Spermatogonial asynchrony in Tex14 mutant mice lacking intercellular bridges.

The existence of cytoplasmic passages between germ cells and their potential function in the control of the spermatogenic process has long been an intriguing question. Evidence of the important role of such structures, known as intercellular bridges (ICB), in spermatogenesis has been implicated by the failure of spermatogenesis in testis-expressed gene 14 (Tex14) mutant mice, which lack the ICBs, to progress past the pachytene spermatocyte stage. Using these Tex14 mutants, the present study evaluated, for the first time, the behavior and synchrony of the spermatogonial lineage in the absence of ICBs. Our data suggest that the absence of these cytoplasmic connections between cells affects the expansion of the undifferentiated type A (Aundiff) spermatogonia compartment and their transition to A1, resulting in a significant numerical reduction of differentiating A1 spermatogonia, but did not interfere with cell amplification during subsequent mitotic steps of differentiating spermatogonia from A1 through intermediate (In). However, beginning at the type B spermatogonia, the synchrony of differentiation was impaired as some cells showed delayed differentiation compared to their behavior in a normal seminiferous epithelium cycle. Thus although spermatogonial development is able to proceed, in the absence of ICBs in Tex14-/- mutants, the yield of cells, specific steps of differentiation, the synchrony of the cell kinetics, and the subsequent progression in meiosis are quantitatively lower than normal.

A switch in regulatory T cells through farm exposure during immune maturation in childhood.

BACKGROUND: Farm exposure protects against development of allergies early in life. At 4.5 years, protection against asthma by farm-milk exposure was partially mediated by regulatory T cells (Tregs). The aim of this study was to investigate the critical time window of the 'asthma-protective' farm effect via Tregs during childhood immune maturation. METHODS: Tregs were assessed longitudinally at 4.5 and 6 years in 111 children (56 farm and 55 reference children) from the PASTURE/EFRAIM birth cohort (flow cytometry). Peripheral blood mononuclear cells were cultured unstimulated (U), with phorbol 12-myristate 13-acetate/ionomycin (PI) or lipopolysaccharide (LPS), and stained for Tregs (CD4+ CD25high FOXP3upper20% ). mRNA expression of Treg/Th1/Th2/Th17-associated cell markers was measured ex vivo. Suppressive capacity of Tregs on effector cells and cytokines was assessed. Detailed questionnaires assessing farm exposures and clinical phenotypes from birth until age 6 years were answered by the parents. RESULTS: Treg percentage before and after stimulation and FOXP3mRNA expression ex vivo decreased from age 4.5 to 6 years (P(U,LPS) < 0.001; P(PI) = 0.051; P(FOXP3) < 0.001). High vs low farm-milk and animal-stable exposure was associated with decreased LPS-stimulated Treg percentage at age 6 years (P(LPS) = 0.045). Elevated LPS-stimulated-Treg percentage at age 6 was associated with increased risk of asthma (aOR = 11.29, CI: 0.96-132.28, P = 0.053). Tregs from asthmatics vs nonasthmatics suppressed IFN-γ (P = 0.015) and IL-9 (P = 0.023) less efficiently. mRNA expression of Th1/Th2/Th17-associated cell markers decreased between 4.5 and 6 years (P < 0.001). CONCLUSIONS: Tregs at the age of 6 years were decreased with farm exposure and increased within asthmatics, opposite to age 4.5 years. This immunological switch defines a critical 'time window' for Treg-mediated asthma protection via environmental exposure before age 6 years.

Predictors of nonresponse to fluid restriction in hyponatraemia due to the syndrome of inappropriate antidiuresis.

BACKGROUND: Fluid restriction (FR), the first-line treatment for hyponatraemia due to the syndrome of inappropriate antidiuresis (SIAD), often does not lead to successful correction of hyponatraemia. Therefore, predictive markers of treatment response are desirable. We evaluated routinely measured serum (s) and urine (u) parameters, s-copeptin and s-mid-regional pro-atrial natriuretic peptide (s-MR-proANP), as possible predictors of FR response. METHODS: In this prospective observational study, we included patients with profound hyponatraemia (s-sodium <125 mmol L-1 ) due to SIAD. Patients were classified as FR responders (increase in s-sodium concentration of >3 mmol L-1 within 24 h) or nonresponders (increase of ≤3 mmol L-1 within 24 h). Initial laboratory parameters were compared between groups with logistic regression analysis. RESULTS: Of 106 SIAD patients analysed, 82 underwent treatment with FR; 48 (59%) patients showed a successful response to FR and 34 (41%) were considered nonresponders. High levels of u-sodium and u-osmolality were significantly associated with nonresponse to FR [odds ratio (OR) 15.0, 95% confidence interval (CI) 2.4-95.8, P = 0.004 and OR 34.8, 95% CI 1.2-1038.8, P = 0.041, respectively). The association of u-sodium and nonresponse remained significant also after adjustment for diuretic use. Lower levels of s-MR-proANP were associated with nonresponse (OR 0.03, 95% CI 0.003-0.3, P = 0.004), whereas s-copeptin was not significantly associated with response to FR. CONCLUSION: Easily measured laboratory parameters, especially u-sodium, correlate with therapeutic response and identify patients most likely to fail to respond to FR. Measurement of these parameters may facilitate early treatment choice in patients with SIAD.

Mid-regional pro-atrial natriuretic peptide and the assessment of volaemic status and differential diagnosis of profound hyponatraemia.

BACKGROUND: Hyponatraemia is common and its differential diagnosis and consequent therapy management is challenging. The differential diagnosis is mainly based on the routine clinical assessment of volume status, which is often misleading. Mid-regional pro-atrial natriuretic peptide (MR-proANP) is associated with extracellular and cardiac fluid volume. METHODS: A total of 227 consecutive patients admitted to the emergency department with profound hypo-osmolar hyponatraemia (Na < 125 mmol L(-1) ) were included in this prospective multicentre observational study conducted in two tertiary centres in Switzerland. A standardized diagnostic evaluation of the underlying cause of hyponatraemia was performed, and an expert panel carefully evaluated volaemic status using clinical criteria. MR-proANP levels were compared between patients with hyponatraemia of different aetiologies and for assessment of volume status. RESULTS: MR-proANP levels were higher in patients with hypervolaemic hyponatraemia compared to patients with hypovolaemic or euvolaemic hyponatraemia (P = 0.0002). The area under the curve (AUC) to predict an excess of extracellular fluid volume, compared to euvolaemia, was 0.73 [95% confidence interval (CI) 0.62-0.84]. Additionally, in multivariate analysis, MR-proANP remained an independent predictor of excess extracellular fluid volume after adjustment for congestive heart failure (P = 0.012). MR-proANP predicted the syndrome of inappropriate antidiuresis (SIAD) versus hypovolaemic and hypervolaemic hyponatraemia with an AUC of 0.77 (95% CI 0.69-0.84). CONCLUSION: MR-proANP is associated with extracellular fluid volume in patients with hyponatraemia and remains an independent predictor of hypervolaemia after adjustment for congestive heart failure. MR-proANP may be a marker for discrimination between the SIAD and hypovolaemic or hypervolaemic hyponatraemia.

Accuracy and clinical outcomes of mandibular reconstruction with a virtually planned deep circumflex iliac artery flap with stock temporomandibular joint prosthesis.

The repair of hemimandibulectomy defects involving the temporomandibular joint (TMJ) is challenging. This study compared the functional outcomes and reconstruction accuracy using a deep circumflex iliac artery (DCIA) flap with and without a virtually planned stock TMJ prosthesis (TMJP) after hemimandibulectomy. Ten patients were assessed: five with a TMJP (TMJP group) and five without (control group). A three-dimensional comparison revealed a mean deviation of 0.11 ± 0.04 mm between the planned and actual DCIA flap with TMJP. The planned and actual TMJP positions differed by 0.56 ± 0.57 mm in height, 0.33 ± 0.24 mm ventrally/dorsally, and 1.18 ± 0.42 mm medially/laterally. Mouth opening, laterotrusion, and midline deviation were significantly greater in the control group than in the TMJP group (P = 0.024, P = 0.008, P = 0.024). The deviation in ventral to dorsal translation for the DCIA flap was slightly higher than reported values in the literature, while height deviation was comparable. Lower deviations in the literature were due to the DCIA flap being used where both TMJs were intact. The in-house virtually planned DCIA flap with stock TMJP yielded results comparable to more expensive patient-specific prostheses.

Flow cytometric measurement of cellular FLICE-inhibitory protein (c-FLIP) in ovarian carcinoma effusions.

OBJECTIVE: The objective of this study was to establish a flow cytometry assay for measuring c-FLIP in serous effusions. In addition, we studied the clinical relevance in ovarian carcinoma effusions of this inhibitor protein in the death receptor signalling pathway of apoptosis. METHODS: Two c-FLIP antibodies were tested using Western blotting and the best performing one was used for titration of c-FLIP expression in a panel of five cell lines, consisting of ovarian carcinoma, breast carcinoma and malignant mesothelioma. The concentration that provided the best signal-to-noise ratio was used for comparison of the performance of three fixation and permeabilization protocols. The best performing protocol was chosen for analysis of 69 ovarian carcinoma effusions. c-FLIP expression was analysed for association with clinicopathological parameters and survival. RESULTS: Rabbit polyclonal c-FLIP by Abcam and the IntraStain kit by Dako performed best. c-FLIP expression was detected in tumour cells in all 69 effusions (expression range 21-100%, median = 80%). No association was found between c-FLIP expression and clinicopathological parameters, including chemoresponse and survival. However, an inverse correlation was found between c-FLIP levels and expression of the previously studied apoptosis marker cleaved caspase-3 (P = 0.029). CONCLUSIONS: An assay for measuring c-FLIP in cytology specimens is presented. c-FLIP is frequently expressed in ovarian carcinoma effusions, but its expression appears to be unrelated to disease aggressiveness.

Influence of bone density, screw size and surgical procedure on orthodontic mini-implant placement - part B: implant stability.

This in vitro study aimed to investigate the influence of bone density, implant size, and surgical procedure on the primary stability (PS) of orthodontic mini-implants (OMIs). In total, 640 OMIs of various sizes (2.0 × 7, 2.3 × 7, 2.0 × 11 and 2.3 × 11 mm) were inserted in the artificial bone of different densities (D1-D4). Placement was performed with an insertion angle of 90° or 60° to the bone surface and in 320 cases without predrilling, which resulted in 64 groups. PS was measured on the basis of implant stability quotient (ISQ) and insertion torque (IT). With regard to all possible influencing parameters, the mean PS differed between 39.20 and 60.00 (ISQ), and 10.00 and 39.00 Ncm (IT). The effect of OMI size and surgical procedure was dependent on bone quality. For example, implant size had less effect in high-density bone and was stronger with decreasing density. Overall, implant length had a greater influence than the diameter, and a high correlation was found among both PS measurement techniques. Therefore, a suitable choice of implant size and surgical protocol with regard to bone density can positively influence PS. In principle, ISQ and IT are suitable for measuring OMI stability.

Evaluation of a novel osteosynthesis plate system for mandibular defects.

Continuity defects of the jaw can be reconstructed with titanium plates or microvascular bone flaps; osteosynthesis plates are necessary for both. In this study we performed a retrospective review of patients treated with Medartis MODUS® Mandible Trauma/Reco 2.0-2.5, TriLock bridging plates, mandibulectomy and soft tissue free flap or reconstruction with a bony free flap and TriLock mandibular plates from the same system from January 2015 to August 2019. The variables recorded were sex, age, diagnosis, radiotherapy, date of implantation, date of explantation or death of patient, size of mandibular defect, Jewer classification of defect, number of screws used, segments of bony reconstruction, screws per segment, plate exposure, plate breakage, and pseudarthrosis. The bridging plate group consisted of 41 patients, while the mandibular plate group consisted of 24 patients. The percentage of plate exposure was 17.07% for the bridging plate group and 4.17% for the mandibular plate group. Plate breakage was 0 in both groups. Pseudarthrosis was 4.17% in the mandibular plate group. In the bridging plate group, an anterolateral thigh flap covered all exposures. Of 7 plate exposures, 4 were found in a C defect. The complication rate of the investigated plates was lower than the complication rates of other plate systems.

"Flipped OR": a modified didactical concept for a surgical clerkship in Oral and Maxillofacial Surgery.

The concept of the clerkship at the Department of Oral and Maxillofacial Surgery was modified into a modern and sustainable teaching model with a flipped classroom approach. The aim was to prove whether these changes increased the students' learning success. For the preparation of the clerkship, an e-learning module with 10 chapters was established. Students were supposed to access one chapter ahead of each day's seminar. A test to assess the gain in knowledge was undertaken before and after participation in the clerkship. An evaluation took place after the test. The results of the test after the clerkship were significantly higher than before (p<0.0001). The evaluation showed that the students were highly satisfied with the course in general, and the e-learning module specifically. In particular, the surgical videos helped them to better understand the operative steps. The clerkship at the Department of Oral and Maxillofacial Surgery was modified into a sustainable blended learning concept with a flipped classroom approach. With these developments, the "flipped operating room" was shown to be effective in improving knowledge and competence among undergraduate dental students.

Aesthetic aspects of adjacent maxillary single-crown implants-influence of zirconia and titanium as implant materials.

The purpose of this retrospective survey was to assess the esthetic aspects of adjacent zirconia and titanium implants in the anterior maxillary area. A total of 40 patients and 109 adjacent implants (17 patients with 47 titanium implants and 23 patients with 62 zirconia implants) was included. The primary aim of this survey was to assess the papillary fill (Jemt score). Additionally, further esthetic aspects were assessed. Papillae were fully present (Jemt score 3) around 39.1% of adjacent zirconia implants and 17.4% of adjacent titanium implants (p<0.01). The papilla deficit was significantly higher 1.64 mm (SD 1.35) around titanium compared to zirconia implants 0.92 mm (SD 0.94, p<0.01). The evaluation of the soft tissue recessions revealed no differences between implant materials, whereas titanium implants presented more visible implant shadows (p<0.01). Zirconia implants had more implants with papillae that filled the entire proximal space compared to titanium implants. Furthermore, titanium implants had a greater frequency of visible implant shadowing than the zirconia implants. Esthetic rules such as the interdental contact area and golden percentage rules did not differ significantly between the titanium and zirconia implants.

Evaluation of the bactericidal effect of cold atmospheric pressure plasma on contaminated human bone: an in vitro study.

The use of cold atmospheric pressure plasma (CAPP) as a bacterial decontaminant for chronic wounds has shown good results. The purpose of this in vitro study was to evaluate the bactericidal effects of CAPP on the cancellous area of the bone. Sterile glass slides and processed sterile human bone allografts 1, 2, 3, and 4mm thick were used for initial contamination and further CAPP treatment. Each block was contaminated with Staphylococcus aureus suspension on one side. Each slide was turned 180° and treated on the reverse side. The bacterial count in colony-forming units (CFU) was then measured and compared with that of a control group, and the bactericidal effects of CAPP in relation to bone density evaluated. A significant reduction in count was measured between treated and untreated groups (groups A-D: p<0.01 and group E: p=0.04). A strong positive linear relation was found between bone density and the S aureus count (r=0.844, p=0.156). Treatment with CAPP had a bactericidal effect on bone structures with a penetration depth of up to 4mm. It might be used for all diseases involving infected bone, and so extends the existing range of treatments.

Genetic mapping of phenylalanyl-sRNA synthetase in Escherichia coli.

Genetic mapping of the structural gene for a phenylalanyl-sRNA synthetase in Escherichia coli was accomplished by the use of mutants with temperature-sensitive or p-fluorophenylalanine-resistant enzymes. The structural gene is located at minute 33 of the Taylor-Thoman map of the E. coli chromosome, closely linked to a structural gene (aroD) for one of the enzymes involved in the biosynthesis of the aromatic amino acids, and distant from the known locations of other aminoacyl-sRNA synthetase genes.

Structure of Ribosomal Subunits of M. vannielii: Ribosomal Morphology as a Phylogenetic Marker.

On the basis of ribosomal morphology, it has been proposed that the sulfur-metabolizing archaebacteria constitute a group (the eocytes) with a phylogenetic importance equal to that of the eubacteria, archaebacteria, and eukaryotes. It has been further proposed that eocytes should be given kingdom status. Ribosomal subunits from the methanogenic archaebacterium Methanococcus vannielii were examined by electron microscopy, and their structures were compared to those of other archaebacterial, eubacterial, and eukaryotic ribosomes. 30S subunits from M. vannielii showed the elongated contour and the one-third to two-thirds partition characteristic of such subunits. In addition, the angled asymmetric projections of those subunits showed a squarish base and a beak on the head. 50S subunits from M. vannielii were seen in both crown and kidney views. In crown views, the L1 protuberance was frequently pronounced and split; an incision below this protuberance and a protrusion at the base of the particle were also observed. Although previous studies suggested that certain of these structural features were found exclusively in ribosomes from sulfur-metabolizing archaebacteria, these new results indicate that such features also occur in ribosomes from a typical methanogenic archaebacterium and thus may not be reliable phylogenetic markers.

Geographical prevalence, risk factors and impact of hepatitis B and C after renal transplantation.

BACKGROUND: Hepatitis B (HBV) and hepatitis C (HCV) virus infections are major risk factors affecting long-term morbidity and mortality after renal transplantation. Hepatitis prevalence is subject to geographical variations. OBJECTIVE: To compare and analyze the geographical prevalence, risk factors and impact of HBV and HCV infection in multinational cohorts of renal transplant recipients. METHODS: From 1989 - 2002, data on 12,856 kidney transplant recipients in 37 countries were collected within the prospective MOST (Multinational Observational Study in Transplantation). Subgroup analyses of hepatitis-related prevalence, risk factors and impact were conducted on patients whose HBV and HCV status was available at time of transplantation. Countries were substratified according to population prevalence of > or = 5% HBV or > or = 10% HCV. RESULTS: The prevalence of HBV was 2.9%, of HCV 8.7% and of HBV together with HCV 0.4%. Risk factors for hepatitis infection in renal transplant recipients were long dialysis time, retransplantation and blood transfusions. At each study endpoint up to 5 years after transplantation, no significant differences in graft function were observed, although the 1-year acute rejection rate tended to be lower in HCV+ patients. At 5 years post-transplant, there were no differences between the subgroups and regions regarding infections, post-transplant diabetes mellitus or malignancies including PTLD. CONCLUSIONS: Overall, HCV infections are more prevalent than HBV. Despite large geographical differences in prevalence, HBV and HCV status did not appear to have a significant impact on renal graft function, infections, malignancies and post-transplant diabetes mellitus up to 5 years after renal transplantation throughout the MOST countries.

Selenoprotein synthesis: an expansion of the genetic code.

A number of enzymes employ the unusual amino acid selenocysteine as part of their active site because of its high chemical reactivity. Selenocysteine is incorporated into these proteins co-translationally: biosynthesis occurs on a specific tRNA and insertion into a growing polypeptide is directed by a UGA codon in the mRNA. In E. coli, this requires a specific translation factor. Selenocysteine thus represents a unique expansion of the genetic code.

Improvement of anatomical knowledge and surgical skills in head and neck region - An interdisciplinary hands-on course for clinical students.

INTRODUCTION: Anatomical knowledge and manual skills are required for every surgical procedure. During the regular study the students only have few opportunities to practice their surgical skills actively. To improve this situation, an interdisciplinary hands-on-course for head and neck anatomy and surgery has been set up at the RWTH Aachen University. MATERIALS AND METHODS: The new course has been devised for one week with a full-time schedule. A special anatomical region has been studied each day. After an anatomical lecture, dissections under tutorial instructions took place. According to the anatomical region, a clinical lecture was given. Afterwards, surgical techniques were demonstrated and put into practice on fresh cadaver heads. To check the students' knowledge and the knowledge acquisition during the course, participants had to pass a pre- and post-test. The course was finished with an anonymous written evaluation of the course and an open feedback. RESULTS: The evaluations revealed a very high satisfaction of the students with the course. The post-test showed significant better results in anatomical and clinical knowledge than the pre-test. The mean result of the test was raised from 6.8 to 10.0 (p < 0.001) for the anatomical questions and from 5.9 to 10.5 (p < 0.001) for the clinical questions. CONCLUSION: The new interdisciplinary hands-on course is an effective method to consolidate anatomical knowledge and to link this awareness to a better understanding of head and neck surgery. The students improve their manual skills and get more interested and more open-minded for oral and maxillofacial surgery.

Diagnostic accuracy of measurement methods to assess non-adherence to immunosuppressive drugs in kidney transplant recipients.

Valid assessment of immunosuppressive therapy non-adherence (NAH) is vital: NAH is associated with negative transplantation outcomes. We studied the diagnostic accuracy of assay, patient self-reports and clinicians' collateral reports and composite adherence scores using electronic monitoring (EM) as a reference standard. This cross-sectional study included a convenience sample of 249 adult kidney transplant recipients (Ktx) (female: 43.4%; mean age 53.6 [SD: 12.7], median 7 years [IQR: 9 years] post-Ktx). NAH was assessed using EM over 3 months (i.e. reference standard), assays of cyclosporine, tacrolimus, mycophenolat-mofetil, patients' self-reports and clinicians' collateral reports. The constructed composite adherence score included assay, self-reports and collateral reports. NAH's prevalence across the measurement methods was EM: 17.3%; assay: 33% (cyclosporine: 25.8%; tacrolimus: 35.1%; mycophenolat-mofetil: 40.2%); self-report: 12.4%; collateral reports: 24.9% and composite adherence score: 38.9%, respectively. The composite adherence score and collateral reports showed the highest and lowest sensitivities to NAH (72.1% and 15.8%, respectively). Specificity was highest for collateral reports of at least three clinicians (93.1%). Likelihood ratio of a positive test was 2.74 for composite adherence score. No measures showed high sensitivity alongside high specificity. Combining measures increased diagnostic accuracy, indicating the relevance of combined measures for clinical and research purposes.