Factors Predicting Prolonged Mechanical Ventilation in Guillain-Barré Syndrome.

BACKGROUND: Up to 30% of patients with Guillain-Barré syndrome require mechanical ventilation and 5% die due to acute complications of mechanical ventilation. There is a considerable group of patients that will need prolonged mechanical ventilation (considered as >14 days) and should be considered for early tracheostomy. The objective of this study is to identify risk factors for prolonged mechanical ventilation. METHODS: We prospectively analyzed patients with Guillain-Barré diagnosis with versus without prolonged mechanical ventilation. We considered clinical and electrophysiological characteristics and analyzed factors associated with prolonged mechanical ventilation. RESULTS: Three hundred and three patients were included; 29% required mechanical ventilation. When comparing the groups, patients with prolonged invasive mechanical ventilation (IMV) have a lower score on the Medical Research Council score (19.5 ± 16.2 vs 27.4 ± 17.5, p = 0.03) and a higher frequency of dysautonomia (42.3% vs 19.4%, p = 0.037), as well as lower amplitudes of the distal compound muscle action potential (CMAP) of the median nerve [0.37 (RIQ 0.07-2.25) vs. 3.9 (RIQ1.2-6.4), p = <0.001] and ulnar nerve [0.37 (RIQ0.0-3.72) vs 1.5 (RIQ0.3-6.6), p = <0.001], and higher frequency of severe axonal damage in these nerves (distal CMAP ≤ 1.0 mV). Through binary logistic regression, severe axonal degeneration of the median nerve is an independent risk factor for prolonged IMV OR 4.9 (95%CI 1.1-21.5) p = 0.03, AUC of 0.774, (95%CI 0.66-0.88), p = < 0.001. CONCLUSIONS: Severe median nerve damage is an independent risk factor for prolonged mechanical ventilation.

Alzheimer-like cell death after vanadium pentoxide inhalation.

Vanadium (V) toxicity depends on its oxidation state; it seems that vanadium pentoxide (V2O5) is the most toxic to the living cells. It has been reported that oral administration induces changes in motor activity and learning; in rats, I.P. administration increases lipid peroxidation levels in the cerebellum and the concentration of free radicals in the hippocampus and cerebellum. Mice that inhaled V2O5 presented a reduced number of tubulin+ in Leydig and Sertoli cells; it has also been reported that inhaled V2O5 induces loss of dendritic spines, necrosis, and hippocampus neuropil alterations; considering the direct consequence of the interaction of V with cytoskeletal components, makes us believe that V2O5 exposure could cause neuronal death in the hippocampus similar to that seen in Alzheimer disease. This work aimed to determine pyramidal hippocampal CA1 cytoskeletal alterations with Bielschowsky stain in rats exposed to V2O5. Male Wistar rats inhaled 0.02 M of V2O5 one h two times a week for two and six months. We found that rats, which inhaled V2O5 reached 56,57% of dead neurons after six months of inhalation; we recognize strong argyrophilic and collapsed somas and typical flame-shaped in all V-exposed rats hippocampus CA1 compared to controls. We also observe somatodendritic distortions. Axons and dendrites displayed thick dark bands replaced by noticeable thickening and nodosities and the cytoskeleton fibrillary proteins' linear traces. Our findings suggest that V2O5 inhalation induces Alzheimer-like cell death with evident cytoskeletal alterations.

Effect of the use of vitamin "E" in the diluent on the viability of ram sperm.

The sperm membrane is damaged in cryopreservation processes; this damage can be minimized using antioxidants such as vitamin E. The objective of the present study was to evaluate the effect of the addition of vitamin "E" on the viability of ram sperm during preservation processes. Two experiments were carried out; in the first, 32 ejaculates were used, which after evaluation were divided into two aliquots for processing; the first received Triladyl + vitamin "E" (T + E), and the second received only Triladyl (T); these aliquots were cooled and stored at 5 °C for 24 h. The viability (sperm motility, integrity, and membrane permeability) was evaluated at 0 and 24 h after dilution. In the second experiment, the same procedure was performed as experiment 1, except that the samples were also frozen, and the viability was evaluated at zero and 48 h post-freezing. Dependent variables were analyzed using mixed models in a split plot design. In experiment 1, the integrity and permeability of the sperm membrane was better in the group: "T + E" (P <0.05). In experiment 2, the vitamin significantly improved (P <0.05) the sperm viability. It is concluded that the addition of vitamin "E" improves sperm viability.

Protracted hyperthermia and delayed rhabdomyolysis in ecstasy toxicity: A case report.

RATIONALE: Despite toxicity and unpredictable adverse effects, ecstasy use has increased in the United States. Onset of hyperpyrexia, rhabdomyolysis, disseminated intravascular coagulation (DIC), among other symptoms, occurs within hours of ingestion. Moreover, patients who experience hyperpyrexia, altered mental status, DIC, and multiorgan failure, rarely survive. This case presents a chronic ecstasy user whose symptoms would have predicted mortality. The report demonstrates a patient who experiences protracted hyperthermia, with delayed rhabdomyolysis and DIC. In addition, his peak creatine kinase (CK) of 409,440 U/L was far greater than the expected 30,000 to 100,000 U/L, being the second largest CK recorded in a survivor. PATIENT CONCERNS: This case report presents a 20-year-old man who presented to the emergency department after experiencing a severe reaction to ecstasy. He was a chronic user who took his baseline dosage while performing at a music event. He experienced hyperpyrexia immediately (106.5°F) while becoming stiff and unresponsive. Before emergency medical service arrival, his friends placed cold compresses on the patient and rested him in an ice filled bathtub. DIAGNOSES: Per history from patient's friends and toxicology results, the patient was diagnosed with ecstasy overdose, which evolved to include protracted hyperthermia and delayed rhabdomyolysis. INTERVENTIONS: Due to a Glasgow coma scale score of 5, he was intubated and sedated with a propofol maintenance. Hyperpyrexia resolved (temperature dropped to 99.1°F) after start of propofol maintenance. He was extubated after 24 hours, upon which he experienced hyperthermia (101.4°F at 48 hours), delayed rhabdomyolysis, and DIC (onset at 37 hours). He remained in hyperthermia for 120 hours until carvedilol permanently returned his temperature to baseline. His plasma CK reached a peak of 409,440 U/L at 35 hours. OUTCOMES: After primary management with intravenous fluids, the patient returned to baseline health without any consequences and was discharged after 8 days. A follow-up of 3 months postdischarge revealed no complications or disability. LESSONS: Clinically, the case highlights how physicians should be aware of the unusual time course adverse effects of ecstasy can have. Lastly, as intensity and duration of hyperpyrexia are predictors of mortality, our case indicates maintenance of sedation with propofol and use of oral carvedilol; both are efficacious for temperature reduction in ecstasy toxicity.

Ovarian dynamics of buffalo (Bubalus bubalis) synchronized with different hormonal protocols.

The study evaluates the effect of three hormonal protocols on ovarian dynamics and progesterone (P4) secretion of buffalo (Bubalus bubalis). Twenty-nine pluriparous Murrah buffaloes were used. The protocols were as follows: OVSYNCH (n = 10): 100 µg of gonadorelin (day 0), 500 µg of cloprostenol (day 7), and 100 µg of gonadorelin (day 9). CIDR+EB (intravaginal device (CIDR®) + estradiol benzoate; n = 10): CIDR plus 2 mg of EB (day 0), withdrew of CIDR, 500 µg of cloprostenol (day 7) and 1 mg of EB (day 8). CIDR+eCG (n = 9): CIDR plus 2 mg of EB (day 0), withdrew of CIDR, 500 µg of cloprostenol and 400 IU of eCG (day 7). Follicles were counted with an ultrasound and measured at 0, 24, and 54 h. The maximum follicle diameter and ovulation were evaluated at 70, 80, and 94 h after CIDR withdrew. Estrous was detected per 1 h three times daily. Blood samples were collected on days 0, 7, 10, 15, and 22 to determine P4 concentration. In CIDR+EB protocol, 50% of buffaloes presented estrous, at 69.6 h. All buffaloes ovulated. CIDR+eCG group had the shortest (69 h) ovulation time. No treatment differences for follicular population, maximum follicle diameter, and P4 concentration on days 7 and 10 (P > 0.05) were found. The P4 concentration in OVSYNCH and CIDR+eCG protocols were > 1 ng/ml, on days 15 and 22 (P < 0.05). There was no difference in ovarian activity; however, the P4 secretion was normal in the OVSYNCH and CIDR+eCG protocols compared to the CIDR+EB protocol.

Author Correction: Time-restricted feeding restores muscle function in Drosophila models of obesity and circadian-rhythm disruption.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Time-restricted feeding restores muscle function in Drosophila models of obesity and circadian-rhythm disruption.

Pathological obesity can result from genetic predisposition, obesogenic diet, and circadian rhythm disruption. Obesity compromises function of muscle, which accounts for a majority of body mass. Behavioral intervention that can counteract obesity arising from genetic, diet or circadian disruption and can improve muscle function holds untapped potential to combat the obesity epidemic. Here we show that Drosophila melanogaster (fruit fly) subject to obesogenic challenges exhibits metabolic disease phenotypes in skeletal muscle; sarcomere disorganization, mitochondrial deformation, upregulation of Phospho-AKT level, aberrant intramuscular lipid infiltration, and insulin resistance. Imposing time-restricted feeding (TRF) paradigm in which flies were fed for 12 h during the day counteracts obesity-induced dysmetabolism and improves muscle performance by suppressing intramuscular fat deposits, Phospho-AKT level, mitochondrial aberrations, and markers of insulin resistance. Importantly, TRF was effective even in an irregular lighting schedule mimicking shiftwork. Hence, TRF is an effective dietary intervention for combating metabolic dysfunction arising from multiple causes.

Reproductive behavior in hair sheep rams under tropical conditions.

The reproductive efficiency of sheep herds depends to a great extent on the ram. Male reproductive evaluation allows to select for the best and eliminate those with reproductive problems. The objective was to evaluate the effect of breed and age group on the reproductive behavior of hair sheep rams in the tropics of Mexico. Pelibuey (n = 42), Blackbelly (n = 30), Dorper (n = 44), and Katahdin (n = 30) rams of two age groups: young (n = 74, 1-1.5 years old) and adult (n = 72, 2-4 years old) were evaluated. Serving capacity (10-min duration) and breeding soundness evaluation (BSE) tests were carried out in each ram. In the first test, rams were classified as suitable and unsuitable, and in the BSE test, they were classified as satisfactory, questionable, or unsatisfactory. The response variables were analyzed using chi-square test or analyses of variance (ANOVA). ANOVA included the fixed effects of breed, age group, and their interaction. In the serving capacity test, 79.5% of the rams were considered suitable, with no breed differences (P > 0.05). Adult rams (90.3%) had the highest proportion of suitable rams (P < 0.05). In the BSE test, 80.2% of the rams were satisfactory; only breed being significant (P < 0.05). Pelibuey breed had the highest proportion of satisfactory rams (91.4%). Breed × age interaction was no significant for any trait. After serving capacity and BSE tests, a high proportion of rams was found not suitable for reproduction (36.3%), which is expected to cause low fertility in the flock.

Effect of season on follicular population, quality and nuclear maturation of bovine oocytes under tropical conditions.

The aim was to determine the effect of season of the year and the presence of a corpus luteum (CL) on follicular population (FP) and the quality of the oocytes, and of season on nuclear maturation of the bovine oocytes under tropical conditions. Three seasons were evaluated: hot-dry (March-June), hot-humid (July-October) and fresh-humid (November-February). In a first study, 1112 bovine ovaries were obtained from a local slaughterhouse. Follicles were classified as small (≤4mm), middle (4.1-8mm) and large (≥8.1mm); and the maximum diameter of the follicle (MDF) and CL (MDCL) were also recorded. The oocytes were collected by aspiration and classified as viable (grade I and II) and damaged (grade III and IV). In the second study, 2261 viable oocytes were matured in vitro, and then fixed and stained with Lacmoid to classify the stage of development as mature (metaphase II), immature or degenerate. Data were analyzed using analysis of variance and chi-square procedures. The largest FP of large follicles (0.67), MDF (1.18mm), MDCL (1.87mm), and the highest proportion of viable oocytes (34.19%) were obtained during the hot-humid season (P<0.05). The ovaries without CL had the greatest FP (10.34) with more viable oocytes (24.44%). The highest proportion of mature oocytes (76.92%) was also obtained in the hot-humid season. In conclusion, season influenced FP, MDF, MDCL, and the quality and nuclear maturation of oocytes. The presence of a CL in the ovary resulted in a decrease of FP and viability of oocytes.

Type 2 diabetes mellitus BALB/c mice are more susceptible to granulomatous amoebic encephalitis: Immunohistochemical study.

Granulomatous amoebic encephalitis (GAE) is a chronic, difficult to resolve infection caused by amphizoic amoebae of the genus Acanthamoeba, which in most cases occurs in immunosuppressed persons or with chronic diseases such as diabetes. In this study, we describe the early events of A. culbertsoni infection of GAE in diabetic mice model. Diabetes was induced in male BALB/c mice, with a dose of streptozotocin (130 mg/kg). Healthy and diabetic mice were inoculated via intranasal with 1 × 106 trophozoites of A. culbertsoni. Then were sacrificed and fixed by perfusion at 24, 48, 72 and 96 h post-inoculation, the brains and nasopharyngeal meatus were processed to immunohistochemical analysis. Invasion of trophozoites in diabetic mice was significantly greater with respect to inoculated healthy mice. Trophozoites and scarce cysts were immunolocalized in respiratory epithelial adjacent bone tissue, olfactory nerve packets, Schwann cells and the epineurium base since early 24 h post-inoculation. After 48 h, trophozoites were observed in the respiratory epithelium, white matter of the brain, subcortical central cortex and nasopharyngeal associated lymphoid tissue (NALT). At 72 h, cysts and trophozoites were immunolocalized in the olfactory bulb with the presence of a low inflammatory infiltrate characterized by polymorphonuclear cells. Scarce amoebae were observed in the granular layer of the cerebellum without evidence of inflammation or tissue damage. No amoebas were observed at 96 h after inoculation, suggesting penetration to other tissues at this time. In line with this, no inflammatory infiltrate was observed in the surrounding tissues where the amoebae were immunolocalized, which could contribute to the rapid spread of infection, particularly in diabetic mice. All data suggest that trophozoites invade the tissues by separating the superficial cells, penetrating between the junctions without causing cytolytic effect in the adjacent cells and subsequently reaching the CNS, importantly, diabetes increases the susceptibility to amoebae infection, which could favor the GAE development.

In vivo CNS infection model of Acanthamoeba genotype T4: the early stages of infection lack presence of host inflammatory response and are a slow and contact-dependent process.

This study was developed in order to describe the early morphological events observed during the invasion of two pathogenic strains of Acanthamoeba (genotype T4); A. castellanii and A. culbertsoni, at the olfactory meatus and cerebral, pulmonary, renal, hepatic and splenic tissues levels, an in vivo invasion study. Histological and immunohistochemical description of the events at 24, 48, 72, and 96 h postintranasal inoculations of BALB/c mice was performed. A. castellanii showed a higher invasion rate than A. culbertsoni, which was only able to reach lung and brain tissue in the in vivo model. The current study supports previous evidence of lack of inflammatory response during the early stages of infection. Acanthamoeba invasion of the CNS and other organs is a slow and contact-dependent process. The early morphological events during the invasion of amoebae include the penetration of trophozoites into different epithelia: olfactory, respiratory, alveolar space, and renal tubule, which resemble the process of amoebae invasion described in corneal tissue. The data suggest that after reaching the nasal epithelium, trophozoites continued invasion, separating and lifting the most superficial cells, then migrating and penetrating between the cell junctions without causing a cytolytic effect on adjacent cells. These results reaffirm the idea that contact-dependent mechanisms are relevant for amoebae of Acanthamoeba genus regardless of the invasion site.

Reproductive evaluation of red deer (Cervus elaphus) stags under tropical conditions / Evaluación reproductiva del ciervo rojo (Cervus elaphus) macho bajo condiciones tropicales / Avaliação reprodutiva do cervo rojo (Cervus elaphus) macho sob condições tropicais

Background: evaluation of reproductive traits of red deer is important to understand its performance. Objective: to evaluate seminal traits of red deer (Cervus elaphus) at three different stages of the breeding season (beginning, middle, and end) and to relate semen quality traits with pregnancy rate of hinds. Methods: scrotal circumference, semen volume, mass motility, individual motility, sperm concentration, morphology, and intact acrosomes were evaluated in seven stags. After evaluation, each of five stags was bred to 23 to 30 hinds. Pregnancy diagnosis was carried out using ultrasonography 45 days after the breeding season. Data were analyzed using the Student's t and chi-square tests, and simple correlation procedures. Results: scrotal circumference was reduced 5.4 cm (p<0.05) from the beginning to the end of the reproductive season, although semen volume was similar at the three different stages of the season. Sperm concentration (194 vs. 622.7/10(6)), mass motility (1.6 vs. 2.8), individual motility (28.6 vs. 63.3%), and intact acrosome (52.7 vs. 75.5%) were greater (p<0.05) at the middle of the breeding season in comparison with values found at the beginning. Percentage of abnormal spermatozoa was similar at the beginning and middle of the breeding season (p>0.05). No spermatozoa were found in stags at the end of the breeding season. Pregnancy rates were similar among bucks (p>0.05), ranging from 80% to 91.3%, and there was no relationship between pregnancy rate and semen traits. Conclusions: there was a clear seasonality of semen traits of red deer and no relationship between semen traits and pregnancy rate.

Antecedentes: la evaluación reproductiva de los ciervos es de suma importancia para entender su comportamiento reproductivo bajo el sistema en el cual se producen. Objetivo: evaluar algunos rasgos seminales del venado rojo (Cervus elaphus) en diferentes momentos de la estación reproductiva (inicio, medio y final) y relacionar la calidad seminal con la tasa de preñez de las hembras. Métodos: se utilizaron siete machos para medir la circunferencia escrotal, volumen seminal, motilidad masal, motilidad individual, concentración de espermatozoides, morfología y acrosomas intactos. Después de la evaluación, cinco machos fueron apareados con 23 a 30 hembras cada uno. El diagnóstico de preñez se realizó por ultrasonografía, 45 días después de la estación de apareamiento. Los datos se analizaron mediante las pruebas de t de Student y chi cuadrado, y procedimientos de correlación simple. Resultados: la circunferencia escrotal se redujo 5,4 cm (p<0,05) del inicio al final de la estación reproductiva, aunque el volumen de semen fue similar en los tres momentos de la estación reproductiva. La concentración de espermatozoides (194 vs 622,7/106), motilidad masal (1,6 vs 2,8), motilidad individual (28,6 vs 63,3%) y acrosomas intactos (52,7 vs 75,5%) fueron mejores (p<0,05) en la mitad de la estación reproductiva que en comparación con los valores del inicio de la misma. El porcentaje de espermatozoides anormales fue similar al inicio y en la mitad de la estación reproductiva (p>0,05). No se encontraron espermatozoides al final de la estación reproductiva. Las tasas de preñez fueron similares entre machos (p>0,05) variando de 80 a 91,3%, y no se encontró relación entre la tasa de preñez y los rasgos seminales. Conclusiones: hubo una clara estacionalidad de los rasgos seminales del venado rojo bajo las condiciones tropicales y una falta de relación entre los rasgos seminales y la tasa de preñez.

Antecedentes: a avaliação reprodutiva do cervo rojo é muito importante para entender seu comportamento reprodutivo no sistema em que eles são produzidos. Objetivo: avaliar algumas caraterísticas do sêmen do cervo rojo (Cervus elaphus) em diferentes momentos da estação reprodutiva (inicio, meio e final) e relacionar a qualidade seminal com a taxa de prenhez das fêmeas. Métodos: foram utilizados sete machos para medir a circunferência escrotal, volume seminal, motilidade masal, motilidade individual, concentração de espermatozóides, morfologia e acrossomas intactos. Depois da avaliação, cinco machos foram acasalados com 23-30 fêmeas cada um. El diagnóstico de gestação foi realizado por ultrassonografia, 45 dias depois da estação de acasalamento. Os dados foram analisados usando provas de t de Student e chi-quadrado, e procedimentos de correlação simples. Resultados: a circunferência escrotal se reduz 5,4 cm (p<0,05) do inicio ao final da estação reprodutiva, ainda o volume de sêmen foi similar nos três momentos da estação reprodutiva. A concentração de espermatozoides (194 vs 622,7/10(6)), motilidade masal (1,6 vs 2,8), motilidade individual (28,6 vs 63,3%) e acrossomas intactos (52,7 vs 75,5%) foram melhores (p<0,05) na parte meia em comparação com os valores do inicio da estação reprodutiva. A porcentagem de espermatozoides anormais foi similar ao inicio e em meio da estação reprodutiva (p>0,05). No se encontraram espermatozoides ao final da estação reprodutiva. As taxas de prenhez foram similares entre machos (p>0,05) variando de 80 a 91,3%, e não se encontrou relação entre a taxa de prenhez e os rasgos seminais. Conclusões: houve uma clara estacionalidade dos rasgos seminais do cervo rojo sob as condiciones tropicais e uma falta de relação entre os rasgos seminais e a taxa de prenhez.

Resveratrol potentiates effects of simvastatin on inhibition of rat ovarian theca-interstitial cells steroidogenesis.

BACKGROUND: Polycystic ovary syndrome (PCOS) is characterized by ovarian enlargement, hyperplastic theca compartment and increased androgen production due to, at least in part, excessive expression of several key genes involved in steroidogenesis. Previously, our group has demonstrated that simvastatin, competitive inhibitor of 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA reductase), a rate-limiting step of the mevalonate pathway, reduces rat-theca interstitial cell steroidogenesis by inhibiting Cyp17a1 gene expression, the key enzyme of the androgen biosynthesis pathway. Recently, we demonstrated that resveratrol, a bioflavonoid abundant in red grapes, decreases rat theca-interstitial cell steroidogenesis and this suppressive effect is mediated through mechanisms independent of the mevalonate pathway. The present study evaluated the effect of combining simvastatin and resveratrol treatments on rat theca-interstitial cell steroidogenesis. METHODS: Rat theca-interstitial cells isolated from 30 day-old female rats were cultured for up to 48 h with or without simvastatin (1 µM) and/or resveratrol (3-10 µM). Steroidogenic enzymes gene expression was evaluated by quantitative real time PCR and steroid levels were measured by liquid chromatography-mass spectrometry. Comparisons between groups were performed using ANOVA and Tukey test. RESULTS: Resveratrol potentiated inhibitory effects of simvastatin on androstenedione and androsterone production in theca-interstitial cells. This suppressive effect correlated with profound inhibition in Cyp17a1 mRNA expression in the presence of a combination of resveratrol and simvastatin. CONCLUSIONS: The present findings indicate that resveratrol potentiates the simvastatin-induced inhibitory effect on theca-interstitial cell androgen production, raising the possibility of development of novel treatments of PCOS.

Comparison of effects of different statins on growth and steroidogenesis of rat ovarian theca-interstitial cells.

Statins are competitive inhibitors of 3-hydroxy-3-methyl-glutaryl-CoA reductase, the rate-limiting enzyme of the cellular production of cholesterol and other products of the mevalonate pathway. Statins exert hepatic and extrahepatic effects, modulating the function of various tissues and organs, including ovaries. Previously, we have demonstrated that simvastatin inhibited cellular proliferation and reduced androgen production by ovarian theca-interstitial cells. The above actions are of translational relevance to the most common endocrine disorder among women in reproductive age: polycystic ovary syndrome. However, different statins may have distinctly different profiles of effects on cholesterol and androgens. The present study was designed to compare the effects of several statins on growth and steroidogenesis of rat theca-interstitial cells. The cells were incubated in the absence (control) or in the presence of simvastatin, lovastatin, atorvastatin, or pravastatin. Assessment of effects of statins on cell growth was carried out by evaluation of DNA synthesis and by estimation of the number of viable cells. Effects on steroidogenesis were evaluated by quantification of steroid production and expression of mRNA for the key enzyme regulating androgen production: Cyp17a1. Among tested statins, simvastatin exerted the greatest inhibitory effects on all tested parameters. The rank order of the effects of the tested statins is as follows: simvastatin > lovastatin > atorvastatin ≥ pravastatin. While the lipophilicity is likely to play a major role in determining the ability of statins to act on nonhepatic cells, other factors unique to individual cell types are also likely to be relevant.

The combination of oral L-DOPA/rimonabant for effective dyskinesia treatment and cytological preservation in a rat model of Parkinson's disease and L-DOPA-induced dyskinesia.

Parkinson's disease is the second most prevalent neurodegenerative disease in the world. Its treatment is limited so far to the management of parkinsonian symptoms with L-DOPA (LD). The long-term use of LD is limited by the development of L-DOPA-induced dyskinesias and dystonia. However, recent studies have suggested that pharmacological targeting of the endocannabinoid system may potentially provide a valuable therapeutic tool to suppress these motor alterations. In the present study, we have explored the behavioral (L-DOPA-induced dyskinesias severity) and cytological (substantia nigra compacta neurons and striatum neuropil preservation) effects of the oral coadministration of LD and rimonabant, a selective antagonist of CB1 receptors, in the 6-hydroxydopamine rat model of Parkinson's disease. Oral coadministration of LD (30 mg/kg) and rimonabant (1 mg/kg) significantly decreased abnormal involuntary movements and dystonia, possibly through the conservation of some functional tyrosine hydroxylase-immunoreactive dopaminergic cells, which in turn translates into a well-preserved neuropil of a less denervated striatum. Our results provide anatomical evidence that long-term coadministration of LD with cannabinoid antagonist-based therapy may not only alleviate specific motor symptoms but also delay/arrest the degeneration of striatal and substantia nigra compacta cells.

Resveratrol is not as effective as physical exercise for improving reproductive and metabolic functions in rats with dihydrotestosterone-induced polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a reproductive and metabolic disorder associated with obesity and insulin resistance that often precedes the development of type-2 diabetes. Rats continuously exposed to dihydrotestosterone from prepuberty display typical reproductive and metabolic PCOS characteristics including anovulation, polycystic ovaries, insulin resistance, and obesity. Our aim was to investigate if resveratrol improves reproductive and metabolic functions in PCOS rats. The effect was compared to exercise. Control and PCOS rats were treated with vehicle or resveratrol (400 mg · kg(-1) · day(-1)) for 5-6 weeks. Another group of PCOS rats received vehicle treatment and exercised for 5-6 weeks. Insulin sensitivity was determined by euglycemic-hyperinsulinemic clamp. The glucose infusion rate was lower in the PCOS-vehicle group compared to control-vehicle rats (P < 0.05). Exercise increased insulin sensitivity compared with PCOS-vehicle rats (P < 0.05), but resveratrol did not. Resveratrol treatment and exercise resulted in smaller adipocytes, upregulated estrogen-related receptor α gene expression in subcutaneous fat, and improved estrus cyclicity in the previously acyclic PCOS rats. Although resveratrol had positive effects on adiposity and cyclicity in a similar manner to exercise, resveratrol does not seem to be a good candidate for treating insulin resistance associated with PCOS because no improvement in insulin sensitivity was observed in PCOS rats on normal chow.

Resveratrol potentiates effect of simvastatin on inhibition of mevalonate pathway in human endometrial stromal cells.

CONTEXT: Growth of endometriotic lesions in rodent model of endometriosis is inhibited by resveratrol, a natural polyphenol with antiproliferative and antiinflammatory properties, and simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) activity. OBJECTIVE: The objective of the investigation was to study the mechanism of action of resveratrol and its interactions with simvastatin, focusing on cholesterol biosynthesis and HMGCR gene expression and protein activity in primary cultures of human endometrial stromal (HES) cells. METHODS: HES cells were obtained from healthy volunteers. Biosynthesis of cholesterol was assessed by measuring the conversion of [(14)C]acetate to [(14)C]cholesterol. HMGCR mRNA transcripts were quantified by real-time PCR, protein expression by Western blot analysis, and enzyme activity by measuring the conversion of [3-(14)C]3-hydroxy-3-methyl-glutaryl-coenzyme A to [(14)C]mevalonic acid lactone in HES cell microsomes. RESULTS: Resveratrol inhibited cholesterol biosynthesis, HMGCR mRNA, and enzyme activity. Simvastatin inhibited cholesterol biosynthesis and enzyme activity but increased HMGCR mRNA and protein expression. Resveratrol potentiated the inhibitory effects of simvastatin on cholesterol biosynthesis and HMGCR enzyme activity and abrogated the stimulatory effects of simvastatin on HMGCR mRNA transcripts and protein expression. CONCLUSIONS: Resveratrol inhibits key steps of the mevalonate pathway by mechanisms that are partly complementary to and partly comparable with simvastatin via reducing both expression and activity of HMGCR. A combination of resveratrol and simvastatin may be of potential clinical relevance to development new treatments of human endometriosis.

Effects of simvastatin on retinoic acid system in primary human endometrial stromal cells and in a chimeric model of human endometriosis.

CONTEXT: Retinoic acid (RA) may promote survival or apoptosis of cells, depending on the levels of binding proteins: apoptosis-inducing cellular RA binding protein 2 (CRABP2), and cell survival-promoting fatty acid binding protein 5 (FABP5). Increased cellular uptake of retinol and altered actions of RA related to reduced expression of CRABP2 may contribute to the development of endometriosis. Recently statins have been shown to inhibit growth of human endometrial stromal (HES) cells and to reduce the number and size of endometriotic implants in experimental models of this disorder. OBJECTIVE: The objective of the study was to determine whether effects of simvastatin on HES cells and experimental endometriotic implants are related to the modulation of the RA system. METHODS: Effects of simvastatin and RA on proliferation and apoptosis of HES cells were evaluated. Expression of stimulated by RA 6 (STRA6), CRABP2, and FABP5 was determined by real-time PCR and Western blotting. Effects of simvastatin were also evaluated in a nude mouse model of human endometriosis. RESULTS: Simvastatin potentiated an inhibitory effect of RA on growth of HES cells. In HES cells, simvastatin induced expression of STRA6 and CRABP2 but not FABP5. Similarly, simvastatin treatment of nude mice bearing human endometrial xenografts led to an increased expression of CRABP2 and STRA6 proteins in ectopic lesions. CONCLUSIONS: Simvastatin interacts with the RA system, inducing the expression of the key protein regulating the uptake of retinol (STRA6) and the expression of apoptosis-promoting CRABP2. These effects may contribute to cooperative apoptosis-inducing effects of simvastatin and RA and support the examination of these compounds in the treatment of endometriosis.

Letrozole increases ovarian growth and Cyp17a1 gene expression in the rat ovary.

OBJECTIVE: To evaluate the effects of letrozole on ovarian size and steroidogenesis in vivo, as well as on proliferation and steroidogenesis of theca-interstitial cells alone and in coculture with granulosa cells using an in vitro model. DESIGN: In vivo and in vitro studies. SETTING: Research laboratory. ANIMAL(S): Immature Sprague-Dawley female rats. INTERVENTION(S): In vivo effects of letrozole were studied in intact rats receiving either letrozole (90-day continuous-release SC pellets, 400 µg/d) or placebo pellets (control group). In in vitro experiments, theca cells were cultured alone or in coculture with granulosa cells in the absence or presence of letrozole. MAIN OUTCOME MEASURE(S): Deoxyribonucleic acid synthesis was determined by thymidine incorporation assay; steroidogenesis by mass spectrometry; and steroidogenic enzyme messenger RNA (mRNA) expression by polymerase chain reaction. RESULT(S): In vivo, letrozole induced an increase in ovarian size compared with the control group and also induced a profound increase of androgen, LH levels, and Cyp17a1 mRNA expression. Conversely, a decrease in Star, Cyp11a1, and Hsd3b1 transcripts was observed in letrozole-exposed rats. In vitro, letrozole did not alter either theca cell proliferation or Cyp17a1 mRNA expression. Similarly, letrozole did not affect Cyp17a1 transcripts in granulosa-theca cocultures. CONCLUSION(S): These findings suggest that letrozole exerts potent, but indirect, effect on growth of rat ovary and dramatically increases androgen levels and Cyp17a1 mRNA expression, the key enzyme regulating the androgen biosynthesis pathway. The present findings reveal novel mechanisms of action of letrozole in the rat ovary.

Effects of resveratrol on growth and function of rat ovarian granulosa cells.

OBJECTIVE: To evaluate the effects of resveratrol on growth and function of granulosa cells. Previously, we demonstrated that resveratrol exerts profound proapoptotic effects on theca-interstitial cells. DESIGN: In vitro study. SETTING: Research laboratory. ANIMAL(S): Immature Sprague-Dawley female rats. INTERVENTION(S): Granulosa cells were cultured in the absence or presence of resveratrol. MAIN OUTCOME MEASURE(S): DNA synthesis was determined by thymidine incorporation assay, apoptosis by activity of caspases 3/7, cell morphology by immunocytochemistry, steroidogenesis by mass spectrometry, antimüllerian hormone (AMH), and vascular endothelial growth factor (VEGF) expression by polymerase chain reaction and Western blot. RESULT(S): Resveratrol induced a biphasic effect on DNA synthesis, whereby a lower concentration stimulated thymidine incorporation and higher concentrations inhibited it. Additionally, resveratrol slightly increased the cell number and modestly decreased the activity of caspases 3/7 with no effect on cell morphology or progesterone production. However, resveratrol decreased aromatization and VEGF expression, whereas AMH expression remained unaltered. CONCLUSION(S): Resveratrol, by exerting cytostatic but not cytotoxic effects, together with antiangiogenic actions mediated by decreased VEGF in granulosa cells, may alter the ratio of theca-to-granulosa cells and decrease vascular permeability, and therefore may be of potential therapeutic use in conditions associated with highly vascularized theca-interstitial hyperplasia and abnormal angiogenesis, such as those seen in women with polycystic ovary syndrome.