Ca2+ signalling is critical for autoantibody-induced blistering of human epidermis in pemphigus.

BACKGROUND: Pemphigus is a severe bullous autoimmune skin disease. Pemphigus foliaceus (PF) is characterized by antidesmoglein (Dsg) 1 IgG causing epidermal blistering; mucosal pemphigus vulgaris (mPV) by anti-Dsg3 IgG inducing erosions in the mucosa; and mucocutaneous pemphigus vulgaris (PV) by affecting both, with autoantibodies targeting Dsg1 and Dsg3. OBJECTIVES: To characterize the Ca2+ flux pathway and delineate its importance in pemphigus pathogenesis and clinical phenotypes caused by different antibody profiles. METHODS: Immunoprecipitation, Ca2+ flux analysis, Western blotting, immunofluorescence staining, dissociation assays and a human skin ex vivo model were used. RESULTS: PV IgG and PF IgG, but neither Dsg3-specific monoclonal antibody (AK23) nor mPV IgG, caused Ca2+ influx in primary human keratinocytes. Phosphatidylinositol 4-kinase α interacts with Dsg1 but not with Dsg3. Its downstream target - phospholipase-C-γ1 (PLC) - was activated by PV IgG and PF IgG but not AK23 or mPV IgG. PLC releases inositol 1,4,5-trisphosphate (IP3) causing IP3 receptor (IP3R) activation and Ca2+ flux from the endoplasmic reticulum into the cytosol, which stimulates Ca2+ release-activated channels (CRAC)-mediated Ca2+ influx. Inhibitors against PLC, IP3R and CRAC effectively blocked PV IgG and PF IgG-induced Ca2+ influx; ameliorated alterations of Dsg1 and Dsg3 localization, and reorganization of keratin and actin filaments; and inhibited loss of cell adhesion in vitro. Finally, inhibiting PLC or IP3R was protective against PV IgG-induced blister formation and redistribution of Dsg1 and Dsg3 in human skin ex vivo. CONCLUSIONS: Ca2+ -mediated signalling is important for epidermal blistering and dependent on the autoantibody profile, which indicates different roles for signalling complexes organized by Dsg1 and Dsg3. Interfering with PLC and Ca2+ signalling may be a promising approach to treat epidermal manifestations of pemphigus.

[Two neutrophilic dermatoses and one multiple myeloma : A case report]. / Zwei neutrophile Dermatosen und ein multiples Myelom : Ein Fallbericht.

Pyoderma gangraenosum and subcorneal pustulosis are neutrophilic diseases. Different neutrophilic dermatoses can very rarely occur together in one patient, even with a latent period. Furthermore, there seems to be a direct association between IgA gammopathy and neutrophilic dermatosis. Therefore, a gammopathy must always be clarified in the presence of a neutrophilic dermatosis.

H1-antihistamine-refractory chronic spontaneous urticaria: it's worse than we thought - first results of the multicenter real-life AWARE study.

BACKGROUND: Most data on chronic spontaneous urticaria (CSU) originate from highly selected patient populations treated at specialized centres. Little is known about CSU patient characteristics and the burden of CSU in routine clinical practice. AWARE (A World-wide Antihistamine-Refractory chronic urticaria patient Evaluation) is an ongoing global study designed to assess chronic urticaria in the real-life setting. OBJECTIVE: To describe the baseline characteristics of the first 1539 German AWARE patients with H1-antihistamine-refractory CSU. METHODS: This prospective non-interventional study included patients (18-75 years) with a diagnosis of H1-antihistamine-refractory CSU for > 2 months. Baseline demographic and disease characteristics, comorbidities, and pharmacological treatments were recorded. Quality of life (QoL) was assessed using the dermatology life quality index (DLQI), chronic urticaria QoL questionnaire (CU-Q2 oL), and angioedema QoL questionnaire (AE-QoL, in cases of angioedema). Previous healthcare resource utilization and sick leave data were collected retrospectively. RESULTS: Between March and December 2014, 1539 patients were assessed in 256 sites across Germany. The percentage of females, mean age, and mean body mass index were 70%, 46.3 years, and 27 kg/m2 , respectively. The mean urticaria control test score was 7.9, one in two patients had angioedema, and the most frequent comorbidities were chronic inducible urticaria (CIndU; 24%), allergic rhinitis (18.2%), hypertension (18.1%), asthma (12%), and depression (9.5%). Overall, 57.6% of patients were receiving at least one pharmacological treatment including second-generation H1-antihistamines (46.3%), first-generation H1-antihistamines (9.1%), and corticosteroids (15.8%). The mean DLQI, total CU-Q2 oL, and total AE-QoL scores were 8.3, 36.2, and 46.8, respectively. CSU patients reported frequent use of healthcare resources, including emergency services (29.7%), general practitioners (71.9%), and additional allergists or dermatologists (50.7%). CONCLUSIONS AND CLINICAL RELEVANCE: This study reveals that German H1-antihistamine-refractory CSU patients have high rates of uncontrolled disease, angioedema, and comorbid CIndU, are undertreated, have impaired QoL, and rely heavily on healthcare resources.

Prevalence of type I sensitization to alpha-gal in forest service employees and hunters.

BACKGROUND: The production of IgE molecules specific to the carbohydrate galactose-α-1,3-galactose (alpha-gal) is known to induce delayed anaphylaxis against mammalian meat. Tick bites constitute the primary sensitization source, as ticks transfer alpha-gal in their saliva to a host during a bite. The reported prevalence of alpha-gal-specific IgE (alpha-gal-sIgE) positivity varies between different populations from diverse geographic regions. OBJECTIVE: To investigate the prevalence of alpha-gal-sIgE positivity in a population of forest service employees who are highly exposed to ticks in comparison with a residential population and a historic sample. METHODS: A cross-sectional study evaluating 300 forest service employees and hunters from southwest Germany was performed. Alpha-gal-sIgE levels were assessed by ImmunoCAP assay. The prevalence of alpha-gal-sIgE-positive individuals was compared with a matched cohort composed of a residential population and blood samples from forest service employees collected 15 years ago. RESULTS: In the study population, the prevalence of alpha-gal-sIgE-positive (≥0.10 kUA /L) individuals was 35.0%, whereas the prevalence of individuals with alpha-gal-sIgE levels ≥0.35 kUA /L was 19.3%. Alpha-gal-sIgE positivity was associated with total IgE levels and recent tick bites. Mammalian meat-induced delayed anaphylaxis was found in 8.6% of the participants with alpha-gal-sIgE levels ≥0.35 kUA /L. For forest service employees and hunters, the odds ratio for alpha-gal-sIgE positivity was 2.48 compared to the residential population. The prevalence of alpha-gal-sIgE positivity in the current and historic cohort was comparable. CONCLUSION: Forest service employees and hunters compose a population with a high prevalence of alpha-gal-sIgE positivity and carry a considerable risk of red meat allergy.

[Modern allergy diagnostic procedures and their clinical application]. / Moderne Allergiediagnostik und ihre klinische Anwendung.

Due to the increasing incidence of allergies, the importance of allergy diagnostic procedures is growing. In addition to a patient's history and prick, intracutaneous and patch testing, serological testing is an important diagnostic procedure. In recent years, tremendous advances have been made in the area of in vitro allergy tests. In particular, it is possible to predict severity and risk management of food allergies with component-based IgE diagnostic procedures. Even new allergy syndromes have been elucidated at the molecular level.

Evaluation of the effect of GnRH agonist on menstrual reverse in breast cancer cases treated with cyclophosphamide.

UNLABELLED: Twenty-five percent of breast cancer cases are detected during premenopausal period and the number of young women suffering from breast cancer is increasing in the world, especially in Iran. Preservation of fertility and ovarian function leads to improved quality of life of these patients. The aim of this study was to evaluate the effect of gonadotropin releasing hormone (GnRH) agonist on menstrual reverse in breast cancer cases treated with cyclophosphamide regimen. MATERIALS AND METHODS: This randomized clinical trial (RCT) was conducted on 42 adenocarcinoma cases. Mean age of patients was 37 +/- 5 years (range 25 to 45). Primary stages to Stage II (T2N1M0) whose histology reports were negative ER/PR were enrolled in this study. All the enrolled patients were candidates for cyclophosphamide (600 mg/m2), adriamycin (60 mg/m2), and taxoter (75 mg/m2) chemotherapy regimens. RESULTS: Spontaneous menstrual reverse occurred in 90.5% of patients receiving diphereline at three to six months after treatment which occurred in 33.3% of control cases. In control group, 14.3% (three cases) had oligomenorrhea and hypomenorrhea during chemotherapy and 19%(four cases) had spontaneous menstrual reverse at three to six months. It should be noted that there was a significant difference between controls and cases (p < 0.001). This difference was insignificant in cases younger than 35 years (p < 0.594). In 100% of patients older than 35 years who received diphereline, spontaneous menstrual reverse occurred during six months after chemotherapy, but this occurred in only 20% of controls (p < 0.001). Mean serum level of follicle stimulating hormone (FSH) and luteinizing hormone (LH) during and at three months after therapy was significantly lower in cases in comparison with controls, but serum level of estradiol was significantly more in cases three months after chemotherapy (p < 0.001). CONCLUSION: GnRH agonists significantly improve ovarian function andfertility. They also lead to spontaneous menstrual reverse in negative ER/PR breast cancer cases.

MRI in hypertrophic mono- and polyneuropathies.

Different conditions that may lead to enlarged nerves or nerve roots include hereditary motor and sensory neuropathy (HMSN), neurofibromatosis (NF) type 1, chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), and intraneural perineurioma. Differential diagnosis of hypertrophic mono- and polyradiculopathies remains challenging but is important because of different treatments and prognosis. Magnetic resonance imaging (MRI) can identify the hypertrophic nerve segments and guide a fascicular biopsy. A fascicular biopsy will often be necessary for precise diagnosis.

Comparing letrozole with medroxyprogesterone acetate (MPA) as hormonal therapy for simple endometrial hyperplasia without atypia in adult and middle-aged women.

BACKGROUND: The aim of this survey was to compare the effect of letrozole with medroxyprogesterone acetate (MPA) in treatment of simple endometrial hyperplasia to preserve fertility in young women. MATERIALS AND METHODS: Forty-five patients referred to Shahid Sadoughi gynecology clinics from 2009 until 2011 who suffered from abnormal vaginal bleeding or endometrial thickness, that underwent curettage with diagnosis of simple endometrial hyperplasia without atypia were enrolled. The patients were divided randomly into two groups. First group including 22 women receive ten mg MPA, for ten days during a month for three months. All cases were followed by interview, endometrial curetage, and vaginal sonography. Serum level of estradiol was checked before and after treatment. At the end of the study, biopsy was retaken in 41 patients. All the patients were under observation by two gynecologists. RESULTS: Age range of patients was 20 to 42 years. Mean body mass index (BMI) in the MPA and letrozole groups was 29.13 +/- 4.8 and 25.42 +/- 4.2, respectively. Fifty and 34.8 percent of cases had history of obesity or polycystic ovarian syndrome (PCOS) in MPA and letrozole groups, respectively. Forty-one selected cases (20 of the MPA and 21 of the letrozole groups) continued the treatment for three months. The endometrial thicknesses decreased in both groups. Serum estradiol level also decreased in both groups. The most common complication in the MPA and letrozole groups was headache (27.3%) and flashing and dizziness, respectively. The side-effects were reported less in the letrozole group and the most common ones in this group were dizziness and flashing. DISCUSSION: In women suffering from simple endometrial hyperplasia without atypia, letrozole can lead to decrease of serum estradiol level and endometrial thickness like MPA. In both groups, there was no simple hyperplasia report in curettage report following treatment. It should be noted that there was an incomplete response to treat case with pathology of disordered proliferative type. CONCLUSION: Letrozole is a good therapeutic option in simple endometrial hyperplasia without atypia: cases candidate for medical treatment. To confirm the effect and safety of letrozole, more studies with larger samples are recommended.

A comparison of the effect of levonorgestrel IUD with oral medroxyprogesterone acetate on abnormal uterine bleeding with simple endometrial hyperplasia and fertility preservation.

OBJECTIVE: Endometrial hyperplasia is clinically important, because it can lead to abnormal uterine bleeding (AUB) which itself can precede endometrial cancer. Endometrial carcinoma is the most common malignancy of the female genital tract, occurring in about 75%-85% younger, perimenopausal women as endometrial hyperplasia. The treatment is hysterectomy or hormone therapy with progesterone. The aim of this study was, therefore, to compare the effect of levonorgestrel intrauterine device (LNG-IUD) with medroxyprogesterone acetate (MPA) on simple endometrial hyperplasia for fertility preservation. MATERIALS AND METHODS: Forty women in reproductive age (22-47 years) with AUB with endometrial biopsies confirming simple hyperplasia, were enrolled in this study and then randomly divided into two groups. All patients presented with designed special checklist which was filled with satisfaction. Complete history and physical examination especially blood pressure (BP), body mass index (BMI), breast examination, bimanual vaginal examination, and transvaginal sonography (to measure the thickness of endometrial and exclude the other pathologic lesions) were performed. In the first group, treatment was performed with MPA (20 mg/daily) for ten days and in other group with LNG-IUD was prescribed. After three months, transvaginal sonography and biopsy of endometrium were done. The status of AUB and side-effects of two methods,along with the rate of satisfactory were evaluated. RESULT: The findings showed the significant differences in the treatment of simple hyperplasia between two groups (LNG-IUD group vs. MPA group) (p < 0.047). Recovery of AUB in the group LNG was enhanced (p < 0.047). Endometrial thickness was reduced in both groups (p < 0.001), but further reduction in LNG group was seen. Also, LNG was tolerated more than MPA. Side-effects of MPA were more and reached significance (p < 0.003). The rate of satisfaction with LNG was higher than MPA and reached significance (p < 0.048). CONCLUSION: The results of this study show that LNG-IUD is more effective than MPA in treatment of simple endometrial hyperplasia and can be helpful in young women who want to preserve their fertilities.

Biomaterials / bioinks and extrusion bioprinting.

Bioinks are formulations of biomaterials and living cells, sometimes with growth factors or other biomolecules, while extrusion bioprinting is an emerging technique to apply or deposit these bioinks or biomaterial solutions to create three-dimensional (3D) constructs with architectures and mechanical/biological properties that mimic those of native human tissue or organs. Printed constructs have found wide applications in tissue engineering for repairing or treating tissue/organ injuries, as well as in vitro tissue modelling for testing or validating newly developed therapeutics and vaccines prior to their use in humans. Successful printing of constructs and their subsequent applications rely on the properties of the formulated bioinks, including the rheological, mechanical, and biological properties, as well as the printing process. This article critically reviews the latest developments in bioinks and biomaterial solutions for extrusion bioprinting, focusing on bioink synthesis and characterization, as well as the influence of bioink properties on the printing process. Key issues and challenges are also discussed along with recommendations for future research.

Latex agglutination test based on single-chain Fv recombinant antibody fragment.

Recombinant antibodies have been proposed as invaluable tools for various therapeutic and diagnostic purposes. Here, we describe the development of a novel latex agglutination test (LAT) using single-chain Fv recombinant antibody fragment for the detection of K99(+) enterotoxigenic Escherichia coli strains. For the production of a single-chain Fv antibody fragment (scFv) against the major colonization factor (FanC) of K99 antigen, the scFv gene was integrated into a bacterial expression vector under the control of T7 promoter. After high-level expression of soluble scFv (approximately 50 mg/l) in flask cultivation of E. coli DE3 and purification, scFv was immobilized on different latex particles, and then, these sensitized beads were used in LAT. Results obtained with our latex reagents revealed that the recombinant antibody-coated particles were able to give a good agglutination signal with purified antigen, intact cells displaying this protein and clinical specimens. The strength of agglutination of scFv-coated beads for antigen was comparable to that of polyclonal anti-K99-coated particles. However, the assay proved to be simple and rapid, similar to conventional LATs, and owing to more convenient and economical production of recombinant antibodies, they can be considered as a useful reagent for replacing monoclonal antibodies in LATs.

An Absorbing Boundary Condition for the Lattice Boltzmann Method Based on the Perfectly Matched Layer.

The Lattice Boltzmann Method (LBM) is a well established computational tool for fluid flow simulations. This method has been recently utilized for low Mach number computational aeroacoustics. Robust and nonreflective boundary conditions, similar to those used in Navier-Stokes solvers, are needed for LBM-based aeroacoustics simulations. The goal of the present study was to develop an absorbing boundary condition based on the perfectly matched layer (PML) concept for LBM. The derivation of formulations for both two and three dimensional problems are presented. The macroscopic behavior of the new formulation is discussed. The new formulation was tested using benchmark acoustic problems. The perfectly matched layer concept appears to be very well suited for LBM, and yielded very low acoustic reflection factor.

The role of pre-operative infection in the recurrence of thyroglossal duct cyst.

OBJECTIVES: To evaluate the role of pre-operative infection in thyroglossal duct cyst recurrence and post-operative complications of Sistrunk procedure. METHODOLOGY: Medical records of 55 patients undergoing thyroglossal duct cyst excision from 1998 to 2005 were reviewed. The following data was collected: age at operation, gender, surgical technique, presence and length of infection prior to operation, time interval between resolution of infection and operation, recurrence, management of recurrence, and complications. RESULTS: Forty-two patients (76.3%) had no recurrence following Sistrunk procedure, while 13 patients (23.7%) underwent one or more additional procedures due to recurrence. Our results indicate that 61.5% (8 of 13) of those with a failed Sistrunk had pre-operative thyroglossal cyst infection that lasted more than 6 months. In 92.3% of patients with recurrence, the interval between resolution of infection and time of operation was less than one month. CONCLUSION: Effective treatment of thyroglossal duct cyst infection, at least one month pre-operatively, is warranted to reduce post-operative recurrence rate.

Comparison the inflammatory effects of early supplemental parenteral nutrition plus enteral nutrition versus enteral nutrition alone in critically ill patients.

BACKGROUND AND THE PURPOSE OF THE STUDY: It is believed that enteral nutrition (EN) support is the preferred route as compared to parenteral nutrition (PN). Critically ill patients on EN receive less than 60% of their metabolic requirements. To meet patients' calorie goal addition of PN to EN was proposed. This study was conducted to determine whether supplemental PN have any difference with EN alone in regard to inflammatory indices. METHODS: Twenty patients were randomized to either receive EN alone or EN+PN for 7 days. Pre albumin and inflammatory indices including interleukin IL-1, IL-6 and tumor necrosis factor-α (TNF-α) were measured on days of 0, 3,7. Also Sequential Organ Failure Assessment (SOFA) score and Therapeutic Intervention Scoring System-28 (TISS-28) score were calculated on days of 0, 3 and 7. RESULTS AND MAJOR CONCLUSION: IL-1, IL-6 and TNF-α did not show significant difference between two interventions. Pre-albumin was increased from baseline by 9% and 81% in EN and EN+PN groups respectively but it did not reach to statistical significance. SOFA score did not show significant difference. TISS score was higher in EN+PN group on days of 3 and 7. No difference was found between EN and EN+PN regimens in regard to inflammation, while severity of illness may not change with these regimens, nursing workload increases with implementation of supplemental PN.