Epigenetic-focused CRISPR/Cas9 screen identifies (absent, small, or homeotic)2-like protein (ASH2L) as a regulator of glioblastoma cell survival.

BACKGROUND: Glioblastoma is the most common and aggressive primary brain tumor with extremely poor prognosis, highlighting an urgent need for developing novel treatment options. Identifying epigenetic vulnerabilities of cancer cells can provide excellent therapeutic intervention points for various types of cancers. METHOD: In this study, we investigated epigenetic regulators of glioblastoma cell survival through CRISPR/Cas9 based genetic ablation screens using a customized sgRNA library EpiDoKOL, which targets critical functional domains of chromatin modifiers. RESULTS: Screens conducted in multiple cell lines revealed ASH2L, a histone lysine methyltransferase complex subunit, as a major regulator of glioblastoma cell viability. ASH2L depletion led to cell cycle arrest and apoptosis. RNA sequencing and greenCUT&RUN together identified a set of cell cycle regulatory genes, such as TRA2B, BARD1, KIF20B, ARID4A and SMARCC1 that were downregulated upon ASH2L depletion. Mass spectrometry analysis revealed the interaction partners of ASH2L in glioblastoma cell lines as SET1/MLL family members including SETD1A, SETD1B, MLL1 and MLL2. We further showed that glioblastoma cells had a differential dependency on expression of SET1/MLL family members for survival. The growth of ASH2L-depleted glioblastoma cells was markedly slower than controls in orthotopic in vivo models. TCGA analysis showed high ASH2L expression in glioblastoma compared to low grade gliomas and immunohistochemical analysis revealed significant ASH2L expression in glioblastoma tissues, attesting to its clinical relevance. Therefore, high throughput, robust and affordable screens with focused libraries, such as EpiDoKOL, holds great promise to enable rapid discovery of novel epigenetic regulators of cancer cell survival, such as ASH2L. CONCLUSION: Together, we suggest that targeting ASH2L could serve as a new therapeutic opportunity for glioblastoma. Video Abstract.

The Effect of Lavandula stoechas on Wound Healing in an Experimental Diabetes Model.

INTRODUCTION: Diabetic foot is a consequential and dangerous complication of diabetes, contributing to decreased quality of life, escalated hospitalizations, and increased mortality rates. Using an experimental model of diabetes, this study aims to investigate the effect of Lavandula stoechas on wound healing. METHODS: A total of 35 albino Wistar rats, 250-350 grams in weight, were used. The rats were divided into five groups, seven rats in each group. Of these, 21 rats were induced with 50 mg/kg streptozotocin (STZ) to mimic the diabetic condition. Additionally, 14 rats remained non-diabetic and served as the control group. The diabetic rats were further divided into three subgroups. The non-diabetic group was split into two subgroups based on the dressing materials used (allicin, physiological serum, and control). Wound dimensions were assessed on Days 0, 7, 14, and 21. Biopsies were taken from the wound sites at the same time. RESULTS: There were significant differences between groups on Days 7, 14, and 21. The percentage of healing was highest in the Lavandula Stoechas group on Days 7, 14, and 21. Microscopic examination of the biopsies supported accelerated wound healing on Days 7 and 14. Reduced mononuclear cell density and increased hair follicle and adipose tissue development were observed in the DM (diabetes mellitus)-Lavandula Stoechas group on Day 7. On Day 14, the DM-Lavandula Stoechas group increased collagen levels and hair follicles. Similarly, the non-DM-Lavandula Stoechas group showed reduced bullae, dermal edema, and intraepithelial edema on Day 7. This was followed by increased fibroblast levels on Day 14. CONCLUSIONS: In conclusion, this study provides compelling evidence for the potential of Lavandula stoechas extract in the enhancement of diabetic wound healing. The multiple interactions revealed here highlight the need for further investigation into the underlying mechanisms. A cost-effective use of Lavandula stoechas opens up promising prospects in managing diabetic foot healing. This warrants additional research and clinical translation.

EPIKOL, a chromatin-focused CRISPR/Cas9-based screening platform, to identify cancer-specific epigenetic vulnerabilities.

Dysregulation of the epigenome due to alterations in chromatin modifier proteins commonly contribute to malignant transformation. To interrogate the roles of epigenetic modifiers in cancer cells, we generated an epigenome-wide CRISPR-Cas9 knockout library (EPIKOL) that targets a wide-range of epigenetic modifiers and their cofactors. We conducted eight screens in two different cancer types and showed that EPIKOL performs with high efficiency in terms of sgRNA distribution and depletion of essential genes. We discovered novel epigenetic modifiers that regulate triple-negative breast cancer (TNBC) and prostate cancer cell fitness. We confirmed the growth-regulatory functions of individual candidates, including SS18L2 and members of the NSL complex (KANSL2, KANSL3, KAT8) in TNBC cells. Overall, we show that EPIKOL, a focused sgRNA library targeting ~800 genes, can reveal epigenetic modifiers that are essential for cancer cell fitness under in vitro and in vivo conditions and enable the identification of novel anti-cancer targets. Due to its comprehensive epigenome-wide targets and relatively high number of sgRNAs per gene, EPIKOL will facilitate studies examining functional roles of epigenetic modifiers in a wide range of contexts, such as screens in primary cells, patient-derived xenografts as well as in vivo models.

First case report of Pacinian corpuscle hyperplasia following complex regional pain syndrome.

Pacinian disorders are exceedingly rare, and the exact pathogenesis is still unknown. The most common symptoms are pain, sensory changes, and a visible or palpable mass, and diagnosis is usually made by pathological examination after the excision of the painful nodule. In this case report, we present the case of a 49-year-old male with Pacinian corpuscle hyperplasia located on the metacarpophalangeal joint, emerging at the same hand of the patient two years after the treatment due to complex regional pain syndrome (CRPS). To the best of our knowledge, this is the first case report revealing the association of CRPS with hyperplasia of Pacinian corpuscles.

A supportive donor nerve for long-term facial paralysis: Anatomical analysis of the posterior auricular nerve and micro-anatomical comparison with zygomatic nerve.

BACKGROUND: The posterior auricular nerve (PAN) is an inspiring candidate for the additional axonal source in long-term facial paralysis to improve the functional results of the cross-facial nerve (FN) graft technique. However, no studies have analyzed the PAN's axonal load and its microscopic anatomy to assess its utilization in facial reanimation. The present study aims to examine the anatomical and microscopic features of the PAN to analyze its feasibility as a donor nerve. METHODS: The bilateral facial side of 14 fresh frozen adult human cadavers was examined for the study. The PAN's anatomical course was recorded, and nerve specimens from the PAN and zygomatic nerve (ZN) were obtained to compare their microscopic anatomy and axon counts using a light microscope and transmission electron microscope. RESULTS: The PAN's average branching distance and its course length were 5.8 ± 2.69 mm and 59.2 ± 5.85, respectively. The mean number of myelinated axons was 600.28 ± 69.97 in the PAN and 728.85 ± 166.31 in the ZN. This difference between the two nerves was statistically significant (p = 0.002). However, considering the gender variable, the mean axon counts of PAN and ZN were statistically similar for face sides and their average. Furthermore, the ultrastructural anatomy of both nerves was similar in electron microscopic evaluation. CONCLUSIONS: The present study confirms that the PAN is a proper candidate to be a supportive donor nerve due to its isolated site, consistent anatomical course, convenient ultrastructural anatomy as well as axonal load.

Evaluation of versatility and outcomes of the first dorsal metacarpal artery flap in thumb defects.

BACKGROUND: It is clinically vital to determine the best technique to reconstruct thumb defects with satisfactory esthetic and functional outcomes. We aimed to quantitatively present the safety, versatility, limitations, advantages, and functional results of the first dorsal metacarpal artery flap (FDMAF) and evaluate its outcomes in thumb defect reconstruction by comparing it with the other current surgical options. METHODS: A total of 21 patients underwent thumb defect reconstruction. They were evaluated with the following parameters: Etiology, age, timing of reconstruction, flap vitality, Semmes-Weinstein monofilament (SWM) test, static two-point discrimination (2PD) test, pain, cortical reorientation, cold intolerance questionnaire, and Michigan hand outcomes questionnaire. Their functional outcomes were evaluated by comparing their scores with the other current surgical options published in the literature. RESULTS: The mean follow-up period was 22.3 months. The mean pain score of the flap was 0.4±0.6 and no patient had pain in the donor area (range, 0-10). The sensory outcome was 'good' (8.6 mm) based on the static 2PD test. The mean SWM test score was 4.02 g. Patient satisfaction was 4.6 according to the Michigan hand outcomes questionnaire (range, 0-5). The cold intolerance questionnaire scores showed that the patients had mild cold intolerance (mean, 10.5; range 0-100). Complete cortical reorientation was seen in 81% of the patients. CONCLUSION: Restoration of the innervation of thumb defects is possible with the FDMAF with high satisfaction rates based on our results. The absence of the microsurgical vessel or nerve repair facilitates the surgery, shortens its duration, and reduces morbidity, reserving the microsurgical options for more complicated cases. Therefore, it can be concluded that the FDMAF is an effective flap with great modifications for complicated thumb defects but surgeons should consider their clinical outcomes and prolonged surgery when choosing which technique to be used.

Aneurysmal Bone Cyst of Pisiform Bone: Diagnosis and Treatment of a Rare Entity: Case Report and a Review of the Literature.

Aneurysmal bone cyst (ABC) is a benign expansile bone tumor without metastasis capability. Only 3-4% of ABCs occur in the hand and they mainly take place in metaphysis' of long bones like metacarpals or phalanges. Carpal ABCs have been reported as individual case reports in the literature due to rarity. A patient presented with pain in her right wrist. Magnetic resonance imaging revealed a well circumscribed one cm sized mass in the pisiform bone that resembled an aneurysmal bone cyst. Total pisiformectomy was performed. Treatment options are total excision or curettaging in ABCs. But rarity of these lesions may delay the diagnosis process for the inexperienced surgeon.

Gout tophus in transferred tendons after 33 years: A case report.

Surgical site gout is an extremely rare complication that is difficult to diagnose, particularly in patients without a history of gout. A 57-year-old male patient was admitted with no previous history of gout, complaining of surgical site gout located at the junction where flexor carpi ulnaris tendon was transferred to extensor digitorum communis tendon after 33 years of the initial surgery. The patient was presented with a progressive swelling over the last three months which was located on the dorsoulnar side of the right wrist joint. Magnetic resonance imaging revealed an iso/hypointense mass. During the excisional biopsy, retained non-absorbable suture materials were observed within the mass. Histopathological examination result was reported as a typical gout tophus. No recurrence was observed after 18 months of follow-up. In conclusion, surgical site gout may be observed at transferred tendons years after the initial surgery.

IS BODY MASS INDEX AND OBESITY SURGERY MORTALITY SCORE IMPORTANT IN PERIOPERATIVE COMPLICATIONS OF LAPAROSCOPIC SLEEVE GASTRECTOMY BEFORE DISCHARGE?

BACKGROUND: Morbid obesity surgery and related complications have increased with time. AIM: To evaluate the relationship between perioperative complications before discharge and preoperative body mass index and obesity surgery mortality score in laparoscopic sleeve gastrectomy. METHOD: 1617 patients who met the inclusion criteria were evaluated retrospectively. The patients were examined in terms of demographic data, presence of comorbidities, whether there were complications or not, type of complications and obesity surgery mortality score. RESULTS: Complications were seen in 40 patients (2.5%) and mortality wasn't seen in the early postoperative period before discharge. The mean age of patients with complications was 36.3±10.02 years (19-57) and without complications 34.12±9.54 (15-64) years. The preoperative mean BMI values of patients with and without complications were 45.05±3.93 (40-57) kg/m2 and 44.8±3.49 (35-67) kg/m2 respectively. According to BMI groups 40-45 kg/m2, 45-50 kg/m2 and 50 and over, there was not any statistical significance seen in three groups in terms of complication positivity and major-minor complication rates. There was not any statistical significance seen between the patients with and without major-minor complications and obesity surgery mortality score. CONCLUSION: There was not any relation between perioperative laparoscopic sleeve gastrectomy complication rates before discharge and BMI and obesity surgery mortality scores.

A Diagnostic Dilemma for Acute Abdomen: Sclerosing Encapsulating Peritonitis (Abdominal Cocoon Syndrome); A Retrospective Cohort Study.

BACKGROUND: Sclerosing encapsulating peritonitis (SEP) is a rare cause of acute abdomen and can be easily misdiagnosed. Preoperative diagnosis of the SEP can be performed with preoperative imaging studies. We aimed to evaluate the clinical features of ileus cases who were diagnosed with primary or secondary SEP in the last five years. METHODS: This retrospective cohort study evaluated the patients who were admitted with ileus or acute abdomen symptoms to the Emergency Department of Elazig Training and Research Hospital and underwent surgery by the same surgical team of General Surgery Department between January 2014 and January 2019. Patients who were diagnosed with primary or secondary SEP were included. The demographic data, clinical presentation, whether the disease was primary or secondary, the treatment options performed and mortality rates were evaluated. RESULTS: SEP was observed in 11 of the patients. Ten patients underwent surgery (90.9%), and one patient (9.1%) was treated conservatively. Of the patients, six had secondary SEP (54.5%) and five had primary SEP (45.5%). In total, five patients were female (45.5%) and six were male (54.5%). The median age of the patients was 35 years (24-69). The median age of the patients with primary disease was 48 (29-69) years, while the median age of patients with secondary disease was 34.5 (24-64) years. One patient expired in the postoperative 8th hour. CONCLUSION: SEP should be considered in the case of recurrent abdominal pain attacks, especially in patients undergoing peritoneal dialysis, and it should be known that the mortality rate is high when misdiagnosed.

Comparative assessment of fingertip replantation in paediatric and adult patients within a single institution.

The purpose of this retrospective study was to compare the outcomes of paediatric and adult fingertip replantation within a single institution. Our retrospective study found no significant difference in the survival rate between the paediatric (10/12) and adult (22/26) groups. At 6 months follow-up, there was no significant difference in sensory recovery between both groups, as measured with Semmes-Weinstein testing, but a significant difference in mean static two-point discrimination testing values between the paediatric (4.0 mm) and adult (6.2 mm) groups. Moreover, the mean time for regaining sensation was faster in paediatric patients (1.3 months) as compared with adult patients (4.1 months). Five children and four adults received erythrocyte transfusion. Paediatric fingertip replantation has similar survival rates, faster and better sensory nerve recovery as compared with adults despite a higher erythrocyte transfusion rate. Although technically demanding, paediatric fingertip replantation is recommended, whenever possible, because of the good outcomes achievable.Level of evidence: IV.

The effects of sponges soaked with chlorhexidine gluconate and metronidazole on safety of colonic anastomosis in an experimental model of peritonitis.

BACKGROUND: The present study aims to evaluate the use of the chlorhexidine gluconate and metronidazole impregnated compresses concerning anastomosis safety in the left colonic anastomosis in the presence of peritonitis. METHODS: This study was conducted on 21 Wistar-Albino-rats divided into three equal groups. After median laparotomy, the whole layer of the left colon was cut 2 cm over the pelvic peritoneum. The faeces were spread around the injury for fecal contamination. Then, fasia and skin were closed with 3/0 silk. After one day period, relaparatomy was performed. The abdomen was cleared isotonic sodium chloride with impregnated material before starting colonic anastomosis in the first group and then double layer colonic anastomosis was performed. In the second Group-II, abdomen was cleared with the metronidazole impregnated compresses then double layer colonic anastomosis was performed. In the group-III, abdomen was cleared with the chlorhexidine gluconate impregnated compresses then double layer colonic anastomosis was performed. Tissue hydroksiproline levels and anastomosis bursting pressures were measured and histopathologic findings on the anastomosis line were evaluated on the postoperative tenth day by performing relaparatomy. RESULTS: The highest anastomosis bursting pressure was found in Group-III (p<0.05). The highest tissue hydroksiproline level was found in Group-III (p<0.005 Group I-III, Group II-III). When histopathologic findings were evaluated by comparing the three groups in this study, the healing of the intestine tissue score was statistically insignificant between group-II and III, for both group-II and III, healing score was statistically significant higher than Group-I (p<0.05 Group I-III and Group I-II). CONCLUSION: Cleaning the abdomen before the anastomosis using antibacterial soaked material increased resection safety in the presence of peritonitis and anastomosis safety in primary anastomosis.

Papillary intralymphatic angioendothelioma: An extremely rare tumor.

Papillary intralymphatic angioendothelioma (PILA), an intralymphatic lesion of vascular ducts, is an extremely rare tumor. It is generally encountered as an unexpected pathology following excision of a vascular skin lesion. Re-excision is the commonly preferred treatment option once the pathological diagnosis is established. In this article, we present a 12-year-old male patient with a PILA treated with re-excision, skin grafting, and sentinel lymph node biopsy. The patient was symptom-free at annual follow-up. A sentinel lymph node biopsy during re-excision might be a good option in the surgical management of PILA. Frequent examination of lymph nodes and skin lesion also might be beneficial during the follow-ups.

Is body mass index and obesity surgery mortality score important in perioperative complications of laparoscopic sleeve gastrectomy before discharge? / Os índices de mortalidade, de massa corporal e da cirurgia da obesidade são importantes nas complicações perioperatórias de gastrectomia vertical laparoscópica antes da alta?

ABSTRACT Background: Morbid obesity surgery and related complications have increased with time. Aim: To evaluate the relationship between perioperative complications before discharge and preoperative body mass index and obesity surgery mortality score in laparoscopic sleeve gastrectomy. Method: 1617 patients who met the inclusion criteria were evaluated retrospectively. The patients were examined in terms of demographic data, presence of comorbidities, whether there were complications or not, type of complications and obesity surgery mortality score. Results: Complications were seen in 40 patients (2.5%) and mortality wasn't seen in the early postoperative period before discharge. The mean age of patients with complications was 36.3±10.02 years (19-57) and without complications 34.12±9.54 (15-64) years. The preoperative mean BMI values of patients with and without complications were 45.05±3.93 (40-57) kg/m2 and 44.8±3.49 (35-67) kg/m2 respectively. According to BMI groups 40-45 kg/m2, 45-50 kg/m2 and 50 and over, there was not any statistical significance seen in three groups in terms of complication positivity and major-minor complication rates. There was not any statistical significance seen between the patients with and without major-minor complications and obesity surgery mortality score. Conclusion: There was not any relation between perioperative laparoscopic sleeve gastrectomy complication rates before discharge and BMI and obesity surgery mortality scores.

RESUMO Racional: A cirurgia da obesidade mórbida e complicações relacionadas aumentaram com o tempo. Objetivo: Avaliar a relação entre as complicações perioperatórias antes da alta e o índice de massa corporal pré-operatório e o escore de mortalidade da cirurgia de obesidade na gastrectomia vertical laparoscópica. Método: 1.617 pacientes que atenderam aos critérios de inclusão foram avaliados retrospectivamente. Os pacientes foram examinados quanto aos dados demográficos, presença de comorbidades, ocorrência ou não de complicações, tipo de complicações e escore de mortalidade cirúrgico da obesidade. Resultados: Complicações foram observadas em 40 pacientes (2,5%) e mortalidade não foi observada no período pós-operatório imediato antes da alta. A média de idade dos pacientes com complicações foi de 36,3±10,02 anos (19-57) e sem complicações de 34,12±9,54 (15-64) anos. Os valores médios de IMC pré-operatórios dos pacientes com e sem complicações foram 45,05±3,93 (40-57) kg/m2 e 44,8±3,49 (35-67) kg/m2, respectivamente. De acordo com os grupos de IMC 40-45 kg/m2, 45-50 kg/m2 e 50 e mais, não houve qualquer significância estatística observada em três grupos em termos de positividade de complicações e taxas de complicações maiores-menores. Não houve significância estatística entre os pacientes com e sem complicações maiores e menores e o escore de mortalidade da cirurgia de obesidade. Conclusão: Não houve qualquer relação entre as taxas de complicações da gastrectomia vertical laparoscópica perioperatória antes da alta e os escores de mortalidade da cirurgia de obesidade e IMC.

A micropillar-based microfluidic viscometer for Newtonian and non-Newtonian fluids.

In this study, a novel viscosity measurement technique based on measuring the deflection of flexible (poly) dimethylsiloxane (PDMS) micropillars is presented. The experimental results show a nonlinear relationship between fluid viscosity and the deflection of micropillars due to viscoelastic properties of PDMS. A calibration curve, demonstrating this nonlinear relationship, is generated, and used to determine the viscosity of an unknown fluid. Using our method, viscosity measurements for Newtonian fluids (glycerol/water solutions) can be performed within 2-100 cP at shear rates γ = 60.5-398.4 s-1. We also measured viscosity of human whole blood samples (non-Newtonian fluid) yielding 2.7-5.1 cP at shear rates γ = 120-345.1 s-1, which compares well with measurements using conventional rotational viscometers (3.6-5.7 cP). With a sensitivity better than 0.5 cP, this method has the potential to be used as a portable microfluidic viscometer for real-time rheological studies.

Oberg-Manske-Tonkin Classification of Congenital Upper Extremity Anomalies: The First Report From Turkey.

PURPOSE: Although the Oberg-Manske-Tonkin (OMT) classification has been recommended by the International Federation of Societies for Surgery of the Hand owing to some insufficiencies of the Swanson classification system, it has not achieved a universal adoption by hand surgeons. In this study, we hypothesize that the OMT classification can be used easily to classify congenital upper extremity anomalies. We also aim to make epidemiological analysis of congenital upper extremity anomalies with the OMT classification and to compare the applicability of the OMT and the Swanson classifications. METHODS: We retrospectively analyzed 711 patients and 833 extremities operated on between 2012 and 2017. Photographs, plain x-rays, and brief medical histories of the patients were evaluated by 4 plastic surgeons. Two independent evaluations were made by each surgeon in 1-month interval using these classification systems. RESULTS: Total number of upper extremity anomalies recorded was 1050. Of the 711 patients operated on, 122 had bilateral anomalies. The anomalies were identified in 833 extremities because many extremities had more than a single diagnosis. We were able to classify all of the anomalies within the OMT classification. The OMT classification gives better reliability results compared with the Swanson classification according to intrarater and interrater reliabilities. CONCLUSIONS: Compared with the Swanson classification system based on phenotypic evaluation of the extremity, the OMT classification system is easier to apply and the association of the anomaly with the embryologic origin during evaluation is possible. We believe that multiple studies from different centers will boost the international acceptance of the OMT classification.

Drug Repositioning Screen on a New Primary Cell Line Identifies Potent Therapeutics for Glioblastoma.

Glioblastoma is a malignant brain cancer with limited treatment options and high mortality rate. While established glioblastoma cell line models provide valuable information, they ultimately lose most primary characteristics of tumors under long-term serum culture conditions. Therefore, established cell lines do not necessarily recapitulate genetic and morphological characteristics of real tumors. In this study, in line with the growing interest in using primary cell line models derived from patient tissue, we generated a primary glioblastoma cell line, KUGBM8 and characterized its genetic alterations, long term growth ability, tumor formation capacity and its response to Temozolomide, the front-line chemotherapy utilized clinically. In addition, we performed a drug repurposing screen on the KUGBM8 cell line to identify FDA-approved agents that can be incorporated into glioblastoma treatment regimen and identified Topotecan as a lead drug among 1,200 drugs. We showed Topotecan can induce cell death in KUGBM8 and other primary cell lines and cooperate with Temozolomide in low dosage combinations. Together, our study provides a new primary cell line model that can be suitable for both in vitro and in vivo studies and suggests that Topotecan can offer promise as a therapeutic approach for glioblastoma.

Nesprin-1 impact on tumorigenic cell phenotypes.

The largest protein of the nuclear envelope (NE) is Nesprin-1 which forms a network along the NE interacting with actin, Emerin, Lamin, and SUN proteins. Mutations in the SYNE1 gene and reduction in Nesprin-1 protein levels have been reported to correlate with several age related diseases and cancer. In the present study, we tested whether Nesprin-1 overexpression can reverse the malignant phenotype of Huh7 cells, a human liver cancer cell line, which carries a mutation in the SYNE1 gene resulting in reduced Nesprin-1 protein levels, has altered nuclear shape, altered amounts and localization of NE components, centrosome localization and genome stability. Ectopic expression of a mini-Nesprin-1 led to an improvement of the nuclear shape, corrected the mislocalization of NE proteins, the centrosome positioning, and the alterations in the DNA damage response network. Additionally, Nesprin-1 had a profound effect on cellular senescence. These findings suggest that Nesprin-1 may be effective in tumorigenic cell phenotype correction of human liver cancer.