Exogenous Nicotinamide Adenine Dinucleotide Attenuates Postresuscitation Myocardial and Neurologic Dysfunction in a Rat Model of Cardiac Arrest.

OBJECTIVES: To investigate the therapeutic potential and underlying mechanisms of exogenous nicotinamide adenine dinucleotide+ on postresuscitation myocardial and neurologic dysfunction in a rat model of cardiac arrest. DESIGN: Thirty-eight rats were randomized into three groups: 1) Sham, 2) Control, and 3) NAD. Except for the sham group, untreated ventricular fibrillation for 6 minutes followed by cardiopulmonary resuscitation was performed in the control and NAD groups. Nicotinamide adenine dinucleotide+ (20 mg/kg) was IV administered at the onset of return of spontaneous circulation. SETTING: University-affiliated research laboratory. SUBJECTS: Sprague-Dawley rats. INTERVENTIONS: Nicotinamide adenine dinucleotide+. MEASUREMENTS AND MAIN RESULTS: Hemodynamic and myocardial function were measured at baseline and within 4 hours following return of spontaneous circulation. Survival analysis and Neurologic Deficit Score were performed up to 72 hours after return of spontaneous circulation. Adenosine triphosphate (adenosine triphosphate) level was measured in both brain and heart tissue. Mitochondrial respiratory chain function, acetylation level, and expression of Sirtuin3 and NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 9 (NDUFA9) in isolated mitochondrial protein from both brain and heart tissue were evaluated at 4 hours following return of spontaneous circulation. The results demonstrated that nicotinamide adenine dinucleotide+ treatment improved mean arterial pressure (at 1 hr following return of spontaneous circulation, 94.69 ± 4.25 mm Hg vs 89.57 ± 7.71 mm Hg; p < 0.05), ejection fraction (at 1 hr following return of spontaneous circulation, 62.67% ± 6.71% vs 52.96% ± 9.37%; p < 0.05), Neurologic Deficit Score (at 24 hr following return of spontaneous circulation, 449.50 ± 82.58 vs 339.50 ± 90.66; p < 0.05), and survival rate compared with that of the control group. The adenosine triphosphate level and complex I respiratory were significantly restored in the NAD group compared with those of the control group. In addition, nicotinamide adenine dinucleotide+ treatment activated the Sirtuin3 pathway, down-regulating acetylated-NDUFA9 in the isolated mitochondria protein. CONCLUSIONS: Exogenous nicotinamide adenine dinucleotide+ treatment attenuated postresuscitation myocardial and neurologic dysfunction. The responsible mechanisms may involve the preservation of mitochondrial complex I respiratory capacity and adenosine triphosphate production, which involves the Sirtuin3-NDUFA9 deacetylation.

Galectin-3 in septic acute kidney injury: a translational study.

BACKGROUND: Galectin-3 (Gal-3) is a pleiotropic glycan-binding protein shown to be involved in sepsis and acute kidney injury (AKI). However, its role has never been elucidated in sepsis-associated AKI (S-AKI). We aimed to explore Gal-3's role and its potential utility as a therapeutic target in S-AKI. METHODS: In 57 patients admitted to the intensive care unit (ICU) with sepsis, serum Gal-3 was examined as a predictor of ICU mortality and development of AKI. In a rat model of S-AKI induced by cecal ligation and puncture (CLP), 7-day mortality and serum Gal-3, Interleukin-6 (IL-6), and creatinine were examined at 2, 8, and 24 hours (h) post-CLP. Two experimental groups received the Gal-3 inhibitor modified citrus pectin (P-MCP) at 400 mg/kg/day and 1200 mg/kg/day, while the control group received water only (n = 18 in each group). RESULTS: Among 57 patients, 27 developed AKI and 8 died in the ICU. Serum Gal-3 was an independent predictor of AKI (OR = 1.2 [95% CI 1.1-1.4], p = 0.01) and ICU mortality (OR = 1.4 [95% CI 1.1-2.2], p = 0.04) before and after controlling for age, AKI, and acute physiology and chronic health evaluation (APACHE II) score. In the CLP rat experiment, serum Gal-3 peaked earlier than IL-6. Serum Gal-3 was significantly lower in both P-MCP groups compared to control at 2 h post-CLP (400 mg: p = 0.003; 1200 mg: p = 0.002), and IL-6 was significantly lower in both P-MCP groups at all time points with a maximum difference at 24 h post-CLP (400 mg: p = 0.015; 1200 mg: p = 0.02). In the Gal-3 inhibitor groups, 7-day mortality was significantly reduced from 61% in the control group to 28% (400 mg P-MCP: p = 0.03) and 22% (1200 mg P-MCP: p = 0.001). Rates of AKI per RIFLE criteria were significantly reduced from 89% in the control group to 44% in both P-MCP groups (400 mg: p = 0.007; 1200 mg: p = 0.007). CONCLUSIONS: This translational study demonstrates the importance of Gal-3 in the pathogenesis of S-AKI, and its potential utility as a therapeutic target.

Vacuum therapy ameliorates erectile dysfunction in bilateral cavernous nerve crush rats by inhibiting apoptosis and activating autophagy.

Postprostatectomy erectile dysfunction (pPED) remains a current problem despite improvements in surgical techniques. Vacuum therapy is clinically confirmed as a type of pPED rehabilitation. However, its underlying mechanisms are incompletely understood. Recently, autophagy and apoptosis were extensively studied in erectile dysfunction resulting from diabetes, senescence, and androgen deprivation but not in the context of pPED and vacuum therapy. Therefore, this study was designed to investigate the roles of autophagy and apoptosis in pPED and vacuum therapy. Twenty-four adult male Sprague-Dawley rats were randomly divided into three groups: the control group, bilateral cavernous nerve crush (BCNC) group, and BCNC + vacuum group. After 4 weeks of treatment, intracavernosal pressure was used to evaluate erectile function. Real-time quantitative polymerase chain reaction, western blot, and immunohistochemistry were used to measure the molecular expression. TdT-mediated dUTP nick-end labeling staining was used to assess apoptosis. Transmission electron microscopy was used to observe autophagosomes. After treatment, compared with those of the BCNC group, erectile function and cavernosal hypoxia had statistically significantly improved (P < 0.05). Apoptosis and the relative protein expression of B-cell lymphoma-2-associated X and cleaved Caspase3 were decreased (P < 0.05). Autophagy-related molecules such as phosphorylated unc-51-like autophagy-activating kinase 1 (Ser757) and p62 were decreased. Beclin1, microtubule-associated protein 1 light chain 3 A/B, and autophagosomes were increased (P < 0.05). Besides, the phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin signaling pathway, as a negative regulator of autophagy to some degree, was inhibited. This study revealed that vacuum therapy ameliorated pPED in BCNC rats by inhibiting apoptosis and activating autophagy.

Modeling Burn Progression Using Comb Burns: The Impact of Thermal Contact Duration on Model Outcomes.

BACKGROUND: Burn progression is a phenomenon that remains poorly characterized. The mechanisms of burn conversion are not completely understood, and consequently, both predictive diagnostic tools and interventions are limited. The rat comb burn model is a commonly used approach to study horizontal burn conversion. However, there is significant variability in how the model is performed. Skin contact duration, comb device heating method, comb heating duration, amount of pressure applied, the weight of the comb, and associated depth of burn are all variables that are heterogeneous in studies utilizing the model. MATERIALS AND METHODS: Here, contact duration was examined to determine the impact the duration of burn delivery has on the conversion of interspaces in this model. Data from multiple experiments consisting of 10, 15, 20, 30, 40, and 45 s comb burns were compiled and assessed. Burns were made using combs heated in a 100°C dry bath and then monitored for 2 d. Interspace viability was assessed by digital and laser doppler imaging and biopsy procurement. RESULTS: Laser Doppler Imaging and viable interspace measurements showed that as burn duration increased, the percentage of the viable interspace and interspace perfusion decreased. Additionally, a contact time of 30 s or greater was required to result in 100% interspace conversion. CONCLUSIONS: These results demonstrate a need to better characterize and potentially standardize the rat comb burn model to reduce variation and maintain it as a valuable tool for controlled studies of the pathophysiology of burn wound progression.

Diffusion Tensor Tractrography Visualizes Partial Nerve Laceration Severity as Early as 1 Week After Surgical Repair in a Rat Model Ex Vivo.

BACKGROUND: Previous studies in our laboratory have demonstrated that a magnetic resonance imaging method called diffusion tensor imaging (DTI) can differentiate between crush and complete transection peripheral nerve injuries in a rat model ex vivo. DTI measures the directionally dependent effect of tissue barriers on the random diffusion of water molecules. In ordered tissues such as nerves, this information can be used to reconstruct the primary direction of diffusion along fiber tracts, which may provide information on fiber tract continuity after nerve injury and surgical repair. METHODS: Sprague-Dawley rats were treated with different degrees of partial transection of the sciatic nerve followed by immediate repair and euthanized after 1 week of recovery. Nerves were then harvested, fixed, and scanned with a 7 Tesla magnetic resonance imaging to obtain DTIand fiber tractography in each sample. Additional behavioral (sciatic function index, foot fault asymmetry) and histological (Toluidine blue staining) assessments were performed for validation. RESULTS: Tractography yielded a visual representation of the degree of injury that correlated with behavioral and histological evaluations. CONCLUSIONS: DTI tractography is a noninvasive tool that can yield a visual representation of a partial nerve transection as early as 1 week after surgical repair.

A Practical Noncontact Model to Create Standardized Experimental Burn Wounds of Any Thickness: Blue Beam Laser Pointer for Burn Induction.

The objective of this study was to describe a predictable and easy-to-use model that can create standardized burn wounds. A 450-nm 1000-mW blue beam laser pointer was used to create burn wounds on the dorsal skin of 24 Sprague Dawley rats. Twelve distinct areas of dorsal skin were pulsed for 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, and 23 seconds with the help of a punched plastic card template. Three groups of 8 animals were killed immediately after on the third day and on the seventh day of the procedure and tissue samples were taken for histological evaluation and measurements. A second-degree burn was obtained in all animals with 3 and 5 seconds of laser application on the same day, third day, and seventh day measurements. Seven seconds of application resulted in a burn depth of 84.87% of dermis on the application day which deepened to involve the whole dermal layer on the third and seventh day. Nine seconds and longer application times resulted in third-degree burn wounds. Burn induction with blue beam laser pointer is an easy-to-use, predictable and safe model to create a standardized burn wound of desired thickness.

Differential Effects of Caffeine on Motor and Cognitive Outcomes of Penetrating Ballistic-Like Brain Injury.

This study assessed the effect of caffeine on neurobehavioral recovery in the WRAIR penetrating ballistic-like brain injury (PBBI) model. Unilateral frontal PBBI was produced in the right hemisphere of anesthetized rats at moderate (7%-PBBI) or severe (10%-PBBI) injury levels. Animals were randomly assigned to pretreatment groups: acute caffeine (25 mg/kg CAF gavage, 1 h prior to PBBI), or chronic caffeine (0.25 g/L CAF drinking water, 30 days prior to PBBI). Motor function was evaluated on the rotarod at fixed-speed increments of 10, 15, and 20 RPM. Cognitive performance was evaluated on the Morris water maze. Acute caffeine showed no significant treatment effect on motor or cognitive outcome. Acute caffeine exposure prior to 10%-PBBI resulted in a significantly higher thigmotaxic response compared to vehicle-PBBI groups, which may indicate caffeine exacerbates post-injury anxiety/attention decrements. Results of the chronic caffeine study revealed a significant improvement in motor outcome at 7 and 10 days post-injury in the 7%-PBBI group. However, chronic caffeine exposure significantly increased the latency to locate the platform in the Morris water maze task at all injury levels. Results indicate that chronic caffeine consumption prior to a penetrating TBI may provide moderate beneficial effects to motor recovery, but may worsen the neurocognitive outcome.

Evaluating the Effects of a Topical Preparation with Dexpanthenol, Silbiol, Undecylenic Acid, and Lidocaine on Palatal Mucosa Wound Healing in a Rat Model

Background: Postoperative complications occur after periodontal plastic surgeries, but an ideal treatment to overcome them has not been found yet. Aims: To evaluate the effects of topically applied Oral-norm gel on the healing of excisional wounds. Study Design: Animal experiment. Methods: Excisional wounds with a diameter of 3 mm were made in the center of the palatal mucosa of 63 Sprague Dawley rats. Seven animals were sacrificed at time 0. The remaining rats were divided into two groups: a test group in which the topical Oral-norm gel was applied three times a day and a control group in which nothing was applied. Seven animals in each group were sacrificed at 3, 7, 14, and 21 days. Mean wound surface area was measured photographically, while wound healing and width were evaluated microscopically. Results: The mean wound surface area decreased significantly after 3 days in both groups (p<0.001). Between days 3 and 7, the mean wound surface area decreased from 6.62 (2.85) to 0.83 (1.62) mm2 in the control group and 5.07 (0.88) to 1.42 (1.67) mm2 in the test group. The wound width decreased significantly on day 7 in both groups (p<0.001), with no further changes by day 14. Both groups had a significant increase in inflammation and vascularization on day 3 (p<0.001), with a reduction thereafter. No significant differences in macroscopic and microscopic measurements were observed between the groups at any time point (p>0.05). Conclusion: The Oral-norm gel has no positive healing effects in the palatal mucosa of rats.

Effects on Neurons and Hippocampal Slices by Single and Multiple Primary Blast Pressure Waves From Detonating Spherical Cyclotrimethylenetrinitramine (RDX) Explosive Charges.

Threshold shock-impulse levels required to induce cellular injury and cumulative effects upon single and/or multiple exposures are not well characterized. Currently, there are few in vitro experimental models with blast pressure waves generated by using real explosives in the laboratory for investigating the effects of primary blast-induced traumatic brain injury. An in vitro indoor experimental platform is developed using real military explosive charges to accurately represent battlefield blast exposure and to probe the effects of primary explosive blast on dissociated neurons and tissue slices. Preliminary results indicate that physical insults altered membrane permeability, impacted cellular viability, created axonal beadings, and led to synaptic protein loss in hippocampal slice cultures. Injuries from blast under the conditions that were examined did not appear to cause immediate or sustained damage to the cells. Three consecutive primary blasts failed to disrupt the overall cellular integrity in the hippocampal slice cultures and produced a unique type of pathology comprised with distinct reduction in synaptic proteins before cellular deterioration set in. These observed changes might add to the challenges in regard to enhancing our understanding of the complex biochemical and molecular mechanisms caused by primary blast-induced injury.

Operation Brain Trauma Therapy: 2016 Update.

Operation brain trauma therapy (OBTT) is a multi-center, pre-clinical drug and biomarker screening consortium for traumatic brain injury (TBI). Therapies are screened across three rat models (parasagittal fluid percussion injury, controlled cortical impact [CCI], and penetrating ballistic-like brain injury). Operation brain trauma therapy seeks to define therapies that show efficacy across models that should have the best chance in randomized clinical trials (RCTs) and/or to define model-dependent therapeutic effects, including TBI protein biomarker responses, to guide precision medicine-based clinical trials in targeted pathologies. The results of the first five therapies tested by OBTT (nicotinamide, erythropoietin, cyclosporine [CsA], simvastatin, and levetiracetam) were published in the Journal of Neurotrauma. Operation brain trauma therapy now describes preliminary results on four additional therapies (glibenclamide, kollidon-VA64, AER-271, and amantadine). To date, levetiracetam was beneficial on cognitive outcome, histology, and/or biomarkers in two models. The second most successful drug, glibenclamide, improved motor function and histology in CCI. Other therapies showed model-dependent effects (amantadine and CsA). Critically, glial fibrillary acidic protein levels predicted treatment effects. Operation brain trauma therapy suggests that levetiracetam merits additional pre-clinical and clinical evaluation and that glibenclamide and amantadine merit testing in specific TBI phenotypes. Operation brain trauma therapy has established that rigorous, multi-center consortia could revolutionize TBI therapy and biomarker development.

Trends of reactive hyperaemia responses to repetitive loading on skin tissue of rats - Implications for pressure ulcer prevention.

Tissue recovery is important in preventing tissue deterioration, which is induced by pressure and may lead to pressure ulcers (PU). Reactive hyperaemia (RH) is an indicator used to identify people at risk of PU. In this study, the effect of different recovery times on RH trend is investigated during repetitive loading. Twenty-one male Sprague-Dawley rats (seven per group), with body weight of 385-485 g, were categorised into three groups and subjected to different recovery times with three repetitive loading cycles. The first, second, and third groups were subjected to short (3 min), moderate (10 min), and prolonged (40 min) recovery, respectively, while fixed loading time and pressure (10 min and 50 mmHg, respectively). Peak hyperaemia was measured in the three cycles to determine trends associated with different recovery times. Three RH trends (increasing, decreasing, and inconsistent) were observed. As the recovery time is increased (3 min vs. 10 min vs. 40 min), the number of samples with increasing RH trend decreases (57% vs. 29% vs. 14%) and the number of samples with inconsistent RH trend increases (29% vs. 57% vs. 72%). All groups consists of one sample with decreasing RH trend (14%). Results confirm that different recovery times affect the RH trend during repetitive loading. The RH trend may be used to determine the sufficient recovery time of an individual to avoid PU development.

The Effect of Pycnogenol on Wound Healing in Diabetic Rats.

Pycnogenol (PYC), an extract of pine bark, is known to have photoprotective, antimicrobial, antioxidant, and anti-inflammatory properties. An in vivo study was conducted to evaluate the effects of PYC treatment on wound healing in 48 adult male Sprague-Dawley rats, of which 24 were injected with a single dose of alloxan to induce diabetes. Three (3) excisional skin wounds (1.3 cm x 1.3 cm x 2 mm) were created in each healthy and diabetic animal. One (1) wound in each animal was left untreated, 1 was treated daily with a cleanser (ethacridine lactate) and covered with silver sulfadiazine (SSD), and 1 was treated with PYC powder (30 mg). After measuring wound size, 6 animals from both groups were sacrificed on days 3, 7, 14, and 21 and tissue samples were taken for histopathological evaluation of acute and chronic inflammation, granulation tissue, fibroblast maturation, collagen deposition, epithelialization, and neovascularization using a scoring system of 0 = none, 1 = mild, 2 = moderate, and 3 = abundant. Because the wounds created were not uniform in size within and among the animals, healing was expressed as a percentage of the initial wound size for each animal. Data were compared using 2-way analysis of variance; histopathological lesion scores were reported in median values in univariate analysis, with P <.05 denoting statistical significance. The mean initial wound surface area was 1.69 ± 0.44 cm². On day 21, the average reduction in wound size was lower in diabetic than in healthy rats (47.42% versus 50.91%, P <.0001) and, in both groups combined, the average reduction was 45.73% in untreated, 48.73% in cleanser/SSD-treated, and 58.03% in PYC-treated wounds (P <.0001). Wound size reduction was also significantly different between PYC and the cleanser/SSD treatment depending on the rats' health status (P <.0001): 49.68% and 47.84% using cleanser/SSD and 56.17% and 49.84% using PYC in healthy and diabetic rats, respectively. After 3 weeks, wound size for the healthy rats had decreased more than in the diabetic rats (mean 50.91% versus 47.42%). Although reepithelialization was complete in both groups by day 21, complete neovascularization was evident in the healthy rats but not in the diabetic rats. Overall, compared to the untreated control wounds, treatments with cleanser/SSD and PYC were equally effective in lowering acute and chronic inflammation scores on days 7 and 21. In diabetic rat wounds, collagen deposition and neovascularization scores were higher in wounds treated with PYC than cleanser/SSD-treated wounds (1.5 versus 1.0 and 2.0 versus 1.5, respectively). PYC appears to be a viable option to accelerate wound healing. Further in vivo and human research is warranted.

Establishment of a new anal sphincter injury model in rats based on cardiotoxin.

BACKGROUND/PURPOSE: Mainstream models for anal sphincter injury use large animals. We developed a simple and stable anal sphincter injury model in a small animal (i.e., rats) to obtain manometry measurements by using a miniaturized probe and applying cardiotoxin. METHODS: The histological structure of the anal canal was evaluated by using manometry in normal rats (n=40). We damaged the internal and external anal sphincters by locally administering snake poison (cardiotoxin; 20 uM, 100µL 8 points). We evaluated the anal canal function through manometry measurements (n=5) and examined the histology using hematoxylin-eosin staining (at each time point, n=3; total n=15). RESULTS: The manometry parameters and structure of the anal canal of normal rats were similar to those of humans, because rats have resting pressure, rectoanal reflex in the manometry, and an external and internal anal sphincter. After inducing injury, the following findings were observed: rhythmic wave loss and a remarkable reduction in the anal sphincter resting pressure; and local bleeding and advanced infiltration of the inflammatory cells (day 1) and the loss of muscle fibers (day 3). CONCLUSION: This new rat model will contribute to increasing the knowledge on the anal canal.

Effects of systemic erythropoietin on ischemic wound healing in rats.

Results of in vivo studies have shown erythropoietin (EPO) is associated with anti-inflammatory, anti-apoptotic, and cell protective effects on wound healing. These effects are dose-dependent. The aim of this study was to evaluate whether the duration of EPO treatment affects the healing process in the ischemic wound. Forty-two (42) Sprague-Dawley rats were anesthetized, wounded with H-shaped flaps, and randomized to 2 groups; Group 1 received 400 u/kg/day EPO and Group 2 received a saline solution, both via intraperitoneal injection following the wounding. All substances were administered once daily at the same time for up to 10 days after surgery. At days 3, 5, and 10, 7 rats from each group were sacrificed. Skin samples were stained with hematoxylin/eosin, viewed under an optical microscope at 10X and 40X magnification, and analyzed by blinded investigators for re-epithelialization, neovascularization amount and maturation of granulation tissue, inflammatory cells, and ulcer healing using an evaluation scale where 0 = none; 1 = partial; 2 = complete, but immature/thin: and 4 = complete and mature. Blood hemoglobin and hematocrit levels also were measured. Data were analyzed using ANOVA one-way test (P <0.05). Hemoglobin and hematocrit levels rose while subsequent doses of EPO were administered over time, accompanied by a transient surge in healing on day 5, when differences in healing scores were significant. Flap necrosis, ulceration, and abscess were noted on post-wounding day 10 near the pedicle. The study showed EPO therapy can improve wound healing early in the post-wounding period but can reduce wound healing after post-injury treatment day 5. Further research is necessary, particularly to establish how EPO influences the microcirculation and rheology.

Tratamiento con ultrasonido continuo de alta intensidad en nervios isquiáticos de rata (Sprague Dawley) lesionados por compresión / Treatment with continuous high-intensity ultrasound on rat sciatic nerve (Sprague Dawley) and compression injuries

Actualmente las patologías compresivas de nervios espinales son de alta incidencia. Como una forma de contribuir con fundamentos morfológicos a la aplicación de ultrasonido terapéutico se realizó la presente experiencia, que se refiere a la reparación de los nervios espinales luego de una injuria compresiva. Para esto se utilizaron 9 ratas Sprague Dawley las cuales fueron separadas en tres grupos con 3 ratas cada uno: A) control sano, B) control lesionado y C) aplicación de ultrasonido terapéutico de 1w/cm2. Las ratas de los grupos B y C fueron anestesiadas y se les aisló quirúrgicamente el nervio isquiático, el cual fue pinzado durante 45 segundos con una fuerza constante de 40N. La compresión se realizó a 10mm sobre su bifurcación distal, luego se desinfectó y suturó. 24 horas después de la compresión las ratas del grupo C fueron irradiadas con un equipo de ultrasonido terapéutico, utilizando 1w/cm2 de intensidad con frecuencia de 3 MHZ y un cabezal de 0,5cm2, durante 1 minuto y en 10 días seguidos. 28 días después de la irradiación se extrajeron los nervios isquiáticos y fueron sometidos a técnicas histológicas de rutina con tinción de HE y VG. Se realizó el diagnóstico histopatológico y la morfometría donde se midió: grosor del perineuro, perímetro de los núcleos de neurilemocitos, perímetro del axón mielínico, perímetro de la mielina, ancho del núcleo del fibroblastocito y largo del núcleo del fibroblastocito. Los resultados permiten concluir que el ultrasonido de modalidad continua estimula la regeneración del nervio espinal lesionado por compresión.

Currently, the pathologies of spinal nerves are of high incidence. As a way to contribute to morphological grounds the application of therapeutic ultrasound was used in the repair of spinal nerves after a compressive injury. 9 tests were carried out using Sprague Dawley rats which were separated into three groups with 3 rats each: A) healthy control, B) injured control and C) the application of therapeutic ultrasound 1w/cm2. The rats in groups B and C were anesthetized and sciatic nerve was surgically isolated, which was clamped for 45 seconds with constant force of 40N. Compression was performed on the distal bifurcation 10mm, then disinfected and sutured. 24 hrs after compression, rats in group C were irradiated with a therapeutic ultrasound equipment, using 1w/cm2 of intensity with frequency of 3 MHz and a head of 0.5 cm2 for 1 minute, for 10 days. 28 days after irradiation sciatic nerve was removed and subjected to routine histological staining with HE and VG. Histopathological diagnosis was made and morphometry which measured: perineum thickness, perimeter of the nuclei of neurolemocyte cell, perimeter of myelinated axons, perimeter of myelin, the fibroblastocyte core width and length of the fibroblastocyte nucleus. The results show that the continuous mode ultrasound stimulates the regeneration of injured spinal nerve compression.

Effects of ischemia/reperfusion on beta cells of pancreas and protective effects of melatonin treatment / Efectos de la isquemia/reperfusión sobre células beta del páncreas y efectos protectores del tratamiento de melatonina

Oxygen free radicals are considered to be important components involved in the pathophysiological tissue alterations observed during ischemia-reperfusion (I/R). In this study, we investigated the putative protective effects of melatonin treatment on pancreatic I/R injury. Sprague Dawley male rats were subjected to 30 min of pancreatic pedicle occlusion followed by 90 min reperfusion. Melatonin (10 mg/kg. s.c) was administrated 30 min prior to ischemia or I/R application. At the end of the reperfusion periods, rats were decapitated. Pancreatic samples were taken for transmission electron microscopy. The results indicated that ischemia created b cell damage as evidenced by dilatation between the nucleus inner and outer membrane and degeneration on islets of Langerhans cells, was reversed by melatonin treatment. As melatonin administration reversed these microscopic damage, it seems likely that melatonin protects pancreatic tissue against oxidative damage.

Los radicales libres del oxígeno son considerados como uno de los componentes más importantes que participan en las alteraciones fisiopatológicas del tejido durante la isquemia-reperfusión (I/R). En este estudio, se investigó el supuesto efecto protector del tratamiento de melatonina sobre la lesión pancreática I/R. Ratas Sprague Dawley machos fueron sometidas a 30 minutos de oclusión del pedículo pancreático seguido de 90 minutos de reperfusión. La melatonina (10 mg/kg) fue administrada 30 minutos antes de la isquemia o de la aplicación I/R. Al finalizar los periodos de reperfusión, las ratas fueron decapitadas. Fueron tomadas muestras pancreáticas para el análisis en microscopía electrónica de transmisión. Los resultados indicaron que la isquemia ocasionó daño en las células beta demostrado por la dilatación entre el núcleo interior y la membrana exterior y la degeneración de los islotes de células pancreáticas, los que fueron revertidos por el tratamiento de melatonina. Como la administración de melatonina revirtió estos daños microscópicos, parece probable que ella proteja al tejido pancreático contra el daño oxidativo.

Investigation of wound healing activity of Lantana camara L. in Sprague dawley rats using a burn wound model.

Summary: Lantana camara is used in herbal medicine for the treatment of skin itches, as an antiseptic for wounds, and externally for leprosy and scabies. The objective of our study is to investigate burn wound healing activity of the leaf extract of L.camara in rats. The animals were divided into two groups of 6 each. The test group animals were treated with the ethanol extract of L. camara (100 mg kg-1 day-1) topically and the control group animals were left untreated. Healing was assessed by the rate of wound contraction, period of epithelialization. Antimicrobial activities of the extract against the specific microorganisms were assessed. The extract showed antimicrobial activity against Staphylococcus aureus, Klebsiella Pneumoniae and E.coli. Extract treated wounds were healed in about 21 days which is not distinct from the controls. Our data suggest that L.camara has antimicrobial activity but not wound healing promoting activity on burn wound. Industrial relevance: Extensive work has been done on the L. camara and demonstrated the antimicrobial and fungicidal activity of its chemical constituents. Lantana oil is used for the treatment of skin itches and as an antiseptic for wounds. It has been used in folk medicine for the treatment of cancers, chicken pox, measles, ulcers, swellings, eczema. Our earlier work showed the healing activity on excision wound model. However, there is no data to support the wound healing activity of L. camara on burn wound. Hence, we have conducted the present study to explore the wound healing activity and the antimicrobial activities of L. camara against the specific microorganisms which generally infect burn wound.

Cecropia peltata L(Cecropiaceae) has wound-healing potential: a preclinical study in a Sprague Dawley rat model

The plant Cecropia peltata is used in traditional medicine to treat a variety of disorders. The objective of the study presented in this report was to screen the extracts of this plant leaf for its wound-healing properties based on its traditional use for wound healing. Aqueous and ethanol extracts were prepared for topical and oral administration. The dose studied was 150 mgkg-1day-1 for 10 days, using the excision wound model in rats. Carboxymethyl cellulose (1 per cent) was used as control in topical and oral route studies. Animals were randomized to treatment or control, the experiment being done with prior ethical approval from the university. Wound areas were measured. On day 11, tissue was excised to determine the contents of protein, hydroxyproline, and hexosamine. Wound areas reduced statistically significantly in all treatment groups compared to respective controls (P < .001). Biochemistry and tissue histology outcomes were consistent with changes in the treatment groups. No differences were detected within the treatment groups. The study permits the conclusion that Crecopia peltata has wound-healing potential.

Apoptosis of spinal interneurons induced by sciatic nerve axotomy in the neonatal rat is counteracted by nerve growth factor and ciliary neurotrophic factor.

We have previously shown that not only motoneurons and dorsal root ganglion cells but also small neurons, presumably interneurons in the spinal cord, may undergo apoptotic cell death as a result of neonatal peripheral nerve transection in the rat. With the aid of electron microscopy, we have here demonstrated that apoptosis in the spinal cord is confined to neurons and does not involve glial cells at the survival time studied (24 hours). To define the relative importance of the loss of a potential target (motoneuron) and a potential afferent input (dorsal root ganglion cell) for the induction of apoptosis in interneurons in this situation, we have compared the distributions and time courses for TUNEL labeling, which detects apoptotic cell nuclei, in the L5 segment of the spinal cord and the L5 dorsal root ganglion after sciatic nerve transection in the neonatal (P2) rat. In additional experiments, we studied the effects on TUNEL labeling of interneurons after treatment of the cut sciatic nerve with either ciliary neurotrophic factor (CNTF) to rescue motoneurons or nerve growth factor (NGF) to rescue dorsal root ganglion cells. The time courses of the TUNEL labeling in motoneurons and interneurons induced by the lesion show great similarities (peak at 8-48 hours postoperatively), whereas the labeling in dorsal root ganglion cells occurs later (24-72 hours). Both CNTF and NGF decrease the number of TUNEL-labeled interneurons, but there is a regional difference, in that CNTF preferentially saves interneurons in deep dorsal and ventral parts of the spinal cord, whereas the rescuing effects of NGF are seen mainly in the superficial dorsal horn. The results are interpreted as signs of a trophic dependence on both the target and the afferent input for the survival of interneurons neonatally.