CLCNKB mutations causing mild Bartter syndrome profoundly alter the pH and Ca2+ dependence of ClC-Kb channels

ClC-Kb, a member of the ClC family of Cl^? channels/transporters, plays a major role in the absorption of NaCl in the distal nephron. CLCNKB mutations cause Bartter syndrome type 3, a hereditary renal salt-wasting tubulopathy. Here, we investigate the functional consequences of a Val to Met substitution at position 170 (V170M, α helix F), which was detected in eight patients displaying a mild phenotype. Conductance and surface expression were reduced by ~40–50 %. The regulation of channel activity by external H⁺ and Ca²⁺ is a characteristic property of ClC-Kb. Inhibition by external H⁺ was dramatically altered, with pK_H shifting from 7.6 to 6.0. Stimulation by external Ca²⁺ on the other hand was no longer detectable at pH 7.4, but was still present at acidic pH values. Functionally, these regulatory modifications partly counterbalance the reduced surface expression by rendering V170M hyperactive. Pathogenic Met170 seems to interact with another methionine on α helix H (Met227) since diverse mutations at this site partly removed pH sensitivity alterations of V170M ClC-Kb. Exploring other disease-associated mutations, we found that a Pro to Leu substitution at position 124 (α helix D, Simon et al., Nat Genet 1997, 17:171–178) had functional consequences similar to those of V170M. In conclusion, we report here for the first time that ClC-Kb disease-causing mutations located around the selectivity filter can result in both reduced surface expression and hyperactivity in heterologous expression systems. This interplay must be considered when analyzing the mild phenotype of patients with type 3 Bartter syndrome.     

Synovial chondromatosis of the temporomandibular joint with glenoid fossa erosion: Disk preservation for spontaneous anatomical recovery

Synovial chondromatosis (SC) of the temporomandibular joint is a pseudoneoplastic condition characterized by benign cartilaginous metaplasia of synovial tissue mesenchymal residues with intra-articular nodule formation. TMJ involvement is rare. Interposition of loose bodies in the articular space can generate pressure, leading to glenoid fossa erosion with intracranial extension.The aim of this study was to present six SC cases with intracranial extension treated using a surgical procedure.All the patients were treated with open surgery. The superior compartment of the TMJ was opened widely to carefully remove the metaplasic mass. Temporal synovectomy was then performed. Attention was paid to preserving the integrity of the articular disc. The exposed dura mater was also preserved. No material was used to reconstruct the gap in the glenoid fossa.A 1-year follow-up showed no swelling or pain. Patients demonstrated good recovery of mouth opening, with improvement over previous mouth limitations. Morphological studies, performed using MRI and CT, showed complete anatomical recovery of the TMJ and total bone reconstruction of the glenoid fossa.Simple removal of intra-articular nodules, with TMJ arthroplasty and articular disk preservation, represents an efficient treatment option for full anatomical and functional recovery in synovial chondromatosis of the temporomandibular joint with glenoid fossa erosion of less than 1 cm².     

The ontogeny of the endocrine pancreas in the fetal/newborn baboon

Erratic regulation of glucose metabolism including hyperglycemia is a common condition in premature infants and is associated with increased morbidity and mortality. The objective of this study was to examine histological and ultrastructural differences in the endocrine pancreas in fetal (throughout gestation) and neonatal baboons. Twelve fetal baboons were delivered at 125 days (d) gestational age (GA), 140d GA, or 175d GA. Eight animals were delivered at term (185d GA); half were fed for 5 days. Seventy-three nondiabetic adult baboons were used for comparison. Pancreatic tissue was studied using light microscopy, confocal imaging, and electron microscopy. The fetal and neonatal endocrine pancreas islet architecture became more organized as GA advanced. The percent areas of α-β-δ-cell type were similar within each fetal and newborn GA (NS) but were higher than the adults (P<0.05) regardless of GA. The ratio of β cells within the islet (whole and core) increased with gestation (P<0.01). Neonatal baboons, which survived for 5 days (feeding), had a 2.5-fold increase in pancreas weight compared with their counterparts killed at birth (P=0.01). Endocrine cells were also found in exocrine ductal and acinar cells in 125, 140 and 175d GA fetuses. Subpopulation of tissue that coexpressed trypsin and glucagon/insulin shows the presence of cells with mixed endo-exocrine lineage in fetuses. In summary, the fetal endocrine pancreas has no prevalence of a α-β-δ-cell type with larger endocrine cell percent areas than adults. Cells with mixed endocrine/exocrine phenotype occur during fetal development. Developmental differences may play a role in glucose homeostasis during the neonatal period and may have long-term implications.     

Regular and platform switching: bone stress analysis varying implant type

Purpose: This study aimed to evaluate stress distribution on peri‐implant bone simulating the influence of platform switching in external and internal hexagon implants using three‐dimensional finite element analysis. Materials and Methods: Four mathematical models of a central incisor supported by an implant were created: External Regular model (ER) with 5.0 mm × 11.5 mm external hexagon implant and 5.0 mm abutment (0% abutment shifting), Internal Regular model (IR) with 4.5 mm × 11.5 mm internal hexagon implant and 4.5 mm abutment (0% abutment shifting), External Switching model (ES) with 5.0 mm × 11.5 mm external hexagon implant and 4.1 mm abutment (18% abutment shifting), and Internal Switching model (IS) with 4.5 mm × 11.5 mm internal hexagon implant and 3.8 mm abutment (15% abutment shifting). The models were created by SolidWorks software. The numerical analysis was performed using ANSYS Workbench. Oblique forces (100 N) were applied to the palatal surface of the central incisor. The maximum (σ_max) and minimum (σ_min) principal stress, equivalent von Mises stress (σ_vM), and maximum principal elastic strain (ε_max) values were evaluated for the cortical and trabecular bone. Results: For cortical bone, the highest stress values (σ_max and σ_vm) (MPa) were observed in IR (87.4 and 82.3), followed by IS (83.3 and 72.4), ER (82 and 65.1), and ES (56.7 and 51.6). For ε_max, IR showed the highest stress (5.46e‐003), followed by IS (5.23e‐003), ER (5.22e‐003), and ES (3.67e‐003). For the trabecular bone, the highest stress values (σ_max) (MPa) were observed in ER (12.5), followed by IS (12), ES (11.9), and IR (4.95). For σ_vM, the highest stress values (MPa) were observed in IS (9.65), followed by ER (9.3), ES (8.61), and IR (5.62). For ε_max, ER showed the highest stress (5.5e‐003), followed by ES (5.43e‐003), IS (3.75e‐003), and IR (3.15e‐003). Conclusion: The influence of platform switching was more evident for cortical bone than for trabecular bone, mainly for the external hexagon implants. In addition, the external hexagon implants showed less stress concentration in the regular and switching platforms in comparison to the internal hexagon implants.     

Phase II study of sorafenib in patients with relapsed or refractory lymphoma

The safety and activity of the multikinase inhibitor sorafenib were investigated in patients with relapsed or refractory lymphoproliferative disorders who received sorafenib (400 mg) twice daily until disease progression or appearance of significant clinical toxicity. The primary endpoint was overall response rate (ORR). Biomarkers of sorafenib activity were analysed at baseline and during treatment. Thirty patients (median age, 61 years; range, 18-74) received a median of 4 months of therapy. Grade 3-4 toxicities included hand/foot skin reactions (20%), infections (12%), neutropenia (20%) and thrombocytopenia (14%). Two patients achieved complete remission (CR), and two achieved partial remission (PR) for an ORR of 13%. Stable disease (SD) and progressive disease (PD) was observed in 15 (50%) and 11 patients (37%), respectively. The median overall survival (OS) for all patients was 16 months. For patients who achieved CR, PR and SD, the median time to progression and OS was 5 and 24 months, respectively. Compared with patients with PD, responsive patients had significantly higher baseline levels of extracellular signal-regulated kinase phosphorylation and autophagy and presented a significant reduction of these parameters after 1 month of therapy. Sorafenib was well tolerated and had a clinical activity that warrants development of combination regimens.     

Stress amplifies lung tissue mechanics, inflammation and oxidative stress induced by chronic inflammation

ABSTRACT Background: Mechanisms linking behavioral stress and inflammation are poorly understood, mainly in distal lung tissue. Objective: We have investigated whether the forced swim stress (FS) could modulate lung tissue mechanics, iNOS, cytokines, oxidative stress activation, eosinophilic recruitment, and remodeling in guinea pigs (GP) with chronic pulmonary inflammation. Methods: The GP were exposed to ovalbumin or saline aerosols (2×/wk/4wks, OVA, and SAL). Twenty-four hours after the 4th inhalation, the GP were submitted to the FS protocol (5×/wk/2wks, SAL-S, and OVA-S). Seventy-two hours after the 7th inhalation, lung strips were cut and tissue resistance (Rt) and elastance (Et) were obtained (at baseline and after OVA and Ach challenge). Strips were submitted to histopathological evaluation. Results: The adrenals' weight, the serum cortisol, and the catecholamines were measured. There was an increase in IL-2, IL-5, IL-13, IFN-γ, iNOS, 8-iso-PGF2α, and in %Rt and %Et after Ach challenge in the SAL-S group compared to the SAL one. The OVA-S group has had an increase in %Rt and %Et after the OVA challenge, in %Et after the Ach and in IL-4, 8-iso-PGF2α, and actin compared to the OVA. Adrenal weight and cortisol serum were increased in stressed animals compared to nonstressed ones, and the catecholamines were unaltered. Conclusion & clinical relevance: Repeated stress has increased distal lung constriction, which was associated with an increase of actin, IL-4, and 8-iso-PGF2α levels. Stress has also induced an activation of iNOS, cytokines, and oxidative stress pathways.