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.

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.

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.

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.

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.

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.

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.

alpha-Trifluoromethyl-beta-alanyl glycine (F3MBAG): a novel mammalian metabolite of trifluridine (F3TdR).

19F NMR spectroscopy has been used to study further the metabolism of 5-trifluoromethyl-2'-deoxyuridine (trifluridine; F3TdR). The synthesis and characterization of alpha-trifluoromethyl-beta-alanyl glycine (F3MBAG), a putative new metabolite of F3TdR, are now reported. This study describes ex vivon and in vivo detection of F3MBAG and other previously reported metabolites of trifluridine, using 19F NMR spectroscopy, in male BALB/C mice bearing EMT-6 tumors. A parallel 19F NMR spectroscopic study was also performed on rats dosed with F3TdR, to observe the qualitative pattern of F3TdR metabolism in another species. Unexpectedly, 5-trifluoromethyl-5,6-dihydroxyuracil (DOHF3T), alpha-trifluoromethyl-beta-ureidopropionic acid (F3MUPA) and fluoride, which result from the metabolic degradation of F3TdR and which were detected in various biological samples from mice dosed with F3TdR, could not be identified in rat urine or in homogenized tissue extracts. The presence of these metabolites in intact tissues is uncertain since in this study 19F NMR spectroscopy of these samples always displayed a broad resonance "hump" across the range of chemical shifts that would encompass these metabolites. No clear explanation for the loss of spectroscopic resolution in this region has been rationalized. N-Carboxy-alpha-trifluoromethyl-beta-alanine (F3MBA-CO2), alpha-trifluoromethyl-beta-alanyl alanine (F3MBAA) and N-acetyl-alpha-trifluoromethyl-beta-alanine (Ac-F3MBA) were synthesized and characterized, but were not detected as metabolites in any of the biological specimens examined.

Detection of new metabolites of trifluridine (F3TdR) using 19F NMR spectroscopy.

The metabolism of 5-trifluoromethyl-2'-deoxyuridine (trifluridine, F3TdR) in male BALB/C mice has been studied by 19F NMR spectroscopy. Contrary to previous reports, a number of fluorinated metabolites were observed in urine, whole livers and blood samples taken from mice after i.p. injection of F3TdR. The present study describes the identification of two new metabolites in mouse urine using the 19F NMR technique. The NMR of crude urine showed the presence of F3TdR 5-trifluorothymine (F3T), the newly-identified metabolites, 5-trifluoromethyl-5,6-dihydrouracil (DHF3T) and 5-trifluoromethyl-5,6-dihydroxyracil (DOHF3T), and several new, as yet unidentified fluorinate metabolites. These two new metabolites were characterized by comparison to authentic compounds prepared synthetically from F3T.

Synthesis and antitumour effect of the melanogenesis-based antimelanoma agent N-propionyl-4-S-cysteaminylphenol.

Chemotherapy of malignant melanoma is still a great challenge, as no effective drugs are available. The development of melanogenesis-based drugs is a promising area of research because melanogenesis is a unique biochemical pathway operating only in melanoma cells (and their normal counterparts) so that the tumour can be targeted. We have been using cysteinylphenol, a sulphur-containing analogue of tyrosine, and derivatives for that purpose. N-Acetyl-4-S-cysteaminylphenol was found to have the best antimelanoma effect in cell culture systems and in mice bearing B16 melanoma tumours. It also caused depigmentation of the skin, suggesting the possibility of use as a hypopigmenting agent. To improve the efficiency of the drug, we thought of replacing the acetyl group in N-acetyl-4-S-cysteaminylphenol with a propionyl group in the hope that increased hydrophobicity would increase the cellular uptake of the drug. N-Propionyl-4-S-cysteaminylphenol was synthesized by condensing 4-hydroxythiophenol with 2-ethyl-2-oxazoline. The drug showed both cytostatic and cytocidal effects in a human melanotic melanoma cell line. The drug was found to be a good depigmenting agent for the black hair follicles of C57 black mice when given s.c. for 14 days. A 10-day treatment with N-propionyl-4-S-cysteaminylphenol at 300 mg/kg body weight reduced the growth rate of B16 melanoma s.c. tumours in mice by 36%. The propionyl derivative was found to increase the life span of mice bearing melanoma more effectively than did the acetyl derivative.

Plexogenic angiopathy in pulmonary intralobar sequestrations: pathogenetic mechanisms.

Vascular remodeling to form plexiform (glomoid) lesions is a little-known manifestation of intralobar pulmonary sequestration. We describe histologic and immunohistochemical features of these lesions in resected specimens from three subjects aged 3, 19, and 28 years. The vascular changes, which included medial and intimal thickening, angioblastic proliferation, plexiform lesions, and dilation lesions, occurred in a setting of hypoxia, chronic inflammation, and high pressure and flow via a systemic arterial supply. We demonstrated strong immunoreactivity of the angioblastic tissue and the plexiform lesions with antibodies to muscle actin, alpha-smooth muscle actin, and vimentin, and weak to absent reactivity with antibody to desmin. We suggest that in these sequestrations plexiform lesions develop via angioblastic proliferation at arterial branch points and that dilation lesions develop from subsequent expansion of distal anastomoses.

Cardiopulmonary effects of fenoterol and salbutamol aerosols.

Comparative evaluation of the bronchodilator and cardiovascular effects of cumulative dose of fenoterol and salbutamol (albuterol) aerosols was carried out in an open crossover study on 15 patients with chronic stable reversible airways obstruction. On a weight for weight basis, there was no significant difference in the bronchodilator efficacy of fenoterol and salbutamol; though the heart rates after fenoterol were significantly higher than after salbutamol. Ventricular dysrhythmias requiring premature cessation of the study in four patients were seen with fenoterol; whereas no significant ventricular dysrhythmias were observed with salbutamol. It is concluded that salbutamol has lesser cardiostimulatory effects than fenoterol.

Double-blind crossover study of nedocromil sodium in partially reversible chronic obstructive airways disease.

The efficacy of 8 weeks of treatment with 4 mg of nedocromil sodium 4 times daily in the management of partially reversible chronic airflow obstruction was evaluated in a randomized, double-blind, placebo-controlled crossover trial at Repatriation General Hospital, Western Australia. Sixty-seven patients with a mean forced expiratory volume in 1 s (FEV1) of 0.99 L, which improved by 21.8 percent after beta 2-adrenergic bronchodilator, took part in the full 24-week study. After a 4-week run-in period, in addition to their regular therapy comprised of inhaled or oral bronchodilators and steroids (which remained unchanged), patients were randomly allocated to receive treatment with nedocromil sodium or placebo for 8 weeks. This was followed by a 4-week washout, and then the patients were given the alternative test treatment to that given in the first period for another 8 weeks. Compared with placebo, patients receiving nedocromil sodium had improvement in their morning and evening peak flow rates, symptom scores for cough, sleep disturbance, and overall treatment efficacy. Hence, treatment with nedocromil sodium should be considered for patients with partially reversible chronic airflow obstruction not fully controlled with bronchodilators and steroids.