Genome-wide mRNA profiling identifies X-box-binding protein 1 (XBP1) as an IRE1 and PUMA repressor.

Accumulation of misfolded proteins in ER activates the unfolded protein response (UPR), a multifunctional signaling pathway that is important for cell survival. The UPR is regulated by three ER transmembrane sensors, one of which is inositol-requiring protein 1 (IRE1). IRE1 activates a transcription factor, X-box-binding protein 1 (XBP1), by removing a 26-base intron from XBP1 mRNA that generates spliced XBP1 mRNA (XBP1s). To search for XBP1 transcriptional targets, we utilized an XBP1s-inducible human cell line to limit XBP1 expression in a controlled manner. We also verified the identified XBP1-dependent genes with specific silencing of this transcription factor during pharmacological ER stress induction with both an N-linked glycosylation inhibitor (tunicamycin) and a non-competitive inhibitor of the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) (thapsigargin). We then compared those results to the XBP1s-induced cell line without pharmacological ER stress induction. Using next-generation sequencing followed by bioinformatic analysis of XBP1-binding motifs, we defined an XBP1 regulatory network and identified XBP1 as a repressor of PUMA (a proapoptotic gene) and IRE1 mRNA expression during the UPR. Our results indicate impairing IRE1 activity during ER stress conditions accelerates cell death in ER-stressed cells, whereas elevating XBP1 expression during ER stress using an inducible cell line correlated with a clear prosurvival effect and reduced PUMA protein expression. Although further studies will be required to test the underlying molecular mechanisms involved in the relationship between these genes with XBP1, these studies identify a novel repressive role of XBP1 during the UPR.

Regulation of the abundance of Y-family polymerases in the cell cycle of budding yeast in response to DNA damage.

Y-family DNA polymerases mediate DNA damage tolerance via translesion synthesis (TLS). Because of the intrinsically error-prone nature of these enzymes, their activities are regulated at several levels. Here, we demonstrate the common regulation of the cellular abundance of Y-family polymerases, polymerase eta (Pol eta), and Rev1, in response to DNA damage at various stages of the cell cycle. UV radiation influenced polymerase abundance more when cells were exposed in S-phase than in G1- or G2-phases. We noticed two opposing effects of UV radiation in S-phase. On one hand, exposure to increasing doses of UV radiation at the beginning of this phase increasingly delayed S-phase progression. As a result, the accumulation of Pol eta and Rev1, which in nonirradiated yeast is initiated at the S/G2-phase boundary, was gradually shifted into the prolonged S-phase. On the other hand, the extent of polymerase accumulation was inversely proportional to the dose of irradiation, such that the accumulation was significantly lower after exposure to 80 J/m2 in S-phase than after exposure to 50 J/m2 or 10 J/m2. The limitation of polymerase accumulation in S-phase-arrested cells in response to high UV dose was suppressed upon RAD9 (but not MRC1) deletion. Additionally, hydroxyurea, which activates mainly the Mrc1-dependent checkpoint, did not limit Pol eta or Rev1 accumulation in S-phase-arrested cells. The results show that the accumulation of Y-family TLS polymerases is limited in S-phase-arrested cells due to high levels of DNA damage and suggest a role of the Rad9 checkpoint protein in this process.

Mean Platelet Volume in Crohn's Disease Patients Predicts Sustained Response to a 52-Week Infliximab Therapy: A Pilot Study.

BACKGROUND: The loss of response to infliximab in Crohn's disease (CD) patients is currently a major clinical problem. Recently, mean platelet volume (MPV) has been proposed as a new biomarker of CD activity. Here, we hypothesized that MPV may be used as cheap and efficient biomarker of subclinical inflammation during 52-week therapy in CD patients responding to infliximab induction treatment. AIMS: The aim of study was to establish whether MPV at baseline and pre-infusion at week 14 are good predictors of sustained response after week 14 in CD patients undergoing 52-week infliximab therapy. METHODS: A retrospective study of 30 adult CD patients who underwent a 52-week course of treatment with infliximab and achieved response at week 14 to induction treatment was performed. The association between MPV, baseline disease parameters and maintained clinical response or remission during infliximab therapy was assessed. RESULTS: Higher MPV at week 14 was observed in CD patients with sustained response to infliximab after week 14 than in patients with loss of response (p = 0.0019). In patients with loss of response to maintenance infliximab treatment, lower ΔMPV between baseline and week 14 was calculated (p = 0.0003). MPV > 10.3 fl at week 14 predicts sustained response with 67 % sensitivity and 80 % specificity. ΔMPV between baseline and week 14 >0.4 fl predicts sustained response with 87 % sensitivity and 93 % specificity. CONCLUSION: MPV at week 14 and ΔMPV between baseline and week 14 are good predictors of sustained response to infliximab maintenance treatment in CD patients.

Codon bias and the folding dynamics of the cystic fibrosis transmembrane conductance regulator.

Synonymous or silent mutations are often overlooked in genetic analyses for disease-causing mutations unless they are directly associated with potential splicing defects. More recent studies, however, indicate that some synonymous single polynucleotide polymorphisms (sSNPs) are associated with changes in protein expression, and in some cases, protein folding and function. The impact of codon usage and mRNA structural changes on protein translation rates and how they can affect protein structure and function is just beginning to be appreciated. Examples are given here that demonstrate how synonymous mutations alter the translational kinetics and protein folding and/or function. The mechanism for how this occurs is based on a model in which codon usage modulates the translational rate by introducing pauses caused by nonoptimal or rare codons or by introducing changes in the mRNA structure, and this in turn influences co-translational folding. Two examples of this include the multidrug resistance protein (p-glycoprotein) and the cystic fibrosis transmembrane conductance regulator gene (CFTR). CFTR is also used here as a model to illustrate how synonymous mutations can be examined using in silico predictive methods to identify which sSNPs have the potential to change protein structure. The methodology described here can be used to help identify "non-silent" synonymous mutations in other genes.

Can biologic treatment induce cutaneous focal mucinosis?

Skin mucinosis is a rare skin disease which clinically manifests as firm papules and waxy nodules. We report a case of a 66-year-old female psoriatic patient who developed skin mucinosis during biological therapy. Because of a previous lack of response to the local and conventional systemic treatment of psoriasis, the patient received biological therapy (infliximab from June 2008 to May 2009 - initial clinical improvement and loss of treatment effectiveness in the 36(th) week of the therapy; adalimumab from June 2009 to January 2010 - lack effectiveness; ustekinumab from March 2012 to the present). Throughout 2 months we observed a manifestation of the skin mucinosis as well-demarcated, yellow and brown, papulo-nodular lesions of 5-10 mm in diameter, localized on the back. Histopathological examination with alcian blue staining demonstrated mucin deposits in the dermis. On the basis of clinical and histopathological findings, the diagnosis of cutaneous focal mucinosis was established. We present the case because of the extremely rare occurrence of the disease. Scarce literature and data suggest that there is an association between focal mucinosis and thyroid dysfunction, as well as possible adverse effects of biological therapy with TNF-α antagonists.

Genome-wide mRNA profiling identifies RCAN1 and GADD45A as regulators of the transitional switch from survival to apoptosis during ER stress.

Endoplasmic reticulum (ER) stress conditions promote a cellular adaptive mechanism called the unfolded protein response (UPR) that utilizes three stress sensors, inositol-requiring protein 1, protein kinase RNA-like ER kinase, and activating transcription factor 6. These sensors activate a number of pathways to reduce the stress and facilitate cell survival. While much is known about the mechanisms involved that modulate apoptosis during chronic stress, less is known about the transition between the prosurvival and proapoptotic factors that determine cell fate. Here, we employed a genetic screen that utilized three different pharmacological stressors to induce ER stress in a human-immortalized airway epithelial cell line, immortalized human bronchial epithelial cells. We followed the stress responses over an 18-h time course and utilized real-time monitoring of cell survival, next-generation sequencing, and quantitative real-time PCR to identify and validate genes that were upregulated with all three commonly employed ER stressors, inhibitor of calpain 1, tunicamycin, and thapsigargin. growth arrest and DNA damage-inducible alpha (GADD45A), a proapoptotic factor, and regulator of calcineurin 1 (RCAN1) mRNAs were identified and verified by showing that small interfering RNA (siRNA) knockdown of GADD45A decreased CCAAT-enhancer-binding protein homologous protein (a.k.a DDIT3), BCL2-binding component 3 (a.k.a. BBC3), and phorbol-12-myristate-13-acetate-induced protein 1 expression, 3 proapoptotic factors, and increased cell viability during ER stress conditions, whereas siRNA knockdown of RCAN1 dramatically decreased cell viability. These results suggest that the relative levels of these two genes regulate cell fate decisions during ER stress independent of the type of ER stressor.

Aspects of ageing on professional opera singer's voice - preliminary findings.

OBJECTIVES: The main goals of this study were to assess the most significant morphological changes and acoustic measures for the ageing process of a professional opera singer's voice. DESIGN: This investigation included 58 healthy professional opera singers, who were compared with 42 young opera singers from the control group. METHODS: All participants underwent a voice assessment protocol: ENT specialist examination and speech therapist evaluation. Acoustic parameters and subjective observations were obtained and analysed. R esults: Fundamental frequency (F0) level was distinctly decreased in the case of older female singers, but F0 in older male singers had stable levels in comparison to that in younger singers. Older singers were found not to have substantially different values of jitter than younger ones. Maximal phonation time (MPT) was longer in the older women's group when compared to the younger singers, but not relatively different in the men's group. Shimmer value presented no age-related change. Morphological changes seem to correlate with the age of subjects. C onclusions: The main characteristic of voice change with age was a decreased F0 level among older female professional singers and rather stable F0 levels in male singers. This study gives preliminary results on the ageing of voice in the population of professional opera singers.

Silent sinus syndrome: combined sinus surgery and orbital reconstruction - report of 15 cases.

BACKGROUND: Silent sinus syndrome (SSS) is defined as spontaneous, painless enophthalmos, hypoglobus with orbital floor resorption and maxillary sinus collapse on the ipsilateral side. Different methods of orbital floor reconstruction have been proposed. AIMS/OBJECTIVES: The purpose was to analyse the results of combined endoscopic sinus surgery (ESS) and reconstruction using orbital floor implant of 15 patients with SSS and to present recent histological findings. MATERIALS AND METHODS: Retrospective case review of 15 patients with SSS treated in clinic between 2007 and 2017. RESULTS: Eleven women and four men presented with unilateral, spontaneous enophthalmos. Averaged duration of enophthalmos was 10.7 months. On affected side, mean enophthalmos was 2.6 mm and hypoglobus 2.7 mm. Computed tomography imaging (CT) imaging showed maxillary sinus opacification on the affected side in every case, and the orbital floor was displaced downwards in all cases. In total, 13 patients underwent simultaneous ESS and rebuilding of orbital floor with a titanium implant. Statistical analysis confirmed significant differences for pre- and postoperative measure of enophthalmos and hypoglobus. CONCLUSION AND SIGNIFICANCE: Implementation of titanium implants is the reliable method of reconstruction that allows good aesthetic result, shorter time of procedure with an excellent long-term outcome and satisfactory patient's tolerance.

Lacrimal gland pleomorphic adenoma: a review of 52 cases, 15-year experience.

BACKGROUND: Lacrimal gland (LG) tumours are rare neoplasms. Pleomorphic adenoma (PA) is the most common histologic variant, representing ∼20% of all LG tumours. PA tends to recur leading to great morbidity. AIMS: We carried out this study to share our experience and provide recent data on the clinical aspects, radiologic findings, management and outcome after treatment. METHODS: We realised a hospital-based retrospective case review including data collected from 52 patients, managed over 15 years. RESULTS: The mean age was 39.8 ± 2 years. Unilateral painless proptosis was the most constant sign (51.9%). CT-scan showed predominantly isodense lesions (96.2%), with regular borders (94.2%), measuring on average 2 cm. MRI showed isointense lesions on T1-weighted images in 96.2% of cases. No pre-operative biopsy was done. Surgical management was mainly external lateral orbitotomy (94.2%). There was no recurrence, considering a mean follow-up period of 12.6 years. Size of tumour at the time of diagnosis increased with age (r = +0.36, p = .01). CONCLUSIONS: Clinical and radiologic characteristics are consistent with literature. Older patients seem to present larger tumours. We believe that biopsy is not necessary if appropriate imaging is done. Complete, intact resection is generally sufficient to minimise the risk of recurrence.

CT-based Morphometric Analysis of Professional Opera Singers' Vocal Folds.

PURPOSE AND STUDY DESIGN: Our work aimed to study the relationship between the length of vocal folds and classical voice type among professional opera singers. Also, the present paper attempts to assess if individual features such as height and body mass index are associated with the vocal folds length (VFL) and the vocal range. Our research compared VFL and voice type by evaluation of neck computed tomography imaging of professional opera singers obtained between 2009 and 2015. Vocal range was measured after scanning procedure by a speech-language pathologist. The results were used to compare VFL with vocal range, voice type, and individual features of every singer. MATERIAL AND METHODS: The group included 93 opera singers (female: 31 sopranos, 15 mezzos, and 5 contraltos; male: 17 tenors, 16 baritones, 6 basses, and 3 countertenors). Different values such as VFL, vocal range, and individuals' features (height, weight, body mass index) were analyzed statistically. RESULTS: Some significant differences and correlation were found. The VFL was diversified between classical voice types. VFL for soprano, mezzo-soprano, and bass was significantly different from every other type of voice. The vocal range has been shown to have a negative correlation with VFL value, which reflects the narrowest vocal range among bass voices. The analysis confirmed a linear correlation between VFL and individual features of the body such as height and body mass index. CONCLUSION: Presented data support the hypothesis that there are significant differences of VFL, vocal range, and body morphology between professional singers of different voice classification.

DNA polymerase ι is acetylated in response to SN2 alkylating agents.

DNA polymerase iota (Polι) belongs to the Y-family of DNA polymerases that are involved in DNA damage tolerance through their role in translesion DNA synthesis. Like all other Y-family polymerases, Polι interacts with proliferating cell nuclear antigen (PCNA), Rev1, ubiquitin and ubiquitinated-PCNA and is also ubiquitinated itself. Here, we report that Polι also interacts with the p300 acetyltransferase and is acetylated. The primary acetylation site is K550, located in the Rev1-interacting region. However, K550 amino acid substitutions have no effect on Polι's ability to interact with Rev1. Interestingly, we find that acetylation of Polι significantly and specifically increases in response to SN2 alkylating agents and to a lower extent to SN1 alkylating and oxidative agents. As we have not observed acetylation of Polι's closest paralogue, DNA polymerase eta (Polη), with which Polι shares many functional similarities, we believe that this modification might exclusively regulate yet to be determined, and separate function(s) of Polι.

Surgical treatment in children with otosclerosis and congenital stapes fixation: our experience and outcome.

AIM: To evaluate outcomes of stapes surgery in children with congenital stapes fixation and juvenile otosclerosis. METHODS: A retrospective chart review was performed from 1987 to 2013 to identify patients under 18 years old who underwent a stapes surgery. Patients' age, gender, pre- and postoperative audiograms, intraoperative findings including aetiology of stapes fixation, prosthesis type, and complications were analysed. RESULTS: 18 children (6 - 17 years old), all with bilateral conductive hearing loss were identified and 34 stapes surgeries were performed (two patients underwent surgery only on one side). The cause of fixation included juvenile otosclerosis in 88% and congenital stapes fixation in 12%. The mean pre-operative air-bone gap (ABG) was 36,24 dB (SD: 10,86) compared to a postoperative mean ABG of 7,74 (SD: 3,3) (p < 0.000). The profound sensorineural hearing loss was not observed in long-term follow-up. CONCLUSIONS: Paediatric stapes surgery has comparable results to stapedectomy in adults regardless of the cause of stapes fixation; however, the better hearing outcome was observed for cases of juvenile otosclerosis rather than congenital stapes fixation.

microRNA single polynucleotide polymorphism influences on microRNA biogenesis and mRNA target specificity.

microRNAs (miRNAs) are nowadays recognized as an essential component of gene regulatory networks. Furthermore, deregulation of miRNAs expression often contributes to human pathologies. Recently, a substantial number of single nucleotide polymorphism (SNPs) and rare mutations within pri-, pre- and mature miRNA sequences have been reported. These miRNA SNPs have often been associated with human disease. However, due to the complexity of miRNA biogenesis and the genome-wide functional effects of miRNAs, the determination of biological consequences of these miRNA SNPs remains challenging. Despite an increasing number of reports linking miRNA SNPs with human pathologies, few reports have analyzed the mechanism by which miRNA-SNPs contribute to disease pathogenesis. In this review, we discuss how single polynucleotide polymorphisms in miRNAs genes may influence miRNAs expression and function and thus potentially alter disease pathogenesis.

Mean platelet volume as a possible biomarker of tumor progression in rectal cancer.

BACKGROUND: Rectal cancer is a major cause of death and an early diagnosis is crucial in decreasing mortality. Previous studies found a relation between chronic inflammation and increased risk of rectal cancer. It was shown that mean platelet volume (MPV) level might be a marker of subclinical inflammatory process in gastrointestinal tract. OBJECTIVE: To determinate whether MPV could be a useful biomarker of tumor progression in the rectal cancer. METHODS: One hundred and three patients with rectal cancer who underwent surgical resection of tumor were enrolled in the study. The control group consisted of 98 healthy subjects. RESULTS: The association between MPV, tumor stage and clinical status were assessed. The analysis proved that pre-operative MPV level was significantly lower in rectal cancer vs. healthy individuals (10.65 ± 0.79 vs. 11.41 ± 0.76 fL; p < 0.001).Receiver-operating characteristic curve analysis suggested 11.3 as the cut-off value for MPV (sensitivity = 83%; specificity = 54%; AUC = 0.745). Surgical resection of tumor resulted in the increase of the MPV level with statistical significance (10.65 ± 0.79 fL vs. 11.21 ± 0.82 fL; p < 0.001). No relationship was found between the post-operative MPV level in cancer patients and control subjects. CONLUSION: MPV level may be potentially useful and easily available biomarker for monitoring subclinical inflammation related to rectal cancer and predicting tumor progression.

Prevention and therapeutic strategies of thromboembolic events in patients with inflammatory bowel diseases: a report of three cases.

Crohn's disease and ulcerative colitis belong to a group of inflammatory bowel diseases (IBD). IBD are characterized by a chronic character of inflammatory process and overlapping immunological abnormalities, which, along with therapeutic strategies are currently available, underlie an increased risk of venous thromboembolic events (VTE). The most common sites of VTE in IBD patients are deep venous thrombosis (DVT) and pulmonary embolism (PE). These complications are particularly important in clinical practice due to a very high mortality rate. Therefore, an early diagnosis of new IBD cases and the control of inflammatory process are thought to play a crucial role in risk reduction for thromboembolic events. Despite considerable evidence supporting the association between IBD and VTE, there is still a lack of recognition of this risk, with dangerous consequences for patients. In this paper authors report three cases of VTE in IBD patients and discuss the most relevant clinical studies found in MEDLINE, Cochrane Library and EMBASE regarding its prevention and management.

Comfort During Digital Rectal Examination - Patient Preference.

UNLABELLED: The sex and age of the physician performing the digital rectal examination (DRE) procedure is one of the parameters influencing patients' comfort. It is postulated that the stress related to DRE during admission to the surgical ward may affect the compliance. The aim of the study was to characterize patients' preferences according to their sex, age, socioeconomic status and according to DRE-related variables. MATERIAL AND METHODS: Patients admitted to the Department of General and Colorectal Surgery at Medical University in Lódz between October 2014 and June 2015 were asked to complete a questionnaire regarding their sex, age, ethnic background, socioeconomic status and preference for a physician performing the DRE during admission to the surgical ward. RESULTS: The study enrolled 225 patients, 52% (n=117) women and 48% (n=108) men. Most patients 73.3% (n=244) expressed no preference for sex of the physician performing the DRE during admission, while 22.7% (n=51) preferred a same-sex physician. Analysis showed that the age, female sex, lower education status, small amount of comorbidities and number of hospitalizations, and no previous colonoscopy experience were all associated with preference for a same-sex physician. CONCLUSIONS: In our study most patients expressed no preference for sex of the physician performing the DRE during admission to the surgical ward. However, over one-tenth of patients reported such preferences. Most of these patients preferred a same-sex physician. It is important to offer these patients the choice of physician performing the DRE. Addressing patients' preferences may improve the atmosphere in the clinical environment, reduce stress, and facilitate better treatment.

Correlations between skin lesions induced by anti-tumor necrosis factor-α and selected cytokines in Crohn's disease patients.

AIM: To investigate the correlation between the appearance of skin lesions and concentration of interleukin (IL)-17A, IL-23 and interferon-γ (IFN-γ) in Crohn's disease (CD) patients during anti-tumor necrosis factor-α (TNF-α) therapy METHODS: A prospective study included 30 adult patients with CD of Caucasian origin (19 men and 11 women; mean age ± SD 32.0 ± 8.6 years) during biological therapy with anti-TNF-α antibodies from January 2012 to March 2013. Eighteen patients were treated with infliximab, seven with adalimumab and five with certolizumab. Inclusion criteria were exacerbation of the underlying disease, Crohn's Disease Activity Index over 300 and the ineffectiveness of previously used non-biological therapies. Patients with a history of psoriasis, atopic dermatitis and other autoimmune skin lesions were excluded from the study. The control group consisted of 12 healthy subjects. A diagnostic survey was carried out, blood tests and careful skin examination were performed, and the serum levels of IL-17, IL-23 and IFN-γ were measured using an enzyme-linked immunosorbent assays technique. Dermatoses that have developed in the course of biological therapy in patients who had no pre-existing skin lesions of similar character were qualified as skin lesions induced by anti-TNF-α therapy. RESULTS: Skin manifestations occurred in 18 of CD patients during the anti-TNF-α therapy (60%), in the average time of 10.16 ± 3.42 mo following the beginning of the 52-wk treatment cycle. Skin lesions observed in CD patients during biological therapy included psoriasiform lesions (44.4%), and eczema forms lesions (22.2%). In CD patients with drug induced skin lesions significantly higher levels of hemoglobin (13.3 ± 1.5 g/dL vs 10.8 ± 1.9 g/dL, P = 0.018) and hematocrit (39.9% ± 4.5% vs 34.3% ± 5.4%, P = 0.01), as well as a significantly lower level of platelets (268 ± 62 × 10(3)/µL vs 408 ± 239 × 10(3)/µL, P = 0.046) was observed compared with CD patients without skin manifestations. The concentrations of IL-17A and IL-23 in CD patients with skin lesions developed under anti-TNF-α therapy were significantly higher compared to those in patients without lesions (IL-17A: 39.01 ± 7.03 pg/mL vs 25.71 ± 4.90 pg/mL, P = 0.00004; IL-23: 408.78 ± 94.13 pg/mL vs 312.15 ± 76.24 pg/mL, P = 0.00556). CONCLUSION: Skin lesions in CD patients during biological therapy may result from significantly increased concentrations of IL-17A and IL-23, which are strongly associated with TNF-α/Th1 immune pathways.