Design and cadaveric validation of a novel device to quantify knee stability during total knee arthroplasty.

The success of total knee arthroplasty depends, in part, on the ability of the surgeon to properly manage the soft tissues surrounding the joint, but an objective definition as to what constitutes acceptable postoperative joint stability does not exist. Such a definition may not exist due to lack of suitable instrumentation, as joint stability is currently assessed by visual inspection while the surgeon manipulates the joint. Having the ability to accurately and precisely measure knee stability at the time of surgery represents a key requirement in the process of objectively defining acceptable joint stability. Therefore, we created a novel sterilizable device to allow surgeons to measure varus-valgus, internal-external, or anterior-posterior stability of the knee during a total knee arthroplasty. The device can be quickly adjusted between 0 deg and 90 deg of knee flexion. The device interfaces with a custom surgical navigation system, which records the resultant rotations or translations of the knee while the surgeon applies known loads to a patient's limb with a handle instrumented with a load cell. We validated the performance of the device by having volunteers use it to apply loads to a mechanical linkage that simulated a knee joint; we then compared the joint moments calculated by our stability device against those recorded by a load cell in the simulated knee joint. Validation of the device showed low mean errors (less than 0.21 ± 1.38 Nm and 0.98 ± 3.93 N) and low RMS errors (less than 1.5 Nm and 5 N). Preliminary studies from total knee arthroplasties performed on ten cadaveric specimens also demonstrate the utility of our new device. Eventually, the use of this device may help determine how intra-operative knee stability relates to postoperative function and could lead to an objective definition of knee stability and more efficacious surgical techniques.

Mast cells are required for the development of renal fibrosis in the rodent unilateral ureteral obstruction model.

Mast cells are associated with inflammation and fibrosis. Whether they protect against or contribute to renal fibrosis is unclear. Based on our previous findings that mast cells can express and secrete active renin, and that angiotensin (ANG II) is profibrotic, we hypothesized that mast cells play a critical role in tubulointerstitial fibrosis. We tested this hypothesis in the 14-day unilateral ureteral obstruction (UUO) model in rats and mast cell-deficient (MCD) mice (WBB6F1-W/Wv) and their congenic controls (CC). In the 14-day UUO rat kidney, mast cell number is increased and they express active renin. Stabilizing mast cells in vivo with administration of cromolyn sodium attenuated the development of tubulointerstitial fibrosis, which was confirmed by measuring newly synthesized pepsin-soluble collagen and blind scoring of fixed trichrome-stained kidney sections accompanied by spectral analysis. Fibrosis was absent in UUO kidneys from MCD mice unlike that observed in the CC mice. Losartan treatment reduced the fibrosis in the CC UUO kidneys. The effects of mast cell degranulation and renin release were tested in the isolated, perfused kidney preparation. Mast cell degranulation led to renin-dependent protracted flow recovery. This demonstrates that mast cell renin is active in situ and the ensuing ANG II can modulate intrarenal vascular resistance in the UUO kidney. Collectively, the data demonstrate that mast cells are critical to the development of renal fibrosis in the 14-day UUO kidney. Since renin is present in human kidney mast cells, our work identifies potential targets in the treatment of renal fibrosis.

A Rac-cGMP signaling pathway.

The small GTPase Rac and the second messenger cGMP (guanosine 3',5'-cyclic monophosphate) are critical regulators of diverse cell functions. When activated by extracellular signals via membrane signaling receptors, Rac executes its functions through engaging downstream effectors such as p21-activated kinase (PAK), a serine/threonine protein kinase. However, the molecular mechanism by which membrane signaling receptors regulate cGMP levels is not known. Here we have uncovered a signaling pathway linking Rac to the increase of cellular cGMP. We show that Rac uses PAK to directly activate transmembrane guanylyl cyclases (GCs), leading to increased cellular cGMP levels. This Rac/PAK/GC/cGMP pathway is involved in platelet-derived growth factor-induced fibroblast cell migration and lamellipodium formation. Our findings connect two important regulators of cellular physiological functions and provide a general mechanism for diverse receptors to modulate physiological responses through elevating cellular cGMP levels.

The broad homeostatic role of natriuretic peptides.

A brief non-inclusive review on natriuretic peptides (NP), their receptors, and their main functional properties is presented. The three main NP, atrial (ANP), brain (BNP) and C-type (CNP) are considered. Guanylyl cyclase receptors modulate all the known systemic effects of NP. Clearance receptors determine the metabolic disposal of NP and in this manner regulate their plasma levels and/or local tissue concentrations. Structure-function properties, and homeostatic properties of NP receptors are presented. ANP, which plays a major role in pressure-volume homeostasis, is discussed in relationship to its effects on renal hemodynamic and excretory functions, inhibition of the renin-angiotensin-aldosterone system, vasorelaxant, and third-spacing action. For BNP special attention is directed to its role as a negative modulator of ventricular remodeling, in view of its anti-hypertrophic, anti-fibrotic and anti-inflammatory effects in the heart. The major effect of CNP in promoting vertebral and longitudinal bone growth is briefly addressed. Finally, emphasis is placed on the recent discovery that ANP affects fat metabolism in humans due to its powerful lipolytic action.

The broad homeostatic role of natriuretic peptides / O amplo papel homeostático dos peptídeos natriuréticos

A brief non-inclusive review on natriuretic peptides (NP), their receptors, and their main functional properties is presented. The three main NP, atrial (ANP), brain (BNP) and C-type (CNP) are considered. Guanylyl cyclase receptors modulate all the known systemic effects of NP. Clearance receptors determine the metabolic disposal of NP and in this manner regulate their plasma levels and/or local tissue concentrations. Structure-function properties, and homeostatic properties of NP receptors are presented. ANP, which plays a major role in pressure-volume homeostasis, is discussed in relationship to its effects on renal hemodynamic and excretory functions, inhibition of the renin-angiotensin-aldosterone system, vasorelaxant, and third-spacing action. For BNP special attention is directed to its role as a negative modulator of ventricular remodeling, in view of its anti-hypertrophic, anti-fibrotic and anti-inflammatory effects in the heart. The major effect of CNP in promoting vertebral and longitudinal bone growth is briefly addressed. Finally, emphasis is placed on the recent discovery that ANP affects fat metabolism in humans due to its powerful lipolytic action.

Este trabalho apresenta uma breve revisão parcial sobre os peptídeos natriuréticos (NP), seus receptores e suas principais propriedades funcionais. Serão discutidos os três principais NP: atrial (ANP), cerebral (BNP) e tipo-C (CNP). Os receptores guanilil-ciclase modulam todos os efeitos sistêmicos conhecidos dos NP. Receptores de clareamento determinam o catabolismo dos NP e, desta maneira, regulam seus níveis plasmáticos e/ou sua concentração tecidual. As propriedades do tipo estrutura-função e homeostáticas dos receptores de NP são apresentadas. O ANP, que tem um importante papel na homeostase pressão-volume, é discutido em relação aos seus efeitos sobre a hemodinâmica renal e funções de excreção, inibição do sistema renina-angiotensina-aldosterona, vaso-relaxamento e ação no terceiro espaço. Quanto ao BNP, especial atenção é focada no seu papel como um modulador negativo da remodelação ventricular, em vista de seus efeitos anti-hipertróficos, anti-fibróticos e anti-inflamatórios no coração. O principal efeito do CNP em promover crescimento ósseo vertebral e longitudinal é discutido brevemente. Finalmente, enfatiza-se a recente descoberta de que o ANP afeta o metabolismo de gorduras em humanos, devido à sua poderosa ação lipolítica.

Csk mediates G-protein-coupled lysophosphatidic acid receptor-induced inhibition of membrane-bound guanylyl cyclase activity.

Natriuretic peptides (NPs) are involved in many physiological processes, including the regulation of vascular tone, sodium excretion, pressure-volume homeostasis, inflammatory responses, and cellular growth. The two main receptors of NP, membrane-bound guanylyl cyclases A and B (GC-A and GC-B), mediate the effects of NPs via the generation of cGMP. NP-stimulated generation of cGMP can be modulated by intracellular processes, whose exact nature remains to be elucidated. Thus, serum and lysophosphatidic acid (LPA), by unknown pathways, have been shown to inhibit the NP-induced generation of cGMP. Here we report that the nonreceptor-tyrosine-kinase Csk is an essential component of the intracellular modulation of atrial natriuretic peptide (ANP)-stimulated activation of GC-A. The genetic deletion of Csk (Csk(-)(/)(-)) in mouse embryonic fibroblasts blocked the inhibitory effect of both serum and LPA on the ANP-stimulated generation of cGMP. Moreover, using a chemical rescue approach, we also demonstrate that the catalytic activity of Csk is required for its modulatory function. Our data demonstrate that Csk is involved in the control of cGMP levels and that membrane-bound guanylyl cyclases can be critically modulated by other receptor-initiated intracellular signaling pathways.