Different chemical proteomic approaches to identify the targets of lapatinib.

The process of identifying the protein targets and off-targets of a biologically active compound is of great importance in modern drug discovery. Various chemical proteomics approaches have been established for this purpose. To compare the different approaches, and to understand which method would provide the best results, we have chosen the EGFR inhibitor lapatinib as an example molecule. Lapatinib derivatives were designed using linkers with motifs, including amino (amidation), alkyne (click chemistry) and the diazirine group (photo-affinity). These modified lapatinib analogues were validated for their ability to inhibit EGFR activity in vitro and were shown to pull down purified recombinant EGFR protein. In all of the approaches evaluated here, we identified EGFR as the main protein target from the lysate of immortalised cell line expressing EGFR, thus validating its potential use to identify unknown protein targets. Taken together, the results reported here give insight into the cellular activities of lapatinib.

The Benzothiazine Core as a Novel Motif for DNA-Binding Small Molecules.

A new series of 4H-1,3-benzothiazine dyes were prepared and fully characterized in an aqueous medium. Benzothiazine salts were synthesized either through the classical synthetic pathway using Buchwald-Hartwig amination or through economical and environmentally friendly electrochemical synthesis. The latest synthetic approach employs successful electrochemical intramolecular dehydrogenative cyclization of N-benzylbenzenecarbothioamides to form 4H-1,3-benzothiazines. 4H-1,3-Benzothiazines were evaluated as novel DNA/RNA probes. Through the use of several methods such as UV/vis spectrophotometric titrations, circular dichroism and thermal melting experiments, the binding of four benzothiazine-based molecules to polynucleotides was examined. Compounds 1 and 2 acted as DNA/RNA groove binders, thus suggesting the potential of these compounds as novel DNA/RNA probes. This is a proof-of-concept study and will be expanded to include SAR/QSAR studies.

Clinical assessment of disease severity in recurrent aphthous stomatitis.

BACKGROUND: Recurrent aphthous stomatitis (RAS) is one of the most common oral mucosal diseases in many parts of the world. However, there is very limited published clinical evidence for the therapies used in this condition. This could be partly due to the difficulty in evaluating the efficacy of oral ulcer treatment objectively. In this paper, we present a method for assessing and monitoring the severity of oral ulcers before and after treatment. METHODS: Six ulcer characteristics, number, size, duration, ulcer-free period, site and pain, were used to generate an ulcer severity score (USS). The scores for 223 RAS patients were determined, and 79 were scored again after 3-month therapy with topical betamethasone. RESULTS: The scores for the minor RAS group were between 18 and 43 (mean 29.2 ± 5.3). The mean score in the major ulcers group (range: 28-60, mean 39.9 ± 6.1) was significantly greater than in the minor group (P < 0.001). The herpetiform recurrent ulcers score range was wide (range: 18-57, mean 36.6 ± 8.4). The mean severity score decreased significantly after treatment (P < 0.001). CONCLUSIONS: The USS was indicative of the disease activity in recurrent oral ulceration. It helped in assessing the efficacy of therapy, as the change in the numerical score reflected the change in ulcer severity in response to treatment. This tool may well prove to be of value in clinical management, research and in clinical trials.

Single stage turn in perforator infraorbital artery island flap for nasal ala reconstruction.

BACKGROUND: The reconstruction of fullthickness nasal alla defects is challenging procedure. Use of local flaps is acceptable approach. Flap based on infraorbital artery could be used for primary reconstruction of nasal ala defects. METHODS: The prospective study include consecutive series of 15 patients with advanced skin carcinoma of the nasal ala and medial cheek staged T4 by TNM, in whom the turn in infraorbital flap was used. The patient characteristics, type of carcinoma and complications were analyzed. RESULTS: The turn in infraorbital flap was used mostly in male patients (80%), mean age 64 years. The basal cell skin carcinoma was found in 60%. Skin layer was skin grafted. All flaps survived, but in one case a partial wound dehiscence in one partial skin graft loss was found, and in two patients partial nasal obstruction occurred. These three complications were solved as secondary procedures under local anesthesia. CONCLUSION: Full-thickness defect of the nasal ala can be properly reconstructed using flap based on infraorbital artery providing exceptional esthetic and functional results, as single stage procedure.

Modularized iCBT­I self-learn training for university staff-prevention and early intervention in the SARS-CoV-2 crisis: A pilot study.

Background: Insomnia is a widespread disease in adults and has a high prevalence rate. As sleep disturbances are a risk factor concerning mental and physical health, prevention and early intervention are necessary. Thus, the aim of this study was to implement a self-learning prevention and early intervention training for university staff members. We adapted an established cognitive behavioral therapy for insomnia (CBT-I) intervention as an online version for use during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) crisis. Methods: Development and adaptation procedure of the internet-based CBT­I (iCBT-I) prevention and early intervention training is described. Sessions and topics are shown in detail. The Online Sleep Prevention and Treatment Acceptance questionnaire (OSTA) and the Online Sleep Prevention and Treatment Feedback questionnaire (OSTF) were used to assess acceptance. Sleep problems of university staff members were assessed using the Pittsburgh Sleep Questionnaire (PSQI). Results: The online-adapted version consisted of seven modules. Contents of sessions and topics were implemented based on video clips. Drawings were added to information regarding sleep and sleep hygiene as well as addressing stress and cognitions. In all, 15 individuals participated in this pilot study. The new iCBT­I self-learning prevention training was well accepted. In addition, participants scored the online version as helpful based on the OSTA. Prior to online training, 89% of the participants reported impaired sleep quality or insomnia symptoms, and 56% had a PSQI score over 10. After training 78% of participants showed reduced sleep problems according to PSQI and 56% reached clinically significant enhancement. In addition, after training 44% were healthy sleepers. Discussion: This is the first iCBT­I prevention and early intervention training for university staff members. The training by participants was very well accepted and they scored the videos as very helpful. Sleep problems decreased after online training. However, further studies with larger samples and more sleep-related assessment strategies, e.g., actigraphy and sleep log, are necessary.

lncRNA Spehd Regulates Hematopoietic Stem and Progenitor Cells and Is Required for Multilineage Differentiation.

Long non-coding RNAs (lncRNAs) show patterns of tissue- and cell type-specific expression that are very similar to those of protein coding genes and consequently have the potential to control stem and progenitor cell fate decisions along a differentiation trajectory. To understand the roles that lncRNAs may play in hematopoiesis, we selected a subset of mouse lncRNAs with potentially relevant expression patterns and refined our candidate list using evidence of conserved expression in human blood lineages. For each candidate, we assessed its possible role in hematopoietic differentiation in vivo using competitive transplantation. Our studies identified two lncRNAs that were required for hematopoiesis. One of these, Spehd, showed defective multilineage differentiation, and its silencing yielded common myeloid progenitors that are deficient in their oxidative phosphorylation pathway. This effort not only suggests that lncRNAs can contribute to differentiation decisions during hematopoiesis but also provides a path toward the identification of functional lncRNAs in other differentiation hierarchies.

Vascularisation of pectoralis maior myocutaneous flap: anatomical study in human fetuses and cadavers.

Pectoral major flap is one of most frequently used flaps in head and neck reconstruction. The wide attractiveness of this flap is based on secure vascular stalk made from pectoral branch of thoracoacromial artery. The aim of study was to analyze the variations in vascular supply of pectoral major muscle. The investigation was performed on 30 fetuses from 20 to 36 gestation week both gender. For determining vascular network the samples injected with barium sulphate were subjected to Spalteholtz technique, and on 30 fresh cadavers where and we performed injection of Metilen blue 1% in pectoral branch of thoracoacromial artery. In 87% we found one pectoral branch and in 13 % two branches for pectoral muscle were found. Before entering the muscle, pectoral branch is dividing in two terminal branches (67%). In all cases lateral part of pectoral muscle is supplied by branch of lateral thoracic artery. The average area of dyed skin was 13 x 20 cm in region between III and VI rib and from parasternal line to anterior axillar line. The average distance of most distal part of dyed skin to midclavicular point was 24 cm. The pectoral branch of thoracoactomial artery obtains main blood supply for pectoral major muscle mass and despite of anatomical variations can be defined as main pedicle for musculocutaneous flap. Dyed skin confirms that pectoral major flap has defined skin territory sufficient for reconstructions in head and neck area.

Treatment of Scabies and Pediculosis in Health Education Publications and Folk Medicine of Eastern Croatia - Slavonija, Baranja, and Western Srijem County.

Scabies and pediculosis are common parasitic infestations of the skin and hair, manifesting with intense pruritus and effectively treated with modern medications. Because of the attached social stigma linking it with poverty and poor hygiene, patients will often attempt alternative folk-based remedies before confiding in their physicians. We conducted a comprehensive bibliographical study of historic folk literature and interviewed 70 individuals experienced in everyday application of folk medicine in order to categorize available remedies and provide a modern, scientific comment on their effectiveness and dangers. Compositions containing sulfur, copper sulfate, petroleum, coal, tar, and highly alkaline soaps and washing solutions undoubtedly have scabicidal and pediculicidal properties, but they are used either in high concentrations with greater possibility of intoxication and irritation or lower concentrations with questionable therapeutic benefit. These remedies, extracted from historical-cultural frameworks, are poorly adapted to modern standards and can lead to side-effects and complications. Physicians today have to be aware of the reasons their patients seek alternative remedies and know the substances and procedures they may use in self-healing, so as to be able to provide the help that may be needed if those complications occur.

lncRNA requirements for mouse acute myeloid leukemia and normal differentiation.

A substantial fraction of the genome is transcribed in a cell-type-specific manner, producing long non-coding RNAs (lncRNAs), rather than protein-coding transcripts. Here, we systematically characterize transcriptional dynamics during hematopoiesis and in hematological malignancies. Our analysis of annotated and de novo assembled lncRNAs showed many are regulated during differentiation and mis-regulated in disease. We assessed lncRNA function via an in vivo RNAi screen in a model of acute myeloid leukemia. This identified several lncRNAs essential for leukemia maintenance, and found that a number act by promoting leukemia stem cell signatures. Leukemia blasts show a myeloid differentiation phenotype when these lncRNAs were depleted, and our data indicates that this effect is mediated via effects on the MYC oncogene. Bone marrow reconstitutions showed that a lncRNA expressed across all progenitors was required for the myeloid lineage, whereas the other leukemia-induced lncRNAs were dispensable in the normal setting.

Second look procedure for large burn defect by banana peel pericranial flap based on one artery.

INTRODUCTION: Scalp and calvarial defects may result from trauma, thermal or electrical burns, resection of benign or malignant tumors, infections or radionecrosis. Reconstruction of large scalp defects is a demanding procedure. The reconstructive "ladder" are applicable to scalp and calvarial defects reconstruction. CASE OUTLINE: A 68-year-old female was admitted to our clinic due to the nine-day old scalp burn wound, incurred under unclear circumstances. Third degree burn wound affected the left frontal-parietal, temporal and part of the occipital region with carbonification of the whole left ear lobe.The treatment was carried out in two stages. Radical full thickness necrectomy of the scalp was performed, the defect margins were curetted to the active bleeding, and the ear lobe was amputated.The defect sized 23 x 15 cm was reconstructed using the"banana peel"transposition galea-cutaneous flap from the remainder of the scalp, which was based only on the right occipital artery.Two months after the surgery the appearance was satisfactory, and all wounds were healed. CONCLUSION: Designing of large-scale flaps is very hazardous, especially in elderly people. Scalp reconstruction based on one artery has to be planned in detail and performed when the possibility of complication is reduced to minimum. Our case report underlines possible reconstruction as delayed procedure even with the exposed bone (second look procedure), as well as the reconstruction of half scalp with the local flap based on one pericranial artery.

Comparative analysis of autodermal graft and polypropylene mesh use in large incisional hernia defects reconstruction.

BACKGROUND: Large defects of the abdominal wall caused by incisional hernia still represent a challenging problem in plastic, reconstructive, and abdominal surgery. For their successful tension-free repair a proper selection of reconstructive material is essential. In the last decades, the use of synthetic meshes was dominant while biological autodermal grafts were rarely used. The aim of the study was to comparatively analyse efficacy and safety of autodermal graft and polypropylene mesh in surgical treatment of large abdominal wall defects. METHODS: This prospective comparative clinical study enroled 40 patients surgically treated for large incisional hernia repair in a 10-year period. The patients were divided into two equal groups consisting of 20 subjects and treated either by biological autodermal graft or by synthetic polypropylene mesh. The surgical techniques of reconstruction, duration of surgery, the occurrence of early, minor, and major (severe) and delyed complications and hospital stay were analysed. The average follow-up took 2 years. RESULTS: Statistically significant differences in demographic characteristics of patients and in size of defects were not found. The surgical technique of reconstruction with an autodermal graft was more complicated. The duration of surgery in patients treated with autodermal grafts was significantly longer. There was no statistically significant difference regarding occurrence of early, minor postoperative complications and hospital stay in our study. Two severe complications were registered in the synthetic mesh group: intestinal obstruction and enterocutaneous fistula. The recurrence rate was 10% in the autodermal graft group and 15% in the group with a synthetic mesh. CONCLUSION: Tension-free repair of large incisional hernia with autodermal grafts was unjustly neglected despite the fact that it is safe and effective. It can be applied in all cases where synthetic mesh are not indicated (presence of infection, immunodeficient patients, after radiotherapy). They are especially important in war surgery and in lack of funds when commercial grafts cannot be purchased.

Sentinel node biopsy for skin melanoma.

BACKGROUND/AIM: Skin melanoma is one of the most malignant diseases with increasing incidence rate. Sentinel node biopsy (SNB) is very important for early detection of metastatic spread. The aim of the study was to analyze the first 40 patients with skin melanoma of 1 to 4 mm Breslow thickness when SNB was indicated. METHODS: The patient characteristics, localization of the primary melanoma as well as histology grade were analyzed. SNB with intraoperative radiocolloid and methylene blue dye detection was performed. RESULTS: Complication rate after SNB was analyzed and seroma was found in 5% of the patients. The therapeutic node dissection was performed in 10 patients with positive sentinel biopsy. The follow-up lasted two years. In five patients the false negative SNB was defined after the mean time of 11 months and the therapeutic dissection was performed. CONCLUSION: SNB in melanoma patients is a useful diagnostic procedure. It is advised for melanoma of 1 to 4 mm Breslow thickness.

Extended orbital exenteration in the treatment of advanced periocular skin cancer with primary reconstruction with a galeacutaneous flap.

Extended orbital exenteration includes the removal of orbital contents together with the surrounding orbital wall(s). Skin cancers (basal cell cancer and squamous cell skin cancer) arising in the periorbital region could present as invasive tumours infiltrating the orbit and orbital walls. We describe the treatment of advanced invasive skin cancers of the periorbital region by extended orbital exenteration. A retrospective consecutive series over a nine-year period, included 21 extended orbital exenterations treated in a tertiary referral centre. The margins of excision were clear in 18. Twenty postoperative defects were reconstructed using galea-skin flaps, and one defect was left to heal by secondary intention. Two patients died of their disease during the three-year follow up. The reconstruction with frontal or frontoparietal galea-skin flap is a suitable option. The technique is versatile and simple, and gives acceptable aesthetic results. The operating time is shorter than that required for microvascular reconstructions, and the complication rate is low. The secondary defect can be closed primarily or by skin grafting. Extended orbital exenteration offers the best chances of cure in the treatment of non-melanotic skin cancers that have infiltrated the orbit and orbital walls.