Comparative effect of composite management of type II diabetes mellitus with induced emesis and Varadi Ghanavati: A pilot clinical study.

INTRODUCTION: Ayurveda offers a rational view in the management of Prameha. Shodhana (∼bio-purification) is advocated as the first line of treatment to eliminate Kapha, which is followed by palliative treatment to further control disease pathology. Evaluation of such composite treatment has rarely been published. The present study was conducted to assess the comparative efficacy and safety of three interventions in the management of Prameha (Diabetes mellitus type II (T2DM)) - (a) Vamana (induced emesis) and Varadi Ghanavati (oral Ayurvedic medication), (b) Varadi Ghanavati, and (c) glibenclamide. METHODS: It was open labelled, comparative, randomised, prospective, three arm pilot study. A total 49 patients, newly diagnosed or known cases of Prameha (T2DM) were randomised in three groups. The first group (Vamana) received Vamana followed by Varadi Ghanavati (500 mg (mg) thrice daily before food for 12 weeks). The second (Varadi) group received Varadi Ghanavati without prior Vamana, while, the control group received glibenclamide 5 mg twice daily before food for 12 weeks. Patients were evaluated at intervals of each four weeks. Assessment was done on changes seen in blood sugar (BSL) fasting (F), postprandial (PP), glycosylated haemoglobin, lipid profile, body mass index (BMI) and clinical symptoms. RESULTS: Patients in all groups showed better glycaemic control as compared to baseline. Mean BSLF before and after treatment were 166.20 ± 78.39 mg/dL and 125.00 ± 58.77 mg/dL in Vamana group (p < 0.01), 163.60 ± 59.34 mg/dL and 127.73 ± 37.94 mg/dL in Varadi group (p < 0.05), while 163.00 ± 59.03 mg/dL and 129.40 ± 41.91 mg/dL in control (p < 0.05). Similarly, BSL PP values before and after treatment were 254.47 ± 99.59 mg/dL and 178.47 ± 68.45 mg/dL in Vamana group (p < 0.01), 233.93 ± 68.95 mg/dL and 185.20 ± 56.73 mg/dL in Varadi group (p < 0.05), 239.80 ± 77.10 mg/dL and 182.53 ± 42.14 mg/dL in control (p < 0.05). Glycosylated haemoglobin, mean values before and after treatment were 8.23 ± 1.71% and 7.18 ± 1.96% in Vamana group (p < 0.05), 7.83 ± 1.21% and 6.75 ± 1.13% in Varadi group (p < 0.05), 7.17 ± 1.12% and 6.60 ± 0.65% in glibenclamide group (p < 0.05). Comparative evaluation in the three groups showed that there was a statistically significant reduction (p < 0.001) in total cholesterol, low-density lipoproteins (LDL) and BMI in Vamana group as compared the other groups. No adverse event was observed. CONCLUSION: Along with better glycaemic control, composite treatment can reduce deranged lipids and BMI, which can help in better management of Prameha (T2DM). Vamana and Varadi Ghanavati can be administered safely. CLINICAL TRIAL REGISTRATION NO: CTRI/2017/10/010127.

Conservative management of acute prolapsed inter-vertebral disc with ayurveda: A case report.

Acute prolapsed inter-vertebral disc (IVDP) is a painful condition that requires immediate treatment by conservative or surgical management. Though majority of patients show remission in symptoms with conservative treatment, regression of herniated disc with non-surgical management has been rarely reported. A 46 years old female patient with acute and severe low back pain, disability and radiating pain towards right lower extremity came to our hospital. Oswestry Disability Index (ODI) score of the patient was 94% indicating bed-ridden condition. MRI of lumbar spine showed diffuse posterior disc bulge between fourth and fifth lumbar vertebra indenting right traversing nerve root and inferior displacement of extruded disc along the body of fifth lumbar vertebra. She was treated according to treatment explained in Ayurveda. She received oral medications, application of medicated oils, fomentation and medicated enema (Basti). After treatment of seven and half months, the patient showed good remission in pain, stiffness and radiculopathy. ODI score reduced to 9% that indicates minimal disability. Follow up MRI showed non significant compression of the nerve root and gross reduction in the inferior displacement of extruded disc. Acute IVDP can be successfully conserved using Ayurveda treatment. The Panchakarma procedures and medicines used in the treatment need further evaluation.

Application of tele-podiatry in diabetic foot management: A series of illustrative cases.

BACKGROUND AND AIMS: Telemedicine had been proposed as a tool to manage diabetes, but its role in management of diabetic foot ulcer is still evolving. The COVID-19 pandemic and related social restrictions have necessitated the use of telemedicine in the management of diabetic foot disease (tele-podiatry), particularly of patients classified as low-risk. MATERIALS AND METHODS: We present a report of three cases of varied diabetic foot problems assessed during the present pandemic using different forms of telemedicine for triaging, management of low-risk cases and for follow-up. RESULTS: Tele-podiatry was effective in the management of low-risk subjects with diabetic foot ulcer, and also useful in referral of high-risk subjects for hospital/clinic visit, facilitating proper management. It also helped in the follow-up of the cases. CONCLUSION: Telemedicine is a good screening tool for diagnosing and managing low-risk subjects with diabetic foot problems, and also enables a triaging system for deciding on hospital visits and hospitalization. Telemedicine offers several benefits in the management of diabetic foot disease, although it also has some limitations. Based on our experience during the pandemic, we recommend its judicious use in the triaging of patients of diabetic foot disease and management of low-risk cases. Future innovation in technology and artificial intelligence may help in better tele-podiatry care in the time to come.

Obesity and its Link to Undiagnosed Diabetes Mellitus and Hypertension in Rural Parts of Western India.

BACKGROUND: Obesity and overweight are becoming major health concerns worldwide. Hence, we studied the association between overweight and obesity with new-onset diabetes and hypertension in a selected rural population. METHODOLOGY: Community health workers made house-to-house visits, inviting adults >20 years of age who were at a higher risk of diabetes, from a predefined rural area of Maharashtra, to visit a mobile diabetes clinic operating in a hub and spoke manner. Sociodemographic data and anthropometric measurements were recorded. BMI and waist circumference was classified according to the WHO recommended cutoffs for Asians. Subjects with capillary blood fasting glucose of ≥126 mg/dL or random glucose of ≥200 mg/dL by glucometer were diagnosed as diabetes and blood pressure of ≥140/90 mmHg by sphygmomanometer were diagnosed as hypertension. Subjects with a known history of diabetes mellitus and hypertension were excluded. RESULTS: Out of 29,324 total population, 16.5% of subjects were overweight and 26.4% were obese. Mean ± SD of BMI of the participants was 22.9 ± 4.1 kg/m2 in males and 22.4 ± 4.2 kg/m2 in females. Around 35% of males and 30.5% of females had a high waist circumference of ≥90 cm and ≥80 cm, respectively, 20.5% of subjects had newly diagnosed hypertension, and 11.4% of subjects had newly diagnosed diabetes mellitus. The occurrence of newly diagnosed hypertension and diabetes showed an increasing trend with increasing BMI. CONCLUSION: Our community-based screening suggested a high prevalence of overweight and obesity in rural India. There was a high prevalence of newly diagnosed hypertension and diabetes in this population.

A clinical study to evaluate efficacy and safety of AHPL/AYTAB/0313 tablet in subjects suffering from osteoarthritis of knee(s).

BACKGROUND: Osteoarthritis (OA) is the most common form of arthritis with unsatisfactory treatment outcomes. OBJECTIVES: To evaluate efficacy and safety of AHPL/AYTAB/0313 tablet in subjects with OA of knee joint. STUDY DESIGN: Prospective, open-label, single-arm clinical study conducted in daily clinical practice setting. METHOD: Subjects were advised to take 2 AHPL/AYTAB/0313 tablets twice daily orally after meals for 180 days. 48 subjects completed the study. The primary endpoints were changes in mean visual analogue scale (VAS) pain score and WOMAC score. Secondary endpoints were quality of life, time to walk 50 feet, knee joint swelling, use of analgesic drug as rescue medicine, and safety parameters. RESULTS: At baseline visit, the mean index knee joint pain (VAS) score was 82.29 ± 15.19, which reduced significantly to 19.38 ± 13.75 on day 180. The mean WOMAC combined score at baseline was 39.94 ± 11.67, which reduced significantly to 09.58 ± 05.77 (76.0%) on day 180. The mean WOMAC pain score at baseline was 09.65 ± 02.91, which reduced significantly to 02.06 ± 01.46 on day 180. The mean WOMAC stiffness score at baseline was 03.48 ± 01.58, which reduced significantly to 00.63 ± 01.08 on day 180. The mean WOMAC difficulty score at baseline was 26.81 ± 09.63, which reduced significantly to 06.90 ± 04.78 on day 180. The mean walking time to walk 50 feet reduced significantly by 40% on day 180. Not a single subject was known to have knee joint swelling from 150 days onwards. Only 5 subjects were using analgesic as rescue medicine on day 180. Twenty-six subjects had adverse events (AEs). Most of the AEs were not associated with the study medication. Vitals and all the safety laboratory parameters were within normal limits both at baseline and on day 180. CONCLUSION: "AHPL/AYTOP/0113" tablet is safe and significantly effective in reducing pain, swelling, and stiffness of knee joints and improving mobility of knee joints in patients with OA. CTRI registration No. is CTRI/2015/09/006177.

Evaluation of Cyavanaprasa on Health and Immunity related Parameters in Healthy Children: A Two Arm, Randomized, Open Labeled, Prospective, Multicenter, Clinical Study.

CONTEXT: Cyavanaprasa (CP) is an Ayurvedic immune booster formulation that confers vigor and vitality while delaying the ageing process. Benefits of CP have been studied widely in adult population. OBJECTIVES: Current study assessed beneficial effects of CP on health and immunity related parameters in healthy children. METHODS: This study was a 6 month long two armed, randomized, open labeled, prospective clinical study. School going healthy children between ages of 5-12 years were randomized to receive orally daily either CP (approx. 6 g) followed by a cup of milk (100 - 200 ml) or cup of milk only twice a day while continuing with their normal/routine diet. Results were analyzed based on number of episodes, severity, duration of illness (infections and allergies) and number of absent days due to illness during the study duration and changes in levels of energy, physical fitness, strength, stamina and quality of life in children which were recorded in subject diary by their parents/Legally Acceptable Representative (LAR). RESULTS: 702 participants were randomized, out of which 627 completed the study (CP n = 313; Control n = 314). Results of immunity (episodes of infections or allergy related conditions) showed more than 2 times protection from immunity related illness in CP Group as compared to the control. CP also showed better percentage improvement in energy levels, physical fitness, strength, stamina and quality of life assessed through KIDSCREEN QOL-27 questionnaires in children. CONCLUSION: Regular consumption of CP for a period of six months could significantly improve immunity, energy levels, physical fitness, strength, stamina and quality of life in school going healthy children. STUDY REGISTRATION: Clinical Trail Registry of India vide CTRI/2015/02/005574, Dated 24 February 2015.

Effect of panchakarma and Ayurvedic treatment in postpartum rheumatoid arthritis (amavata): A case study.

Dream of a mother is to get involved actively in upbringing of child, which is impeded if she is suffering from painful condition like rheumatoid arthritis (RA) in postpartum phase. It causes physical incapacity and psychological trauma as well. Present case is a patient who developed RA one month after full term delivery by caesarean section. In view of symptoms, she was diagnosed as case of amavata. She received Ayurvedic treatment - Simhanada guggulu, Pratapalankeshwara rasa, Dashamoola katutraya kashaya and combination of Swarnabhupati rasa, Tapyadi loha, Mahavatavidhvansa, Chopachini (Smilax china), Shunthi (Zinziber officinale) and Guduchi (Tinospora cordifolia) for four months and course of kala basti (medicated enema) along with application of medicated oil (Vishagharbha taila abhyanga) and sudation (bashpa sweda) for ten days. Complete remission was seen after treatment for four months. The patient was free from oral analgesics. RA test titer that was 160 international units per milliliter (IU/ml) before treatment showed marked reduction (28.12 IU/ml) after 75 days of treatment and later dropped in normal range (6.1 IU/ml). Normal milestones were seen in the child receiving breast feeding. Application of Ayurvedic principles showed excellent results in this case where modern medical management options were limited due to lactation.

Therapeutic treatment with a novel hypoxia-inducible factor hydroxylase inhibitor (TRC160334) ameliorates murine colitis.

BACKGROUND AND AIM: Mucosal healing in inflammatory bowel disease (IBD) can be achieved by improvement of intestinal barrier protection. Activation of hypoxia-inducible factor (HIF) has been identified as a critical factor for barrier protection during mucosal insult and is linked with improvement in symptoms of colitis. Although prophylactic efficacy of HIF hydroxylase inhibitors in murine colitis have been established, its therapeutic efficacy in clinically relevant therapeutic settings have not been established. In the present study we aim to establish therapeutic efficacy of TRC160334, a novel HIF hydroxylase inhibitor, in animal models of colitis. METHODS: The efficacy of TRC160334 was evaluated in two different mouse models of colitis by oral route. A prophylactic efficacy study was performed in a 2,4,6-trinitrobenzene sulfonic acid-induced mouse model of colitis representing human Crohn's disease pathology. Additionally, a therapeutic efficacy study was performed in a dextran sulfate sodium-induced mouse model of colitis, a model simulating human ulcerative colitis. RESULTS: TRC160334 treatment resulted in significant improvement in disease end points in both models of colitis. TRC160334 treatment resulted into cytoprotective heatshock protein 70 induction in inflamed colon. TRC160334 successfully attenuated the rate of fall in body weight, disease activity index, and macroscopic and microscopic scores of colonic damage leading to overall improvement in study outcome. CONCLUSION: Our findings are the first to demonstrate that therapeutic intervention with a HIF hydroxylase inhibitor ameliorates IBD in disease models. These findings highlight the potential of TRC160334 for its clinical application in the treatment of IBD.

TRC210258, a novel TGR5 agonist, reduces glycemic and dyslipidemic cardiovascular risk in animal models of diabesity.

BACKGROUND: Patients with diabesity have a significantly increased risk of developing cardiovascular disease. Therefore, therapy addressing the multiple metabolic abnormalities linked with diabesity and leading to further reduction of cardiovascular risk is highly desirable. Activation of the TGR5 receptor holds therapeutic potential for diabesity. In the present study, we evaluated the efficacy of TRC210258, a novel TGR5 agonist, in clinically relevant animal models of diabesity. METHODS: A novel small molecule, TRC210258 (N-(4-chlorophenyl)-2-(4-fluorophenoxy)-N-methylimidazo (1, 2-a) pyrimidine-3-carboxamide), was synthesized. The in vitro TGR5 receptor activation potential of TRC210258 was assessed by cyclic adenosine monophosphate (cAMP) assay and cAMP-responsive element reporter assay using cells overexpressing the human TGR5 receptor. The effect of TRC210258 on glucagon-like peptide-1 release was evaluated in vitro using a human enteroendocrine cell line. The effect of TRC210258 on energy expenditure and glycemic control was evaluated in high-fat diet-induced obese mice. Additionally, the effect of TRC210258 on dyslipidemic parameters was determined in high fat-fed hamsters. RESULTS: TRC210258 demonstrated potent TGR5 agonist activity, with enhanced glucagon-like peptide-1 release and energy expenditure. Treatment with TRC210258 resulted in better glycemic control and improved parameters of dyslipidemia such as plasma triglyceride, low-density lipoprotein cholesterol, and non-high-density lipoprotein cholesterol levels. Treatment with TRC210258 also improved emerging dyslipidemic cardiovascular risk parameters, including remnant cholesterol and triglyceride clearance. CONCLUSION: This study highlights the potential of TRC210258, a novel TGR5 agonist, to improve dyslipidemic cardiovascular risk beyond glycemic control in patients with type 2 diabetes.

Treatment with a novel hypoxia-inducible factor hydroxylase inhibitor (TRC160334) ameliorates ischemic acute kidney injury.

BACKGROUND: Hypoxia-inducible factor (HIF) transcriptional system plays a central role in cellular adaptation to low oxygen levels. Preconditional activation of HIF and/or expression of its individual target gene products leading to cytoprotection have been well established in hypoxic/ischemic renal injury. Increasing evidence indicate HIF activation is involved in hypoxic/ischemic postconditioning of heart, brain and kidney. Very few studies evaluated the potential benefits of postischemia HIF activation in renal injury employing a pharmacological agent. We hypothesized that postischemia augmentation of HIF activation with a pharmacological agent would protect renal ischemia/reperfusion injury. For this, TRC160334, a novel HIF hydroxylase inhibitor, was used. METHODS: TRC160334, a novel HIF hydroxylase inhibitor, was synthesized. Ability of TRC160334 for stabilization of HIF-α and consequent HIF activation was evaluated in Hep3B cells. Efficacy of TRC160334 was evaluated in a rat model of ischemia/reperfusion-induced AKI. Two different treatment protocols were employed, one involved treatment with TRC160334 before onset of ischemia, the other involved treatment after the reperfusion of kidneys. RESULTS: TRC160334 treatment results in stabilization of HIF-α leading to HIF activation in Hep3B cells. Significant reduction in renal injury was observed by both treatment protocols and remarkable reduction in serum creatinine (23 and 71% at 24 and 48 h, respectively, p < 0.01) was observed with TRC160334 treatment applied after reperfusion. Urine output was significantly improved up to 24 h by both treatment protocols. CONCLUSION: The data presented here provide pharmacologic evidence for postischemia augmentation of HIF activation by TRC160334 as a promising and clinically feasible strategy for the treatment of renal ischemia/reperfusion injury.

Adverse pregnancy outcomes in rural Maharashtra, India (2008-09): a retrospective cohort study.

BACKGROUND: The study was carried out to record adverse pregnancy outcomes and to obtain information about sex ratio at birth in rural especially tribal areas in the State of Maharashtra, India. Although the tribal population is considered vulnerable to innumerable adversities, regretfully information about pregnancy wastage among them is not available. About 10% population of the state is tribal. The study of sex ratio at birth was planned as the overall sex ratio and child sex ratio had declined in the state. METHODS: The cohort of antenatal cases registered in rural areas of Maharashtra in the calendar year 2008 was followed up to study the pregnancy outcomes. A retrospective study was carried out from October 2009 to August 2010. The outcomes of all the registered antenatal cases were recorded by the Auxiliary Nurse Midwives. The summary sheets were obtained by Block Medical Officers. The data was entered at the block level by trained data entry operators in specially designed web-based software. Adverse pregnancy outcome was categorized in two groups abortions and stillbirths. RESULTS: About 1.1 million registered pregnancies were followed up. In the state 5.34% registered pregnancies ended in abortions. In tribal PHCs the relative risk of spontaneous abortion and induced abortion was 0.91 and 0.38 respectively. It was also revealed that about 1.55% pregnancies culminated in stillbirth. The relative risk of stillbirths in tribal PHCs was 1.33. The sex ratio at birth in the state was 850. The ratio was 883 in the tribal PHCs. Correlation was observed between sex ratio at birth and induced abortion rate. CONCLUSIONS: The study indicates that women from tribal PHCs are exposed to higher risk of adverse pregnancy outcome in the form of stillbirths. In non-tribal areas high induced abortion rate and poor sex ratio at birth is observed. These two indicators are correlated. The correlation may be explained by the unscrupulous practice of sex selective abortion.

Delayed intervention in experimental stroke with TRC051384--a small molecule HSP70 inducer.

Induction of HSPs is a natural response of stressed cells that protects against many insults including acute ischemia. TRC051384, a novel compound belonging to substituted 2-propen-1-one class is a potent inducer of heat shock protein 70 (HSP70). The aim of this study was to investigate the ability of TRC051384 in reducing neuronal injury and disability upon delayed treatments (4 and 8 hours post ischemia onset) in a rat model of transient cerebral ischemia. Focal cerebral ischemia was produced in rats by occluding the MCA using the intra luminal suture technique. Rats subjected to 2 hours focal cerebral ischemia were administered by intra-peritoneal route, TRC051384 or vehicle every 2 hours for 48 hours, from 4th hour or 8th hour after onset of ischemia. Progression of infarct and edema was assessed up to 48 hours post ischemic insult using magnetic resonance imaging and the neurological disability and survival studied till 7 days. Here we show for the first time that treatment with TRC051384 significantly reduces stroke associated neuronal injury (87% reduction in area of penumbra recruited to infarct, and 25% reduction in brain edema) and disability in a rat model of transient ischemic stroke even when administered 8 hours post onset of ischemia. Significant improvement in survival (50% by day 2 and 67.3% by day 7) was observed with TRC051384 treatment initiated at 4 hours after ischemia onset. Induction of HSP70 by TRC051384 involves HSF1 activation and results in elevated chaperone and anti-inflammatory activity. These results show that TRC051384 has the potential to be developed as a novel pharmacological agent for the treatment of ischemic stroke.

A multicenter community study on the efficacy of double-fortified salt.

BACKGROUND: Iron and iodine deficiencies affect more than 30% of the world's population. Typical Indian diets contain adequate amounts of iron, but the bioavailability is poor. This serious limiting factor is caused by low intake of meat products rich in heme iron and intake of phytates in staple foods in the Indian diet, which inhibits iron absorption. OBJECTIVE: To test the stability of double-fortified salt (DFS) during storage and to assess its efficacy in improving the iron and iodine status of the communities. METHODS: The stability of both iodized salt and DFS during storage for a 2-year period was determined. The bioefficacy of DFS was assessed in communities covering three states of the country for a period of 1 year. This was a multicenter, single-blind trial covering seven clusters. The experimental group used DFS and the control group used iodized salt. The salts were used in all meals prepared for family members, but determination of hemoglobin by the cyanmethemoglobin method was performed in only two or three members per family, and not in children under 10 years of age (n = 393 and 436 in the experimental and control groups, respectively). The family size was usually four or five, with a male: female ratio of 1:1, consisting of two parents with two or three children. Hemoglobin was measured at baseline, 6 months (midpoint), and 12 months (endpoint). Urinary iodine was measured in only one cluster at baseline and endpoint. All the participants were dewormed at baseline, 6 months, and 12 months. RESULTS: The iron and iodine in the DFS were stable during storage for 2 years. Over a period of 1 year, there was an increase of 1.98 g/dL of hemoglobin in the experimental group and 0.77 g/dL of hemoglobin in the control group; the latter increase may have been due to deworming. The median urinary iodine changed from 200 microg/dL at baseline to 205 microg/dL at the end of the study in the experimental group and from 225 microg/dL to 220 microg/dL in the control group. There was a statistically significant (p < .05) improvement in the median urinary iodine status of subjects who were iodine deficient (urinary iodine < 100 microg/L) in both the experimental and the control groups, a result showing that DFS was as efficient as iodized salt in increasing urinary iodine from a deficient to sufficient status. There was a statistically significant increase (p < .05) in hemoglobin in all seven clusters in the experimental group compared with the control. CONCLUSIONS: The iron and iodine in the DFS are stable in storage for 2 years. The DFS has proved beneficial in the delivery of bioavailable iron and iodine.

Rac1 regulates the release of Weibel-Palade Bodies in human aortic endothelial cells.

BACKGROUND: The release of Weibel-Palade Bodies (WPB) is a form of endothelial cell activation. But the signal transduction pathway leading to WPB release is not yet defined. We hypothesized that small G-protein rac1 and reactive oxygen species (ROS) mediate the ligand induced release of Weibel-Palade Bodies. METHODS: We tested this hypothesis by using wild-type and mutant adenoviral rac1 expression vectors, and by manipulating the production and destruction of superoxide and hydrogen peroxide in human aortic endothelial cells (HAEC). RESULTS: Thrombin (1.0 Unit, 30 min) induced the increase of WPB release by 3.7-fold in HAEC, and that H2O2 (0.1 mmol/L, 30 min) induced by 4.5-fold. These results correlated with thrombin-stimulated activation of rac-GTP binding activity by 3.5-fold, and increase of ROS production by 3.4-fold. The dominant negative adenoviral rac-N17 gene transfer dramatically inhibited the release of WPB by 64.2% (control) and 77.3% (thrombin-stimulation), and decreased ROS production by 65.5% (control) and 83.6% (thrombin-stimulation) compared with non-infected cells, respectively. Anti-oxidants, catalase and N-acetyl-cysteine significantly decreased the release of WPB by 34% and 79% in control cells, and further decreased by 63.6% and 46.7% in rac-N17 transferred cells compared with non-infected cells. We also confirmed that rac1 was located upstream of ROS in the WPB release pathway. CONCLUSIONS: Small G-protein rac1 medicates ligand-induced release of Weibel-Palade Bodies in human aortic endothelial cells, and the signal pathway of WPB release is a rac1-dependent ROS regulating mechanism.

Wall stiffness suppresses Akt/eNOS and cytoprotection in pulse-perfused endothelium.

Increased steady shear stress stimulates nitric oxide synthase (eNOS) in part by Akt-dependent phosphorylation. Arteries in vivo are exposed to pulse perfusion (PP) combining phasic shear with stretch. In compliant vessels, enhancing PP lowers vascular tone by stimulating eNOS; whereas in aged, stiff arteries, flow-mediated dilation declines and PP is a prominent risk factor. Here, we tested the hypothesis that reduced wall distensibility alters PP-induced eNOS/Akt mechano-signaling. Bovine aortic endothelial cells cultured within distensible tubes were exposed to physiological nonreversing steady or PP (7 dynes/cm(2) mean shear, pulse pressure 0 or 90 mm Hgx2 hours) in a custom servo-system. In compliant tubes, PP doubled Akt phosphorylation above nonpulsatile flow levels, whereas P-Akt declined to static levels from PP in stiffer tubes. eNOS phosphorylation (S-1179) similarly increased with PP in compliant tubes but was nearly undetectable with increased PP in stiffer tubes. After PP, brief exposure of cells to ultraviolet irradiation (oxidant stress) and subsequent culture revealed cytoprotection in compliant tubes but diffuse cytotoxicity and cell detachment in stiffer tubes. NOS inhibition by L-NAME converted compliant-tube post-UV behavior to that of stiffer tubes. These data provide novel evidence that wall compliance can directionally mediate endothelial Akt/eNOS phosphorylation and mechano-signaling. This may help explain increased vascular risks resulting from artery stiffening.

Constitutive activation of rac1 results in mitochondrial oxidative stress and induces premature endothelial cell senescence.

OBJECTIVE: Oxidative stress has been implicated in cellular senescence and vascular aging. We determined the role and mechanism of the small GTPase rac1 in vascular endothelial cell senescence. METHODS AND RESULTS: Adenoviral-mediated expression of the constitutively active allele of rac1 (rac1V12) in human umbilical vein endothelial cells resulted in mitochondrial oxidative stress with induction of biochemical, molecular, and morphological features of senescence. Suppression of mitochondrial oxidative stress abrogated rac1-induced premature senescence. Rac1V12 expression also resulted in an increase in endothelial ceramide levels. Moreover, premature endothelial cell senescence induced by an exogenous cell-permeable ceramide analog was not suppressed by inhibiting endogenous rac1 signaling. Finally, in human umbilical vein endothelial cells that had undergone replicative senescence, rac1 was not activated, and expression of the dominant-negative rac1 allele (rac1N17) did not suppress mitochondrial oxidative stress. CONCLUSIONS: These findings paint a picture in which the constitutive activation of rac1, via the generation of ceramide, results in mitochondrial oxidative stress and premature endothelial cell senescence. However, they speak against a role for endogenous rac1 activation in the induction of mitochondrial oxidative stress associated with replicative senescence of endothelial cells.