The BRAF and NRAS mutation prevalence in dermoscopic subtypes of acquired naevi reveals constitutive mitogen-activated protein kinase pathway activation.

BACKGROUND: Acquired naevi can have unique dermoscopic patterns that correspond to distinct microanatomical growth patterns. Previous studies on acquired naevi stratified according to dermoscopic pattern focused on the frequency of somatic BRAF mutations, whereas NRAS mutations remained to be elucidated. OBJECTIVES: To investigate the BRAF and NRAS mutation prevalence and activation of the mitogen-activated protein kinase (MAPK) pathway in distinct dermoscopic subtypes of acquired naevi. METHODS: Common mutations present in BRAF and NRAS were assessed in 40 globular, reticular and peripheral rim of globules (PG) subtypes of acquired naevi from 27 participants (19 male, 8 female; mean age 46·7 years) selected from 1261 eligible volunteers. Mutations were determined using the highly sensitive and quantitative QX200 droplet digital™ polymerase chain reaction (ddPCR) system. RESULTS: The BRAF V600E (c.1799T>A or c.1799_1800delTGinsA) and BRAF V600K mutations were detected in 85% (n = 34/40) of naevi. All BRAF wild-type naevi (15%; n = 6/40) harboured an NRAS codon 12/13 or 61 mutation. BRAF mutations were present in 92% (n = 12/13) of globular and 100% (n = 12/12) of PG naevi, whereas reticular naevi were 67% (n = 10/15) BRAF- and 33% (n = 5/15) NRAS-mutant (P = 0·037). CONCLUSIONS: We discovered that 100% of the assessed acquired naevi had either a BRAF or NRAS mutation. Using sensitive techniques capable of single-cell mutation detection, it is likely that all acquired naevi will be mutated for BRAF or NRAS. Because both of these mutations are prevalent in distinct dermoscopic naevus subsets, our study supports the role of the MAPK pathway in the development of benign melanocytic proliferations, indicating that additional genomic events besides somatic mutations in BRAF or NRAS are required for melanoma development.

Clinical audit training improves undergraduates' performance in root canal therapy.

AIM: To evaluate the effectiveness of clinical audit-feedback cycle as an educational tool in improving the technical quality of root canal therapy (RCT) and compliance with record keeping performed by dental undergraduates. METHODS: Clinical audit learning was introduced in Year 3 of a 5-year curriculum for dental undergraduates. During classroom activities, students were briefed on clinical audit, selected their audit topics in groups of 5 or 6 students, and prepared and presented their audit protocols. One chosen topic was RCT, in which 3 different cohorts of Year 3 students conducted retrospective audits of patients' records in 2012, 2014 and 2015 for their compliance with recommended record keeping criteria and their performance in RCT. Students were trained by and calibrated against an endodontist (κ ≥ 0.8). After each audit, the findings were reported in class, and recommendations were made for improvement in performance of RCT and record keeping. Students' compliance with published guidelines was presented and their RCT performances in each year were compared using the chi-square test. RESULTS: Overall compliance with of record keeping guidelines was 44.1% in 2012, 79.6% in 2014 and 94.6% in 2015 (P = .001). In the 2012 audit, acceptable extension, condensation and the absence of mishap were observed in 72.4, 75.7% and 91.5%; in the 2014 audit, 95.1%, 64.8% and 51.4%; and in 2015 audit, 96.4%, 82.1% and 92.8% of cases, respectively. In 2015, 76.8% of root canal fillings met all 3 technical quality criteria when compared to 48.6% in 2014 and 44.7% in 2012 (P = .001). CONCLUSION: Clinical audit-feedback cycle is an effective educational tool for improving dental undergraduates' compliance with record keeping and performance in the technical quality of RCT.

Influenza B outbreak in female psychiatric ward of Hospital Kuala Lumpur, Malaysia.

Influenza outbreaks in tropical countries are rarely reported. This article reports four cases of influenza within a psychiatric ward of a tertiary hospital in Malaysia. These were patients with severe mental illness who were involuntarily admitted and did not show the classical triad of influenza-like-illness (ILI) at the beginning. However, severe respiratory complications developed requiring intubation. Referral and cooperation with the infectious disease team was initiated to help manage the outbreak while continuing psychiatric treatment. Incidences of influenza among hospitalised psychiatric patients should be treated seriously with immediate multidisciplinary approach to prevent severe unwanted complications.

[Risk factors for acute rejection in living-donor kidney transplant recipients in China: a subgroup analysis of a multi-center, registry study].

Objective: To evaluate pre-and early post-transplantation risk factors for acute rejection(AR) in kidney recipients. Methods: This subgroup analysis of a multi-center registry study was conducted on living-donor kidney transplant recipients in China with 10 years of follow-up. This study analyzed 1 255 recipients including 921 males(73.4%) and with a mean age of (33±10)years. Data from patients were first analyzed with univariate analysis and then multivariate analysis was used for finding out the potential risk factors of AR. Results: A total of 106(8.4%) patients were suspected with AR after kidney transplantation, while 1 149 patients were considered as non-AR. Multivariable analysis demonstrated a significant influence of recipient age and cold ischemia time(CIT) on the occurrence of AR(OR: 0.956, 95% CI: 0.923-0.990; OR: 1.006, 95% CI: 1.002-1.011, respectively). The frequency of severe infection was significantly higher in the AR group than non-AR group(38.7% vs 10.8%; P<0.000 1). The occurrence of new-onset diabetes mellitus and tumors was similar in the two groups. Conclusions: Recipient age and CIT are risk factors for AR after living-donor kidney transplantation. Reducing CIT and intensive management of younger recipient could benefit kidney transplant patients.

A randomized, phase IIa exploratory trial to assess the safety and preliminary efficacy of LEO 43204 in patients with actinic keratosis.

BACKGROUND: LEO 43204 is a novel ingenol derivative in development for the treatment of actinic keratosis. OBJECTIVES: To compare the safety and preliminary efficacy of three doses of LEO 43204 with ingenol mebutate in actinic keratoses (AKs). METHODS: Patients with at least three visible, discrete, nonkeratotic AKs on four separate selected treatment areas on the forearms received LEO 43204 gel (0·025%, 0·05% and 0·075%) and ingenol mebutate 0·05% gel, by investigator-blinded, randomized allocation, for 2 consecutive days. Patients were assessed at 8 weeks. Primary outcomes included maximum composite local skin response (LSR) score and adverse events (AEs). Secondary outcomes included a reduction in the number of visible AKs. RESULTS: Forty patients completed the trial. For all treatments, mean LSR scores peaked at week 1, and were below baseline by week 8. Mean maximum composite LSR scores for LEO 43204 0·025%, 0·05% and 0·075% were 9·2 (Dunnett adjusted P = 0·02), 10·1 (Dunnett adjusted P = 0·90) and 11·2 (Dunnett adjusted P < 0·01), respectively, vs. ingenol mebutate 0·05% gel (10·0). The most frequent AEs across all treatments were application site pruritus, burning sensation and tenderness. Mean reduction in the number of AKs was comparable for ingenol mebutate and the two lowest doses of LEO 43204 (71·9-73·1%), but LEO 43204 0·075% gave a significantly larger reduction (81·8%; Dunnett adjusted P = 0·04). CONCLUSIONS: LEO 43204 had a similar safety profile to ingenol mebutate and a dose-response relationship for LSRs was demonstrated. The highest LEO 43204 dose (0·075%) significantly reduced the AK count when compared with ingenol mebutate.

Histopathology and reflectance confocal microscopy features of photodamaged skin and actinic keratosis.

BACKGROUND: Actinic keratosis (AK) usually co-exists in areas of severe photodamage, but the clinical applicability of reflectance confocal microscopy (RCM) in diagnosing AK currently depends on a set of parameters yet to be defined in comparison to photodamaged skin (PD). OBJECTIVE: To correlate the RCM features of PD and AK with histopathology. METHODS: Twenty participants with a mean age of 64 years and skin phototype I and II were studied. RCM was performed on two PD and one AK within a field of 25 cm2 on the left dorsal forearm, followed by shave biopsies. Blinded evaluation of the histopathological and RCM images using established parameters in AK were performed retrospectively in consensus with an expert confocalist, correlated with the histopathological diagnosis by a board-certified dermatopathologist. RESULTS: A total of 57/60 areas were included. There were 43/57 (75%) and 14/57 (25%) histopathologically confirmed PD and AK respectively. Individual corneocytes, stratum corneum disruption, dermal inflammatory cells, increased vascularity/dilated vessels and solar elastosis were detected in PD and AK upon histopathology and RCM. The features in favour of AK were parakeratosis, hyperkeratosis, more severe keratinocyte pleomorphism and architectural disruption, and the presence of epidermal inflammatory cells. PD also demonstrated keratinocyte pleomorphism and architectural disruption though this was generally less severe than AK. A small subset of PD exhibited a comparable degree of keratinocyte pleomorphism and architectural disruption to the AKs in the cohort. CONCLUSIONS: The viable epidermis demonstrates PD and AK to be part of a disease continuum corresponding to field cancerization. Individual corneocytes, stratum corneum disruption, dermal inflammatory cells, increased vascularity/dilated vessels and solar elastosis may be present in PD; whereas, parakeratosis and hyperkeratosis may represent the key to distinguishing AK from PD using RCM. The significance of epidermal inflammatory cells in the RCM diagnosis of AK remains to be elucidated.

Lack of clinical significance of the ImmuKnow(TM)-Cylex assay for the detection of cellular immune function in patients with renal cell carcinoma.

This study aimed to explore the clinical value of the CD4(+) T cell ATP levels in patients with renal cell carcinoma through the application of the ImmuKnow(TM)-Cylex(®) assay. We recruited 104 patients with renal cancer who had undergone surgery at Fuzhou General Hospital from March 2009 to June 2012, and were subsequently treated by dendritic cell and cytokine-induced killer cell bio-therapy or interferon-α therapy. The changes in CD4(+) T cell ATP levels were detected at the perioperative period and at 10 days, 1 month, 3 months, and 1 year after the surgery using the ImmuKnow assay. In addition, the differences in ATP levels in different therapy groups were compared and the prognosis conditions were analyzed. Our results demonstrated that no significant difference in the ATP levels occurred at different time points; furthermore, there were no obviously different ATP levels between the different therapy groups, and the ATP levels were found to have no clinical significance for the assessment of renal cancer prognosis. Overall, this study suggested that CD4(+) T cell ATP levels as detected by the ImmuKnow assay have no obvious clinical value in patients with renal cancer.

Foot posture is associated with morphometry of the peroneus longus muscle, tibialis anterior tendon, and Achilles tendon.

The aim of this study was to investigate the association between foot type and the morphometry of selected muscles and tendons of the lower limb. Sixty-one healthy participants (31 male, 30 female; aged 27.1 ± 8.8 years) underwent gray-scale musculoskeletal ultrasound examination to determine the anterior-posterior (AP) thickness of tibialis anterior, tibialis posterior, and peroneus longus muscles and tendons as well as the Achilles tendon. Foot type was classified based on arch height and footprint measurements. Potentially confounding variables (height, weight, hip and waist circumference, rearfoot and ankle joint range of motion, and levels of physical activity) were also measured. Multiple linear regression models were used to determine the association between foot type with muscle and tendon morphometry accounting for potentially confounding variables. Foot type was significantly and independently associated with AP thickness of the tibialis anterior tendon, peroneus longus muscle, and Achilles tendon, accounting for approximately 7% to 16% of the variation. Flat-arched feet were associated with a thicker tibialis anterior tendon, a thicker peroneus longus muscle, and a thinner Achilles tendon. Foot type is associated with morphometry of tendons that control sagittal plane motion of the rearfoot; and the peroneus longus muscle that controls frontal plane motion of the rearfoot. These findings may be related to differences in tendon loading during gait.

Gene expression signature analysis and protein-protein interaction network construction of spinal cord injury.

BACKGROUND: The aim of this study was to investigate the gene expression profile of thoracic propriospinal neurons between post-injury rat and controls. MATERIALS AND METHODS: Microarray dataset GSE20907 was downloaded from GEO database, including 12 Spinal Cord Injury (SCI) rat and 12 controls. Student's t test was employed to identified differentially expressed genes with a fold-change > 1.2. Then, we used DAVID to perform functional enrichment analysis to uncover dysfunctional biological processes and molecular signatures database (MsigDB) to find any potential relationship between SCI gene expression signature and other published gene expression signature. Protein-Protein interaction (PPI) network was constructed by STRING and visualized in Cytoscape. Functional analysis of the hub protein was performed by BinGO. RESULTS: The maximum change of gene expression profile was found at 3-days post injury and immune response was found upregulated all tested time points. Interestingly, genes upregulated 2-weeks post injury was found significantly overlapped with genes upregulated in brains from Alzheimer's disease patients. Protein interaction network analysis found that LYN, PTPN6 and SMAD1 could be of great value for further investigation. CONCLUSIONS: It could be inferred that understanding the underlying molecular mechanism post injury, especially at early moment, may provide novel insight for development of therapeutics strategy.

One year results of preoperative single bolus ATG-Fresenius induction therapy in sensitized renal transplant recipients.

UNLABELLED: Sensitization in kidney transplantation is associated with more acute rejections, inferior graft survival, and an increase in delayed graft function. This study was designed to evaluate the efficacy and safety of preoperative single bolus antithymocyte globulin (ATG) induction therapy in sensitized renal transplant recipients. METHODS: Fifty-six cadaveric donor kidney transplant recipients were divided into two groups: Group I (nonsensitized group, n = 30) and group II (sensitized group, PRA>10%, n = 26). ATG was given as a single preoperative bolus induction therapy to group II (ATG IV; 9 mg/kg). The group I patients were treated with mycophenolate mofetil preoperatively as induction therapy. The basic immunosuppressive regimen included tacrolimus (FK-506) or cyclosporine, mycophenolate mofetil, and prednisolone. After hospital discharge, patients were followed on a routine outpatient basis for 12 months. RESULTS: Acute rejection episodes (ARE) occurred in 20% (6/30) of group I and 15.38% (4/26) of group II patients (P = NS). Infections occurred in eight patients (26.7%) as 11 episodes (36.7%), averaging 1.4 episodes per infected patient in group 1, and 6 patients (23.1%) for a total of 10 episodes (38.5%), averaging 1.7 episodes per infected patient, in group II (P = NS). Occurrence of side effects and hospital stay were almost comparable in the two groups. No delayed graft function was observed in either group. The 12-month actuarial patient and graft survival were 100% in Group I and II. CONCLUSION: A preoperative single bolus ATG induction therapy was an effective and safe therapeutic measure, yielding an acceptable acute rejection rate in presensitized renal transplant recipients.

Strength of donor-specific antibodies with the use of Luminex single-antigen beads is a reliable predictor of acute rejection in living-relative kidney recipients.

BACKGROUND: Luminex single-antigen assay is a promising technique for determination of donor-specific antibodies (DSAs) in renal transplantation. METHODS: Serum samples from living-relative renal recipients before and after renal transplantation were examined with the use of Luminex single-antigen assay. The impact of de novo DSAs on the early clinical outcomes of renal allograft was analyzed. RESULTS: Of the 61 patients included, 15 patients (24.6%) presented de novo DSA (4 class I, 4 class II, and 7 both classes) after transplantation, and the average (median) cumulative strength of DSA was 1,283 (range, 0-10,802) mean fluorescence index (MFI) for class I versus 1,324 (range, 0-14,985) MFI for class II (P > .05). Twelve (19.7%) of the 61 patients experienced a clinical/subclinical acute rejection (AR) episode within the 1st 2 years after transplantation. A clinical/subclinical AR episode was diagnosed in 40% of DSA(+) patients and 13.0% of DSA(-) patients (P < .05). DSA(+) patients who developed an AR episode had a higher mean cumulative MFI value (8,118.3 ± 5,287.4; range, 1,785-14,985) than patients who did not develop an AR episode (3,283.7 ± 2,601.0; range, 786-8,113; P < .05). Serum creatinine levels in the DSA(+) group were significantly higher than in the DSA(-) group at 12 and 24 months after transplantation. The graft survival rates at 2 years in the DSA(+) and DSA(-) groups were not different (86.7% vs 91.3%; P > .05). CONCLUSIONS: Patients with de novo DSAs at high strength may suffer a high risk of developing an AR episode. Therefore, careful monitoring of de novo DSAs with the use of Luminex single-antigen beads may help in early intervention for AR in renal allografts and to minimize renal damage caused by the antibodies.

Preoperative intravenous tramadol versus ketorolac for preventing postoperative pain after third molar surgery.

The objective of this study was to compare the analgesic efficacy of a single-dose of preoperative intravenous tramadol versus ketorolac in preventing pain after third molar surgery. Sixty-four patients undergoing elective third molar surgery were randomly assigned into one of the two groups (32 in each group): Group I received tramadol 50 mg, and Group 2 received ketorolac 30 mg intravenously preoperatively before the surgery. After injection of the study drugs, a standard intravenous sedation technique was administered and the impacted third molars were removed under local anaesthetic. The difference in postoperative pain was assessed by four primary end-points: pain intensity as measured by a 100-mm visual analogue scale hourly for 12 h, median time to rescue analgesic, postoperative acetaminophen consumption, and patient's global assessment. Throughout the 12-h investigation period, patients reported significantly lower pain intensity scores in the ketorolac versus tramadol group (P = 0.05, Mann-Whitney U-test). Patients also reported significantly longer median time to rescue analgesic (9.0 h versus 7.0 h, P = 0.007, log rank test), lesser postoperative acetaminophen consumption (P = 0.02, Mann-Whitney U-test) and better global assessment (P = 0.01, chi2 test) for the ketorolac versus tramadol group. Preoperative intravenous ketorolac 30 mg is more effective than tramadol 50 mg in the prevention of postoperative dental pain.

A prospective randomized crossover study of the preemptive analgesic effect of nitrous oxide in oral surgery.

OBJECTIVE: Preliminary animal data has shown that nitrous oxide has a preemptive analgesic effect on postoperative pain. Whether a similar effect occurs in humans is not established. In this prospective randomized crossover study, we investigated the effect of preincisional versus postincisional nitrous oxide on postoperative oral surgical pain.Study design The trial was a crossover study where 36 patients had each of their symmetrical impacted mandibular third molars randomly scheduled for removal in 2 sessions. Each of the 36 patients acted as his or her own control; one side of the jaw was allocated randomly to receive nitrous oxide preoperatively (pretreated side) and the other side postoperatively (posttreated side). The pretreated side received 50% nitrous oxide preoperatively for 20 minutes and 100% oxygen postoperatively for 20 minutes as placebo. The posttreated side received 100% oxygen preoperatively for 20 minutes and 50% nitrous oxide postoperatively for 20 minutes. The difference in postoperative pain between the pretreated and posttreated sides was assessed by 4 primary end-points: pain intensity as measured by a 100-mm visual analog scale (VAS) hourly for 8 hours, time to first analgesic, total analgesic consumption during the first 48 hours, and a 5-point categorical patient global assessment scale (0=poor, 1=fair, 2=good, 3=very good, and 4=excellent). RESULTS: The VAS scores did not differ between the 2 sides at any time (P=.50): neither did the time to first analgesic (P=.8), amount of total analgesic consumption (P=.77), and patient's global assessment differ (P=.63). CONCLUSION: Our results do not support the preliminary animal data that nitrous oxide has a preemptive analgesic effect for postoperative pain. 50% nitrous oxide administered preoperatively for 20 minutes has no preemptive analgesic effect on postextraction pain.

Comparison of patient-controlled sedation with propofol and alfentanil for third molar surgery--preliminary results of a pilot study.

Propofol and alfentanil are commonly used for sedation and pain control. A preliminary study to compare the usefulness of these drugs was carried out in ten healthy patients requiring bilateral wisdom tooth surgery. The operations were done in two appointments with the patient receiving a different drug on each occasion. Anxiety levels were recorded on visual analogue scales pre and post-operatively. Both agents caused a decrease in anxiety scores, with propofol causing a more significant reduction. Vomiting and nausea with alfentanil was noted in three patients. Propofol also had an amnesic effect which alfentanil did not have. In conclusion, propofol would appear to be the drug of choice within the limitations of this pilot study.

Use of sedation in dentistry.

Fear and avoidance of dental treatment are major deterrents to oral health. Sedation can be used to control both the patient's fear and anxiety so that proper dental care can be provided for these patients. The purpose of this article is to discuss the use of sedation in dentistry and to provide a recommendation on the requirements and medico-legal aspects of sedation for the practitioner interested in incorporating sedation into their practice.

Differential expression of splice variant and wild-type parkin in sporadic Parkinson's disease.

BACKGROUND: Altered splicing of parkin under cellular stress could lead to changes in gene expression and altered protein activity. The causative role of parkin in sporadic Parkinson's disease (PD) is unknown. OBJECTIVES: We described a parkin splice variant (SV) in the substantia nigra and leukocytes of sporadic PD patients. Using a case control methodology, we investigated the exon 4 SV (E4SV) and wild-type parkin expression in the leukocytes of sporadic PD patients and healthy individuals. METHODS/RESULTS: We identified a parkin E4SV in the substantia nigra and leukocytes of sporadic PD patients and controls by reverse transcriptase-polymerase chain reaction (PCR). The exon 4 (122 bp) deletion resulted in a reading frame shift over the junction of exons 3-5 and a stop codon (tga) 17 bp downstream from exon 3. The translated truncated protein was associated with a total loss of the two-RING finger functional domain. Utilizing TaqMan real-time PCR with probes located across the junction of exons 3-4 or 3-5, we demonstrated an over-expression of E4SV/wild-type parkin ratio in the leukocytes of sporadic PD patients compared to age-, gender-, and race-matched controls (p<0.0005). A multivariate regression analysis demonstrated that the ratio of E4SV/wild-type parkin expression increased with age in PD patients, but this was not observed in the controls (p<0.0005). CONCLUSION: The relative expression of E4SV/wild type parkin was increased in sporadic PD compared to healthy controls. Based on our observations, further functional studies to determine the pathophysiologic role of E4SV in sporadic PD patients will be of importance.

The novel human MOST-1 (C8orf17) gene exhibits tissue specific expression, maps to chromosome 8q24.2, and is overexpressed/amplified in high grade cancers of the breast and prostate.

AIMS: To elucidate genes that participate in the process of oncogenesis, primers based on the E6 genes of genital human papillomaviruses (HPVs) were used to amplify potential expressed sequence tags (ESTs) from the MOLT-4 T lymphoblastic leukaemia cell line. METHODS: Using the polymerase chain reaction (PCR) with human papillomavirus E6 gene primers, an EST from the MOLT-4 T lymphoblastic leukaemia cell line was amplified. Via rapid amplification of cDNA ends (RACE) and cycle sequencing from MOLT-4 and fetal lung cDNA libraries, overlapping cDNAs of 2786 bp and 2054 bp of the corresponding novel human intronless gene designated MOST-1 (for MOLT-4 sequence tag-1) were characterised and assigned the symbol C8orf17 by the HUGO Nomenclature Committee. RESULTS: Both cDNAs contained a potential open reading frame (ORF) of 297 bp incorporating a methionine codon with an ideal Kozak consensus sequence for translation initiation, and encoding a putative hydrophilic polypeptide of 99 amino acids. Although reverse transcription PCR (RT-PCR) demonstrated MOST-1 expression in all 19 cancer and two normal cell lines tested, differential expression was seen in only nine of 16 normal tissues tested (heart, kidney, liver, pancreas, small intestine, ovary, testis, prostate, and thymus). A 388 bp fragment was amplified from the NS-1 mouse myeloma cell line, the sequence of which was identical to that within the MOST-1 ORF. The MOST-1 gene was mapped by fluorescent in situ hybridisation to chromosome 8q24.2, a region amplified in many breast cancers and prostate cancers, which is also the candidate site of potential oncogene(s) other than c-myc located at 8q24.1. Analysis of paired biopsies of invasive ductal breast cancer and adjacent normal tissue by semiquantitative and real time RT-PCR revealed average tumour to normal ratios of MOST-1 expression that were two times greater in grade 3 cancers than in grade 1 and 2 cancers. Quantitative real time PCR of archival prostatic biopsies displayed MOST-1 DNA values that were 9.9, 7.5, 4.2, and 1.4 times higher in high grade carcinomas, intermediate grade carcinomas, low grade carcinomas, and benign hyperplasias, respectively, than in normal samples. CONCLUSIONS: These data suggest a role for MOST-1 in cellular differentiation, proliferation, and carcinogenesis.