative stress and antioxidant parameters were determined in plasma. The structural changes of the myocardium were observed in paraffin embedded slices contrasted using Goldner's trichrome staining. RESULTS: A statistically significant change in serum oxidative stress biomarkers, including nitric oxide, malondialdehyde, total oxidant status, as well as in the tested antioxidant molecules and total antioxidant capacity were observed in group 3 compared to group 2. Also, rats of group 3 showed an obvious improvement in inflammatory infiltration and repair of necrotic areas through collagen proliferation (proliferation of fibrous connective tissue) compared to group 2. CONCLUSION: Mofette had a beneficial effect on the balance between oxidative stress and antioxidant status following experimentally induced myocardial ischemia.

Osseointegration of zirconium dental implants three months after insertion in rabbit femur. Histopathological study.

Implantology is a highly researched field with a constant concern in finding and studying new implant materials. Lately, zirconium has become a very attractive alternative to the detriment of titanium, but the research results were sometimes contradictory. Thus, we considered as opportune to study the osseointegration of zirconium dental implants in rabbit femur, three months after insertion. The biological material was represented by five rabbits and the experimental protocol was approved by the Ethics Committee of the University of Oradea, Romania. The implants (zirconium ceramic dental implants: 5 mm length, 2.6 mm diameter) were inserted in the femur under controlled conditions, after creating a bone defect. The animals received the appropriate postoperative care. Three months later, the implantation area was harvested and processed for histological examination. The assessment of the osseointegration process of the zirconium implants showed that they were very well tolerated by the host organism that did not trigger any rejection processes. Approximately 80% of the compromised bone was replaced with newly formed bone in advanced stages of remodeling and consolidation. The proliferated bone near the implants acquired a structure similar to the rabbit diaphyseal bone, but with higher density and size of the osteons. The stage reached by the osseointegration process three months after the insertion of the implants, ensures a good consolidation of the implants that supports the prosthetic structures, which are to be built on them.

ElectronixTutor: an intelligent tutoring system with multiple learning resources for electronics.

BACKGROUND: The Office of Naval Research (ONR) organized a STEM Challenge initiative to explore how intelligent tutoring systems (ITSs) can be developed in a reasonable amount of time to help students learn STEM topics. This competitive initiative sponsored four teams that separately developed systems that covered topics in mathematics, electronics, and dynamical systems. After the teams shared their progress at the conclusion of an 18-month period, the ONR decided to fund a joint applied project in the Navy that integrated those systems on the subject matter of electronic circuits. The University of Memphis took the lead in integrating these systems in an intelligent tutoring system called ElectronixTutor. This article describes the architecture of ElectronixTutor, the learning resources that feed into it, and the empirical findings that support the effectiveness of its constituent ITS learning resources. RESULTS: A fully integrated ElectronixTutor was developed that included several intelligent learning resources (AutoTutor, Dragoon, LearnForm, ASSISTments, BEETLE-II) as well as texts and videos. The architecture includes a student model that has (a) a common set of knowledge components on electronic circuits to which individual learning resources contribute and (b) a record of student performance on the knowledge components as well as a set of cognitive and non-cognitive attributes. There is a recommender system that uses the student model to guide the student on a small set of sensible next steps in their training. The individual components of ElectronixTutor have shown learning gains in previous decades of research. CONCLUSIONS: The ElectronixTutor system successfully combines multiple empirically based components into one system to teach a STEM topic (electronics) to students. A prototype of this intelligent tutoring system has been developed and is currently being tested. ElectronixTutor is unique in its assembling a group of well-tested intelligent tutoring systems into a single integrated learning environment.

Comparative immunohistochemical assessment of the effect of repetitive anesthesia with isoflurane and sevoflurane on rat liver / Avaliação imunohistoquímica comparativa do efeito da anestesia repetitiva com isoflurano e sevoflurano sobre o fígado de rato

Abstract Background and objectives: Inhalation anesthetics are used in human, as well as veterinary medical practice. In the present study we investigated the effect of isoflurane and sevoflurane on rat hepatocytes. Methods: A total of 40 Wistar female rats were used in this study. Animals were divided in groups of 5 rats. Groups IM, SM served as control groups. Groups I1, I2, I3 were used to study isoflurane and S1, S2, S3 for sevoflurane study. They were anesthetized 3 times, for 2 h long, at 2 days interval with a concentration of: 1.5% isoflurane (I1, I2, I3) and 2% sevoflurane (S1, S2, S3). The oxygen supply throughout the anesthesia was 1 L O2/min. Groups IM, IS, I1, S1 were sacrificed immediately after the last anesthesia. Groups I2, S2 were sacrificed 6 h after the last anesthesia, and groups I3, S3, 24 h post-anesthesia. Liver samples were harvested to highlight caspase-3 in apoptotic hepatocytes. Results: Following isoflurane administration, there were less than 1% cells in apoptosis highlighted in rat livers from groups IM, I1 and I2. At 24 h post-anesthesia (group I3), a small number of apoptotic hepatocytes was highlighted (around 3.23% cells in apoptosis), with a strictly periacinar disposition, randomly distributed in a small number of hepatic lobules. After sevoflurane administration, less than 1% apoptotic hepatocytes were identified at all control moments throughout the study. Conclusions: The results suggest that the anesthetics do not present a considerable hepatotoxicity. The comparative assessment of the two anesthetics shows that sevoflurane is superior to isoflurane.

Resumo Justificativa e objetivos: Anestésicos inalatórios são usados em humanos e também na prática médica veterinária. No presente estudo investigamos o efeito de isoflurano e sevoflurano em hepatócitos de rato. Métodos: Foram usados neste estudo 40 ratos Wistar fêmeas. Os animais foram divididos em grupos de cinco. Os grupos IM e SM serviram como controle. Os grupos I1, I2 e I3 foram usados para o estudo de isoflurano e os grupos S1, S2 e S3 para o estudo de sevoflurano. Os ratos foram anestesiados três vezes, durante duas horas em intervalos de dois dias, com uma concentração de 1,5% de isoflurano (I1, I2, I3) e 2% de sevoflurano (S1, S2, S3). O fornecimento de oxigênio durante a anestesia foi de 1 L O2/min. Os grupos IM, IS, I1 e S1 foram sacrificados imediatamente após a última anestesia. Os grupos I2 e S2 foram sacrificados seis horas após a última anestesia e os grupos I3 e S3 foram sacrificados 24 horas após a anestesia. Amostras dos fígados foram colhidas para ressaltar a caspase-3 em hepatócitos apoptóticos. Resultados: Após a administração de isoflurano, havia menos de 1% das células em apoptose em destaque nos fígados dos ratos dos grupos IM, I1 e I2. Às 24 horas após a anestesia (grupo I3), um pequeno número de hepatócitos apoptóticos foi destacado (3,23% de células em apoptose), com uma disposição estritamente periacinar, distribuídos aleatoriamente em um pequeno número de lóbulos hepáticos. Após a administração do sevoflurano, menos de 1% de hepatócitos apoptóticos foi identificado em todos os momentos de controle ao longo do estudo. Conclusões: Os resultados sugerem que os anestésicos não apresentam uma hepatotoxicidade considerável. A avaliação comparativa dos dois anestésicos mostra que sevoflurano é superior ao isoflurano.