A split-mouth randomized controlled trial to compare the rate of canine retraction after a soft tissue procedure compared against a corticotomy procedure for accelerated tooth movement.

Background and Aim: Various methods to accelerate the orthodontic tooth movement have been used, among which corticotomy is considered to be the most common one. The suggested reasoning for such acceleration was the regional acceleratory phenomenon (RAP). Since the RAP is a property of both the hard and soft tissues, we designed a soft tissue flap procedure to compare the effects with the conventional corticotomy procedure. A split-mouth study was conducted where the two procedures were assessed in a single participant. Patients and Methods: The total sample size was calculated to be 40 with 20 participants in each group. The rate of tooth movement was the primary outcome measure, and the secondary outcomes were dentoalveolar changes, which were studied in both the conventional corticotomy and the flap-only procedure based on a cone-beam computed tomography (CBCT) wherein the alveolar bone density (BD) around canines, tipping, and rotational changes in canines, premolars, and molars were assessed. Results: Corticotomy resulted in greater canine angulation, lesser canine rotation and premolar rotation, and greater molar rotation compared with flap elevation, but these differences were statistically insignificant. Conclusion: Though the corticotomy resulted in higher BD, the differences were statistically insignificant. There was no significant difference in the rate of space closure assessed by the two techniques compared.

Type-2 diabetic rat heart: The effect of kolaviron on mTOR-1, P70S60K, PKC-α, NF-kB, SOD-2, NRF-2, eNOS, AKT-1, ACE, and P38 MAPK gene expression profile.

It has been established that genetic factors partially contribute to type-2 diabetes and vascular disease development. This study determined the effect of kolaviron on the expression profile of genes associated with the insulin signaling pathway and involved in regulating glucose and lipid metabolism, oxidative stress, inflammation, vascular functions, pro-survival and the apoptosis pathway in the heart of type-2 diabetic rats. After induction and confirmation of type-2 diabetes seven days after, the rats were treated with kolaviron for twenty-eight days before being euthanized. Organs were harvested and stored at - 80 °C in a biofreezer. Total RNA was extracted from the ventricle, reverse transcribed to cDNA followed by a real-time quantitative polymerase chain reaction (RT-qPCR) analysis of the expression of mTOR-1, P70S60K, PKC-α, NF-kB, SOD-2, NRF-2, eNOS, AKT-1, ACE, p38 MAPK and the reference gene (GAPDH), after which they were normalized/standardized. The results show an increase in the relative mRNA expression of mTOR/P70S60K/PKCα /P38MAPK/NF-KB/ACE and a decrease in the relative mRNA expression of NRF2/SOD/AKT/eNOS in the heart of the diabetic rats. Nevertheless, kolaviron modulated the expression profile of these genes, which suggest a therapeutic effect and target for vascular dysfunction and complications in type-2 diabetes through the activation of the NRF-2/AKT-1/eNOS signaling pathway and suppression of the NF-kB/PKC signaling pathway.

Cardioprotective Effects and in-silico Antioxidant Mechanism of L-Ergothioneine in Experimental Type-2 Diabetic Rats.

BACKGROUND: Diabetic cardiotoxicity is commonly associated with oxidative injury, inflammation, and endothelial dysfunction. L-ergothioneine (L-egt), a diet-derived amino acid, has been reported to decrease mortality and risk of cardiovascular injury, provides cytoprotection to tissues exposed to oxidative damage, and prevents diabetes-induced perturbation. OBJECTIVE: This study investigated the cardioprotective effects of L-egt on diabetes-induced cardiovascular injuries and its probable mechanism of action. METHODS: Twenty-four male Sprague-Dawley rats were divided into non-diabetic (n = 6) and diabetic groups (n = 18). Six weeks after the induction of diabetes, the diabetic rats were divided into three groups (n = 6) and administered distilled water, L-egt (35mg/kg), and losartan (20mg/kg) by oral gavage for six weeks. Blood glucose and mean arterial pressure (MAP) were recorded pre-and post-treatment, while biochemical, ELISA, and RT-qPCR analyses were conducted to determine inflammatory, injury-related and antioxidant biomarkers in cardiac tissue after euthanasia. Also, an in-silico study, including docking and molecular dynamic simulations of L-egt toward the Keap1- Nrf2 protein complex, was done to provide a basis for the molecular antioxidant mechanism of Legt. RESULTS: Administration of L-egt to diabetic animals reduced serum triglyceride, water intake, MAP, biomarkers of cardiac injury (CK-MB, CRP), lipid peroxidation, and inflammation. Also, Legt increased body weight, antioxidant enzymes, upregulated Nrf2, HO-1, NQO1 expression, and decreased Keap1 expression. The in-silico study showed that L-egt inhibits the Keap1-Nrf2 complex by binding to the active site of Nrf2 protein, thereby preventing its degradation. CONCLUSION: L-egt protects against diabetes-induced cardiovascular injury via the upregulation of the Keap1-Nrf2 pathway and its downstream cytoprotective antioxidants.

Kolaviron attenuates cardiovascular injury in fructose-streptozotocin induced type-2 diabetic male rats by reducing oxidative stress, inflammation, and improving cardiovascular risk markers.

The prevalence of cardiovascular disease among type-2 diabetic patients has become a source of major concern world over. This study explored the protective effect of kolaviron, a bioflavonoid, against oxidative cardiovascular injury in fructose- streptozotocin-induced type 2 diabetic male Sprague Dawley rats. After acclimatization, induction, and confirmation of type-2 diabetes, kolaviron was administered for 28days, after which the animals were anesthetized with Isofor and euthanized. Blood from each rat were collected, and blood samples were then centrifuged for serum and plasma. Cardiac troponin I (cTnI), creatine kinase myocardial band (CK-MB), Creatine phosphokinase (CK), and insulin levels were immediately determined in serum, while remaining samples (serum, plasma, and organs) were stored in the bio-freezer at - 80 °C and 10% formalin for enzyme-link immunosorbent assay (ELISA), biochemical, molecular, and histopathological studies. The results show that type-2 diabetes induction with fructose and streptozotocin led to increased blood glucose levels, decreased insulin levels and cardiac antioxidant enzyme activities, increased malondialdehyde levels, cardiac biomarkers and pro-inflammatory cytokines levels, resulted in abnormal lipid profile, increased blood pressure and angiotensin-converting enzyme (ACE) activity, and decreased plasma endothelial nitric oxide synthase (eNOS) concentration. The histopathological examination of the cardiac tissue revealed severe lesion, hypertrophy, and myofibrils degeneration. However, administration of kolaviron for 28days remarkably improved these conditions. Hence the result from the study validates the potency of kolaviron, and suggests it could serve as an alternative to existing remedy in ameliorating or protecting against cardiovascular injury in type-2 diabetes.

L-ergothioneine and metformin alleviates liver injury in experimental type-2 diabetic rats via reduction of oxidative stress, inflammation, and hypertriglyceridemia.

Type-2 diabetes (T2D) is associated with liver toxicity. L-ergothioneine (L-egt) has been reported to reduce toxicity in tissues exposed to injury, while metformin is commonly prescribed to manage T2D. Hence, this study evaluates the hepatoprotective role of L-egt, with or without metformin, in T2D male rats. A total of 36 adult male Sprague-Dawley rats were randomly divided into non-diabetic (n = 12) and diabetic (n = 24) groups. After induction of diabetes, animals were divided into six groups (n = 6) and treated orally either with deionized water, L-egt (35 mg/kg bodyweight (bwt)), metformin (500 mg/kg bwt), or a combination of L-egt and metformin for 7 weeks. Body weight and blood glucose were monitored during the experiment. Thereafter, animals were euthanized and liver tissue was excised for biochemical, ELISA, real-time quantitative PCR, and histopathological analysis. L-egt with or without metformin reduced liver hypertrophy, liver injury, triglycerides, oxidative stress, and inflammation. Also, L-egt normalized mRNA expression of SREBP-1c, fatty acid synthase, nuclear factor kappa B, transforming growth factor ß1, nuclear factor erythroid 2-related factor 2, and sirtuin-1 in diabetic rats. Furthermore, co-administration of L-egt with metformin to diabetic rats reduced blood glucose and insulin resistance. These results provide support to the therapeutic benefits of L-egt in the management of liver complications associated with T2D.

L-ergothioneine and its combination with metformin attenuates renal dysfunction in type-2 diabetic rat model by activating Nrf2 antioxidant pathway.

L-ergothioneine (L-egt) is a bioactive compound recently approved by the food and drug administration as a supplement. L-egt exerts potent cyto-protective, antioxidant and anti-inflammatory properties in tissues exposed to injury, while metformin is a first-line prescription in type-2 diabetes. Therefore, the present study investigated the protective effect of L-egt alone, or combined with metformin, on renal damage in a type-2 diabetic (T2D) rat model. T2D was induced in male Sprague-Dawley rats using the fructose-streptozotocin rat model. L-egt administration, alone or combined with metformin, began after confirming diabetes and was administered orally for seven weeks. After the experiment, all animals were euthanized by decapitation, blood samples were collected, and both kidneys were excised. Biochemical analysis, Enzyme-link Immunoassay (ELISA), Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR), western blotting, and histological analyses were done to evaluate various biomarkers and structural changes associated with renal damage. Untreated diabetic rats showed loss of kidney functions characterized by increased serum creatinine, blood urea nitrogen, proteinuria, triglycerides, lipid peroxidation, inflammation, and decreased antioxidant enzymes. Histological evaluation showed evidence of fibrosis, mesangial expansion, and damaged basement membrane in the nephrons. However, L-egt alleviates these functional and structural derangements in the kidney, while co-administration with metformin reduced hyperglycemia and improves therapeutic outcomes. Furthermore, L-egt treatment significantly increased the expression of major antioxidant transcription factors, cytoprotective genes and decreased the expression of inflammatory genes in the kidney. Thus, combining L-egt and metformin may improve therapeutic efficacy and be used as an adjuvant therapy to alleviate renal damage in type-2 diabetes.

Postnatally induced metabolic and oxidative changes associated with maternal high-fat consumption were mildly affected by Quercetin-3-O-rutinoside treatment in rats.

Oxidative stress is usually associated with prolonged intake of high-fat diet (HFD). However, little is known about the impact of maternal HFD on endogenous modulation of antioxidant-defence-enzyme-network, its link to adverse fetal growth and overall effects of Quercetin-3-o-rutinoside (QR) supplementation. Sprague-Dawley rats were initially assigned to normal diet (ND) or HFD for 8 weeks and mated. Post-conception, rats were further divided into four groups, of which two groups had diets supplemented with QR while others continued with their respective diets until delivery. Measurements include food and water consumption, physical parameters (body weight, body mass index (BMI) and fur appearance), oral glucose tolerance, lipid profiles, and placental/liver oxidative changes. We observed that water consumption was significantly increased in dams fed HFD without marked differences in food intake, body weight, BMI and glucose tolerance. Surprisingly, offspring of HFD-fed dams had reduced body weight marked by delayed fur appearance compared to the ND offspring. In dams, there were alterations in lipid profile. Lipid peroxidation was increased in the placenta and liver of gestational day (GD) 19 HFD-fed dams and their postnatal day (PND) 21 male offspring. There was evidence of HFD-induced nitrosative stress in dams and PND28 female offspring. Adaptive defence indicate decreased placenta and liver superoxide dismutase (SOD) levels as well as differential changes in total antioxidant capacity (TAC) and catalase (CAT) activity in HFD treated dams and their progenies. Overall, the results indicate that intrauterine metabolic alterations associated with maternal high-fat consumption may induce oxidative challenge in the offspring accompanied by mild developmental consequences, while QR supplementation has little or no beneficial effects.

Characterization of influx and efflux silicon transporters and understanding their role in the osmotic stress tolerance in finger millet (Eleusine coracana (L.) Gaertn.).

Over the last decade, silicon (Si) has been widely accepted as a beneficial element for plant growth. The advantages plant derives from the Si are primarily based on the uptake and transport mechanisms. In the present study, the Si uptake regime was studied in finger millet (Eleusine coracana (L). Gaertn.) under controlled and stress conditions. The finger millet can efficiently uptake Si and accumulate it by more than 1% of dry weight in the leaf tissues, thus categorized as a Si accumulator. Subsequent evaluation with the single root assay revealed a three-fold higher Si uptake under osmatic stress than control. These results suggest that Si alleviated the PEG-induced stress by regulating the levels of osmolytes and antioxidant enzymes. Further, to understand the molecular mechanism involved in Si uptake, the Si influx (EcoLsi1 and EcoLsi6) and efflux transporters (EcoLsi2 and EcoLsi3) were identified and characterized. The comparative phylogenomic analysis of the influx transporter EcoLsi1 with other monocots revealed conserved features like aromatic/arginine (Ar/R) selectivity filters and pore morphology. Similarly, Si efflux transporter EcoLsi3 is highly homologous to other annotated efflux transporters. The transcriptome data revealed that the expression of both influx and efflux Si transporters was elevated due to Si supplementation under stress conditions. These findings suggest that stress elevates Si uptake in finger millet, and its transport is also regulated by the Si transporters. The present study will be helpful to better explore Si derived benefits in finger millet.

Evaluation of maternal high-fat diet and Quercetin-3-O-rutinoside treatment on the reproductive profile of diet naïve male offspring.

BACKGROUND: Male infertility and reproductive dysfunctions have become major global health problems. Although several causative factors have been attributed to this challenge, of importance are alterations in maternal-foetal environment, diet-induced transcriptional changes and dysregulation in chemical signaling via hypothalamic-gonadal axis. AIM: The present study investigated the impact of maternal high-fat diet (HFD) consumption and the putative role of Quercetin-3-O-rutinoside on reproductive functions of male offspring rats at critical developmental stages with a quest to unravel the underpinned molecular changes. MATERIALS AND METHODS: Fifty-six pregnant rats (previously fed normal diet ND) or 45% HFD) were maintained on supplemented chow (150 mg/kg QR) - ND/QR, HFD/QR throughout gestation. Subsequently, dams (n = 7) and offspring (n = 6) were sacrificed at post-natal day (PND) 21, 28 and 35, respectively, and the blood, placenta, hypothalamus (HT), and testicular samples were processed for molecular analysis of Gonadotropin-releasing hormone (GnRH), Luteinizing hormone (LH), testosterone, chemerin, chemokine-like receptor 1 (CMKLR1), tumour necrosis factor α (TNF-α), interleukin 1ß (IL-1ß) and nuclear factor kappa B (NF-κB). KEY FINDINGS: We observed a significant decrease in GnRH level in the HFD group at PND21 and PND28 in male offspring and treatment with QR significantly reduced GnRH. There was a significant reduction in LH levels in the HFD group at PND 21 in the male offspring accompanied by a significant decrease in testosterone level at PND 28 and PND35 which appears to be age dependent. In the HT, Chemerin and CMKLR1 was significantly upregulated in the HFD group at PND 21 and PND 35 respectively while CMKLR1 was significantly downregulated in the HFD group of the placenta and testis at PND 21. TNF-α, IL-1ß and NF-κB were also expressed in the placenta, HT and testis at PND 21. SIGNIFICANCE: Male fertility is affected by maternal HFD consumption while chemerin, CMKLR1 and TNF-α, may play a significant role in male steroidogenesis. Treatment with QR had little or no ameliorative effect on HFD induced alterations in male reproductive functions.

An insight into the role of silicon on retaliation to osmotic stress in finger millet (Eleusine coracana (L.) Gaertn).

Finger millet, a vital nutritional cereal crop provides food security. It is a well-established fact that silicon (Si) supplementation to plants alleviates both biotic and abiotic stresses. However, precise molecular targets of Si remain elusive. The present study attempts to understand the alterations in the metabolic pathways after Si amendment under osmotic stress. The analysis of transcriptome and metabolome of finger millet seedlings treated with distilled water (DW) as control, Si (10 ppm), PEG (15%), and PEG (15%) + Si (10 ppm) suggest the molecular alterations mediated by Si for ameliorating the osmotic stress. Under osmotic stress, uptake of Si has increased mediating the diversion of an enhanced pool of acetyl CoA to lipid biosynthesis and down-regulation of TCA catabolism. The membrane lipid damage reduced significantly by Si under osmotic stress. A significant decrease in linolenic acid and an increase of jasmonic acid (JA) in PEG + Si treatment suggest the JA mediated regulation of osmotic stress. The relative expression of transcripts corroborated with the corresponding metabolites abundance levels indicating the activity of genes in assuaging the osmotic stress. This work substantiates the role of Si in osmotic stress tolerance by reprogramming of fatty acids biosynthesis in finger millet.

The effect of multivitamin-multimineral supplementation on the health status of inbred Wistar and spontaneously hypertensive rat strains.

The nutraceutical industry has proliferated in recent years, with the most popular form of supplementation being the multivitamin-multimineral (MVMM) supplement. In the animal health sector, supplement use has also expanded. The objective of this study was to determine the effects of MVMM supplementation, beneficial or otherwise, on the general health status of the spontaneously hypertensive rat (SHR) strain, an animal model used in hypertension research. A commercially prepared MVMM supplement was given tri-weekly via oral dosing for 8 weeks to two groups of seven adult female SHR and Wistar rats. Their corresponding control groups were dosed with deionised water only. Systolic and diastolic blood pressure, fasting blood glucose, growth rate and food and water intake were measured weekly. At the end of 8 weeks, the animals were euthanased and a full blood profile, urine sodium to potassium ratio, blood urea nitrogen levels and total plasma cholesterol was measured for all groups. The results indicated that growth rate was higher for the SHR supplemented group. Supplementation also decreased diastolic blood pressure in both Wistar and SHR groups and increased red blood cell count and decreased total cholesterol in the SHR group. No adverse effects on the general health status of the animals were observed. MVMM supplementation may therefore be useful in aiding growth and delaying the onset of hypertension and its effects. It may also assist in the longevity of the breeding stock of SHR rats.

In Vitro Callus Induction and Plant Regeneration from Stem Explants of Ceropegia noorjahaniae, a Critically Endangered Medicinal Herb.

An efficient protocol has been developed for in vitro regeneration of a large number of plantlets of Ceropegia noorjahaniae Ansari via indirect organogenesis from stem explants excised from in vitro-germinated seedlings. The callus was efficiently induced from the stem explants using Murashige and Skoog (MS) medium supplemented with auxins and their combinations. The highest number of shoots (16.0 ± 0.2) and shoot length (5.5 ± 0.1 cm) was achieved when the callus was subcultured to MS medium supplemented with 6-benzylaminopurine, BAP (2.0 mg/l) and indole-3-acetic acid, IAA (0.2 mg/l). The in vitro-developed shoots were rooted well in half-strength MS medium supplemented with 1.0 mg/l of indole-3-butyric acid (IBA) and 0.3 mg/l of α-naphthalene acetic acid (NAA). The plantlets were successfully hardened with 82 % survival rate. This is the first report on the regeneration of plants through indirect shoot organogenesis from stem derived calli of C. noorjahaniae.

Micropropagation and non-steroidal anti-inflammatory and anti-arthritic agent boswellic acid production in callus cultures of Boswellia serrata Roxb.

Micropropagation through cotyledonary and leaf node and boswellic acid production in stem callus of a woody medicinal endangered tree species Boswellia serrata Roxb. is reported. The response for shoots, roots and callus formation were varied in cotyledonary and leafy nodal explants from in vitro germinated seeds, if inoculated on Murshige and Skoog's (MS) medium fortified with cytokinins and auxins alone or together. A maximum of 8.0 ± 0.1 shoots/cotyledonary node explant and 6.9 ± 0.1 shoots/leafy node explants were produced in 91 and 88 % cultures respectively on medium with 2.5 µM 6-benzyladenine (BA) and 200 mg l(-1) polyvinylpyrrolidone (PVP). Shoots treated with 2.5 µM IBA showed the highest average root number (4.5) and the highest percentage of rooting (89 %). Well rooted plantlets were acclimatized and 76.5 % of the plantlets showed survival upon transfer to field conditions. Randomly amplified polymorphic DNA (RAPD) analysis of the micropropagated plants compared with mother plant revealed true-to-type nature. The four major boswellic acid components in calluses raised from root, stem, cotyledon and leaf explants were analyzed using HPLC. The total content of four boswellic acid components was higher in stem callus obtained on MS with 15.0 µM IAA, 5.0 µM BA and 200 mg l(-1) PVP. The protocol reported can be used for conservation and exploitation of in vitro production of medicinally important non-steroidal anti-inflammatory metabolites of B. serrata.

Risk of malignancy with long-term immunosuppression in renal transplant recipients.

BACKGROUND: Improvements in immunosuppressive regimens have significantly enhanced patient and graft survival in renal transplant recipients. However, susceptibility to neoplastic disorders is increased as a consequence of prolonged immunosuppression. Available data pertaining to cancer risks in renal transplant recipients have been inconsistent, and much of it is derived from international studies, which may not be truly representative of the United States population. METHODS: We studied a total of 1979 transplants performed in 1739 patients from a single center in the United States with a mean follow-up of 6.1 years, and a total of 9852 person-years' follow-up. RESULTS: The mean age at the time of diagnosis of cancer was 50 years, and the mean interval between transplant and diagnosis of cancer was 95 months. Older patients receiving a transplant had a significantly higher risk for developing cancer as opposed to younger patients (RR 6.2 for >60 years compared with <40 years). When compared with the general population using data from the Surveillance, Epidemiology and End Results (SEER) registry, the overall risk for nonskin malignancies was modestly increased in our transplant recipients, with a standardized incidence ratio (SIR) of 1.4 (P= 0.01). When stratified by age groups, younger age at transplant (<40 years) had the highest SIR, at 2.3 (P < 0.001). Similarly, duration post-transplant >10 years had an SIR of 2.4 (P < 0.001). CONCLUSION: We believe that this study is representative of the United States' renal transplant population, and highlights the need for reduced immunosuppression in the long-term and increased vigilance for cancers in younger patients receiving renal transplantation.

Sepsis in a renal transplant recipient due to Citrobacter braakii.

Cellulitis is usually caused by organisms such as beta-hemolytic streptococci and Staphylococcus aureus. Citrobacter are gram-negative bacilli that can cause opportunistic infections in immunocompromised hosts. They are rarely implicated in skin or soft tissue infections. The genus Citrobacter has been respeciated according to genetic relatedness. Citrobacter braakii refers to the genomospecies 6 of the Citrobacter freundii complex. There are no detailed studies of infections caused by the newly formed specific genetic species. We report a case of C. braakii infection in a renal transplant patient receiving immunosuppressive therapy. The patient's lower extremity cellulitis did not respond to conventional antibiotic therapy. Blood cultures grew C. braakii. Sensitivity studies and treatment with appropriate antibiotics resulted in prompt recovery. Immunosuppressive therapy in renal transplant recipients predisposes to infection by unusual pathogens, and this should be suspected when lack of a clinical response to conventional antibiotics is observed. We believe this is the first reported case of C. braakii cellulitis and bacteremia in a renal transplant recipient.