The Role of Botulinum Neurotoxin A in the Conservative Treatment of Fractures: An Experimental Study on Rats.

This paper explores the role of botulinum neurotoxin in aiding fracture recovery through temporary muscle paralysis. Specifically, it investigates the effects of botulinum neurotoxin-induced paralysis of the sternocleidomastoid muscle on clavicle fractures in rats. The research aims to assess safety, effectiveness, and the impact on fracture healing. Healthy male Albino Wistar rats were divided into four groups: clavicle fracture, botulinum neurotoxin injection, both, and control. Surgeries were conducted under anaesthesia, and postoperatively, animals were monitored for 28 days. Euthanasia and radiological assessment followed, examining fracture healing and muscle changes, while tissues were histopathologically evaluated. The modified Lane-Sandhu scoring system was used for the radiographic evaluation of clavicle fractures, and the results varied from complete healing to nonunion. Histopathological examination at 28 days postfracture showed fibrous tissue, mesenchymal cells, and primary callus formation in all groups. Despite varied callus compositions, botulinum neurotoxin administration did not affect clavicle healing, as evidenced by similar scores to the control group. Several studies have explored botulinum neurotoxin applications in fracture recovery. Research suggests its potential to enhance functional recovery in certain types of fractures. Theoretical benefits include managing muscle spasticity, aiding reduction techniques, and preventing nonunion. However, botulinum neurotoxin's transient effect and nonuniversal applications should be considered. The present study found that botulinum toxin had no clear superiority in healing compared to controls, while histological evaluation showed potential adverse effects on muscle tissue. Further research is essential to understand its risk-benefit balance and long-term effects.

Antioxidant effect of a polyphenol-rich grape pomace extract on motility, viability and lipid peroxidation of thawed bovine spermatozoa.

BACKGROUND: Grape extracts of the Greek species Vitis vinifera possess potent antioxidant properties in vitro. The freeze/thaw process and the preparation of semen during assisted reproductive techniques can adversely affect the functional integrity of spermatozoa. The objective was to assess the effect of three different concentrations (1 µg ml(-1), 2 µg ml(-1) and 5 µg ml(-1)) of a polyphenol-rich grape pomace extract on motility, viability, acrosomal and lipid peroxidation status of thawed bovine spermatozoa after 2 and 4 hrs of incubation. RESULTS: The results indicate that the percentage of "Rapid" spermatozoa remained significantly increased (p <0.05) in the presence of 5 µg ml(-1) of the extract, compared to the control after 2 hrs of incubation. Additionally, the incubation of spermatozoa with 2 µg ml(-1) and 5 µg ml(-1) of the extract for 2 hrs resulted in a significantly better maintenance of viable spermatozoa with intact acrosome (p <0.05). The other parameters did not show statistically significant changes. Moreover, the presence of 2 µg ml(-1) and 5 µg ml(-1) of the extract kept the levels of Malondialdehyde (MDA) production in significantly lower level, compared to the other groups, after 2 hrs and 4 hrs of incubation (p <0.05). Particularly, a dose-dependent effect was noticed after 2 hrs of incubation. CONCLUSIONS: Our results suggest that the grape pomace extract exerts a powerful antioxidant role, by suppressing lipid peroxidation, and provides protection in terms of motility and acrosomal integrity, which are correlated with in vivo fertility. The optimal extract concentration is 5 µg ml(-1).

CCl4 induces tissue-type plasminogen activator in rat brain; protective effects of oregano, rosemary or vitamin E.

The high metabolic rate and relatively low antioxidant defenses of the lipid-rich brain tissue render it highly susceptible to reactive oxygen species (ROS) and oxidative stress, whereas the implication of ROS in the pathogenesis of several diseases in the central nervous system is well-established. The plasminogen activator (PA) system is a key modulator of extracellular proteolysis, extracellular matrix remodeling and neuronal cell signaling and has been implicated in the pathogenesis of these diseases. This study evaluates the role of tissue-type PA (t-PA) in oxidative stress and the protective role of dietary antioxidants in the rat brain. We used the CCl4 experimental model of ROS-induced lipid peroxidation and evaluated the antioxidant effect of oregano, rosemary or vitamin E. CCl4-treated Wistar rats exhibited elevated brain t-PA activity, which was decreased upon long-term administration of oregano, rosemary or vitamin E. PA inhibitor-1 (PAI-1) activity was also slightly elevated by CCl4, but this increase was not affected by the antioxidants. We hypothesize that the CCl4-induced t-PA activity indicates extracellular proteolytic activity that may be linked to neuronal cell death and brain damage. Vitamin E or antioxidants present in oregano or rosemary are effective in inhibiting t-PA elevation and can be considered as a potential protection against neuronal damage.

Endosulfan-induced lipid peroxidation in rat brain and its effect on t-PA and PAI-1: ameliorating effect of vitamins C and E.

Endosulfan provokes systemic toxicity in mammals and induces reactive oxygen species (ROS) and lipid peroxidation (LPO). The brain is susceptible to LPO and several studies implicate ROS and LPO in CNS diseases. Tissue plasminogen activator (t-PA) has been accredited with plasminogen-dependent roles in the CNS, as well as plasminogen-independent functions. The aim of this study was to investigate the activities of t-PA and its inhibitor, plasminogen activator inhibitor-1 (PAI-1) in the adult rat brain, after subchronic endosulfan treatment. Furthermore, the potency of vitamins C and E to attenuate these effects was explored. Endosulfan was administered in Wistar rats either alone or with vitamin C and/or vitamin E. The induced oxidative stress was manifested by induction of LPO as determined by higher malondialdehyde levels. This was accompanied by elevation of t-PA and PAI-1 activities. Vitamins E and C, both well-known for their antioxidant properties, substantially acted in a preventive way and protected the brain from these effects.

Plasminogen activator activity in the porcine oviduct during the oestrous cycle.

The mammalian oviduct is a dynamic tissue, which lies under the influence of ovarian steroids and produces proteins that affect various stages of fertilization and post-fertilization events. In this study, expression of urokinase-type plasminogen activator (u-PA mRNA) and plasminogen activator activity (PAA) were examined in porcine oviducts by reverse transcription and polymerase chain reaction (RT-PCR) and activity assays, respectively. For this purpose, oviducts were collected from Landrace cycling sows and divided into three segments (isthmus, ampulla, infundibulum). Different concentrations of u-PA mRNA were detected in the three segments following the pattern isthmus>ampulla>infundibulum and this pattern was maintained during the oestrous cycle. On the contrary, the highest PAA was measured in the ampulla compared to the isthmus and the infundibulum and the highest ampullary PAA was detected during the first 2 days of the oestrous cycle. The different regulation of u-PA mRNA expression and PAA is probably due to the existence of PA inhibitors. Recent observations suggest that PAI-1, the main inhibitor of PAs, shows greater expression in the isthmus compared to the ampulla and the local generation of plasmin is inhibited. The latter may be related to observations that spermatozoa are quiescent in the isthmus before fertilization. This study supports the suggestion that urokinase-type plasminogen activator has a biological role within the porcine oviduct, especially at or near the time of fertilization.

Effect of plasmin on movement characteristics of ejaculated bull spermatozoa.

Proteolytic enzymes appear to have an essential role in multiple phases of mammalian fertilization. Several observations suggest that the plasminogen activator/plasmin system might also play a role in mammalian fertilization. Movement characteristics of bovine sperm incubated with different concentrations of plasmin were investigated using a computer-assisted automated semen analysis system. Sperm were incubated up to 4h in a modified Tyrode's medium (control) and 0.1, 1, 10 and 100 mU/ml of plasmin. The percentage motile sperm was significantly higher at 0 h for sperm incubated in 1, 10, and 100 mU of plasmin. Relative to sperm incubated in control medium, lateral head displacement (ALH), curvilinear velocity, beat cross frequency, path velocity and straight line velocity (VSL) of sperm treated with 100 mU of plasmin for 0 h were increased. After 2h of incubation, sperm treated with 100 mU of plasmin showed an increase in ALH, but a decrease in VSL, straightness and linearity. The effect of plasmin on most motility parameters appears to be direct since all these parameters were affected at 0 h of incubation. Our results support the notion of hyperactivation of bovine spermatozoa following incubation with different concentrations of plasmin. The present work provides additional information to further characterize motility movement of bovine sperm associated with final preparation for fertilization.