Erector spinae plane block vs. peri-articular injection for pain control after arthroscopic shoulder surgery: a randomised controlled trial.

Interscalene brachial plexus block is the standard regional analgesic technique for shoulder surgery. Given its adverse effects, alternative techniques have been explored. Reports suggest that the erector spinae plane block may potentially provide effective analgesia following shoulder surgery. However, its analgesic efficacy for shoulder surgery compared with placebo or local anaesthetic infiltration has never been established. We conducted a randomised controlled trial to compare the analgesic efficacy of pre-operative T2 erector spinae plane block with peri-articular infiltration at the end of surgery. Sixty-two patients undergoing arthroscopic shoulder repair were randomly assigned to receive active erector spinae plane block with saline peri-articular injection (n = 31) or active peri-articular injection with saline erector spinae plane block (n = 31) in a blinded double-dummy design. Primary outcome was resting pain score in recovery. Secondary outcomes included pain scores with movement; opioid use; patient satisfaction; adverse effects in hospital; and outcomes at 24 h and 1 month. There was no difference in pain scores in recovery, with a median difference (95%CI) of 0.6 (-1.9-3.1), p = 0.65. Median postoperative oral morphine equivalent utilisation was significantly higher in the erector spinae plane group (21 mg vs. 12 mg; p = 0.028). Itching was observed in 10% of patients who received erector spinae plane block and there was no difference in the incidence of significant nausea and vomiting. Patient satisfaction scores, and pain scores and opioid use at 24 h were similar. At 1 month, six (peri-articular injection) and eight (erector spinae plane block) patients reported persistent pain. Erector spinae plane block was not superior to peri-articular injection for arthroscopic shoulder surgery.

An optimal saponification and extraction method to determine carotenoids in avocado.

Quantification of carotenoids in avocado fruit is a great challenge due to their co-extraction with high-oil concentration and the inherent nature of carotenoids to degrade and undergo cis/trans photoisomerization with prolonged extraction times and high temperatures. The study provides an optimised and validated methodology for quantification of carotenoids in the high-oil avocado matrix, with > 93% recovery of all carotenoids tested being significantly greater than previously published. Saponification with 15% KOH for 60 min was optimal for the avocado matrix. For the first time, this study identified that soap, produced during the saponification reaction, resulted in a significant reduction of carotenoid content from the avocado matrix, due to the production of micelles. A significantly higher carotenoid content (3.58 versus 2.0 mg/100 g DW) was able to be extracted after saponification with acidified phosphate buffer, instead of water as reported previously. Carotenoid profiles of five avocado cultivars were identified and quantified.

Molecular and cellular evidence for the alternative lengthening of telomeres (ALT) mechanism in chicken.

Telomere maintenance is an important genetic mechanism controlling cellular proliferation. Normally, telomeres are maintained by telomerase which is downregulated upon cellular differentiation in most somatic cell lineages. Telomerase activity is upregulated in immortalized cells and cancers to support an infinite lifespan and uncontrolled cell growth; however, some immortalized and transformed cells lack telomerase activity. Telomerase-negative tumors and immortalized cells utilize an alternative mechanism for maintaining telomeres termed alternative lengthening of telomeres (ALT). This research explored evidence for the ALT pathway in chicken cell lines by studying nontransformed immortalized cell lines (DF-1 and OU2) and comparing them to a normal (mortal) cell line and a transformed cell line (DT40). The research consisted of molecular and cellular analyses including profiling of telomeric DNA (array sizing and total content), telomerase activity, and expression of genes involved in the telomerase, recombination, and ALT pathways. In addition, an immunofluorescence analysis for an ALT marker, i.e. ALT-associated promyelocytic leukemia bodies (APBs), was conducted. Evidence for ALT was observed in the telomerase-negative immortalized cell lines. Additionally, the APB marker was also found in the other cell systems. The attributes of the chicken provide an additional vertebrate model for investigation of the ALT pathway.

Temperature and Maturity Stages Affect Anthocyanin Development and Phenolic and Sugar Content of Purple-Pericarp Supersweet Sweetcorn during Storage.

Purple-pericarp sweetcorn (PPS) is a novel product, requiring both purple pigment development and maintenance of sweetness. Storage period and temperature had a profound impact on total anthocyanin accumulation (TAC) and sugar content. While TAC remained relatively unchanged during 14-day storage at 4 °C, the first recorded observation of continuing accumulation of anthocyanin and phenolic compounds was concurrent with an increase in purple pigment coverage across the surface of the kernel at 23 °C. TAC in PPS significantly increased, doubling after 14 days at 23 °C. Anthocyanin concentration and kernel coverage were also affected by harvest maturity. The results indicated that biosynthesis of anthocyanins is still occurring during postharvest storage of PPS. A significant decline in sugar concentration was also observed during storage with a greater decline at 23 °C. As anthocyanin accumulation and maintaining sweetness are important factors for sweetcorn, identifying storage temperatures that optimize both quality criteria are required.

Optimisation of extraction procedure and development of LC-DAD-MS methodology for anthocyanin analysis in anthocyanin-pigmented corn kernels.

An ultra-high-performance liquid chromatography-diode array detector-mass spectrometry method was developed for characterisation and quantification of anthocyanin components in complex corn-kernel matrices. The anthocyanin profiles and total anthocyanin content (TAC) of mature seeds of five types of anthocyanin-pigmented corn were reported. Internal standard was used to validate the efficiency of extraction and optimise the liquid extraction procedure for anthocyanins. A total of eighteen anthocyanins were identified and quantified. Cyanidin-based glucosides were the major pigments of purple-pericarp sweetcorn (75.5% of TAC) and blue-aleurone maize (91.6%), while pelargonidin-based glucosides composed the main anthocyanins of reddish-purple-pericarp sweetcorn (61.1%) and cherry-aleurone maize (74.6%). Importantly, previous studies reported the presence of acetylated and succinylated anthocyanins in corn kernels; these compounds were found to be artefact pigments, generated during the extraction process. These crucial findings provide the correct anthocyanin profiles of pigmented corns, and emphasise the importance of using acidified solutions for the extraction of corn-based anthocyanins.

Anthocyanin composition and changes during kernel development in purple-pericarp supersweet sweetcorn.

The current study reports the anthocyanin profile of purple 'supersweet' sweetcorn, recently developed from purple Peruvian maize, and the effect of kernel maturity on anthocyanin accumulation. Twenty anthocyanin compounds, consisting of cyanidin-, peonidin-, and pelargonidin-based glucosides, were identified and quantified in purple- and reddish-purple-pericarp sweetcorn accessions. For the first time, four isomers of cyanidin-3-malonylglucoside, four isomers of pelargonidin-3-malonylglucoside and two to three isomers each of cyanidin-3-dimalonylglucoside, peonidin-3-malonylglucoside and pelargonidin-3-dimalonylglucoside, were identified in the new pigmented sweetcorn. While cyanidin-based glucosides predominated in the purple-pericarp accession, pelargonidin-based glucosides predominated in the reddish-purple accession. Total anthocyanin concentration increased significantly (p < 0.05) during the optimum sweetcorn eating period (23 to 28 DAP) and continued to increase as the kernels further matured (>28 DAP). As kernels continued to mature, pigment coverage across the pericarp progressively increased from a small spot at the stigma end of the kernel, to gradually spreading over the entire kernel.

Late anastomotic breakdown with bevacizumab in colorectal cancers, a case-based review.

BACKGROUND: Bevacizumab is the first angiogenesis inhibitor to be approved for metastatic colorectal cancer. Unfortunately, bevacizumab treatment has been associated with a variety of complications including haemorrhage, poor wound healing and gastrointestinal perforation. Late anastomotic breakdown related to bevacizumab therapy however has rarely been described. CASE REPORT: Here, we present the case of a 56-year-old woman who had a bevacizumab-related anastomotic breakdown 17 months following her primary anastomosis. She initially underwent an emergency Hartmann's procedure and two further laparotomies for significant intra-abdominal haemorrhage. Despite the best efforts of the surgical and intensive care teams, ultimately, the patient passed away. DISCUSSION: There is a small but growing body of literature relating to bevacizumab use and late anastomotic breakdown. It would seem prudent to take extra caution when using bevacizumab in those patients with previous pelvic irradiation, who have a rectal site of anastomosis or have experienced a previous anastomotic leak.

A single nucleotide polymorphism in the coding region of ABL and its effects on sensitivity to imatinib.

We have identified a gene polymorphism (K247R) within or close to the P-loop of BCR-ABL, which leads to the substitution of arginine for lysine. We investigated the allelic frequency of K247R by screening 157 CML patients and 213 healthy blood donors with conventional sequencing, restriction enzyme digest and single strand conformational polymorphism analysis, and found the arginine allele to be rare. Three out of five CML patients with the arginine allele of K247R failed to achieve a major cytogenetic response to imatinib, suggesting that the arginine allele may have reduced sensitivity. However, despite K247R's position in or near to the P-loop, biochemical and cellular assays of imatinib and dasatinib sensitivity showed no alteration compared to wild type. Clinicians should be aware that possession of the arginine allele of K247R does not reflect a mutation that necessitates a change in the therapeutic strategy, unless there are other signs of inadequate response to drug.

A Retrospective Cohort Study of Total Colonic Aganglionosis: Is the Appendix a Reliable Diagnostic Tool?

BACKGROUND: Hirschsprung's disease (HD) is characterized by a lack of ganglion cells in the myenteric and submucosal plexus, associated with increased numbers of acetyl cholinesterase (AChE) positive nerve fibres. In approximately 10% of patients with HD the entire colon will be affected; a condition known as Total Colonic Aganglionosis (TCA). Aganglionosis of the appendix has long been considered to be an important finding in a patient in whom TCA is suspected, but its reliability for diagnosis has seldom been discussed. The aim of our study was to assess the reliability of the appendix as a histological specimen for the diagnosis of TCA, and to evaluate the long-term outcome of TCA. METHODS: A retrospective cohort study was performed of all pathological specimens of patients with confirmed HD in our institution between March 2006 and April 2016. RESULTS: Out of a total of 91 patients identified, 15 patients also had histopathological analysis of the appendix. Nine of these cases were confirmed as having TCA. The remaining 6 patients had HD involving the rest of the bowel up to the ascending colon, with normal ganglion present in the caecum. The appendix was removed in all the 15 cases. All 9 patients with confirmed TCA had aganglionosis of the appendix as well. The remaining 6 cases of short and long segment HD not involving the caecum, demonstrated normal ganglion cells within the appendix. CONCLUSION: Aganglionosis of the appendix is a reliable tool in the diagnosis of TCA. The authors recommend that at the time of levelling biopsies, if aganglionosis extends to the mid-transverse colon, an ileostomy be performed and appendix sent for definitive confirmation of TCA. However, at the time of definitive surgery, a frozen section of pull-through segment of bowel is recommended to confirm the presence of ganglion cells.

A role for FOXO1 in BCR-ABL1-independent tyrosine kinase inhibitor resistance in chronic myeloid leukemia.

Chronic myeloid leukemia (CML) patients who relapse on imatinib due to acquired ABL1 kinase domain mutations are successfully treated with second-generation ABL1-tyrosine kinase inhibitors (ABL-TKIs) such as dasatinib, nilotinib or ponatinib. However, ~40% of relapsed patients have uncharacterized BCR-ABL1 kinase-independent mechanisms of resistance. To identify these mechanisms of resistance and potential treatment options, we generated ABL-TKI-resistant K562 cells through prolonged sequential exposure to imatinib and dasatinib. Dual-resistant K562 cells lacked BCR-ABL1 kinase domain mutations, but acquired other genomic aberrations that were characterized by next-generation sequencing and copy number analyses. Proteomics showed that dual-resistant cells had elevated levels of FOXO1, phospho-ERK and BCL-2, and that dasatinib no longer inhibited substrates of the PI3K/AKT pathway. In contrast to parental cells, resistant cells were sensitive to growth inhibition and apoptosis induced by the class I PI3K inhibitor, GDC-0941 (pictilisib), which also induced FOXO1 nuclear translocation. FOXO1 was elevated in a subset of primary specimens from relapsed CML patients lacking BCR-ABL1 kinase domain mutations, and these samples were responsive to GDC-0941 treatment ex vivo. We conclude that elevated FOXO1 contributes to BCR-ABL1 kinase-independent resistance experienced by these CML patients and that PI3K inhibition coupled with BCR-ABL1 inhibition may represent a novel therapeutic approach.

Age-related mutations and chronic myelomonocytic leukemia.

Chronic myelomonocytic leukemia (CMML) is a hematologic malignancy nearly confined to the elderly. Previous studies to determine incidence and prognostic significance of somatic mutations in CMML have relied on candidate gene sequencing, although an unbiased mutational search has not been conducted. As many of the genes commonly mutated in CMML were recently associated with age-related clonal hematopoiesis (ARCH) and aged hematopoiesis is characterized by a myelomonocytic differentiation bias, we hypothesized that CMML and aged hematopoiesis may be closely related. We initially established the somatic mutation landscape of CMML by whole exome sequencing followed by gene-targeted validation. Genes mutated in ⩾10% of patients were SRSF2, TET2, ASXL1, RUNX1, SETBP1, KRAS, EZH2, CBL and NRAS, as well as the novel CMML genes FAT4, ARIH1, DNAH2 and CSMD1. Most CMML patients (71%) had mutations in ⩾2 ARCH genes and 52% had ⩾7 mutations overall. Higher mutation burden was associated with shorter survival. Age-adjusted population incidence and reported ARCH mutation rates are consistent with a model in which clinical CMML ensues when a sufficient number of stochastically acquired age-related mutations has accumulated, suggesting that CMML represents the leukemic conversion of the myelomonocytic-lineage-biased aged hematopoietic system.

The effect of sustained heavy exercise on the development of pulmonary edema in trained male cyclists.

To determine whether intense, prolonged activity can induce transient pulmonary edema, eight highly trained male cyclists (mean +/- S.D.: age, 26.9 +/- 3.0 years; height, 179.9 +/- 5.7 cm; weight, 76.1 +/- 6.5 kg) performed a 45-min endurance cycle test (ECT). V(O2,max) was determined (4.84 +/- 0.4 L min(-1), 63.7 +/- 2.6 ml min(-1) g(-1)) and the intensity of exercise for the ECT was set at 10% below ventilatory threshold (approximately 76% V(O2, max) 300 +/- 25 W). Pre- and post-exercise pulmonary diffusion (DL(CO)) measurements and magnetic resonance imaging of the lung were made. DL(CO) and pulmonary capillary blood volume (VC) decreased 1h post-exercise by 12% (P = 0.004) and 21% (P = 0.017), respectively, but no significant change in membrane diffusing capacity (DM) was found. The magnetic resonance scans demonstrated a 9.4% increase (P = 0.043) in pulmonary extravascular water 90 min post-exercise. These data support the theory that high intensity, sustained exercise in well-trained athletes can result in transient pulmonary edema.

Replacements in the exposed loop of the T15 antibody VH CDR2 affect carrier recognition of PC-containing pathogens.

A panel of mutant antibodies of the phosphocholine (PC)-binding antibody, T15, was tested for binding to PC-protein, Streptococcus pneumoniae, Trichinella spiralis and Ascaris suum. Relative to wildtype T15, all the mutant antibodies showed differential recognition of the panel of PC-associated antigens. These mutant antibodies contain amino acid replacements in the CDR2 region of the heavy chain variable region, indicating the importance of CDR2 in recognition of carrier determinants. A model of T15 is shown that illustrates the strategic placement of mutations that could allow interaction with determinants associated with PC. A direct implication of this finding is that the T15 antibody combining site accommodates structures larger than phosphocholine and that recognition of associated carrier determinants could be a significant force in shaping the immune response to PC-containing pathogens.

A coiled-coil mimetic intercepts BCR-ABL1 dimerization in native and kinase-mutant chronic myeloid leukemia.

Targeted therapy of chronic myeloid leukemia (CML) is currently based on small-molecule inhibitors that directly bind the tyrosine kinase domain of BCR-ABL1. This strategy has generally been successful, but is subject to drug resistance because of point mutations in the kinase domain. Kinase activity requires transactivation of BCR-ABL1 following an oligomerization event, which is mediated by the coiled-coil (CC) domain at the N terminus of the protein. Here, we describe a rationally engineered mutant version of the CC domain, called CC(mut3), which interferes with BCR-ABL1 oligomerization and promotes apoptosis in BCR-ABL1-expressing cells, regardless of kinase domain mutation status. CC(mut3) exhibits strong proapoptotic and antiproliferative activity in cell lines expressing native BCR-ABL1, single kinase domain mutant BCR-ABL1 (E255V and T315I) or compound-mutant BCR-ABL1 (E255V/T315I). Moreover, CC(mut3) inhibits colony formation by primary CML CD34(+) cells ex vivo, including a sample expressing the T315I mutant. These data suggest that targeting BCR-ABL1 with CC mutants may provide a novel alternative strategy for treating patients with resistance to current targeted therapies.

ß-Catenin is required for intrinsic but not extrinsic BCR-ABL1 kinase-independent resistance to tyrosine kinase inhibitors in chronic myeloid leukemia.

Activation of nuclear ß-catenin and expression of its transcriptional targets promotes chronic myeloid leukemia (CML) progression, tyrosine kinase inhibitor (TKI) resistance, and leukemic stem cell self-renewal. We report that nuclear ß-catenin has a role in leukemia cell-intrinsic but not -extrinsic BCR-ABL1 kinase-independent TKI resistance. Upon imatinib inhibition of BCR-ABL1 kinase activity, ß-catenin expression was maintained in intrinsically resistant cells grown in suspension culture and sensitive cells cultured in direct contact (DC) with bone marrow (BM) stromal cells. Thus, TKI resistance uncouples ß-catenin expression from BCR-ABL1 kinase activity. In ß-catenin reporter assays, intrinsically resistant cells showed increased transcriptional activity versus parental TKI-sensitive controls, and this was associated with restored expression of ß-catenin target genes. In contrast, DC with BM stromal cells promoted TKI resistance, but had little effects on Lef/Tcf reporter activity and no consistent effects on cytoplasmic ß-catenin levels, arguing against a role for ß-catenin in extrinsic TKI resistance. N-cadherin or H-cadherin blocking antibodies abrogated DC-based resistance despite increasing Lef/Tcf reporter activity, suggesting that factors other than ß-catenin contribute to extrinsic, BM-derived TKI resistance. Our data indicate that, while nuclear ß-catenin enhances survival of intrinsically TKI-resistant CML progenitors, it is not required for extrinsic resistance mediated by the BM microenvironment.

A simple method for determining K(A)s of both low and high affinity IgG antibodies.

A rapid and convenient method for measuring affinity constants (K(A)) of IgG antibody-hapten complexes is described. A key advantage of this method is its suitability for quantification of both low and high affinity interactions. A comparison is made of the K(A)s of the low affinity anti-phosphocholine (PC) antibody T15 (K(A) = 2.9 x 10(5) M(-1)) and five heavy chain complementarity determining region 2 (HCDR2) mutant antibodies expressed as IgG2b transfectants. As a demonstration of the general applicability of the technique, colchicine binding to the high affinity monoclonal IgG2a antibody C44 (K(A) = 1.5 x 10(9) M(-1)) is measured also; thus the assay is applicable over a four-log range of affinities. The assay, based upon use of fixed Staphylococcus aureus Cowan I strain cells as an adsorbent for antibody-radiolabeled antigen complexes, is conducted over a range of hapten concentrations at constant antibody concentration. The K(A) is obtained by Scatchard analysis.

Upper airway EMG responses to acute hypoxia and asphyxia are impaired in streptozotocin-induced diabetic rats.

Obstructive sleep apnoea (OSA) is a major clinical disorder that is characterised by multiple episodes of upper airway obstruction due to failure of the upper airway dilator muscles to maintain upper airway patency. The incidence of OSA is high in many endocrine disorders including both insulin-dependent and non-insulin-dependent diabetes but the reasons for this are not known. We wished to test the hypothesis that central respiratory motor output to the upper airway muscles is preferentially impaired in a rat model of diabetes mellitus. Sternohyoid (SH) and diaphragm (DIA) EMG activities were recorded in control and streptozotocin (STZ)-induced diabetic rats during normoxia, hypoxia (7.5% O2 in N2) and asphyxia (7.5% O2 and 3% CO2) under pentobarbitone anaesthesia. SH EMG responses to acute hypoxia and asphyxia were significantly impaired in STZ-induced diabetic rats compared to control animals (+47.1 +/- 5.7 vs. +11.7 +/- 1.9% during hypoxia in control and diabetic animals respectively and +56.5 +/- 7.9 vs. +15.7 +/- 5.0% during asphyxia). However, DIA EMG responses to hypoxia and asphyxia were not different for the two groups. We propose that the higher prevalence of OSA in diabetic patients is related to preferential impairment of cranial motor output to the dilator muscles of the upper airway in response to physiological stimuli.

Measuring problem drinking in first time offenders. Development and validation of the College Alcohol Problem Scale (CAPS).

Research on college drinking continues to justify serious concerns for the psychological, social, and physical well-being of young persons who abuse alcohol. However, despite considerable interest and research in this regard, there are few valid, reliable and clinically useful brief screening instruments available to measure youthful drinking problems. The current study of 315 college students cited their first time for breaking university drinking rules describes the development and validation of the College Alcohol Problem Scale (CAPS) for measuring different psychosocial dimensions of problem drinking in college students. Two related but distinct factors emerged defining Socio-Emotional and Community Problems. These two factors explained almost two thirds of the variance, and showed very good internal reliabilities. MANOVA analysis demonstrated concurrent validity for the CAPS with both a measure of heavy drinking derived from the QFI and a modified version of the MAST. Implications for using the CAPS for identifying potential drinking problems in young persons are emphasized.

The Drinking Context Scale. A confirmatory factor analysis.

The current article examines the development and validation of the Drinking Context Scale through the use of confirmatory factor analysis. The scale measures the self-reported likelihood of excessive drinking across a number of specific social-cognitive drinking contexts. Five-hundred-and-five college students adjudicated for breaking university drinking rules filled out the anonymous questionnaire. Three factors including convivial, intimate, and negative coping contexts were confirmed, and these factors demonstrated good reliability and evidence of concurrent validity with other substance abuse indices, including the Alcohol Use Disorders Identification Test and the College Alcohol Problem Scale. Implications for the DCS as an assessment tool for prevention and early intervention with young people are discussed.

Validating the Alcohol Use Disorder Identification Test with college first-offenders.

Although the Alcohol Use Disorders Identification Test (AUDIT) has been shown to have good validity and reliability with clinical samples, little data has been examined with respect to youthful problem drinkers, particularly college students. Data collected with 312 students cited their first time for breaking university drinking rules was examined to evaluate the factorial validity and internal consistency of the 10-item scale, and also to test the validity of the AUDIT against two scales designed with a previous cohort specifically to measure hazardous (The Drinking Context Scale) and harmful drinking (the College Alcohol Problem Scale) in college students. Overall, results suggest that the AUDIT is a valid and reliable screening device for college students, and could play an important role in assessing youthful problem drinkers for early intervention programming.