Gas Chromatography-Mass Spectrometry Detection of Thymoquinone in Oil and Serum for Clinical Pharmacokinetic Studies.

Thymoquinone (TQ) is the primary component of Nigella sativa L. (NS) oil, which is renowned for its potent hepatoprotective effects attributed to its antioxidant, anti-fibrotic, anti-inflammatory, anti-carcinogenic, and both anti- and pro-apoptotic properties. The aim of this work was to establish a method of measuring TQ in serum in order to investigate the pharmacokinetics of TQ prior to a targeted therapeutic application. In the first step, a gas chromatography-mass spectrometry method for the detection and quantification of TQ in an oily matrix was established and validated according to European Medicines Agency (EMA) criteria. For the assessment of the clinical application, TQ concentrations in 19 oil preparations were determined. Second, two serum samples were spiked with TQ to determine the TQ concentration after deproteinization using toluene. Third, one healthy volunteer ingested 1 g and another one 3 g of a highly concentrated NS oil 30 and 60 min prior to blood sampling for the determination of serum TQ level. After the successful establishment and validation of the measurement method, the highest concentration of TQ (36.56 g/L) was found for a bottled NS oil product (No. 1). Since a capsule is more suitable for oral administration, the product with the third highest TQ concentration (No. 3: 24.39 g/L) was used for all further tests. In the serum samples spiked with TQ, the TQ concentration was reliably detectable in a range between 5 and 10 µg/mL. After oral intake of NS oil (No. 3), however, TQ and/or its derivatives were not detectable in human serum. This discrepancy in detecting TQ after spiking serum or following oral ingestion may be attributed to the instability of TQ in biomatrices as well as its strong protein binding properties. A pharmacokinetics study was therefore not viable. Studies on isotopically labeled TQ in an animal model are necessary to study the pharmacokinetics of TQ using alternative modalities.

Robotic colorectal resection in combination with a multimodal enhanced recovery program - results of the first 100 cases.

PURPOSE: In Germany, colorectal robot-assisted surgery (RAS) has found its way and is currently used as primary technique in colorectal resections at our clinic. We investigated whether RAS can be extensively combined with enhanced recovery after surgery (ERAS®) in a large prospective patient group. METHODS: Using the DaVinci Xi surgical robot, all colorectal RAS from 09/2020 to 01/2022 were incorporated into our ERAS® program. Perioperative data were prospectively recorded using a data documentation system. The extent of resection, duration of the operation, intraoperative blood loss, conversion rate, and postoperative short-term results were analyzed. We documented the postoperative duration of Intermediate Care Unit (IMC) stay and major and minor complications according to the Clavien-Dindo classification, anastomotic leak rate, reoperation rate, hospital-stay length, and ERAS® guideline adherence. RESULTS: One hundred patients (65 colon and 35 rectal resections) were included (median age: 69 years). The median durations of surgery were 167 min (colon resection) and 246 min (rectal resection). Postoperatively, four patients were IMC-treated (median stay: 1 day). In 92.5% of the colon and 88.6% of the rectum resections, no or minor complications occurred postoperatively. The anastomotic leak rate was 3.1% in colon and 5.7% in rectal resection. The reoperation rate was 7.7% (colon resection) and 11.4% (rectal resection). The hospital stay length was 5 days (colon resection) and 6.5 days (rectal resection). The ERAS® guideline adherence rate was 88% (colon resection) and 82.6% (rectal resection). CONCLUSION: Patient perioperative therapy per the multimodal ERAS® concept is possible without any problems in colorectal RAS, leading to low morbidity and short hospital stays.

Depletion of CLL cells by venetoclax treatment reverses oxidative stress and impaired glycolysis in CD4 T cells.

Acquired T-cell dysfunction is characteristic of chronic lymphocytic leukemia (CLL) and is associated with reduced efficacy of T cell-based therapies. A recently described feature of dysfunctional CLL-derived CD8 T cells is reduced metabolic plasticity. To what extend CD4 T cells are affected and whether CD4 T-cell metabolism and function can be restored upon clinical depletion of CLL cells are currently unknown. We address these unresolved issues by comprehensive phenotypic, metabolic, transcriptomic, and functional analysis of CD4 T cells of untreated patients with CLL and by analysis of the effects of venetoclax plus obinutuzumab on the CD4 population. Resting CD4 T cells derived from patients with CLL expressed lower levels of GLUT-1 and displayed deteriorated oxidative phosphorylation (OXPHOS) and overall reduced mitochondrial fitness. Upon T-cell stimulation, CLL T cells were unable to initiate glycolysis. Transcriptome analysis revealed that depletion of CLL cells in vitro resulted in upregulation of OXPHOS and glycolysis pathways and restored T-cell function in vitro. Analysis of CD4 T cells from patients with CLL before and after venetoclax plus obinutuzumab treatment, which led to effective clearance of CLL in blood and bone marrow, revealed recovery of T-cell activation and restoration of the switch to glycolysis, as well as improved T-cell proliferation. Collectively, these data demonstrate that CLL cells impose metabolic restrictions on CD4 T cells, which leads to reduced CD4 T-cell functionality. This trial was registered in the Netherlands Trial Registry as #NTR6043.

The primary stability of two dental implant systems in low-density bone.

Primary stability in low-density bone is crucial for the long-term success of implants. Tapered implants have shown particularly favourable properties under such conditions. The aim of this study was to compare the primary stability of tapered titanium and novel cylindrical zirconia dental implant systems in low-density bone. Fifty implants (25 tapered, 25 cylindrical) were placed in the anterior maxillary bone of cadavers meeting the criteria of low-density bone. The maximum insertion (ITV) and removal (RTV) torque values were recorded, and the implant stability quotients (ISQ) determined. To establish the isolated influence of cancellous bone on primary stability, the implantation procedure was performed in standardized low-density polyurethane foam bone blocks (cancellous bone model) using the same procedure. The primary stability parameters of both implant types showed significant positive correlations with bone density (Hounsfield units) and cortical thickness. In the cadaver, the cylindrical zirconia implants showed a significantly higher mean ISQ when compared to the tapered titanium implants (50.58 vs 37.26; P < 0.001). Pearson analysis showed significant positive correlations between ITV and ISQ (P = 0.016) and between RTV and ISQ (P = 0.035) for the cylindrical zirconia implants; no such correlations were observed for the tapered titanium implants. Within the limitations of this study, the results indicate that cylindrical zirconia implants represent a comparable viable treatment option to tapered titanium implants in terms of primary implant stability in low-density human bone.

[Challenges in neonatal dermatology : An introduction]. / Besonderheiten und Herausforderungen der neonatalen Dermatologie : Eine Einführung in die Problematik.

In order to support dermatologists' interest for the fascinating area of neonatal dermatology, we provide (1) an introduction to the specifics of skin barrier in premature and full-term neonates as well as their clinical implications and (2) an example of age-dependent differential diagnoses and approach to a facial vascular stain in a neonate.

Endoscopic Resection Without Subsequent Ablation Therapy for Early Barrett's Neoplasia: Endoscopic Findings and Long-Term Mortality.

INTRODUCTION: After endoscopic resection (ER) of neoplasia in Barrett's esophagus (BE), it is recommended to ablate the remaining BE to minimize the risk for metachronous disease. However, we report long-term outcomes for a nationwide cohort of all patients who did not undergo ablation of the remaining BE after ER for early BE neoplasia, due to clinical reasons or performance status. METHODS: Endoscopic therapy for BE neoplasia in the Netherlands is centralized in 8 expert centers with specifically trained endoscopists and pathologists. Uniformity is ensured by a joint protocol and regular group meetings. We report all patients who underwent ER for a neoplastic lesion between 2008 and 2018, without further ablation therapy. Outcomes include progression during endoscopic FU and all-cause mortality. RESULTS: Ninety-four patients were included with mean age 74 (± 10) years. ER was performed for low-grade dysplasia (LGD) (10%), high-grade dysplasia (HGD) (25%), or low-risk esophageal adenocarcinoma (EAC) (65%). No additional ablation was performed for several reasons; in 73 patients (78%), the main argument was expected limited life expectancy. Median C2M5 BE persisted after ER, and during median 21 months (IQR 11-51) with 4 endoscopies per patient, no patient progressed to advanced cancer. Seventeen patients (18%) developed HGD/EAC: all were curatively treated endoscopically. In total, 29/73 patients (40%) with expected limited life expectancy died due to unrelated causes during FU, none of EAC. CONCLUSION: In selected patients, ER monotherapy with endoscopic surveillance of the residual BE is a valid alternative to eradication therapy with ablation.

Immediate dental and skeletal influence of distractor position on surgically assisted rapid palatal expansion with or without pterygomaxillary disjunction.

The outcome of surgically assisted rapid palatal expansion (SARPE) can be affected by pterygomaxillary disjunction (PMD) and the distractor position. In this study, SARPE was performed, with or without PMD, in 20 fresh cadaver heads. Transverse expansion was conducted twice using a bone-borne distractor in the anterior and posterior positions, resulting in four groups (n=10). Cone beam computed tomography scans were completed before and after SARPE to evaluate maxillary changes. A comparative anterior decrease and posterior increase in midpalatal opening resulted from SARPE with PMD combined with a posteriorly placed distractor. Significant differences in the internal transverse changes were found between the two SARPE techniques combined with an anterior distractor at the level of the premolars and molars for alveolar ridge width (P=0.040, P=0.024), and at the level of the molars for the dental crown width (P=0.017) and corresponding tooth cusp width (P=0.018). In contrast, using a posteriorly placed distractor led to a significant difference for tooth cusp width only (P=0.050). No statistically significant differences were found between external transverse changes or between distractor positions. PMD is more important in achieving a more uniform and parallel transverse expansion pattern than the distractor position. However, a posterior distractor seems to intensify the effects of PMD.

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.

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.

"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.

Multicentre study of short-course radiotherapy, systemic therapy and resection/ablation for stage IV rectal cancer.

BACKGROUND: The optimal treatment sequence for patients with rectal cancer and synchronous liver metastases remains unclear. The aim of this study was to evaluate the feasibility and effectiveness of short-course pelvic radiotherapy (5 × 5 Gy) followed by systemic therapy and local treatment of all tumour sites in patients with potentially curable stage IV rectal cancer in daily practice. METHODS: This was a retrospective study performed in eight tertiary referral centres in the Netherlands. Patients aged 18 years or above with rectal cancer and potentially resectable liver ± extrahepatic metastases, treated between 2010 and 2015, were eligible. Main outcomes included full completion of treatment schedule, symptom control and survival. RESULTS: In total, 169 patients were included with a median follow-up of 49·5 (95 pr cent c.i. 43·6 to 55·6) months. The completion rate for the entire treatment schedule was 65·7 per cent. Three-year progression-free survival and overall survival (OS) rates were 24·2 (95 per cent c.i. 16·6 to 31·6) and 48·8 (40·4 to 57·2) per cent respectively. Median OS of patients who responded well and completed the treatment schedule was 51·5 months, compared with 15·1 months for patients who did not complete the treatment (P < 0·001). Adequate symptom control of the primary tumour was achieved in 87·0 per cent of all patients. CONCLUSION: Multimodal treatment leads to relief of symptoms in most patients, and is associated with good survival rates in those able to complete the schedule. [Correction added on 12 February 2020, after first online publication: the Conclusion has been reworded for clarity].

ANTECEDENTES: La secuencia óptima de tratamiento en pacientes con cáncer de recto y metástasis hepáticas sincrónicas sigue sin estar clara. El objetivo de este estudio fue evaluar en la práctica diaria la viabilidad y efectividad de la radioterapia pélvica de ciclo corto (5 x 5 Gy) seguida de tratamiento sistémico y tratamiento local de todas las localizaciones del tumor primario en pacientes con cáncer de recto estadio IV potencialmente curables. MÉTODOS: Estudio retrospectivo realizado en ocho centros terciarios de referencia en Holanda. Se consideró elegibles a los pacientes mayores de 18 años con cáncer de recto y metástasis hepáticas ± extrahepáticas potencialmente resecables, que fueron tratados entre 2010 y 2015. Los criterios de valoración principales incluyeron la finalización completa del programa de tratamiento, el control de los síntomas y la supervivencia. RESULTADOS: En total se incluyeron 169 pacientes con una mediana de seguimiento de 50 meses (rango 2-89 meses). La tasa de finalización del programa de tratamiento completo fue del 65,7%. Las tasas de supervivencia libre de progresión a 3 años y supervivencia global (overall survival, OS) fueron 24,2% (i.c. del 95% 16,6-31,6) y 48,8% (i.c. del 95% 40,4-57,2), respectivamente. La mediana de OS de los pacientes que respondieron bien y completaron el programa de tratamiento fue de 51,5 meses, en comparación con 15,1 meses en pacientes que no completaron el tratamiento (P < 0,001). Se logró un control adecuado de los síntomas del tumor primario en el 87,0% de todos los pacientes. CONCLUSIÓN: El tratamiento multimodal consigue paliar los síntomas en la mayoría de los pacientes y se asocia con buenas tasas de supervivencia en aquellos pacientes que pueden completar el programa.

The value of tumor-stroma ratio as predictor of pathologic response after neoadjuvant chemoradiotherapy in esophageal cancer.

BACKGROUND AND PURPOSE: With currently available techniques, the prediction of pathologic complete response after neoadjuvant chemoradiotherapy is insufficient. The tumor-stroma ratio (TSR) has proven to be a predictor of survival for several types of cancer, including esophageal. The aim of this study was to investigate the value of TSR in predicting pathologic response after neoadjuvant chemoradiotherapy in esophageal cancer patients. MATERIALS AND METHODS: Patients with esophageal adenocarcinoma or squamous cell carcinoma who received neoadjuvant chemoradiotherapy followed by a resection were selected. Haematoxylin and eosin (H&E) stained sections of diagnostic biopsies were collected and TSR was independently assessed by two investigators. Patients were categorized in stroma-low (≤50% stroma) and stroma-high (>50% stroma) groups for further analyses. The tumor regression grade (TRG) was assessed on H&E stained sections of the resected primary tumor to determine pathologic response. RESULTS: A total of 94 patients were included in this study, of which 76 patients were categorized as stroma-low and 18 as stroma-high. Forty-two (45%) patients had a major pathologic response (TRG 1-2), whereas 52 (55%) were considered non-responders. After adjustment for gender, tumor type, cT-status and differentiation grade, patients with a stroma-high tumor showed a higher chance of no response compared to patients with a stroma-low tumor (OR 3.57, 95%CI 1.03-12.31, P = 0.04). CONCLUSION: TSR showed to have the potential to aid in the prediction of pathologic response in esophageal cancer patients receiving neoadjuvant chemoradiotherapy. Larger validation studies are necessary before implementing this method in daily practice.

Integrated demultiplexing and amplification of coherent optical combs.

The explosive growth of the internet during the last few decades has been enabled by two complementary innovations in optical communications: the use of multiple optical channels within a single optical fibre, and the increase in the bandwidth of individual channels to hundreds of Gbps. Further increases in overall bandwidth look to be provided by more spectrally efficient optical superchannels that use coherent sub-carriers generated using optical orthogonal frequency division multiplexing (OFDM). Yet, a cost effective way of generating these signals has not been demonstrated. One crucial, but missing piece is an effective means to separate the closely frequency spaced optical sub-carriers from the coherent optical comb before placing information on each sub-carrier, and thus creating the OFDM signal. Here, we demonstrate a flexible strategy implemented in a compact photonic integrated circuit (PIC) that is used to separate and amplify these sub-carriers using on-chip injection locking.

Disrupted regulation of serpinB9 in circulating T cells is associated with an increased risk for post-transplant skin cancer.

Cutaneous squamous cell carcinoma (cSCC) is a serious complication after organ transplantation and patients benefit from an early risk assessment. We hypothesized that functional differences in circulating T cells may represent risk factors for post-transplant cSCC development. Here, we analysed genome-wide DNA methylation of circulating T cells of kidney transplant recipients before the clinical onset of cSCC, to identify differences associated with post-transplant cSCC development. This analysis identified higher DNA methylation of SERPINB9, which is an intracellular inhibitor of granzyme B, a protein that induces apoptosis in target cells. High DNA methylation of SERPINB9 in circulating T cells was confirmed in a second patient cohort during recurrent cSCC, indicating that high SERPINB9 methylation represents a persistent risk factor for cSCC development. At the functional level, the inverse correlation between DNA methylation and messenger RNA expression present in non-cSCC patients was absent in the cSCC patients. Also, a significant difference in serpinB9 protein expression between cSCC patients and non-cSCC patients was observed. It was concluded that disturbed regulation of serpinB9 in circulating T cells represents a novel risk factor for post-transplant cSCC in kidney transplant recipients.

Detection of Rickettsia spp. and Bartonella spp. in Ctenocephalides felis fleas from free-ranging crab-eating foxes (Cerdocyon thous).

Fleas are insects with a worldwide distribution that have been implicated in the transmission of several pathogens. The present study aimed to investigate the presence of Rickettsia spp. (Rickettsiales: Rickettsiaceae) and Bartonella spp. (Rhizobiales: Bartonellaceae) in fleas from free-ranging crab-eating foxes Cerdocyon thous (Linnaeus, 1766) (Carnivora: Canidae) from Rio Grande do Sul, southern Brazil. Fleas were collected manually from animals and used for the molecular detection of Rickettsia spp. and Bartonella spp. Twenty-nine C. thous were sampled in six municipalities. Four foxes were parasitized by 10 fleas, all of which were identified as Ctenocephalides felis (Bouché, 1935) (Siphonaptera: Pulicidae). DNA from Rickettsia felis Bouyer et al., 2001 and Rickettsia asembonensis Maina et al., 2016 were found in three and eight fleas, respectively. In four fleas, DNA of Bartonella sp. was identified. Phylogenetic analysis grouped Bartonella sp. together with other genotypes previously reported in C. felis worldwide. The scenario described in the present study highlights a Neotropical canid parasitized by the invasive cosmopolitan cat flea, which in turn, is carrying potentially invasive vector-borne microorganisms. These findings suggest that C. felis is adapted to wild hosts in wilderness areas in southern Brazil, hypothetically exposing the Neotropical fauna to unknown ecological and health disturbances.

Evaluation of the lingual fracture patterns after bilateral sagittal split osteotomy according to Hunsuck/Epker modified by an additional inferior border osteotomy using a burr or ultrasonic device.

This study was conducted to compare fracture patterns and operation times after sagittal split osteotomy (SSO) by Hunsuck/Epker approach, performed using a burr or ultrasonic device, with and without osteotomy modification. A total of 80 SSOs were performed in fresh human cadavers using a burr or ultrasonic device to investigate the influence of surgical instruments as well as an additional bone cut on the inferior border of the mandible in terms of lingual fracture patterns. The times required for osteotomy and sagittal split were measured, and postoperative cone beam computed tomography images of all splits were analyzed. Without an additional inferior osteotomy, preferred splits according to Hunsuck/Epker were achieved in 35% of cases (7/20) with the burr and 45% (9/20) with the ultrasonic instrument. The inferior modification resulted in a greater number of unwanted fracture patterns in both groups. There was no relationship between the split technique and the fracture pattern (P=0.7854). Statistically significant differences in osteotomy time were observed between burr osteotomy and modified burr osteotomy (P=0.006), as well as modified ultrasonic osteotomy (P<0.001), but not between burr and ultrasonic surgery both without the inferior cut (P=0.36). The bone cut on the inferior border did not improve split control, but rather increased the risk of unwanted fractures and extended the operation time.

The new agreement of the international RIGA consensus conference on nasal airway function tests.

The report reflects an agreement based on the consensus conference of the International Standardization Committee on the Objective Assessment of the Nasal Airway in Riga, 2nd Nov. 2016. The aim of the conference was to address the existing nasal airway function tests and to take into account physical, mathematical and technical correctness as a base of international standardization as well as the requirements of the Council Directive 93/42/EEC of 14 June 1993 concerning medical devices. Rhinomanometry, acoustic rhinometry, peak nasal inspiratory flow, Odiosoft-Rhino, optical rhinometry, 24-h measurements, computational fluid dynamics, nasometry and the mirrow test were evaluated for important diagnostic criteria, which are the precision of the equipment including calibration and the software applied; validity with sensitivity, specificity, positive and negative predictive values, reliability with intra-individual and inter-individual reproducibility and responsiveness in clinical studies. For rhinomanometry, the logarithmic effective resistance was set as the parameter of high diagnostic relevance. In acoustic rhinometry, the area of interest for the minimal cross-sectional area will need further standardization. Peak nasal inspiratory flow is a reproducible and fast test, which showed a high range of mean values in different studies. The state of the art with computational fluid dynamics for the simulation of the airway still depends on high performance computing hardware and will, after standardization of the software and both the software and hardware for imaging protocols, certainly deliver a better understanding of the nasal airway flux.