miRNA expression profile of mucoepidermoid carcinoma.

OBJECTIVES: MicroRNAs (miRNAs) are single-stranded RNAs that have been implicated in cancer initiation and progression and act as tumour suppressors or oncogenes. In this study, miRNA profiling was conducted on the most frequent malignancy of salivary glands, mucoepidermoid carcinoma (MEC), in comparison with normal tissues. MATERIALS AND METHODS: The TaqMan Human miRNA Cards Array was used for the miRNA profiling of MEC and normal tissues. To validate the differentially expressed miRNAs in MEC, we used real-time PCR (qRT-PCR). RESULTS: miR-302a was the most significantly increased miRNA in cancer tissues (p < .05). Here, we demonstrate that the upregulation of miR-302a expression in SGT cell lines induced cancer cell invasion in vitro. CONCLUSIONS: These promising results suggest the need for further studies to establish mir-302a as a marker of invasion and aggressiveness in MEC.

Late responses to adenoviral-mediated transfer of the aquaporin-1 gene for radiation-induced salivary hypofunction.

We evaluated late effects of AdhAQP1 administration in five subjects in a clinical trial for radiation-induced salivary hypofunction (http://www.clinicaltrials.gov/ct/show/NCT00372320?order=). All were identified as initially responding to human aquaporin-1 (hAQP1) gene transfer. They were followed for 3-4 years after AdhAQP1 delivery to one parotid gland. At intervals we examined salivary flow, xerostomic symptoms, saliva composition, vector presence and efficacy in the targeted gland, clinical laboratory data and adverse events. All displayed marked increases (71-500% above baseline) in parotid flow 3-4.7 years after treatment, with improved symptoms for ~2-3 years. There were some changes in [Na+] and [Cl-] consistent with elevated salivary flow, but no uniform changes in secretion of key parotid proteins. There were no clinically significant adverse events, nor consistent negative changes in laboratory parameters. One subject underwent a core needle biopsy of the targeted parotid gland 3.1 years post treatment and displayed evidence of hAQP1 protein in acinar, but not duct, cell membranes. All subjects responding to hAQP1 gene transfer initially had benefits for much longer times. First-generation adenoviral vectors typically yield transit effects, but these data show beneficial effects can continue years after parotid gland delivery.

Role of Runx2 in IGF-1Rß/Akt- and AMPK/Erk-dependent growth, survival and sensitivity towards metformin in breast cancer bone metastasis.

The mechanisms underlying reprogramming of growth factor signaling and metabolic pathways during bone metastasis of breast cancer are not clear. The Runt-related transcription factor (Runx2) regulates cell signaling during mammary epithelial morphogenesis and promotes invasion; therefore, we investigated its role in cell growth and metabolic signaling in bone-seeking breast cancer cells. We performed systemic inoculation of control or Runx2 knockdown invasive MDA-MB-231 cells in NOD/SCID mice, and compared parental and bone-derived variants for phenotypic and molecular alterations. The Runx2 knockdown showed early (0-2 weeks) inhibition of metastatic spread but late (4-6 weeks) outgrowth, suggesting Runx2-dependent bi-phasic response and reprogramming of metastatic cells. The late-stage tumor outgrowth of bone-derived Runx2 knockdown cells was associated with increased insulin-like growth factor- 1Rß (IGF-1Rß) levels. Interestingly, glucose uptake and glycolysis were reduced in the bone-derived Runx2 knockdown cells that could be further reduced by extracellular-regulated protein kinase (Erk1/2) inhibition. Furthermore, the Runx2 knockdown cells displayed activation of AMP-activated protein kinase (AMPKα), the sensor of cellular metabolism. Importantly, the Runx2 knockdown in bone-derived cells resulted in increased sensitivity to both Erk1/2 inhibition and AMPKα activation by PD184161 and metformin, respectively, despite increased IGF-1Rß and AMPKα levels. Our results reveal that Runx2 promotes metastatic spread of mammary tumor cells. The growth of late-stage tumor cells can be targeted by Runx2 knockdown in combination with Mek-Erk1/2 inhibition and metformin treatment.

Postoperative coagulopathy after live related donor hepatectomy: Incidence, predictors and implications for safety of thoracic epidural catheter.

BACKGROUND: Coagulopathy after living donor hepatectomy (LDH) may endanger donor safety during removal of thoracic epidural catheter (TEC). The present study was conducted to evaluate the extent and duration of immediate postoperative coagulopathy after LDH. MATERIALS AND METHODS: A retrospective analysis of perioperative record of LDH over three years was conducted after IRB approval. Variables such as age, gender, BMI, ASA classification, liver volume on CT scan, preoperative and postoperative INR, platelet count (PC) and ALT of each donor for five days was noted. In addition, duration of surgery, remnant as percentage total liver volume (Remnant%), blood loss, day of peak in PC and INR were also noted. Coagulopathy was defined as being present if INR exceeded 1.5 or platelet count fell below 1 × 10 5 /mm 3 on any day. Data was analyzed using SPSS 20 for Windows. Between group comparison was made using the Student 't' test for continuous variables and chi square test for categorical variables. Univariate analysis was done. Multiple logistic regression analysis was used to find independent factor associated with coagulopathy. RESULTS: Eighty four (84) donors had coagulopathy on second day (mean INR 1.9 ± 0.42). Low BMI, % of remnant liver and duration of surgery were independent predictors of coagulopathy. Right lobe hepatectomy had more coagulopathy than left lobe and low BMI was the only independent predictor. There was no correlation of coagulopathy with age, gender, blood loss, presence of epidural catheter, postoperative ALT or duration of hospital stay. High INR was the main contributor for coagulopathy. CONCLUSIONS: Coagulopathy is seen after donor hepatectomy. We recommend removal of the epidural catheter after the fifth postoperative day when INR falls below 1.5.

Comparative evaluation of prophylactic use of pregabalin, gabapentin and diclofenac sodium for prevention of succinylcholine-induced myalgia: a randomized, double-blinded study.

CONTEXT: Succinylcholine a depolarizing muscle relaxant with rapid onset, predictable course and short duration of action is associated with myalgia. AIM: The aim of this study is to evaluate the efficacy of pregabalin, gabapentin and diclofenac on the incidence and severity of succinylcholine-induced myalgia. SETTINGS AND DESIGN: Tertiary Care Teaching Hospital. MATERIALS AND METHODS: A total of 120 patients undergoing laparoscopic cholecystectomy were randomly assigned into three groups: Pregabalin group received 150 mg of pregabalin, gabapentin group received 600 mg of gabapentin and diclofenac group received 100 mg of diclofenac sodium orally 2 h prior to surgery. Anesthesia was induced with fentanyl 3 µg/kg, propofol 2-2.5 mg/kg and succinylcholine 1.5 mg/kg and was maintained with oxygen with sevoflurane in the air and intermittent vecuronium bromide. A blinded observer recorded post-operative pain scores on visual analog scale at different time intervals and myalgia at 24 h. Post-operative pain relief was provided with fentanyl based patient-controlled analgesia. Fentanyl consumption in 24 h was recorded as a primary outcome. STATISTICAL ANALYSIS: Patients' characteristics and total fentanyl consumption were compared using one-way ANOVA followed by post-hoc test. Pain score was compared amongst the groups using Kruskal Wallis test. RESULTS: The myalgia occurred in 15, 14 and 13 patients in pregabalin, gabapentin and diclofenac sodium group respectively (P > 0.85). Patients in diclofenac group had significantly higher fentanyl consumption (674.85 ± 115.58 µg) compared with pregabalin group (601.87 ± 129.57 µg) (95% confidence interval [CI] = 34.8-120.7) and gabapentin group (612.29 ± 105.12 µg) (95% CI = 14.9-170.5). However, there was no significant difference in fentanyl consumption between pregabalin and gabapentin groups (95% CI = -34.8-120.7). There was a significant difference in visual analog score at time points 12, 18 and 24 h among the study groups. CONCLUSION: Pre-treatment with pregabalin, gabapentin and diclofenac had equal efficacy in reducing the incidence and severity of succinylcholine-induced myalgia. However, pre-treatment with pregabalin and gabapentin decreased post-operative pain scores and fentanyl consumption.

Hepatorenal syndrome: a decade later.

Hepatorenal syndrome is a unique form of acute kidney injury seen in patients with acute liver failure or chronic liver disease in absence of any other identifiable cause of renal failure. It is primarily a diagnosis of exclusion. Despite of good pathophysiological understanding and better available therapeutic options for management of hepatorenal syndrome, it is still associated with significant morbidity and mortality. Liver transplantation forms the cornerstone for its management. In this review article, we have attempted to assimilate and summarise the advances made in the previous decade with regards to pathophysiology, classification and management of this entity.

Quadriplegia in a child following adenotonsillectomy.

Neurological deterioration in a child following routine surgery, although rare, has potentially life threatening consequences. We report the case of a child who, following adentonsillectomy, developed quadriplegia and acute respiratory distress due to previously undetected atlanto-axial instability. Patients with atlanto-axial instability often have mild or non-specific symptoms, despite severe cervical cord compression. Subtle manifestations may be ignored or attributed to other disease processes, which render patients with undiagnosed atlanto-axial instability at risk of serious neurological injury during general anaesthesia, particularly at the time of laryngoscopy and tracheal intubation.

Deep sequencing of short RNAs reveals novel microRNAs in minor salivary glands of patients with Sjögren's syndrome.

OBJECTIVES: Sjögren's syndrome is a complex autoimmune disease of the salivary gland with an unknown etiology, so a thorough characterization of the transcriptome would facilitate our understanding of the disease. We use ultradeep sequencing of small RNAs from patients with Sjögren's syndrome and healthy volunteers, primarily to identify and discover novel miRNA sequences that may play a role in the disease. METHODS: Total RNA was isolated from minor salivary glands of healthy volunteers and patients with either high or low salivary flow and sequenced on the SOLiD platform. Prediction of mature miRNAs from the sequenced reads was carried out using miRanalyzer, and expression was validated using Taqman qPCR assays. RESULTS: We validated the presence of six previously unidentified miRNA sequences in patient samples and in several cell lines. One of the validated novel miRNAs shows promise as a biomarker for salivary function. CONCLUSION: Sequencing small RNAs in the salivary gland is largely unprecedented, but here, we show the feasibility of discovering novel miRNAs and disease biomarkers by sequencing the transcriptome.

Intra procedure rupture of intracranial aneurysm during endovascular coiling: neurosurgeons' experience and review of the literature.

OBJECTIVE: The aim of this study was to determine the incidence, risk factors and outcome of intra procedure rupture (IPR) during endovascular coiling of intracranial aneurysms, a neurosurgeons' experience. MATERIALS AND METHODS: The frequency of IPR was studied in 168 aneurysms treated by endovascular coiling in 150 consecutive patients. Aneurysm size, morphology, history of previous subarachnoid hemorrhage (ruptured) and timing of treatment after subarachnoid hemorrhage were the data collected for comparison. RESULTS: Procedure-related rupture during endovascular coiling occurred in five (2.97%) of the 168 aneurysms treated. IPR was the cause for 1.78% treatment-related deaths. Small aneurysm size was the major risk factor for IPR in this series (P < 0.001). CONCLUSIONS: In this study, the frequency of IPR was similar to the reported frequency in the procedures performed by neurointerventionists.

Use of nitinol shape memory alloy staples (NiTi clips) after cervical discoidectomy: minimally invasive instrumentation and long-term results.

BACKGROUND: Anterior cervical discoidectomy with or without fusion is a well established surgical remedy for cervical prolapsed intervertebral disc (PIVD) disease. If fusion is done by an iliac bone graft then internal fixation is commonly used to keep the graft in position. This study was conducted to determine the efficacy and tolerability of shape memory alloys, especially NiTi (nickel titanium) clips in the stabilization of grafts following anterior cervical discoidectomy. METHODS: 133 NiTi clips were applied in 119 patients between January 2002 and December 2008. The patients age ranged from 38-60 years. There were 66 male and 53 females. Various indications for fixation of the spine included degenerated cervical spondylosis with single level PIVD (105) and two level PIVD in 14 patients. The cine mode fluoroscopy confirmed the perioperative correct placement of grafts and clips in all the patients. Follow-up ranged from 2 to 8 years (mean: 4.6 years). RESULTS: Single level discoidectomy was performed in 105 patients and two level disc removal was done in 14 patients. A single NiTi clip was applied in all the cases except for 14 cases of two level PIVD. No procedural complication or adverse reaction to the clip was noted. There was no movement at the operated level in dynamic lateral view X-ray of cervical spine at the 1st postoperative day as well as on follow-up. Graft extrusion was seen in one patient on the 2nd day after surgery and was reoperated. Bony fusion occurred in all patients after 9 - 12 months of surgery. There was no incidence of breakage or dislodgement of the clip from the site where it was inserted. No artifact was noted in cervical MRI done in 33 patients. CONCLUSION: NiTi clips are a simple alternative for cervical spine stabilization after discoidectomy. Their insertion is simple, minimally invasive, does not require any special set of instruments and they are much more economical than other established methods of treatment. These clips are accepted well by human tissue and do not interfere with MRI.

Bladder distension: An unusual cause of reflux of blood and hemodynamic changes (autonomic dysreflexia) during endovascular coiling.

Autonomic dysreflexia due to distended bladder is well known. Reflux of blood during endovascular procedure is also a common observation. It happens due to difference in pressure gradient between arterial pressure and that of infusing solution. Generally it happens when the infusion bottle is empty or the pressure in infusion bottles fall. We present an uncommon situation where distended bladder mechanically resulted in reflux of blood into endovascular catheters as well as alteration in hemodynamic parameters. Both settled once the bladder was empty.

Evaluation of cross-reactive cell-mediated immune responses among human, bovine and porcine adenoviruses.

The absence of preexisting immunity against porcine adenovirus (Ad) serotype 3 (PAd3) and bovine Ad serotype 3 (BAd3) in humans makes them attractive alternatives to human Ad serotype 5 (HAd5) vectors. To determine whether there is significant cross-reactivity among HAd5, BAd3 and PAd3 at the level of cell-mediated immune responses, BALB/c mice were inoculated intraperitoneally with wild-type (WT) or replication-defective (RD) HAd5, BAd3 or PAd3. After 35 days of the first inoculation, cross-reactive CD8+ cytotoxic T cells, as well as CD4+ Th1- and Th2-helper T cells, in the spleen were analyzed by enzyme-linked-immunospot, flow cytometry and cytotoxic T lymphocyte assays. Virus-neutralization assays were used to evaluate humoral cross-reactivity. CD8+ or CD4+ T cells primed with WT or RD HAd5, PAd3 or BAd3 showed significant (P<0.005) reactivity with homologous Ad antigens, whereas only minimal cross-reactivity was observed on stimulation with heterologous Ad antigens. Ad-neutralizing antibodies were found to be homologous Ad specific. Overall, these results suggest that there is no significant immunological cross-reactivity among HAd5, BAd3 and PAd3, thereby supporting the rationale for the use of BAd3 and PAd3 as alternative HAd vectors to circumvent anti-HAd immunity in humans.

Incidence and pattern of intraoperative hemodynamic response to endoscopic third ventriculostomy.

BACKGROUND: In patients undergoing endoscopic third ventriculostomy (ETV), various cardiovascular changes occur in the intraoperative period. AIM: We tried to determine a pattern in these changes and their relation to different surgical steps. MATERIALS AND METHODS: A total of 260 patients were studied over a period of six years. Heart rate and mean arterial pressures were recorded before introduction of the endoscope and thereafter at various stages of the operation. RESULTS: Tachycardia was the predominant observed abnormality in 20% of patients, occurring mostly during manipulations and irrigation in the third ventricle (TV). Bradycardia was seen in 12% of patients, more often during fenestration of the floor of the third ventricle. CONCLUSIONS: Tachycardia observed during ETV may be related to hypothalamic stimulation or a rise in intracranial pressure and bradycardia may be due to stimulation of the hypothalamus or the third cranial nerve. Anticipation of these cardiovascular changes during the relevant steps of the operation can help in taking appropriate corrective action, thus preventing potentially serious complications of ETV.

Effect of combining viscous fibre with lovastatin on serum lipids in normal human subjects.

OBJECTIVES: Soluble fibre supplements are recommended to reduce the levels of low-density lipoprotein cholesterol (LDL-C). Limited information exists on the interaction between fibre and hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins). The purpose of the present study was to evaluate the per se effect of psyllium (10 g/day) and lovastatin (20 mg/day) alone and in combination on serum lipids in normal human volunteers. METHODS: In a 4-week open label, randomised, parallel study, subjects were randomised to receive 20 mg of lovastatin, 10 g of psyllium or 20 mg of lovastatin plus 10 g of psyllium in evening daily. Levels of total cholesterol (TC), LDL-C, high-density lipoprotein cholesterol (HDL-C), TC/LDL-C ratio, LDL-C/HDL-C ratio and triglycerides were determined after 1, 2, 3 and 4 weeks of treatment. RESULTS: The study group comprised 36 adult, male subjects. All treatments were well tolerated, and after 4 weeks the mean LDL-C, TC and TG levels in the group receiving 20 mg of lovastatin plus 10 g of psyllium fell by 30.88%, 26.88% and 26.21% from baseline, compared with 24.78%, 19.55% and 32.88% in the group receiving 20 mg of lovastatin and 3.58%, 2.90% and 10.95% in the group receiving 10 g of psyllium respectively. Although additive effect was observed in the group receiving combination compared with group receiving lovastatin, the observed difference was not statistically significant. No significant changes from baseline in HDL-C levels occurred. CONCLUSIONS: Psyllium soluble fibre should be considered as a safe and well-tolerated dietary supplement option to enhance cholesterol lowering.

Comparative evaluation of myolaxin and EMLA cream for attenuation of venous cannulation pain: a prospective, randomised, double blind study.

We evaluated the efficacy of topical Myolaxin (capsaican ointment, Geno, Mumbai) ointment over EMLA (eutectic mixture of lignocaine, prilocaine; Neon, Goa) cream for attenuating venous cannulation pain in this prospective, randomised, double blind study. Sixty adult patients undergoing elective laparoscopic cholecystectomy were randomly assigned into two equal groups. Group I (EMLA) received EMLA cream, whereas Group II (Myolaxin) received Myolaxin ointment. For both groups the cream was applied at the venous cannulation site (dorsum of the non-dominant hand) one hour prior to venous cannulation and was covered with an occlusive dressing. Following venous cannulation patients were asked if they felt pain during venous cannulation. If the answer was yes, they were asked to rate the severity of venous cannulation pain using a VisualAnalogue Scale (VAS) of 0-10. The incidence of venous cannulation pain was similar between groups: in the EMLA group 65% (18/28) compared to 67% (20/30) in the Myolaxin group (P=0.19). The severity of pain (median VAS with inter quartile range) was also similar between the groups: in the EMLA group 1.5 (3) compared to 1.5 (2) in the Myolaxin group (P=0.46). As the topical application of Myolaxin ointment is cheaper than EMLA and has similar efficacy, it may be a suitable alternative for reducing the incidence and severity of venous cannulation pain.

Achalasia cardia: A study of 113 patients managed with indigenous dilator.

BACKGROUND: Endoscopic dilatation of achalasia cardia is an effective nonsurgical management option. It requires costly pneumatic dilators which are used under fluoroscopic guidance. This study assesses the efficacy and safety of an indigenous pneumatic dilator used without fluoroscopic guidance. METHODS: Over a period of eleven years, 113 patients (69M, 44F) ofachalasia cardia underwent dilatation with indigenous pneumatic dilators without fluoroscopic guidance. The dilatation was performed under endoscopic vision. RESULTS: The procedure was successful in all patients. After six weeks following dilatation, there was significant improvement in the mean dysphagia score 3.63 + 0.61 to 0.53 + 0.93 (P<0.01). The response was still significant (0.78 + 1.03, P <0.05) at the end of one year. Excellent response with single dilatation was seen in 70.7% patients. After two dilatation sessions 92% of patients showed an excellent response. One patient had perforation. There was no mortality. CONCLUSION: Pneumatic dilatation under endoscopic vision without fluoroscopic assistance with the indigenous dilator is very effective and safe for short term treatment of achalasia carida.