Hypoglycemic and antioxidative activity evaluation of phenolic compounds derived from walnut diaphragm produced in Xinjiang.

Walnut diaphragm is defined as a dry wood septum located between the walnut shell and kernel. In this work, seven phenolic compounds from walnut diaphragm were purified and characterized, and their antioxidant activities and mechanisms of hypoglycemia were investigated. Compounds 1-7 were tested for DPPH, ABTS scavenging ability, and FRAP assay to evaluate the antioxidant activity. α-Amylase inhibition assay was introduced to assess the hypoglycemic activity, and the mechanism was investigated by kinetic analysis, CD spectrum, and molecular docking. Compound 6 showed the strongest antioxidant ability, while compound 1 exhibited the strongest inhibition of α-amylase by changing the secondary structure of α-amylase in a mixed competitive inhibition mode. Molecular docking test predicted that the tetrahydropyran part in compound 1 may contribute to its hypoglycemic effect. This study furnishes a new theoretical reference for the utilization and development of walnut diaphragm into a health food with antioxidant and hypoglycemic properties. PRACTICAL APPLICATIONS: The finding of this research may serve as a basis for the subsequent development of walnut diaphragm into instant tea-based health food or added to other food carriers to achieve auxiliary antioxidant and hypoglycemic effects. This study revealed that polyphenolic components were the material basis for the antioxidant and hypoglycemic effects of walnut diaphragm, which could be identified as landmark chemical components for controlling quality standards in the development of walnut diaphragm, thus accelerating the research process of quality standards for walnut diaphragm-related products. Furthermore, the studies on the mechanism of hypoglycemic activity supply more credible data to support the development of walnut diaphragm into a safe and consumer-friendly health food. With abundant resources and clear efficacy, walnut diaphragm has great development prospect and application value.

Acute and Subacute Toxicity and Cytotoxicity of Opuntia Dillenii (Ker-Gawl) Haw. Seed Oil and Its Impact on the Isolated Rat Diaphragm Glucose Absorption.

This study aims to assess the safety of the Opuntia dillenii (Ker-Gawl) haw. seed oil (ODSO) and its effect on the glucose absorption activity of the isolated rat hemidiaphragm. This oil's safety was studied by exploring its acute (doses 1, 3, 5, and 7 mL/kg) and subacute (doses 1 and 2 mL/kg) toxicities in albino mice and Wistar rats, respectively. The safety of the ODSO was also assessed by studying its effect on the HepG2 cell viability in vitro. The effect of ODSO, or combined with the insulin, on the glucose absorption activity of isolated rat hemidiaphragm was evaluated at the dose 1 g/L in vitro. The results demonstrated the safety of ODSO. Indeed, this study showed that this oil does not produce any mortality or signs of toxicity after the single-dose administration in mice. Additionally, the daily intake of the ODSO during four weeks does not induce a significant variation in the biochemical parameters and body weight of rats compared with the control group. Besides, the cell viability of HepG2 did not change in the presence of ODSO. On the other hand, the ODSO increased the glucose absorption activity of the isolated rat hemidiaphragm, and this activity was significantly enhanced when combined with insulin. This study confirms, on one side, the safety of this oil and its efficacy and, on the other side, encourages its potential use as a complement to treat diabetes.

Hyperbaric Oxygen Treatment Following Mid-Cervical Spinal Cord Injury Preserves Diaphragm Muscle Function.

Oxidative damage to the diaphragm as a result of cervical spinal cord injury (SCI) promotes muscle atrophy and weakness. Respiratory insufficiency is the leading cause of morbidity and mortality in cervical spinal cord injury (SCI) patients, emphasizing the need for strategies to maintain diaphragm function. Hyperbaric oxygen (HBO) increases the amount of oxygen dissolved into the blood, elevating the delivery of oxygen to skeletal muscle and reactive oxygen species (ROS) generation. It is proposed that enhanced ROS production due to HBO treatment stimulates adaptations to diaphragm oxidative capacity, resulting in overall reductions in oxidative stress and inflammation. Therefore, we tested the hypothesis that exposure to HBO therapy acutely following SCI would reduce oxidative damage to the diaphragm muscle, preserving muscle fiber size and contractility. Our results demonstrated that lateral contusion injury at C3/4 results in a significant reduction in diaphragm muscle-specific force production and fiber cross-sectional area, which was associated with augmented mitochondrial hydrogen peroxide emission and a reduced mitochondrial respiratory control ratio. In contrast, rats that underwent SCI followed by HBO exposure consisting of 1 h of 100% oxygen at 3 atmospheres absolute (ATA) delivered for 10 consecutive days demonstrated an improvement in diaphragm-specific force production, and an attenuation of fiber atrophy, mitochondrial dysfunction and ROS production. These beneficial adaptations in the diaphragm were related to HBO-induced increases in antioxidant capacity and a reduction in atrogene expression. These findings suggest that HBO therapy may be an effective adjunctive therapy to promote respiratory health following cervical SCI.

Respiratory muscle training decreases diaphragm DNA damage in rats with heart failure.

Respiratory muscle training (RMT) promotes beneficial effects on respiratory mechanics, heart and lung morphological changes, and hemodynamic variables in rats with heart failure (HF). However, the relation between RMT effects and diaphragm oxidative stress remains unclear. Therefore, the aim of this study was to evaluate the RMT effects on diaphragm DNA damage in HF rats. Wistar rats were allocated into 4 groups: sedentary sham (Sed-Sham, n = 8), trained sham (RMT-Sham, n = 8), sedentary HF (Sed-HF, n = 8), and trained HF (RMT-HF, n = 8). The animals underwent a RMT protocol (30 min/day, 5 days/week for 6 weeks), whereas sedentary animals did not exercise. Groups were compared by a two-way ANOVA and Tukey's post hoc tests. In rats with HF, RMT promoted reduction in pulmonary congestion (p < 0.0001) and left ventricular end diastolic pressure (p < 0.0001). Moreover, RMT produced a decrease in the diaphragm DNA damage in HF rats. This was demonstrated through the reduction in the percentage of tail DNA (p < 0.0001), tail moment (p < 0.01), and Olive tail moment (p < 0.001). These findings showed that a 6-week RMT protocol in rats with HF promoted an improvement in hemodynamic function and reduces diaphragm DNA damage.

Kinetics of ventilation-induced changes in diaphragmatic metabolism by bilateral phrenic pacing in a piglet model.

Perioperative necessity of deep sedation is inevitably associated with diaphragmatic inactivation. This study investigated 1) the feasibility of a new phrenic nerve stimulation method allowing early diaphragmatic activation even in deep sedation and, 2) metabolic changes within the diaphragm during mechanical ventilation compared to artificial activity. 12 piglets were separated into 2 groups. One group was mechanically ventilated for 12 hrs (CMV) and in the second group both phrenic nerves were stimulated via pacer wires inserted near the phrenic nerves to mimic spontaneous breathing (STIM). Lactate, pyruvate and glucose levels were measured continuously using microdialysis. Oxygen delivery and blood gases were measured during both conditions. Diaphragmatic stimulation generated sufficient tidal volumes in all STIM animals. Diaphragm lactate release increased in CMV transiently whereas in STIM lactate dropped during this same time point (2.6 vs. 0.9 mmol L-1 after 5:20 hrs; p < 0.001). CMV increased diaphragmatic pyruvate (40 vs. 146 µmol L-1 after 5:20 hrs between CMV and STIM; p < 0.0001), but not the lactate/pyruvate ratio. Diaphragmatic stimulation via regular electrodes is feasible to generate sufficient ventilation, even in deep sedation. Mechanical ventilation alters the metabolic state of the diaphragm, which might be one pathophysiologic origin of ventilator-induced diaphragmatic dysfunction. Occurrence of hypoxia was unlikely.

[Regulatory Effect of Chinese Drugs for Stasis Removing and Collaterals Dredging on the Expres- sions of podocin and CD2AP in Podocyte Slit Diaphragm of Diabetic Nephropathy Rats].

OBJECTIVE: To observe the regulatory effect of Chinese drugs for stasis removing and collaterals dredging (CDSRCD) on the expressions of podocin and CD2AP in podocyte slit diaphragm (SD) of diabetic nephropathy (DN) rats. METHODS: DN rat model was duplicated in 40 male Sprague- Dawley rats by feeding high fat high glucose diet combined with intraperitoneally injecting 1 % streptozoto- cin (STZ, 35 mg/kg). Totally 36 successfully modeled rats were divided into the model group, the CD- SRCD group,- and the irbesartan group according to random digit table, 12 in each group. Besides, anoth- er 10 normal rats were recruited as a normal group. Rats in the CDSRCD group and the irbesartan group were intragastrically fed with CDSRCD and irbesartan respectively. Rats in the normal group and the mod- el group were fed with equal volume of distilled water at the same time. 24 h urine protein quantitation was detected using ELISA at various time points. Body weight (BW) , kidney weight ( KW), kidney index (KI) , fasting blood glucose (FBG) , serum creatinine (SCr), blood urea nitrogen (BUN), and uric acid (UA) in each group were detected after 16 weeks of intervention. The pathomorphological changes of re- nal tissue were observed under light microscope and electron microscope respectively. The protein and mRNA expressions of podocin and CD2AP were detected by Western blot and Real-time PCR respectively. RESULTS: (1) Compared with the normal group, 24 h urine protein quantitation significantly increased at week 4, 8, 12, and 16, respectively (P <0. 01). BW was decreased; KI and levels of FBG, SCr, BUN, and UA all increased after modeling (P <0. 01). Compared with the model group, 24 h urine protein quan- titation significantly decreased in the CDSRCD group and the irbesartan group at week 4, 8, 12, and 16, respectively (P <0. 01). Besides, it was more obviously reduced in the CDSRCD group than in the irbe- sartan group (P <0. 05, P <0.01). BUN level obviously decreased both in the CDSRCD group and the irbesartan group after modeling (P <0. 05, P <0. 01). (2) Results of renal pathology showed that disar- ranged renal structure, obviously thickened basement membrane, severely proliferated mesenteria, widely fused foot processes in the model group. All these pathological changes were attenuated in the CD- SRCD group and the irbesartan group to some degree. (3) Results of Western blot and Real-time PCR showed, compared with the normal group, protein and mRNA expressions of podocin and CD2AP decreased in the model group (P <0. 01). Compared with the model group, protein and mRNA expressions of podocin and CD2AP increased in the CDSRCD group and the irbesartan group (P <0. 01). Protein and mRNA expressions of podocin and CD2AP increased more in the CDSRCD group than in the irbesartan group (P <0. 05). CONCLUSIONS: CDSRCD could protect renal function by lowering urinary protein in DN rats, improve renal pathological changes. Its mechanism might be related to up-regulating mRNA and protein expressions of podocin and CD2AP.

Antidiabetic effects of Mukia maderaspatana and its phenolics: an in vitro study on gluconeogenesis and glucose uptake in rat tissues.

CONTEXT: Traditional medicine is used by over 60% of the world's population for health care. Mukia maderaspatana (L.) M. Roem. (Cucurbitaceae) (Mukia) is extensively used in folklore medicine as an antidiabetic plant. It is rich in phenolics that contribute to its medicinal properties. OBJECTIVE: Mukia extract and phenolics such as quercetin and phloroglucinol are investigated for their in vitro antidiabetic activity. MATERIALS AND METHODS: Quercetin, phloroglucinol, and methanol extract of the dried whole plant (0.25 and 0.5 mg/ml) were studied for the inhibition of gluconeogenesis in rat liver slices and glucose uptake in isolated rat hemi-diaphragm (50 and 100 µg/ml). Phenolics of Mukia were analyzed by HPLC. RESULTS AND DISCUSSION: Glucose (1.2 mg/g/h) was synthesized from pyruvate and the synthesis was completely inhibited by insulin (1 U/ml). Quercetin at 0.25 and 0.5 mg/ml caused 65% and 89% inhibition (0.42 mg/g/h and 0.13 mg/g/h glucose). Addition of insulin did not increase inhibition. Phloroglucinol inhibited 100% glucose production with or without insulin. Mukia (0.25 mg/ml) inhibited gluconeogenesis (0.65 mg/g/h) by 45%, and with insulin, inhibition increased to 50% (0.59 mg/g/h). At 0.5 mg/ml, glucose production was stimulated by1.2-fold, but with insulin it was inhibited by 89% (0.13 mg/g/h glucose). Mukia had no effect on glucose uptake, but potentiated the action of insulin mediated glucose uptake (152.82 ± 13.30 mg/dl/g/30 min) compared with insulin control (112.41 ± 9.14 mg/dl/g/30 min) (p < 0.05). HPLC analysis revealed the presence of phenolics. CONCLUSION: Results provide scientific rationale for the use of Mukia in folk medicine as an antidiabetic nutraceutical.

The effects of inspiratory muscle training on plasma interleukin-6 concentration during cycling exercise and a volitional mimic of the exercise hyperpnea.

It is unknown whether the respiratory muscles contribute to exercise-induced increases in plasma interleukin-6 (IL-6) concentration, if this is related to diaphragm fatigue, and whether inspiratory muscle training (IMT) attenuates the plasma IL-6 response to whole body exercise and/or a volitional mimic of the exercise hyperpnea. Twelve healthy males were divided equally into an IMT or placebo (PLA) group, and before and after a 6-wk intervention they undertook, on separate days, 1 h of 1) passive rest, 2) cycling exercise at estimated maximal lactate steady state power (EX), and 3) volitional hyperpnea at rest, which mimicked the breathing and respiratory muscle recruitment patterns achieved during EX (HYPEX). Plasma IL-6 concentration remained unchanged during passive rest. The plasma IL-6 response to EX was reduced following IMT (main effect of intervention, P = 0.039) but not PLA (P = 0.272). Plasma IL-6 concentration increased during HYPEX (main effect of time, P < 0.01) and was unchanged postintervention. There was no evidence of diaphragm fatigue (measured by phrenic nerve stimulation) following each trial. In conclusion, plasma IL-6 concentration is increased during EX and HYPEX and this occurred in the absence of diaphragm fatigue. Furthermore, IMT reduced the plasma IL-6 response to EX but not HYPEX. These findings suggest that the respiratory muscles contribute to exercise-induced increases in plasma IL-6 concentration in the absence of diaphragm fatigue and that IMT can reduce the magnitude of the response to exercise but not a volitional mimic of the exercise hyperpnea.

Phrenic nerve stimulation protects against mechanical ventilation-induced diaphragm dysfunction in rats.

INTRODUCTION: We investigated a novel application of phrenic nerve stimulation (PNS) in diaphragm dysfunction induced by mechanical ventilation (MV). METHODS: Twenty-one Sprague-Dawley rats were assigned randomly to 3 groups: spontaneous breathing, 18-h controlled MV, and 18-h controlled MV with PNS. Upon completion of the experimental protocol, diaphragm contractility, gene expression of insulin-like growth factor-1 (IGF-1) and ubiquitin ligases, and serum IGF-1 levels were analyzed. RESULTS: Compared with the spontaneously breathing rats, impaired diaphragm contractile function, including force-related properties and force-frequency responses, were pronounced with MV. Furthermore, MV suppressed IGF-1 and induced muscle ring finger 1 mRNA expression in the diaphragm. In contrast, PNS counteracted MV-induced gene expression changes in the diaphragm and restored diaphragm function. CONCLUSIONS: PNS exerted a protective effect against MV-induced diaphragm dysfunction by counteracting altered expression of IGF-1 and ubiquitin ligase in the diaphragm.

Dexamethasone antagonizes the in vivo myotoxic and inflammatory effects of Bothrops venoms.

In the present work we investigated the toxic activities of two Bothrops snake venoms using in vivo and in vitro experimental protocols in mice and tested the protective effect of dexamethasone (DEXA) in different conditions, comparing it with the polyvalent antivenom. We also expanded the investigations on the antiophidic effect of the Eclipta prostrata (EP) crude extract. The administration of Bothrops jararaca and Bothrops jararacussu snake venoms induced muscle damage demonstrated in vivo by the elevation on plasma creatine kinase (CK) activity in mice and by the decrease in CK content in the extensor digitorum longus (EDL) muscle of these animals, and in vitro by the increase in the rate of CK release from the isolated EDL muscle. We also observed inflammatory response following perimuscular injection of B. jararacussu venom (1.0 mg/kg). Treatment with DEXA (1.0 mg/kg) preserved over 50% of the EDL muscle CK content in vivo when evaluated 24 and 72 h after the injection of B. jararacussu venom in mice, and likewise reduced about 20% of the edema induced by this venom. DEXA reduced in 50% the presence of inflammatory cells and their activity in EDL muscle. The EP extract (50 mg/kg) showed similar ability in preventing the induction of edema and the decrease in muscle CK content, and its association with DEXA showed additive effect. EP reduced over 77% of the plasma CK activity induced by the B. jararacussu venom. In the in vitro experiments, DEXA was not able to change the rate of CK release from EDL muscles exposed to 25 µg/mL of B. jararacussu venom, neither to prevent the fall in the amplitude of the indirectly evoked twitch at the phrenic-diaphragm preparation. EP extract showed otherwise a protective effect on these protocols, reaching up to 100% of protection when concentrations of 50.0 and 100.0 µg/mL were used. Altogether our results show that inflammation is at least in part responsible for the tissue damage induced by Bothrops snake venoms, once the steroidal anti-inflammatory drug dexamethasone was able to decrease the myotoxic effects of these venoms, by reducing the inflammatory response to the venom injection.

Grape seed extract neutralizes the effects of Cerastes cerastes cerastes post-synaptic neurotoxin in mouse diaphragm.

This work was undertaken to investigate the toxic activity of the post-synaptic neurotoxic fraction isolated from the venom of the Egyptian sand viper (Cerastes cerastes cerastes), and the ability of grape seed extract to antagonize this effect produced in sublethally intoxicated mice, with an emphasis on ultrastructural features. Light and transmission electron microscopy of diaphragms of intoxicated mice showed myonecrosis, myofiber hypercontraction, sarcomere disorganization, and mitochondrial damage. Alterations in motor neurons and axon terminals were also observed. The toxic activities of C. cerastes cerastes neurotoxin were inhibited by administrating grape seed extract, either before or after intoxication, showing that grape seed extract has protective and therapeutic potential to be used as antivenom.

Anti-diabetic and anti-cataract effects of Chromolaena odorata Linn., in streptozotocin-induced diabetic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Chromolaena odorata Linn., is used in traditional Indian medicine in the treatment of diabetes and eye problems. AIM OF THE STUDY: The present study was designed to investigate the effect of the ethanol extract Chromolaena odorata leaves (ACO) in streptozotocin (STZ; 45 mg/kg, i.v) induced diabetes and cataract in rats. MATERIALS AND METHODS: Different doses of ACO (200 and 400mg/kg) was administered once daily for eight weeks to STZ-induced diabetic rats. To know the mechanism of action of title plant, AUC(glucose), AUC(insulin), Homeostatic Model Assessment (HOMA), insulin tolerance test (ITT) and glucose uptake by rat hemi-diaphragms were carried out. Further, cataract score was taken once in a week upto eight weeks and opacity index was measured. HPLC fingerprinting profiling of ACO was also carried out. RESULTS: Administration of ACO exhibited significant reduction in glucose, HOMA, lipid profiles and significantly improved glucose and insulin tolerance, glycogen content, glucose uptake by skeletal muscle, serum insulin and HDL-c levels. In addition, ACO also decreased oxidative stress by improving endogenous antioxidants. Further, treatment of ACO showed significantly reduced onset and extent of cataract. CONCLUSION: The present data suggested that the treatment of ACO reversed the STZ-induced diabetes and cataract in rats. The observed beneficial effects may be mediated by interacting with multiple targets operating in diabetes mellitus and its complication. Taken together, this study provided the scientific evidence for the traditional use of Chromolaena odorata.

Olive oil improved the impairment of in vitro insulin-stimulated glucose uptake by diaphragm in ovariectomized female Wistar rats.

UNLABELLED: This work was carried out to examine the impact of ovariectomy in female Wistar rats on in vitro basal and insulin-stimulated glucose uptake by diaphragm, and the possible beneficial effects of olive oil supplementation in these rats. METHODS: we studied 21 female Wistar rats aged 12-14 months, divided into three groups: sham-operated control (SHAM), ovariectomized (OVX), and ovariectomized rats supplemented with extra-virgin olive oil (Olive-OVX) orally for 12 weeks; 4 weeks before ovariectomy and 8 weeks after. At the end of the experiment, blood samples were collected for lipid profile assays. Malondialdehyde (MDA) in soleus muscles was assayed. The diaphragms were collected for determination of glucose uptake by the muscles. Soleus muscles and pancreas were processed for histological examination. RESULTS: ovariectomy impaired insulin-stimulated glucose uptake by diaphragm of female rats, induced abnormalities of lipid metabolism, and increased the oxidative stress marker, MDA level in soleus muscles. Olive oil supplementation to ovariectomized rats resulted in an enhancement of insulin-stimulated glucose uptake by diaphragm and a better lipid profile, as well as a decrease in the MDA level in soleus muscles. Histological study in OVX rats revealed a decrease in the size and number of islets of Langerhans in the pancreatic tissue, and decreases in both glycogen content, and in mitochondrial density in the soleus muscles. In Olive-OVX rats the size of islets of Langerhans was normal, and muscle glycogen, and muscle mitochondrial density were increased as compared to OVX rats. CONCLUSION: ovariectomy impaired in vitro insulin-stimulated glucose uptake by diaphragm. The improvement of this impairment by olive oil supplementation could be attributed to its antioxidant properties, its high content of monounsaturated fatty acids, as well as, its enhancement effect on mitochondrial density in skeletal muscles.

Antidiabetic effect of some medicinal plants of Oriental Morocco in neonatal non-insulin-dependent diabetes mellitus rats.

The goal of the present study is to test the effect of water extract (WE) of four medicinal plants used as antidiabetics in Eastern Morocco (Arbutus unedo: Au, Ammoïdes pusilla: Ap, Thymelaea hirsuta: Th, and Urtica dioïca: Ud). These plants are used in cooking to bring out the flavor in a dish or to complement it. The first experiment was realized in order to determine the antidiabetic effect of the WE of these plants during 5 weeks' treatment. Seven groups of Wistar rats were used: Healthy controls, neonatal streptozotocin (n-stz) induced-diabetic rats (90 mg/kg; intraperitoneally [i.p.]), n-stz + tolbutamide (400 mg/l), and 4 groups n-stz + WE of plants (400 mg/l, drink water). The percentages of Plasma glucose lowering effect were, respectively for Au, Ap, Th, Ud and tolbutamide: 31.6 % p<0.01, 27.4 % p<0.05, 38.2 % p<0.01, 13 % and 33.9 % p<0.05 when compared with untreated diabetic controls. In a second experiment, oral glucose tolerance tests were carried out in n-stz induced-diabetic rats. The i.p. administration of the water extract (WE) of Ap and Ud (150 mg/kg) 30 minutes before the glucose overload (2 g/kg) showed a significant reduction glycemia, respectively of 36 % at 60 min (p<0.05) and 50 % at 180 min (p<0.05) after glucose overload compared with controls. In contrast, the effect of WE of Au and Th (150 mg/kg, i.p.) was not significant. The in vitro study of glucose utilization by isolated rat hemidiaphragm suggests that these extracts in combination with insulin potentiate its activity and enhance the utilization of glucose. In conclusion, it seems that these plants possess antidiabetic activity.

Distribution of hypothalamic neurons with orexin (hypocretin) or melanin concentrating hormone (MCH) immunoreactivity and multisynaptic connections with diaphragm motoneurons.

Prior work showed that neurons in the lateral, dorsal, and perifornical regions of the tuberal and mammillary levels of the hypothalamus participate in the control of breathing. The same areas also contain large numbers of neurons that produce either orexins (hypocretins) or melanin concentrating hormone (MCH). These peptides have been implicated in regulating energy balance and physiological changes that occur in transitions between sleep and wakefulness, amongst other functions. The goal of this study was to determine if hypothalamic neurons involved in respiratory control, which were identified in cats by the retrograde transneuronal transport of rabies virus from the diaphragm, were immunopositive for either orexin-A or MCH. In animals with limited rabies infection of the hypothalamus (<10 infected cells/section), where the neurons with the most direct influences on diaphragm motoneurons were presumably labeled, a large fraction (28-75%) of the infected hypothalamic neurons contained orexin-A. In the same cases, 6-33% of rabies-infected hypothalamic cells contained MCH. However, in animals with more extensive infection, where rabies had presumably passed transneuronally through more synapses, the fraction of infected cells that contained orexin-A was lower. The findings from these experiments thus support the notion that hypothalamic influences on breathing are substantially mediated through orexins or MCH.

Effects of acute administration of corticosteroids during mechanical ventilation on rat diaphragm.

RATIONALE: Mechanical ventilation is known to induce ventilator-induced diaphragm dysfunction. Patients submitted to mechanical ventilation often receive massive doses of corticosteroids that may cause further deterioration of diaphragm function. OBJECTIVES: To examine whether the combination of 24 hours of controlled mechanical ventilation with corticosteroid administration would exacerbate ventilator-induced diaphragm dysfunction. METHODS: Rats were randomly assigned to a group submitted to 24 hours of controlled mechanical ventilation receiving an intramuscular injection of saline or 80 mg/kg methylprednisolone, a group submitted to 24 hours of spontaneous breathing receiving saline, or methylprednisolone and a control group. MEASUREMENTS AND MAIN RESULTS: The diaphragm force-frequency curve was shifted downward in the mechanical ventilation group, but this deleterious effect was prevented when corticosteroids were administered. Diaphragm cross-sectional area of type I fibers was similarly decreased in both mechanical ventilation groups while atrophy of type IIx/b fibers was attenuated after corticosteroid administration. The mechanical ventilation-induced reduction in diaphragm MyoD and myogenin protein expression was attenuated after corticosteroids. Plasma cytokine levels were unchanged while diaphragm lipid hydroperoxides were similarly increased in both mechanical ventilation groups. Diaphragmatic calpain activity was significantly increased in the mechanical ventilation group, but calpain activation was abated with corticosteroid administration. Inverse correlations were found between calpain activity and diaphragm force. CONCLUSIONS: A single high dose of methylprednisolone combined with controlled mechanical ventilation protected diaphragm function from the deleterious effects of controlled mechanical ventilation. Inhibition of the calpain system is most likely the mechanism by which corticosteroids induce this protective effect.

Inspiratory muscle training reduces blood lactate concentration during volitional hyperpnoea.

Although reduced blood lactate concentrations ([lac(-)](B)) have been observed during whole-body exercise following inspiratory muscle training (IMT), it remains unknown whether the inspiratory muscles are the source of at least part of this reduction. To investigate this, we tested the hypothesis that IMT would attenuate the increase in [lac(-)](B) caused by mimicking, at rest, the breathing pattern observed during high-intensity exercise. Twenty-two physically active males were matched for 85% maximal exercise minute ventilation (.V(E) max) and divided equally into an IMT or a control group. Prior to and following a 6 week intervention, participants performed 10 min of volitional hyperpnoea at the breathing pattern commensurate with 85% .V(E) max. The IMT group performed 6 weeks of pressure-threshold IMT; the control group performed no IMT. Maximal inspiratory mouth pressure increased (mean +/- SD) 31 +/- 22% following IMT and was unchanged in the control group. Prior to the intervention in the control group, [lac(-)](B) increased from 0.76 +/- 0.24 mmol L(-1) at rest to 1.50 +/- 0.60 mmol L(-1) (P < 0.05) following 10 min volitional hyperpnoea. In the IMT group, [lac(-)](B) increased from 0.85 +/- 0.40 mmol L(-1) at rest to 2.02 +/- 0.85 mmol L(-1) following 10 min volitional hyperpnoea (P < 0.05). After 6 weeks, increases in [lac(-)](B) during volitional hyperpnoea were unchanged in the control group. Conversely, following IMT the increase in [lac(-)](B) during volitional hyperpnoea was reduced by 17 +/- 37% and 25 +/- 34% following 8 and 10 min, respectively (P < 0.05). In conclusion, increases in [lac(-)](B) during volitional hyperpnoea at 85% .V(E) max were attenuated following IMT. These findings suggest that the inspiratory muscles were the source of at least part of this reduction, and provide a possible explanation for some of the IMT-mediated reductions in [lac(-)](B), often observed during whole-body exercise.

Infusions of rocuronium and cisatracurium exert different effects on rat diaphragm function.

OBJECTIVE: Aminosteroidal and benzylisoquinoline neuromuscular blocking agents are used in the intensive care unit to facilitate mechanical ventilation. The use of these agents has been associated with development of critical illness myopathy; however, the relative frequency of myopathy development among agents is not known. The aim of our study was to compare the effects of 24 h infusion of rocuronium or cisatracurium on the diaphragm in mechanically ventilated rats. DESIGN: Randomized, controlled experiment. SETTING: Basic animal science laboratory. SUBJECTS: Male Wistar rats, 14 weeks old. INTERVENTIONS: Rats were divided into four groups to receive either saline, rocuronium (low dose) or cisatracurium (low or high dose). MEASUREMENTS AND RESULTS: After 24 h, in vitro diaphragm tetanic force was decreased after rocuronium (-33% vs. saline), while the force was more preserved after cisatracurium, even in the high-dose group. Cross-sectional areas of the different diaphragm and gastrocnemius fibers were unaltered. Diaphragmatic MURF-1 mRNA was increased after rocuronium (+44% vs. saline), while unchanged in both cisatracurium groups. Calpain activity was increased after rocuronium (+75% vs. saline) and unchanged in the cisatracurium groups. MURF-1 mRNA expression and calpain activity were negatively correlated with diaphragm force. CONCLUSIONS: Cisatracurium infusion during controlled mechanical ventilation exerted less detrimental effects on diaphragm function and proteolytic activity than infusion of rocuronium, even with the higher effective dose. These data suggest that increased calpain activity and increased activation of the ubiquitin proteasome system play a role in the different effects of these agents.

Studies on hypoglycaemic activity of Solanum xanthocarpum Schrad. & Wendl. fruit extract in rats.

Aqueous extract of the fruits of Solanum xanthocarpum Schrad. & Wendl. (Solanaceae) was investigated for hypoglycaemic activity in rats and mice. Screening for the hypoglycaemic activity was assessed on normoglycaemic, alloxan treated hyperglycaemic and glucose loaded rats along with in vitro study on glucose utilization by isolated rat hemidiaphragm. The various haematological and biochemical parameters were also studied. The extract was found to possess significant hypoglycaemic activity when compared with the reference standard glibenclamide. The in vitro study on glucose utilization by isolated rat hemidiaphragm suggests that the aqueous extract may have direct insulin like activity which enhances the peripheral utilization of glucose and have extra pancreatic effect. The toxicity studies report safety usage of the plant extract.

Dietary trans fatty acids alter diaphragm phospholipid fatty acid composition, triacylglycerol content and glucose transport in rats.

The present study evaluates the effect of dietary trans fatty acids on diaphragm phospholipid fatty acid composition, intramyocellular triacylglycerol content and insulin-stimulated glucose uptake in comparison with dietary saturated fatty acids. Male weanling WNIN rats were divided into three groups and fed for 3 months on one of the following diets containing 10 % oil differing in fatty acid composition: control diet, saturated fatty acid diet and trans fatty acid diet. Dietary trans fatty acids increased the intramyocellular triacylglycerols and decreased the ratio of 20 : 4n-6 to 18 : 2n-6 and long-chain PUFA levels (20 %) in diaphragm phospholipids, indicating inhibition of PUFA biosynthesis. However, saturated fatty acids decreased both 18 : 2n-6 and 20 : 4n-6 without change in the ratio. Trans fatty acid-induced alterations in diaphragm phospholipid fatty acid composition and intramyocellular triacylglycerol content were associated with decreased insulin-stimulated glucose transport in the diaphragm. These observations suggest that dietary trans fatty acids decrease diaphragm insulin sensitivity, possibly due to increased intramyocellular triacylglycerol accumulation and decreased long-chain PUFA in phospholipids.