Valeriana jatamansi is an indigenous medicinal plant used in the treatment of a number of diseases. In the present study, chemical composition of the essential oil was determined by GC-MS. Seven major components were identified in Valeriana jatamansi essential oil, namely, β-vatirenene, β-patchoulene, dehydroaromadendrene, β-gurjunene, patchoulic alcohol, β-guaiene, and α-muurolene. Methanolic, aqueous, and chloroform extracts of Valeriana jatamansi roots were also prepared and analyzed for their polyphenols and flavonoid content. Antioxidant activity of essential oil and different extracts of Valeriana jatamansi roots was determined by DPPH radical scavenging and chelation power assay. A linear correlation has been obtained by comparing the antioxidant activity and polyphenols and flavonoid content of the extracts. Results indicated that antioxidant activity of methanolic extract could be attributed to the presence of rich amount of polyphenols and flavonoid. Essential oil of Valeriana jatamansi roots showed moderate antioxidant activity.
To determine if volumetric changes of diffusion-weighted and contrast material–enhanced magnetic resonance (MR) imaging can help assess early tumor response to intraarterial therapy (IAT) in neuroendocrine liver metastasis (NELM).
Materials and Methods
This retrospective single-center comprehensive imaging analysis was performed in compliance with HIPAA and was institutional review board approved. Informed patient consent was waived. Seventy-one patients (39 men; mean age, 62.3 years) with NELM treated with IAT were analyzed retrospectively. MR studies were performed before and 3–4 weeks after therapy. The index lesion was segmented to provide volumetric functional analysis of apparent diffusion coefficient (ADC) and contrast-enhanced MR imaging in the hepatic arterial phase (HAP) and portal venous phase (PVP). Tumor response was defined as increase in volumetric ADC of 15% or greater and decrease in volumetric enhancement of 25% or greater during the HAP or of 50% or greater during the PVP. Patient overall survival was the primary end point after therapy initiation. Univariate analysis included Kaplan-Meier survival curves. The Cox proportional hazards regression model was used to detect interactions between volumetric ADC and contrast-enhanced MR imaging and to calculate the hazard ratio.
There was significant increase in mean volumetric ADC (27%, P < .0001) and significant decrease in mean volumetric enhancement during the HAP (−25.3%, P < .0001) and the PVP (−22.4%, P < .0001) in all patients. Patients who had 15% or greater volumetric ADC increase (n = 49) after therapy had better prognosis than those who had less than 15% increase in volumetric ADC (n = 22) (log-rank test, P < .002). Patients who had 25% or greater decrease in volumetric arterial enhancement (n = 40) or 50% or greater decrease in venous enhancement (n = 18) had better prognosis than those who had less than 25% decrease in volumetric arterial enhancement (n = 31) or less than 50% decrease in venous enhancement (n = 53) (log-rank test, P < .02).
Volumetric functional MR imaging criteria may act as biomarkers of early response, indicating that these criteria may be important to incorporate in future NELM clinical trials.
The binding of all-trans retinoic acid (ATRA) to retinoid receptor-α (RAR-α) relieves transcriptional repression induced by the promyelocytic leukemia–retinoic acid receptor (PML–RAR) oncoprotein. The ATRA molecule contains a cyclohexenyl ring, a polyene chain containing conjugated double alkene bonds, and a terminal carboxyl group. To determine the contributions of these structural components of ATRA to its clinical efficacy, we synthesized three novel retinoids. These consisted of either a modified conjugated alkene backbone with an intact acid moiety (13a) or a modified conjugated alkene backbone and conversion of the acid group to either an ester (13b) or an aromatic amide (13c). Reporter assays demonstrated that compound 13a successfully relieved transcriptional repression by RAR-α, while 13b and 13c could not, demonstrating the critical role of the acid moiety in this binding. However, only ATRA was able to significantly inhibit the proliferation of APL cells while 13a, 13b, or 13c was not. Furthermore, only 13a led to partial non-significant differentiation of NB4 cells, demonstrating the importance of C9–C10 double bonds in differentiation induced CD11 expression. Our results demonstrate that both the acid moiety and conjugated double bonds present in the ATRA molecule are important for its biological activity in APL and have important implications for the design of future novel retinoids.
Acute promyelocytic leukemia (APL); all-trans retinoic acid (ATRA); retinoids
Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.
Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12–1275 CTCs/ml; Lung cancer samples: 10–1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples.
We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.
Rational and Objective
Radioembolization with yttrium-90 microspheres is a therapy that is used for hepatic tumors. 20–30 μm microspheres loaded with Y90 are supposedly occluding tumor vessels at the capillary level. Then, these spheres deliver high-dose radiation to the tumor. However, this theoretical embolic effect has never been appreciated in imaging. Dual-Phase cone-beam computed tomography (DPCBCT) is a multi-phasic intra-procedural scan that uses only one contrast media injection to visualize early (feeding vessel) and delayed (capillary level) tumor enhancement. The purpose of this study was to determine whether there is a micro-embolic effect induced by TheraSpheres® (MDS Nordion, Ottawa, Ontario, Canada) at the capillary level by using DPCBCT imaging.
Materials and Methods
14 patients with 72 carcinoid or neuroendocrine tumors were treated with radioembolization, and all underwent DPCBCT (Allura Xper, Philips Healthcare) imaging before and immediately after radioembolization with TheraSpheres®. Tumor enhancement was measured in each phase by drawing a region of interest within the tumors.
72 tumors were evaluated: average tumor density in the early arterial phase was 241 and 230 Hounsfield units (HU) (p<0.001) before and after radioembolization, respectively; the average density in the delayed arterial phase was 226 and 161 HU (p<0.001) before and after radioembolization, respectively. Average difference in tumor attenuation before and after radioembolization in early arterial and delayed phase was 11 HU and 64 HU (p<0.001), respectively.
The significant decrease in tumor enhancement in the DPCBCT delayed phase after TheraSpheres® injection indicates that there is an appreciable microembolic effect at the tumor capillary bed level.
C-arm Cone Beam CT; Microembolic effect; Radioembolization
Pharynx is a common site of malignancy in the head and neck region. This study presents a series of 94 cases of pharyngeal malignancy conducted at Himalayan Institute of Medical Sciences, Dehradun, Uttarakhand in the Department of Otorhinolaryngology for a period of one year (2009–2010). Mean age at presentation was 56.8 years (age range 18–100 years). Male:Female ratio was 8.4:1.0. Maximum patients belonged to lower socio-economic status as per Kuppuswamy’s classification (2003). Majority of them were farmer (38.2%) by occupation and belonged to rural areas. 90.4% patients had history of tobacco smoking. Dysphagia was the commonest chief complaint. The most common subsite was oropharynx (51.0%) followed by hypopharynx (45.7%). Ulceroproliferative growth was the most common clinical finding. Histopathologically, squamous cell carcinoma (94.6%) was the commonest. CECT was the commonest and most useful radiological investigation done to see the extent of the disease.
Oropharynx malignancy; Pharynx malignancy; Nasopharynx; Hypopharynx malignancy
Urinary incontinence (UI) following prostate radiotherapy is a rare toxicity that adversely affects a patient’s quality of life. This study sought to evaluate the incidence of UI following stereotactic body radiation therapy (SBRT) for prostate cancer.
Between February, 2008 and October, 2010, 204 men with clinically localized prostate cancer were treated definitively with SBRT at Georgetown University Hospital. Patients were treated to 35–36.25 Gray (Gy) in 5 fractions delivered with the CyberKnife (Accuray). UI was assessed via the Expanded Prostate Index Composite (EPIC)-26.
Baseline UI was common with 4.4%, 1.0% and 3.4% of patients reporting leaking > 1 time per day, frequent dribbling and pad usage, respectively. Three year post treatment, 5.7%, 6.4% and 10.8% of patients reported UI based on leaking > 1 time per day, frequent dribbling and pad usage, respectively. Average EPIC UI summary scores showed an acute transient decline at one month post-SBRT then a second a gradual decline over the next three years. The proportion of men feeling that their UI was a moderate to big problem increased from 1% at baseline to 6.4% at three years post-SBRT.
Prostate SBRT was well tolerated with UI rates comparable to conventionally fractionated radiotherapy and brachytherapy. More than 90% of men who were pad-free prior to treatment remained pad-free three years following treatment. Less than 10% of men felt post-treatment UI was a moderate to big problem at any time point following treatment. Longer term follow-up is needed to confirm late effects.
Prostate cancer; SBRT; Urinary incontinence; Expanded prostate index composite; EPIC; CyberKnife; Quality of life
Migration of Ascaris lumbricoides into the gallbladder is rare, unlike ascariasis of the bile duct and when it does occur, treatment is generally by endoscopic or surgical extraction. We describe a case of the successful treatment of gallbladder ascariasis with conservative therapy.
Ascaris lumbricoides; Gallbladder; Acute cholecystitis; Ultrasonography
We report the surgical management of a case of Zenker's diverticulum in a 64 year old man, complicated by metallic nail penetration and surgical scarring due to previous surgery for cervical vertebral trauma
Zenker’s diverticulum; muscular dysmotility; pharyngoesophageal junction
Eosinophilic esophagitis (EoE) is a clinicopathologic entity of increasing worldwide prevalence. IL-5 is essential for eosinophil trafficking and anti-IL-5 therapy decreases esophageal eosinophilia. EoE is associated with a prominent mast cell infiltration.
We investigated whether anti-IL-5 (mepolizumab) treatment reduced esophageal mast cell accumulation in pediatric EoE biopsy specimens from a previous randomized anti-IL-5 trial.
A sub-analysis was completed for children treated with 0.55, 2.5, or 10mg/kg of mepolizumab monthly for 12 weeks followed by no treatment until week 24. Quantitative immunochemistry was used to assess eosinophils, tryptase-positive mast cells, IL-9+ cells, and mast cell-eosinophil couplets prior to and following treatment.
43 patient biopsies had adequate tissue for paired analysis. 40% of subjects responded to anti-IL-5 (defined as <15 eosinophils per hpf following mepolizumab therapy) and 77% of all subjects had decreased numbers of mast cells following anti-IL-5. In responders, epithelial mast cells decreased from 62 to 19 per hpf (p<0.001), were significantly lower than in non-responders following therapy (p<0.05), and correlated with eosinophil numbers (r=0.75, p<0.0001). Mast cells and eosinophils were found in couplets prior to therapy and these were significantly decreased only in responders following anti-IL-5 (p<0.001). Esophageal eosinophils comprised the majority of cells that made the mast cell growth factor IL-9. IL-9+ cells decreased from 102 to 71 per hpf (p<0.001) following anti-IL-5.
Pediatric EoE patients had significantly fewer mast cells, IL-9+ cells, and mast cell-eosinophil couplets in the esophageal epithelium following anti-IL-5 therapy. Since eosinophils were one source of IL-9, they may support esophageal mastocytosis.
Eosinophilic esophagitis; pediatric; eosinophils; mast cells; IL-5; IL-9
Background Osteochondroma is the most common primary bone tumor, composing 35% of benign bone tumors and 9% of all bone tumors; 1.3 to 4.1% of all osteochondromas originate from the spine. A rare differential diagnosis for globus symptoms is an osteochondroma originating from the anterior surface of the axis. We describe a rare case of osteochondroma of the dens resulting in “globus symptoms” (the subjective sensation of a mass in the throat) treated with excision via the high cervical extrapharyngeal approach.
Purpose To discuss the surgical management of this problem, with an emphasis on surgical approach used. The clinical history, examination, and investigations are presented and illustrated, along with clinical patient outcome.
Study Design/Setting This article is a case report of a patient treated at the Department of Trauma and Orthopaedics in an active university teaching hospital.
Methods Case presentation. For the discussion, we used handpicked articles, as well as MEDLINE and PubMed database searches with the keywords “C2,” “dens,” “osteochondroma,” “globus,” “extrapharyngeal approach.”
Results Uncomplicated procedure. Histological analysis confirmed a benign osteochondroma with no evidence of malignancy. The patient underwent an uncomplicated postoperative recovery and was discharged 24 hours after surgery, fully ambulatory and eating and drinking well.
Conclusions The high cervical retropharyngeal approach is safe and reproducible for the excision of osteochondromas or osteophytes of the upper cervical spine.
osteochondroma; c2; dens; extra-pharyngeal; high pharyngeal; cervical spine; approach; excision
This study explored the antioxidant and immunomodulatory potential of ethnomedicinally valuable species, namely, Arisaema jacquemontii of north-western Himalayan region. The tubers, leaves, and fruits of this plant were subjected to extraction using different solvents. In vitro antioxidant studies were performed in terms of chelation power on ferrous ions and FRAP assay. The crude methanol extract of leaves was found to harbour better chelating capacity (58% at 100 μg/mL) and reducing power (FRAP value 1085.4 ± 0.11 μMFe3+/g dry wt.) than all the other extracts. The crude methanol extract was thus further partitioned with solvents to yield five fractions. Antioxidant study of fractions suggested that the methanol fraction possessed significant chelation capacity (49.7% at 100 μg/mL) and reducing power with FRAP value of 1435.4 μM/g dry wt. The fractions were also studied for immune modulating potential where it was observed that hexane fraction had significant suppressive effect on mitogen induced T-cell and B-cell proliferation and remarkable stimulating effect on humoral response by 141% and on DTH response by 168% in immune suppressed mice as compared to the controls. Therefore, it can be concluded that A. jacquemontii leaves hold considerable antioxidant and immunomodulating potential and they can be explored further for the identification of their chemical composition for a better understanding of their biological activities.
Celiac disease (CD) is under-diagnosed in the United States, and factors related to the performance of endoscopy may be contributory.
to identify newly diagnosed patients with CD who had undergone a prior esophagogastroduodenoscopy (EGD) and examine factors contributing to the missed diagnosis.
We identified all patients age ≥18 years whose diagnosis of CD was made by endoscopy with biopsy at our institution (n=316), and searched the medical record for a prior EGD. We compared those patients with a prior EGD to those with without a prior EGD with regard to age at diagnosis and gender, and enumerated the indications for EGD.
Of the 316 patients diagnosed by EGD with biopsy at our center, 17 (5%) had previously undergone EGD. During the prior non-diagnostic EGD, a duodenal biopsy was not performed in 59% of the patients, and ≥4 specimens (the recommended number) were submitted in only 29% of the patients. On the diagnostic EGD, ≥4 specimens were submitted in 94%. The mean age of diagnosis of those with missed/incident CD was 53.1 years, slightly older than those diagnosed with CD on their first EGD (46.8 years, p=0.11). Both groups were predominantly female (missed/incident CD: 65% vs. 66%, p=0.94).
Among 17 CD patients who had previously undergone a non-diagnostic EGD, nonperformance of duodenal biopsy during the prior EGD was the dominant feature. Routine performance of duodenal biopsy during EGD for the indications of dyspepsia and reflux may improve CD diagnosis rates.
Celiac Disease; Endoscopy; Biopsy; Diagnosis
Introduction: Sex chromatin is a plano-convex to triangular DNA mass measuring approximately 1μm in size and lying adjacent to the inner side of nuclear membrane in the somatic cells of the females. There is consistent loss in the sex chromatin percentage in the carcinoma cases in comparison to benign lesions and normal individuals.
Aim: To know the correlation between the sex chromatin status in female breast tumors on paraffin sections, buccal smears and peripheral blood films.
Materials and Methods: The study was conducted on the paraffin sections prepared from carcinoma breast patients from their lumpectomy and mastectomy specimens. Buccal smears and a peripheral blood films were also prepared from each patient.
Discussion: The control group had shown a mean sex chromatin of 54.6±6.73% which was found to be similar to the mean sex chromatin percentage in the fibroadenoma breast cases i.e. 54.91±6.06%. However, the mean sex chromatin in the carcinoma breast cases was markedly reduced i.e. 8.22±6.03%. Maximum no. of fibroadenoma cases (67%) were in the younger age group i.e. 15 to 25 year, while maximum number of carcinoma breast cases (42%) occurred in the 4th and 5th decade.
Conclusion: There is a loss of sex chromatin in cases of carcinoma breast and is associated with poor histological markers. A statistically significant correlation was also found between sex chromatin status and microscopic grading in carcinoma breast. The tumors with higher microscopic grade had lower sex chromatin as compared to those with lower microscopic grading.
Sex chromatin; Fibroadenoma; Carcinoma breast
BACKGROUND & AIMS
Alterations in methylation of protein-coding genes are associated with Barrett’s esophagus (BE) and esophageal adenocarcinoma (EAC). Dys-regulation of noncoding RNAs occurs during carcinogen-esis but has never been studied in BE or EAC. We applied high-resolution methylome analysis to identify changes at genomic regions that encode noncoding RNAs in BE and EAC.
We analyzed methylation of 1.8 million CpG sites using massively parallel sequencing-based HELP tagging in matched EAC, BE, and normal esophageal tissues. We also analyzed human EAC (OE33, SKGT4, and FLO-1) and normal (HEEpic) esophageal cells.
BE and EAC exhibited genome-wide hypomethylation, significantly affecting intragenic and repetitive genomic elements as well as noncoding regions. These methylation changes targeted small and long noncoding regions, discriminating normal from matched BE or EAC tissues. One long noncoding RNA, AFAP1-AS1, was extremely hypomethylated and overexpressed in BE and EAC tissues and EAC cells. Its silencing by small interfering RNA inhibited proliferation and colony-forming ability, induced apoptosis, and reduced EAC cell migration and invasion without altering the expression of its protein-coding counterpart, AFAP1.
BE and EAC exhibit reduced methylation that includes noncoding regions. Methylation of the long noncoding RNA AFAP1-AS1 is reduced in BE and EAC, and its expression inhibits cancer-related biologic functions of EAC cells.
Esophageal Cancer Progression; Tumor Development; Gene Regulation; Noncoding RNA
Vertical and anterioposterior maxillary excesses can be treated with a combination of orthopaedic functional appliances, orthodontics and surgery. Treatment varies according to the age, patient reports for treatment. In patients who are treated with either of the above mentioned treatment modalities, if they require prosthetic replacement on a later date, especially of anterior teeth, prosthetic treatment alone does not give an aesthetic outcome. A partially edentulous, elderly patient with underlying skeletal discrepancy (Class II Skeletal deformity) in relation to 12,11,21,22 was treated with a combination of orthognathic surgery and prosthetic rehabilltation. An orthognathic surgery (leforte I osteotomy) was performed to manage vertical maxillary excess, class II skeletal pattern of maxilla and increased lower third facial height. Dental compensations in the mandibular arch were decompensated surgically with lower subapical osteotomy. Prosthetic restorations of missing anterior teeth were done later, such that facial and dental aesthetics. The records showed that the results were stable 12 months after prosthognathic (prosthodontic and orthognathic) treatment. A team approach enabled the female patient in her fifth decade of life, to receive better function, aesthetics and increased quality of life. Doing prosthetic restorations in patients with underlying skeletal discrepancies may become a challenge , which should be achieved without compromising on final outcome, with a calculated risk benefit ratio.
Prosthognathic; Orthognathic surgery; Fixed prosthesis
Rationale and Objectives
To evaluate the precision and reproducibility of a semi-automatic tumor segmentation software in measuring tumor volume of hepatocellular-carcinoma–(HCC) before the first trans-arterial chemo-embolization–(TACE) on contrast-enhancement magnetic-resonance-imaging–(CE-MRI) and intra-procedural dual-phase C-arm cone-beam computed-tomography–(DP-CBCT) images.
Materials and Methods
19HCCs were targeted in 19patients(one per patient) who underwent baseline diagnostic CE-MRI and an intra-procedural DP-CBCT. The images were obtained from CE-MRI–(arterial-phase of an intra-venous contrast medium injection) and DP-CBCT–(delayed-phase of an intra-arterial contrast medium injection) before the actual embolization. Three readers measured tumor volumes using a semi-automatic 3D-volumetric segmentation software which used a region-growing method employing non-Euclidean radial basis functions. Segmentation time and spatial position were recorded. The tumor volume measurements between images sets were compared using linear-regression and Student t-test, and evaluated with Intraclass-Correlation analysis–(ICC). The inter-rater Dice Similarity Coefficient–(DSC) accessed the segmentation spatial localization.
All 19 HCCs were analyzed. On CE-MRI and DP-CBCT examinations respectively, A) the mean segmented tumor volumes was 87±8cm3[2–873] and 92±10cm3[1–954], with no statistical difference of segmented volumes by readers of each tumor between the two imaging modalities and the mean time required for segmentation was 66±45seconds [21–173] and 85±34seconds[17–214] (p=0.19), B),the ICCs were 0.99 and 0.974, showing a strong correlation among readers, and C) the inter-rater DSCs showed a good to excellent inter-user agreement on the spatial localization of the tumor segmentation–(0.70±0.07 and 0.74±0.05,p=0.07).
This study shows a strong correlation, precision and reproducibility of semi-automatic tumor segmentation software in measuring tumor volume on CE-MRI and DP-CBCT images. The use of the segmentation software on DP-CBCT and CE-MRI can be a valuable and highly accurate tool to measure the volume of hepatic tumors.
Tumor segmentation software; Dice Similarity Coefficient; C-arm cone-beam CT; MRI; Hepatocellular carcinoma; TACE
Nose bleed is the most common rhinological emergency. There are multiple risk factors for the development of epistaxis and it can affect any age group, but it is the elderly population with their associated morbidity who often require more intensive treatment and subsequent admission. Most cases of epistaxis occur in the Little’s area, a location readily accessible and treatable by cautery or anterior nasal packing. However, posterior epistaxis often requires more aggressive measures including posterior nasal packing and endoscopic cauterization. After posterior nasal packing, the two most common therapies for intractable epistaxis are transantral ligation of the internal maxillary artery and percutaneous embolization of the distal internal maxillary artery. However, optimal management of intractable posterior epistaxis remains controversial. We hereby report fourth case of Waldenstrom Macroglobulinemia in English literature, which presented as isolated persistent epistaxis and was treated by therapeutic plasmapheresis.
Epistaxis; Little’s area; Embolization; Plasmapheresis; Waldenstrom macroglobulinemia
OBJECTIVE: This was a single-center, prospective, pilot study aiming to evaluate the impact of pharmacist involvement in the admission medication history and reconciliation process and to quantify discrepancies found by pharmacists when compared to information collected by other health care providers at a pediatric institution.
METHODS: A pharmacist completed a thorough medication history and reconciled discrepancies with the medical team. Discrepancies included incorrect medication, dose, route, frequency; omitted information; missing medications; or any other inconsistencies outside of these categories. Information was documented in the electronic medical record via a standardized template, and pertinent discrepancies were communicated with the medical team.
RESULTS: Of the 100 medication histories included in the study, a total of 309 discrepancies were identified and corrected in the electronic medical record. The median length of time it took pharmacists to complete the medication history process was 15 minutes per patient (interquartile range, 10–20 minutes). Thirty discrepancies were determined as pertinent and were reported as intervened on and communicated to the medical team.
CONCLUSION: This study provides evidence that pharmacist-obtained admission medication histories and reconciliation have the potential to prevent potentially significant adverse drug reactions and have a positive impact on patient care.Index terms admission, history, medication, pharmacist, reconciliation
admission; history; medication; pharmacist; reconciliation
Type 2 diabetes is a complex metabolic disorder characterized by hyperglycemia, impaired glucose tolerance and insulin resistance associated with dyslipidemia and hypertension. The available drugs are not sufficiently efficacious in reducing cardiovascular risk and restoring normal glucose metabolism associated with type 2 diabetes as a mono- or a combination therapy. The present study examined the combined effects of an antihypertensive (S-Amlodipine) and an insulin-sensitizing agent, peroxisome proliferator-activated receptor (PPAR) agonists (Pioglitazone and Ragaglitazar), on cardiovascular risk factors in aged diabetic and insulin-resistant Zucker fa/fa rats.
Following combination treatment for 14 days, blood pressure (BP), serum glucose, total cholesterol and triglycerides were measured. Aortic ring study was conducted to determine the effect of combination treatments on phenylephrine-induced vasoconstriction and acetylcholine (Ach)-induced vasorelaxation.
In combination, S-Amlodipine and Pioglitazone significantly reduced blood glucose (115.1 ± 6.6 vs. 81.7 ± 4.2), BP (184.4 ± 5.0 vs. 155.1 ± 5.0), serum triglycerides (362.5 ± 47.5 vs. 211.1 ± 23.7) and glucose intolerance when compared with vehicle treated Zucker fa/fa rats. Similar results were observed with the combination of S-Amlodipine and Ragaglitazar (Triglycerides, 362.5 ± 47.5 vs. 252.34 ± 27.86; BP, 184.4 ± 5.0 vs. 159.0 ± 8.0) except for serum glucose. ACh-induced vasorelaxation in aortic rings was also superior with both of the combinations compared to individual treatment. Furthermore, there was less body weight gain and food intake with S-Amlodipine and Pioglitazone combination in Zucker fa/fa rats. S-Amlodipine itself caused significant reduction in glucose (115.1 ± 6.6 vs. 89.7 ± 2.7) and BP (184.4 ± 5.0 vs. 156.1 ± 4.0) with improvement in insulin sensitivity observed through oral glucose tolerance test.
The results suggest that a combination of PPAR agonists and S-Amlodipine has partial benefits in improving the cardiovascular risk factors such as reduction in triglyceride levels, associated with chronic type 2 diabetes, and therefore may be utilized as an approach for addressing some of these devastating metabolic syndrome complications.
Calcium channel blockers; Peroxisome proliferator-activated receptors (PPARs); Type 2 diabetes; Dyslipidemia; Hypertension
Genome-wide association studies (GWAS) have identified chromosomal loci that affect risk of coronary heart disease (CHD) independent of classical risk factors. One such association signal has been identified at 6q23.2 in both Caucasians and East Asians. The lead CHD-associated polymorphism in this region, rs12190287, resides in the 3′ untranslated region (3′-UTR) of TCF21, a basic-helix-loop-helix transcription factor, and is predicted to alter the seed binding sequence for miR-224. Allelic imbalance studies in circulating leukocytes and human coronary artery smooth muscle cells (HCASMC) showed significant imbalance of the TCF21 transcript that correlated with genotype at rs12190287, consistent with this variant contributing to allele-specific expression differences. 3′ UTR reporter gene transfection studies in HCASMC showed that the disease-associated C allele has reduced expression compared to the protective G allele. Kinetic analyses in vitro revealed faster RNA-RNA complex formation and greater binding of miR-224 with the TCF21 C allelic transcript. In addition, in vitro probing with Pb2+ and RNase T1 revealed structural differences between the TCF21 variants in proximity of the rs12190287 variant, which are predicted to provide greater access to the C allele for miR-224 binding. miR-224 and TCF21 expression levels were anti-correlated in HCASMC, and miR-224 modulates the transcriptional response of TCF21 to transforming growth factor-β (TGF-β) and platelet derived growth factor (PDGF) signaling in an allele-specific manner. Lastly, miR-224 and TCF21 were localized in human coronary artery lesions and anti-correlated during atherosclerosis. Together, these data suggest that miR-224 interaction with the TCF21 transcript contributes to allelic imbalance of this gene, thus partly explaining the genetic risk for coronary heart disease associated at 6q23.2. These studies implicating rs12190287 in the miRNA-dependent regulation of TCF21, in conjunction with previous studies showing that this variant modulates transcriptional regulation through activator protein 1 (AP-1), suggests a unique bimodal level of complexity previously unreported for disease-associated variants.
Both genetic and environmental factors cumulatively contribute to coronary heart disease risk in human populations. Large-scale meta-analyses of genome-wide association studies have now leveraged common genetic variation to identify multiple sites of disease susceptibility; however, the causal mechanisms for these associations largely remain elusive. One of these disease-associated variants, rs12190287, resides in the 3′untranslated region of the vascular developmental transcription factor, TCF21. Intriguingly, this variant is shown to disrupt the seed binding sequence for microRNA-224, and through altered RNA secondary structure and binding kinetics, leads to dysregulated TCF21 gene expression in response to disease-relevant stimuli. Importantly TCF21 and miR-224 expression levels were perturbed in human atherosclerotic lesions. Along with our previous reports on the transcriptional regulatory mechanisms altered by this variant, these studies shed new light on the complex heritable mechanisms of coronary heart disease risk that are amenable to therapeutic intervention.
Results of investigation of the physicochemical properties of zinc complexes containing substituted phenols as axial ligand having general formula [X-Zn-t(p-CH3) PP] [where X = different phenolates as axial ligand] in impurity-free organic solvent are presented. The four-coordinated zinc porphyrin accepts one axial ligand in 1 : 1 molar ratio to form five-coordinated complex, which is purified by column chromatography and characterized by physicochemical, biological evaluation and TGA/DTA studies. Absorption spectra show two principal effects: a red shift for phenols bearing substituted electron releasing groups (−CH3, −NH2) and blue shift for phenols bearing electron withdrawing groups (−NO2, −Cl) relative to Zn-t(p-CH3) PP, respectively. 1H NMR spectra show that the protons of the phenol ring axially attached to the central metal ion are merged with the protons of the porphyrin ring. Fluorescence spectra show two fluorescence peaks in the red region with emission ranging from 550 nm to 700 nm. IR spectra confirm the appearance of Zn-NPor and Zn-O vibrational frequencies, respectively. According to the thermal studies, the complexes have a higher thermal stability and the decomposition temperature of these complexes depends on the axial ligation. The respective complexes of X-ZnII-t(p-CH3) PP were found to possess higher antifungal activity (up to 90%) and higher in vitro cytotoxicity against human cancer cells lines.
Cell surface growth factor receptors couple environmental cues to the regulation of cytoplasmic homeostatic process including autophagy, and aberrant activation of such receptors is a common feature of human malignancies. Here, we defined the molecular basis by which the epidermal growth factor receptor (EGFR) tyrosine kinase regulates autophagy. Active EGFR binds to the autophagy protein Beclin 1, leading to its multisite tyrosine phosphorylation, enhanced binding to inhibitors, and decreased Beclin 1-associated Class III phosphatidylinositol-3 kinase activity. EGFR tyrosine kinase inhibitor (TKI) therapy disrupts Beclin 1 tyrosine phosphorylation and binding to its inhibitors, and restores autophagy in non-small cell lung carcinoma (NSCLC) cells with a TKI-sensitive EGFR mutation. In NSCLC tumor xenografts, the expression of a tyrosine phosphomimetic Beclin 1 mutant leads to reduced autophagy, enhanced tumor growth, tumor dedifferentiation, and resistance to TKI therapy. Thus, oncogenic receptor tyrosine kinases directly regulate the core autophagy machinery, which may contribute to tumor progression and chemoresistance.
Experience rearranges anatomical connectivity in the brain, but such plasticity is suppressed in adulthood. We examined the turnover of dendritic spines and axonal varicosities in the somatosensory cortex of mice lacking Nogo Receptor 1 (NgR1). Through adolescence, the anatomy and plasticity of ngr1 null mice are indistinguishable from control, but suppression of turnover after age 26 days fails to occur in ngr1−/− mice. Adolescent anatomical plasticity can be restored to one-year old mice by conditional deletion of ngr1. Suppression of anatomical dynamics by NgR1 is cell autonomous, and is phenocopied by deletion of Nogo-A ligand. Whisker removal deprives the somatosensory cortex of experience-dependent input and reduces dendritic spine turnover in adult ngr1−/− mice to control levels, while an acutely enriched environment increases dendritic spine dynamics in control mice to the level of ngr1−/− mice in a standard environment. Thus, NgR1 determines the low set point for synaptic turnover in adult cerebral cortex.