Intranasal delivery of paroxetine: A preclinical study on pharmacokinetics, depressive-like behaviour, and neurochemical sex differences.

Treatment of major depressive disorder remains a major unmet clinical need. Given the advantages of intranasal administration for targeted brain delivery, the present study aimed at investigating the pharmacokinetics of paroxetine, after its intranasal instillation and assessing its potential therapeutic effect on female and male mice subjected to unpredictable chronic mild stress (UCMS) protocol. IN administration revealed direct nose-to-brain paroxetine delivery but dose- and sex-dependent differences. Pharmacokinetics was nonlinear and paroxetine concentrations were consistently higher in plasma and brain of male mice. Additionally, UCMS decreased animal preference for sucrose in both male and female mice following acute (p < 0.01) and chronic stress (p < 0.05), suggesting anhedonia. Both male and female mice exhibited depressive-like behavior in the forced swimming test. UCMS females displayed a significantly longer immobility time and shorter climbing time than the control group (p < 0.05), while no differences were found between male mice. Two weeks of paroxetine intranasal administration reduced immobility time and lengthened climbing and swimming time, approaching values similar to those observed in the healthy control group. The therapeutic effect was stronger on female mice. Importantly, melatonin plasma levels were significantly decreased in female mice following UCMS (p < 0.05), while males exhibited heightened corticosterone levels. On the other hand, treatment with IN paroxetine significantly increased corticosterone and melatonin levels in both sexes compared to healthy mice (p < 0.05). Intranasal paroxetine delivery undoubtedly ameliorated the behavioral despair, characteristic of depressive-like animals. Despite its efficiency in male and female mice subjected to UCMS, females were more prone to this novel therapeutic strategy.

The usefulness of ecotoxicological tools to improve the assessment of water bodies in a climate change reality.

This study aimed to analyse the added value of using ecotoxicological tools to complement and improve the assessment of natural water bodies status, in situations of climate change, with a higher frequency of extreme events as floods or droughts. Four water bodies of streams in the Guadiana Basin (Álamos, Amieira, Lucefécit, Zebro) were studied in 2017 and 2018 and classified based on the Water Framework Directive (WFD) parameters: Biological Quality Element - Phytobenthos (diatoms), General chemical and physicochemical elements, Specific pollutants, and Priority Substances. Complementarily, bioassays (including lethal and sublethal parameters) were carried out with organisms of different trophic levels: (i) the bacteria Aliivibrio fischeri; (ii) the microalgae Pseudokirchneriella subcapitata; (iii) the crustaceans Daphnia magna, Thamnocephalus platyurus and Heterocypris incongruens. A classification system with 5 scores was developed, permitting to classify water bodies from non-toxic (EC50 > 100 %; growth and feeding rate > 80 %; blue) to highly toxic (EC50 < 10 %; growth and feeding rate < 10 %; red). The comparison between the classification based on the WFD parameters and on ecotoxicological endpoints showed similar results for 71 % of the samples, and significant positive Pearson correlations were detected between the diatom-based Specific Polluosensitivity Index (SPI) and EC50V.fisheri, the algae growth rate and Shannon diversity index. These results indicate that when the biological quality elements cannot be used (namely under drought or flooding conditions) the application of ecotoxicological bioassays may be a good alternative. Further, when ecotoxicological parameters were included, an increase of worse classifications (Bad and Poor) was observed, revealing an improvement in the sensitivity of the classification, mainly in presence of specific and priority substances. So, the ecotoxicological analysis appears to provide useful information regarding the potential presence of both known and unknown contaminants at concentrations that cause biological effects (even within the WFD limits), in agreement with several authors that have already suggested its use in biomonitoring.

Vaginal collagen I and III changes after carbon dioxide laser application in postmenopausal women with the genitourinary syndrome: a pilot study.

OBJECTIVE: The aim of the study was to evaluate the clinical response and collagen remodeling in the vaginal wall after three sessions of carbon dioxide (CO2) laser application. METHODS: Fourteen postmenopausal women with vulvovaginal atrophy, aged 45-65 years and sexually active, were evaluated with clinical questionnaires, gynecological examinations and histological techniques before and after 20 weeks of treatment (ClinicalTrials.gov NCT03939078). Treatment consisted of 3-monthly sessions of the CO2 laser. Clinical questionnaires included the Vaginal Health Index, the Female Sexual Function Index and the International Consultation on Incontinence Questionnaires Short Form. Biopsies were taken from the lateral vaginal wall at week 0 (left wall) and week 20 (right wall). Tissue samples were stained with hematoxylin and eosin, Periodic Acid-Schiff, Picrosirius Red Stain and Orcein dyes. Immunohistochemical study was used to quantify collagens I and III in the samples. RESULTS: The mean age was 54.4 ± 4.5 years, and the average time of amenorrhea was 7.6 ± 5.1 years. The Female Sexual Function Index and the Vaginal Health Index Score values increased while the International Consultation on Incontinence Questionnaire Short Form score decreased after the programmed treatment. There was no significant change in vaginal pH. Histological studies showed increases in the total and superficial epithelial cell layers, and type III collagen fibers (from 10.86 ± 7.66 to 16.87 ± 3.96, p < 0.05), and immunohistochemical studies confirmed the significant increase in collagen III. CONCLUSION: Histological findings revealed epithelial atrophy reversal and collagen remodeling of the vaginal wall. Immunohistochemical analysis showed an increase in collagen type III fibers.

Evaluation of the immunomodulatory activity of thalidomide on tumor-associated macrophages in the 4T1 murine metastatic breast cancer model / [Avaliação do efeito imunomodulador da talidomida em macrófagos associados a tumores no modelo murino de câncer de mama metastático 4T1]

The present work evaluated the immunomodulatory effect of thalidomide (Thal) at different doses on tumor-associated macrophages (TAMs) using a mouse model of human breast cancer. Mice were inoculated with 4T1 cells in the left flank and treated with Thal once a day at concentrations of 50, 100, and 150mg/kg body weight from the 5th day until the 28th day of tumor inoculation. The tumors were sized, proliferation index and TAMs count were evaluated in primary tumors and metastatic lungs. In addition, the metastasis rate was evaluated in the lungs. Thal at 150mg/kg significantly decreased tumor growth, proliferation index, and TAMs infiltration in primary tumors. Conversely, a higher number of TAMs and lower proliferation index were observed in metastatic lungs in mice treated with 150mg/kg of Thal. Furthermore, Thal at 150mg/kg significantly decreased the metastatic nodules in the lungs. Our findings demonstrated that Thal treatment considerably decreased the primary tumor and lung metastasis in mice associated with different TAM infiltration effects in these sites.(AU)

No presente trabalho, foi avaliado o efeito imunomodulador de diferentes doses de talidomida em macrófagos associados ao tumor (TAMs), em um modelo murino de câncer de mama. Camundongos foram inoculados com células 4T1, na região do flanco esquerdo, e tratados com talidomida, uma vez ao dia, nas doses de 50, 100 e 150mg/k, por massa corporal, do quinto dia ao 28º dia de inoculação tumoral. Os tumores foram medidos, o índice de proliferação celular e a contagem de TAMs foram avaliados nos tumores primários e nos pulmões com metástases. Além disso, a taxa de metástases pulmonares também foi avaliada. A talidomida na dose de 150mg/kg diminuiu significativamente o crescimento tumoral, o índice de proliferação celular e a infiltração de TAMs nos tumores primários. Por outro lado, maior número de TAMs e menor índice de proliferação celular foram observados nos pulmões metastáticos, em camundongos tratados com 150mg/kg de talidomida. Ademais, a talidomida na dose de 150mg/kg diminuiu significativamente os nódulos metastáticos nos pulmões. Os resultados demonstraram que o tratamento com talidomida diminuiu o crescimento tumoral e as metástases pulmonares em camundongos, associado com diferentes efeitos na infiltração de TAMs nesses locais.(AU)

Occurrence and risk assessment of pesticides in a Mediterranean Basin with strong agricultural pressure (Guadiana Basin: Southern of Portugal).

The study aimed to assess the occurrence and the environmental risk of a group of 51 selected pesticides in the Guadiana Basin (a biodiversity hotspot, in the Mediterranean). The most abundant pesticides were bentazone and 2,4-D, while terbuthylazine together with terbutryn constituted the most ubiquitous pesticides. Eighteen out of the 38 pesticides detected are no longer approved in Europe, and 5 of them are included in the list of priority substances. The risk assessment showed that azinphos ethyl, diflufenican, irganol, imidacloprid, and oxadiazon occurred occasionally, but always in concentrations above their respective ecotoxicological threshold value. Contrary, bentazone, terbuthylazine, and terbutryn presented a high risk in most of the sampled locations and periods. The site-specific risk assessment showed a spatial and temporal pattern, with a higher risk occurring mainly in intermittent streams, in the drought period. The presence of pesticides banned from the EU market since 2009 showed the importance of improving the monitoring process, to identify the main sources of pollution and the fate of these emerging compounds. The results showed the need of implementing actions to improve the sustainable use of pesticides in agricultural areas, working with farmers and management entities to reduce the contamination of aquatic ecosystems. Transboundary water governance is also required to solve potential transboundary contamination problems.

Development and chemical characterization of biodegradable polymeric implants containing sirolimus for the treatment of malignant solid tumors.

The use of sirolimus and its analogs has been evaluated in studies aimed at combating several types of cancer; however, because of the limited bioavailability of the drug, the search for new forms of administration is required. Biodegradable polymeric implants containing sirolimus were developed and assessed as an alternative method of drug administration. Implants containing 25 % (w/w) sirolimus were prepared employing the polymer matrices chitosan, polycaprolactone and poly(lactic-co-glycolic acid) (PLGA) in two proportions: PLGA 50:50 and PLGA 75:25. Thermal analysis techniques such as thermogravimetry and differential scanning calorimetry, combined with x-ray diffraction were used to characterize and evaluate the compatibility of the constituents of the formulation. No incompatibilities were found between the components, but drug amorphization was observed in all samples. Implants made from the polymers chitosan and PCL may accelerate the degradation of SRL when these polymers are dissolved in methanol at 50 °C. HPLC analysis showed that the implant prepared with PLGA 75:25 did not present degradation products and maintained its appropriate drug content, even when dissolved in methanol and heated to 50 °C. Therefore, it represents the most suitable biodegradable polymer for use in implants developed for the treatment of malignant solid tumors. However, it is still necessary to further study the drug effects after amorphization of the crystal and also to perform stability and solubility analysis.

Antiangiogenic activity of a bevacizumab-loaded polyurethane device in animal neovascularization models.

PURPOSE: To evaluate the antiangiogenic activity of bevacizumab-loaded polyurethane using two animal models of neovascularization. METHODS: The percentage of blood vessels was evaluated in a chicken chorioallantoic membrane model (n=42) and in the rabbit cornea (n=24) with neovascularization induced by alkali injury. In each model, the animals were randomly divided into the groups treated with the bevacizumab-loaded polyurethane device, phosphate-buffered-saline (negative control) and bevacizumab commercial solution (positive control). Clinical examination, as well as histopathological and immunohistochemical evaluation, were performed in the rabbit eyes. Microvascular density in hot spot areas was determined in semi-thin sections of corneal tissue by hematoxylin-eosin staining and factor VIII immunohistochemistry. Immunohistochemical analysis was also performed to evaluate VEGF expression. RESULTS: In the evaluated models, the use of bevacizumab (Avastin®) and the bevacizumab-loaded polyurethane device led to similar results with regard to inhibition of neovascularization. In the chorioallantoic membrane model, the bevacizumab-loaded polyurethane device reduced angiogenesis by 50.27% when compared to the negative control group. In the rabbit model of corneal neovascularization, the mean density of vessels/field was reduced by 46.87% on analysis of factor VIII immunohistochemistry photos in the bevacizumab-loaded polyurethane device group as compared to the negative control (PBS) sections. In both models, no significant difference could be identified between the bevacizumab-loaded polyurethane device and the positive control group, leading to similar results with regard to inhibition of neovascularization. CONCLUSIONS: The present study shows that the bevacizumab-loaded polyurethane device may release bevacizumab and inhibit neovascularization similarly to commercial bevacizumab solution in the short-term.

PLGA Implants containing vancomycin and dexamethasone: development, characterization and bactericidal effects.

Post-operative endophthalmitis is an infection and an inflammation of the eye following a surgical procedure. Its treatment is based on drug injections into the eye. However, this treatment can lead to ocular complications. Intraocular implants could substitute the conventional therapy. Poly(lactic-co-glycolic acid) (PLGA) implants comprising on vancomycin and dexamethasone were evaluated as drug delivery system to treat endophthalmitis after cataract surgery. Implants were characterized by drug content uniformity, Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Wide Angle X-ray Scattering (WAXS), Scanning Electron Microscopy (SEM) and in vitro drug release. The bactericidal effect of vancomycin, eluted from the implants, was demonstrated against Staphylococcus aureus and Staphylococcus epidermidis. The drugs were uniformly distributed in the polymer. The analytical techniques revealed the chemical integrity of the drugs incorporated into the polymer and the modification of dexamethasone semi-crystalline nature. Drugs were controlled released from implants; and the eluted vancomycin showed bactericidal effects. In conclusion, PLGA implants containing vancomycin and dexamethasone may represent a therapeutic alternative to treat post-operative endophthalmitis.

Analysis of acyclovir in vitreous humor by a validated HPLC method.

An HPLC-UV method was developed and validated for the determination of acyclovir in vitreous humor. The method was carried out in isocratic mode using 0.02 mol/L acetic acid/methanol (95:5) as mobile phase, a C18 column at 25 degrees C and UV detection at 254 nm. The method was linear (r2> 0.99) over the range of 35-700 microg/mL, precise (RSD <5%), accurate (recovery ranged from 98.18 to 99.64%), robust, selective regarding of the vitreous humor, and robust remaining unaffected by deliberate variations in relevant parameters. The validated HPLC-UV method can be successfully applied to determine acyclovir directly injected into the vitreous cavity of rabbits' eye.

Intravenous hypertonic saline solution (7.5%) and oral electrolytes to treat of calves with noninfectious diarrhea and metabolic acidosis.

OBJECTIVE: The aim of this study was to compare the efficacy of treating osmotic diarrhea and dehydration in calves with hypertonic saline solution (HSS) IV, isotonic electrolyte solution (IES) PO, and a combination of these 2 solutions (HSS + IES). EXPERIMENTAL DESIGN: Eighteen male calves 8-30 days of age were used to evaluate the efficacy of 3 methods of fluid therapy after induction of osmotic diarrhea and dehydration. The diarrhea and dehydration were induced by administration of saccharose, spironolactone, and hydrochlorothiazide for 48 hours. The animals were randomly divided into 3 experimental groups: Group 1: 7.2% hypertonic saline solution-HSS (5 mL/kg IV); Group 2: oral isotonic electrolyte solution IES (60 mL/kg PO); or Group 3: HSS+IES. Clinical signs and laboratory finding observed 48 hours post-induction (Time 0) included diarrhea, dehydration, lethargy, and metabolic acidosis. RESULTS: Calves treated with HSS + IES experienced decreases in hematocrit, total protein concentration, albumin concentration, urea nitrogen concentration, and plasma volume as well as increases in blood pH, blood bicarbonate concentration, and central venous pressure between 1 and 3 hours post-treatment. These findings also were observed in animals treated with IES, however, at a slower rate than in the HSS + IES-treated animals. Animals treated with HSS continued to display signs of dehydration, lethargy, and metabolic acidosis 24 hours post-treatment. CONCLUSION: Treatment with a combination of HSS and IES produced rapid and sustainable correction of hypovolemia and metabolic acidosis in calves with noninfections diarrhea and dehydration.

Fetal calf serum enhances in vitro production of Bos taurus indicus embryos.

The objective of this study was to determine the effect of fetal calf serum (FCS) on the quality of in vitro produced bovine embryos. Cumulus oocyte-complexes (COCs, n = 2 449) recovered by ovum pick-up from Bos taurus indicus donors were randomly assigned to experimental groups. Sperm selected by Percoll gradient was used for in vitro fertilization (insemination = Day 0). In Experiment 1 (n = 1 745 COCs), zygotes were cultured in vitro in Synthetic Oviduct Fluid + 4 mg/mL of bovine serum albumin (BSA), or BSA + 2% FCS (BSA+FCS). In Experiment 2 (n = 704 COCs), the COCs were cultured in SOF + BSA, BSA + 2% FCS, or BSA + 2% FCS on D4 (BSA + FCSD4). In Experiment 1, blastocyst yield (51%) and Quality I blastocysts (41%) at Day 7 were higher (P < 0.05) in the BSA + FCS treatment than in BSA (42 and 30%, respectively). In Experiment 2, blastocyst yield was higher (P < 0.05) in the BSA+FCS (47%) treatment. Quality I blastocyst yield was higher (P < 0.05) for BSA + FCS (34%) and BSA+FCSD4 (32%) compared to the BSA treatment (20%). A total of 820 embryos were transferred, with no significant differences among groups in pregnancy rates. In conclusion, in vitro culture in SOFaaci + BSA + FCS enhanced blastocyst yield and Quality I blastocysts; adding FCS to the culture medium increased the efficiency of IVP of bovine embryos.

Flare-up of hand osteoarthritis caused by zoledronic acid infusion.

Zoledronic acid is effective for osteoporosis at a single annual intravenous dose. It usually causes few adverse effects; the most common are related to acute phase reactions. We reported the case of a 64-year-old woman who presented flare-up of hand osteoarthritis after zoledronic acid infusions. Despite the fact that arthralgia is a common side effect of intravenous bisphosphonates, development of inflammatory signs in osteoarthritic joints is a rare event. We hypothesized that this side effect is caused by a release of cytokines secondary to activation of gamma-delta T lymphocytes.

Solid state evaluation of some thalidomide raw materials.

Thalidomide presents polymorphism and is a problematic drug due to its poor solubility and difficult tablet processability, which is the dosage form available in Brazil. In most cases, the pharmacopoeias specify do not address solid state characterization of drugs precisely. In this work, different thalidomide commercial samples were characterized by infrared spectroscopy, particle size analysis, scanning electron microscopy, and X-ray diffraction. In addition, the polymorphic forms were quantified for Rietveld analysis and their intrinsic dissolution rates were evaluated. The results demonstrated the market availability of different raw materials which lack of homogeneity due to differences related to crystalline constitution, crystal habit and intrinsic dissolution rate.

Disease activity as a major risk factor for osteonecrosis in early systemic lupus erythematosus.

This study Identifies the possible risk factors for osteonecrosis (ON) in a homogenous group of early system lupus erythematosus (SLE). Forty-six consecutive SLE patients (<5 years duration) followed at the Lupus Clinic, were enrolled between 2004 and 2005. An extensive clinical and laboratory evaluation using a standard electronic protocol established since 1999, including osteonecrosis symptoms and appropriate magnetic resonance imaging (MRI), were carried out at 1-6 months intervals. All other asymptomatic for osteonecrosis patients at study entry underwent MRI. ON confirmed by MRI was found in 10 of 46 patients (22%). Age, disease duration, clinical vascular features, frequency of thrombophilia and hypofibrinolysis factors and the lipoprotein profile were comparable in patients with and without osteonecrosis (P > 0.05). Remarkably, the frequency of patients with system lupus erythematosus disease activity index (SLEDAI) > or =8 in the previous year of osteonecrosis clinical diagnosis was significantly higher when compared to patients without this manifestation (60.0% versus 19.4%, P = 0.011), supported by the higher glucocorticoid cumulative dose in the same period (P = 0.045). In contrast, these two parameters evaluated in 13th-24th months preceding osteonecrosis diagnosis were similar in patients with and without osteonecrosis (P > 0.05). In the logistic regression analysis only SLEDAI remained as an independent risk factor for ON (OR = 6.78, CI = 1.05-43.55, P = 0.04). Disease activity in the previous year of ON clinical diagnosis is the main predictor factor for the development of this complication in early SLE.

Safety and pharmacokinetics of an intravitreal biodegradable implant of dexamethasone acetate in rabbit eyes.

The treatment of vitreoretinal diseases is limited and, nowadays, new drug delivery approaches have been reported in order to increase drug bioavailability. The objective of the current study was to determine the pharmacokinetic profile of a biodegradable dexamethasone acetate implant inserted into the vitreous of rabbits and to evaluate its potential signs of toxicity to the rabbits' eyes. The results showed that the intravitreous drug concentration remained within the therapeutic range along the 8-week period of evaluation. The system under study was not toxic to the normal rabbit retina, and no significant increase in intraocular pressure was observed.

Anestesia inalatória em equinos com halotano e óxido nitroso: efeitos sobre os gases sanguíneos e equilíbrio ácido-base / Effects of halothane and nitrous oxide anesthesia on blood gases and acid-base-balance in horses

Estudaram-se os valores do pH, pCO2, HCO3, BE, pO2, concentraçäo e saturaçäo da hemoglobina do sangue arterial e venoso de 10 equinos mantidos anestesiados durante uma hora com halotano e óxido nitroso, o qual foi misturado com o oxigênio na proporçäo de 1:1, para fazer o transporte do halotano. Houve queda na frequência respiratória, provocando aumento da paCO2 e diminuiçäo do pHa, caracterizando uma acidose respiratória. Os dados sobre pHa, paCO2 e HCO3 näo mostraram diferença estatística entre os tempos. Houve uma queda significativa da concentraçäo de hemoglobina após 15 minutos de anestesia e a saturaçäo da hemoglobina encontrou-se abaixo dos limites normais. A comparaçäo dos valores do sangue arterial e venoso demonstra que näo houve diferença significativa da concentraçäo de hemoglobina, enquanto que a saturaçäo da hemoglobina do sangue arterial apresentou diferença significativa. Houve diferença significativa entre os tempos das variáveis frequência do pulso e näo houve diferença significativa na temperatura corporal. A comparaçäo dos dados da gasometria do sangue arterial e venoso revelou diferença significativa do pH, pCO2, e saturaçäo da hemoglobina. Conclui-se que é aconselhável o uso de sangue arterial na realizaçäo de gasometria em equinos

Pneumatically powered vertebral displacement device for dogs.

A pneumatically powered trauma device was developed to produce vertebral displacement and spinal cord injury in dogs. After surgical exposure of the spine, displacement was done without laminectomy or cutting of the intervertebral disc. Trauma resulted in luxation between L1 and L2 or fractures of L1 and L2. Force required to cause displacement was recorded via a load sensing system and ranged between 1,848 and 2,666 newtons. In a few dogs, there was partial return of the displaced segment upon pressure release from the device at posttrauma hour 4. The device was an effective means of producing spinal injuries in dogs, and provides a method for studying spinal column and spinal cord injuries.