Método de ensaio biomecânico para análise da isometricidade na reconstrução do ligamento patelofemoral medial

OBJETIVO: Apresentar um dispositivo biomecânico para o estudo da reconstrução do ligamento patelofemoral medial (LPFM) e sua isometricidade. MÉTODOS: Foi desenvolvido um sistema biomecânico acessível, que permite a aplicação de forças fisiológicas e não fisiológicas no joelho, através de um braço mecânico e aplicação de pesos e contrapesos, possibilitando a execução de diferentes estudos, além de ter um sistema de medidas bastante preciso de aferição de distâncias entre diferentes estruturas para análise dos experimentos. Este artigo descreve a montagem deste sistema, além de sugerir algumas aplicações práticas. Foram estudados seis joelhos de cadáveres. Os joelhos foram preparados em uma máquina de ensaios desenvolvida no Laboratório de Biomecânica do IOT HC FMUSP, que permitiu a avaliação dinâmica do comportamento patelar, quantificando a sua lateralização entre 0 e 120 graus. A diferença entre as distâncias encontradas, com e sem carga, aplicada na patela foram agrupadas segundo o ângulo de fixação do enxerto (0°, 30°, 60° e 90°) e situação do joelho (íntegro, reconstruído e lesado). RESULTADOS: Houve uma tendência em ocorrer menor desvio lateral em ângulos de fixação acima de 30 graus de flexão, principalmente entre os ângulos entre 45° e 60° graus de flexão, após a reconstrução. Para os demais ângulos não houve significância estatística. CONCLUSÃO: O método desenvolvido é uma ferramenta útil para os estudos da articulação patelofemoral, além de ter um sistema de medidas bastante preciso de aferição de distâncias entre diferentes estruturas e permitir a sua utilização em instituições com menos recursos disponíveis.

Estudo anatômico e funcional do complexo ligamentar colateral medial do cotovelo

OBJETIVO: Realizar um estudo anatômico do ligamento colateral medial, um importante estabilizador do cotovelo, em diferentes graus de flexo-extensão do cotovelo. MÉTODOS: Foram dissecados 40 cotovelos para analisar o comportamento funcional das bandas anterior, posterior e transversa do ligamento nas manobras de estresse em valgo do cotovelo durante seu movimento de flexão e extensão em diferentes graus. Determinou-se dois grupos: no GPA foi seccionado, a banda posterior do ligamento, depois a cápsula articular e, por fim, a banda anterior; no GAP, a ordem de dissecação foi inversa. RESULTADOS: No GPA observou-se instabilidade somente na terceira etapa e a média de abertura foi maior entre 50&º e 70&º de flexão de cotovelo; no GAP, a instabilidade apareceu desde a primeira etapa e os graus de flexão com maior instabilidade foram nos mesmos do grupo A. CONCLUSÃO: A banda anterior do ligamento colateral medial do cotovelo é o mais importante estabilizador na instabilidade em valgo do cotovelo e sua atuação principal acontece entre 50º e 70º de flexão do cotovelo. Nível de evidência III, Estudo Diagnóstico - Investigação de um exame para diagnóstico.

Análise do arco longitudinal medial em adolescentes usuárias de calçados de salto alto

O objetivo do estudo foi analisar a influência do calçado de salto alto no arco longitudinal medial (ALM) do pé de adolescentes. Fizeram parte do estudo 82 adolescentes entre 13 e 20 anos, sendo 54 não usuárias (grupo controle - GC) e 28 usuárias (grupo experimental - GE) de calçado de salto alto. Foram obtidas as impressões plantares de ambos os pés para análise do ALM, antes e depois do uso de um calçado de salto alto padronizado por um período de 30 minutos. As impressões plantares foram avaliadas pelo índice de Chippaux-Smirak (ICS) e pelo arco de Cavanagh & Rodgers (ICR). O teste de Shapiro-Wilks foi utilizado para a verificação da normalidade dos dados. Variáveis paramétricas pareadas foram tratadas com o Teste t de Student pareado e as não-paramétricas com o teste de Wilcoxon. As comparações não-pareadas foram realizadas com o teste t de Student para as variáveis paramétricas e o de Mann-Withney para as não-paramétricas, com nível de significância de 0,05. Houve diferença no ALM entre os lados direito e esquerdo apenas no GC antes do uso do calçado. Na comparação entre antes e depois do uso do sapato, notou-se diferença apenas no pé esquerdo do GC pelo ICS. Já entre GC e GE, não houve diferença. Apesar dos resultados não evidenciarem alterações no ALM, deve-se lembrar que esta é uma medida estática, sendo necessários estudos do componente dinâmico e do uso do calçado de salto crônico para correlacionar com os achados deste trabalho.

Fluoxetine exposure during pregnancy and lactation: effects on acute stress response and behavior in the novelty-suppressed feeding are age and gender-dependent in rats

Fluoxetine (FLX) is commonly used to treat anxiety and depressive disorders in pregnant women. Since FLX crosses the placenta and is excreted in milk, maternal treatment with this antidepressant may expose the fetus and neonate to increased levels of serotonin (5-HT). Long-term behavioral abnormalities have been reported in rodents exposed to higher levels of 5-HT during neurodevelopment. In this study we evaluated if maternal exposure to FLX during pregnancy and lactation would result in behavioral and/or stress response disruption in adolescent and adult rats. Our results indicate that exposure to FLX influenced restraint stress-induced Fos expression in the amygdala in a gender and age-specific manner. In male animals, a decreased expression was observed in the basolateral amygdala at adolescence and adulthood; whereas at adulthood, a decrease was also observed in the medial amygdala. A lack of FLX exposure effect was observed in females and also in the paraventricular nucleus of both genders. Regarding the behavioral evaluation, FLX exposure did not induce anhedonia in the sucrose preference test but decreased the latency to feed of both male and female adolescent rats evaluated in the novelty-suppressed feeding test. In conclusion, FLX exposure during pregnancy and lactation decreases acute amygdalar stress response to a psychological stressor in males (adolescents and adults) as well as influences the behavior of adolescents (males and females) in a model that evaluates anxiety and/or depressive-like behavior. Even though FLX seems to be a developmental neurotoxicant, the translation of these findings to human safe assessment remains to be determined since it is recognized that not treating a pregnant or lactating woman may also impact negatively the development of the descendants.

Commissural nucleus of the solitary tract regulates the antihypertensive effects elicited by moxonidine

The rostral ventrolateral medulla (RVLM) contains the presympathetic neurons involved in cardiovascular regulation that has been implicated as one of the most important central sites for the antihypertensive action of moxonidine (an α2-adrenergic and imidazoline agonist). Here, we sought to evaluate the cardiovascular effects produced by moxonidine injected into another important brainstem site, the commissural nucleus of the solitary tract (commNTS). Mean arterial pressure (MAP), heart rate (HR), splanchnic sympathetic nerve activity (sSNA) and activity of putative sympathoexcitatory vasomotor neurons of the RVLM were recorded in conscious or urethane-anesthetized, and artificial ventilated male Wistar rats. In conscious or anesthetized rats, moxonidine (2.5 and 5 nmol/50 nl) injected into the commNTS reduced MAP, HR and sSNA. The injection of moxonidine into the commNTS also elicited a reduction of 28% in the activity of sympathoexcitatory vasomotor neurons of the RVLM. To further assess the notion that moxonidine could act in another brainstem area to elicit the antihypertensive effects, a group with electrolytic lesions of the commNTS or sham and with stainless steel guide-cannulas implanted into the 4th V were used. In the sham group, moxonidine (20 nmol/1 μl) injected into 4th V decreased MAP and HR. The hypotension but not the bradycardia produced by moxonidine into the 4th V was reduced in acute (1 day) commNTS-lesioned rats. These data suggest that moxonidine can certainly act in other brainstem regions, such as commNTS to produce its beneficial therapeutic effects, such as hypotension and reduction in sympathetic nerve activity.

Facilitation of sodium intake by combining noradrenaline into the lateral parabrachial nucleus with prazosin peripherally

Injections of noradrenaline into the lateral parabrachial nucleus (LPBN) increase arterial pressure and 1.8% NaCl intake and decrease water intake in rats treated with the diuretic furosemide (FURO) combined with a low dose of the angiotensin converting enzyme inhibitor captopril (CAP). In the present study, we investigated the influence of the pressor response elicited by noradrenaline injected into the LPBN on FURO+CAP-induced water and 1.8% NaCl intake. Male Holtzman rats with bilateral stainless steel guide-cannulas implanted into LPBN were used. Bilateral injections of noradrenaline (40 nmol/0.2 μl) into the LPBN increased FURO+CAP-induced 1.8% NaCl intake (12.2±3.5, vs., saline: 4.2±0.8 ml/180 min), reduced water intake and strongly increased arterial pressure (50±7, vs. saline: 1±1 mmHg). The blockade of the α1 adrenoceptors with the prazosin injected intraperitoneally abolished the pressor response and increased 1.8% NaCl and water intake in rats treated with FURO+CAP combined with noradrenaline injected into the LPBN. The deactivation of baro and perhaps volume receptors due to the cardiovascular effects of prazosin is a mechanism that may facilitate water and NaCl intake in rats treated with FURO+CAP combined with noradrenaline injected into the LPBN. Therefore, the activation of α2 adrenoceptors with noradrenaline injected into the LPBN, at least in dose tested, may not completely remove the inhibitory signals produced by the activation of the cardiovascular receptors, particularly the signals that result from the extra activation of these receptors with the increase of arterial pressure.

Intrinsic organization of the suprachiasmatic nucleus in the capuchin monkey

The suprachiasmatic nucleus (SCN), which is the main circadian biological clock in mammals, is composed of multiple cells that function individually as independent oscillators to express the self-sustained mRNA and protein rhythms of the so-called clock genes. Knowledge regarding the presence and localization of the proteins and neuroactive substances of the SCN are essential for understanding this nucleus and for its successful manipulation. Although there have been advances in the investigation of the intrinsic organization of the SCN in rodents, little information is available in diurnal species, especially in primates. This study, which explores the pattern of expression and localization of PER2 protein in the SCN of capuchin monkey, evaluates aspects of the circadian system that are common to both primates and rodents. Here, we showed that PER2 protein immunoreactivity is higher during the light phase. Additionally, the complex organization of cells that express vasopressin, vasoactive intestinal polypeptide, neuron-specific nuclear protein, calbindin and calretinin in the SCN, as demonstrated by their immunoreactivity, reveals an intricate network that may be related to the similarities and differences reported between rodents and primates in the literature.

Purinergic and glutamatergic interactions in the hypothalamic paraventricular nucleus modulate sympathetic outflow

P2X receptors are expressed on ventrolateral medulla projecting paraventricular nucleus (PVN) neurons. Here, we investigate the role of adenosine 5′-triphosphate (ATP) in modulating sympathetic nerve activity (SNA) at the level of the PVN. We used an in situ arterially perfused rat preparation to determine the effect of P2 receptor activation and the putative interaction between purinergic and glutamatergic neurotransmitter systems within the PVN on lumbar SNA (LSNA). Unilateral microinjection of ATP into the PVN induced a dose-related increase in the LSNA (1 nmol: 38 ± 6 %, 2.5 nmol: 72 ± 7 %, 5 nmol: 96 ± 13 %). This increase was significantly attenuated by blockade of P2 receptors (pyridoxalphosphate-6-azophenyl-20,40-disulphonic acid, PPADS) and glutamate receptors (kynurenic acid, KYN) or a combination of both. The increase in LSNA elicited by L-glutamate microinjection into the PVN was not affected by a previous injection of PPADS. Selective blockade of non-N-methyl-D-aspartate receptors (6-cyano-7-nitroquinoxaline-2,3-dione disodium salt, CNQX), but not N-methyl-D-aspartate receptors (NMDA) receptors (DL-2-amino-5-phosphonopentanoic acid, AP5), attenuated the ATP-induced sympathoexcitatory effects at the PVN level. Taken together, our data show that purinergic neurotransmission within the PVN is involved in the control of SNA via P2 receptor activation. Moreover, we show an interaction between P2 receptors and non-NMDA glutamate receptors in the PVN suggesting that these functional interactions might be important in the regulation of sympathetic outflow

The cardiovascular actions of fractalkine/CX3CL1 in the hypothalamic paraventricular nucleus are attenuated in rats with heart failure

The paraventricular nucleus (PVN) of the hypothalamus plays an important role in the regulation of sympathetic nerve activity, which is significantly elevated in chronic heart failure (CHF). Fractalkine (FKN) and its cognate receptor, CX3CR1, are constitutively expressed in the central nervous system, but their role and physiological significance are not well known. The aims of the present study were to determine whether FKN plays a cardiovascular role within the PVN and to investigate how the actions of FKN might be altered in CHF. We show that both FKN and CX3CR1 are expressed on neurons in the PVN of rats, suggesting that they may have a physiological function in this brain nucleus. Unilateral microinjection of FKN directly into the PVN of anaesthetized rats elicited a significant dose-related decrease in blood pressure (1.0 nmol, -5 ± 3 mmHg; 2.5 nmol, -13 ± 2 mmHg; 5.0 nmol, -22 ± 3 mmHg; and 7.5 nmol, -32 ± 3 mmHg) and a concomitant increase in heart rate (1.0 nmol, 6 ± 3 beats min(-1); 2.5 nmol, 11 ± 3 beats min(-1); 5 nmol, 18 ± 4 beats min(-1); and 7.5 nmol, 27 ± 5 beats min(-1)) compared with control saline microinjections. In order to determine whether FKN signalling is altered in rats with CHF, we first performed quantitative RT-PCR and Western blot analysis and followed these experiments with functional studies in rats with CHF and sham-operated control rats. We found a significant increase in CX3CR1 mRNA and protein expression, as determined by quantitative RT-PCR and Western blot analysis, respectively, in the PVN of rats with CHF compared with sham-operated control rats. We also found that the blood pressure effects of FKN (2.5 nmol in 50 nl) were significantly attenuated in rats with CHF (change in mean arterial pressure, -6 ± 3 mmHg) compared with sham-operated control rats (change in mean arterial pressure, -16 ± 6 mmHg). These data suggest that FKN and its receptor, CX3CR1, modulate cardiovascular function at the level of the PVN and that the actions of FKN within this nucleus are altered in heart failure