Sex and the Kidney Drug-Metabolizing Enzymes and Transporters: Are Preclinical Drug Disposition Data Translatable to Humans?

Cross-species differences in drug transport and metabolism are linked to poor translation of preclinical pharmacokinetic and toxicology data to humans, often resulting in the failure of new chemical entities (NCEs) during clinical drug development. Specifically, inaccurate prediction of renal clearance and renal accumulation of NCEs due to differential abundance of enzymes and transporters in kidneys can lead to differences in pharmacokinetics and toxicity between experimental animals and humans. We carried out liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based protein quantification of 78 membrane drug-metabolizing enzymes and transporters (DMETs) in the kidney membrane fractions of humans, rats, and mice for characterization of cross-species and sex-dependent differences. In general, majority of DMET proteins were higher in rodents than in humans. Significant cross-species differences were observed in 30 out of 33 membrane DMET proteins quantified in all three species. Although no significant sex-dependent differences were observed in humans, the abundance of 28 and 46 membrane proteins showed significant sex dependence in rats and mice, respectively. These cross-species and sex-dependent quantitative abundance data are valuable for gaining a mechanistic understanding of drug renal disposition and accumulation. Further, these data can also be integrated into systems pharmacology tools, such as physiologically based pharmacokinetic models, to enhance the interpretation of preclinical pharmacokinetic and toxicological data.

ASPSCR1::TFE3 Drives Alveolar Soft Part Sarcoma by Inducing Targetable Transcriptional Programs.

Alveolar soft part sarcoma (ASPS) is a rare mesenchymal malignancy driven by the ASPSCR1::TFE3 fusion. A better understanding of the mechanisms by which this oncogenic transcriptional regulator drives cancer growth is needed to help identify potential therapeutic targets. In this study, we characterized the transcriptional and chromatin landscapes of ASPS tumors and preclinical models, identifying the essential role of ASPSCR1::TFE3 in tumor cell viability by regulating core transcriptional programs involved in cell proliferation, angiogenesis, and mitochondrial biology. ASPSCR1::TFE3 directly interacted with key epigenetic regulators at enhancers and promoters to support ASPS-associated transcription. Among the effector programs driven by ASPSCR1::TFE3, cell proliferation was driven by high levels of cyclin D1 expression. Disruption of cyclin D1/CDK4 signaling led to a loss of ASPS proliferative capacity, and combined inhibition of CDK4/6 and angiogenesis halted tumor growth in xenografts. These results define the ASPS oncogenic program, reveal mechanisms by which ASPSCR1::TFE3 controls tumor biology, and identify a strategy for therapeutically targeting tumor cell-intrinsic vulnerabilities. Significance: The ASPSCR1::TFE3 fusion propels the growth of alveolar soft part sarcoma  by activating transcriptional programs that regulate proliferation, angiogenesis, mitochondrial biogenesis, and differentiation and can be therapeutically targeted to improve treatment.

From waste to energy storage: post-consumer waste expanded polystyrene/rGO composite as a high performance self-standing electrode for coin cell supercapacitors.

This research work aims to explore the potential usage of post-consumer waste expanded polystyrene (EPS) for the fabrication of self-standing electrodes by incorporating reduced graphene oxide (rGO) into it via a facile cost-effective mechanical mixing process. The π-π interaction between the expanded polystyrene and rGO is evidenced from FT-IR and Raman analysis. The elevated thermal stability of the EPS/rGO composite from thermogravimetric analysis (TGA) further confirms the interconnection between the rGO and EPS. This π-π stacking interaction between the rGO and the polystyrene molecules present in the polymer matrix enable the composite material to be interconnected throughout which is beneficial for the charge transport process. The symmetric coin cell supercapacitor fabricated using the EPS/rGO composite electrode can be operated with a high operating voltage of 1.6 V in aqueous KOH and Na2SO4 electrolytes. The devices fabricated with KOH and Na2SO4 electrolytes deliver an areal capacitance of 11.9 mF cm-2 and 10 mF cm-2 at the discharge current density of 0.1 mA cm-2. Further, the devices fabricated with the KOH and Na2SO4 electrolytes demonstrated remarkable rate capability of 87.1% and 99.5% after 10 000 continuous charge discharge cycles. This facile method of preparation without consuming energy or polluting the environment is a novel approach which can be scaled-up to large-scale fabrication of self-standing plastic electrodes for low-cost energy storage applications.

Surgical outcomes of traumatic C2 body fractures: a retrospective analysis.

BACKGROUND: C2 vertebral body fractures are relatively rare fractures with no defined management protocol and outcomes. The aim of the study was to evaluate the clinical and radiological outcomes of C2 body fractures. METHODS: The study was conducted at the Department of Neurosurgery, Nizam's Iinstitute of Medical Sciences, Hyderabad, India, following clearance from the Institutional Ethical Committee. The data of all patients with traumatic C2 body fracture who were managed at our Department between January 2008 and January 2019 were retrieved from the database. Functional status of the patients was assessed by Neck Disability Index while pain was assessed by VAS at follow-up after at least 6 months. Fusion and regional kyphotic angles (O-C2 and C2-C7) were assessed for radiological outcome. RESULTS: There were a total of 16 patients with isolated C2 body fractures in the defined time period. The male (N.=11): female (N.=5) distribution was 2.2:1. Ten patients had road traffic accidents while the remaining 6 had history of fall from height. Only 3 patients presented with neurological deficits. Benzel type 3 fracture pattern distribution was the most common fracture pattern (type 1=1, type 2=5, type 3=9). Of these, 10 were operated and 6 were managed conservatively. The VAS and NDI values improved significantly in all at follow-up when compared to values at presentation (P=0.001). Time to return to work was significantly shorter in those treated with surgical intervention (mean: 2.9±0.87) (P=0.001). Fusion was achieved in all the patients in both groups. Mean O-C2 angle at follow-up was 21.13±10.1. Mean O-C2 angle was significantly decreased in non-surgical group at follow-up (P=0.039) but no significant reduction was observed in surgical group. CONCLUSIONS: The management of C2 body fractures needs to be individualized, reserving surgical management for fractures requiring fragment retrival or restitution of alignment if facets were fractured.

Sagittal realignment of the subaxial spine following occipito-cervical fusion.

BACKGROUND: Sagittal imbalance after spine surgery may be a major source of pain and disability and this holds good even for highly mobile cervical region. However, very few studies have addressed the issue of the impact on adjacent and distant spinal segments following occipito-cervical fixations. The aim was to analyze the changes in the sagittal alignment of subaxial spine following occipito-cervical fixations. METHODS: A total of 24 consecutive patients who underwent O-C2 fixation, with at least 2 years follow-up were retrospectively reviewed for changes in the parameters of sagittal alignment using mid-sagittal CT scan. Sagittal parameters, McGregor's line, O-C2 angle, C2-C7 angle, O-C7 angle and T1 slope were measured in pre- and at the final follow-up using Auto CAD software 2010 version (AutoDesk, Mill Valley, CA, USA); moreover, statistical analysis was done by using SPSS Statistics for Windows (IBM Corp., Armonk, NY, USA) version 21.0. RESULTS: The mean values were as follows: 1) MG slope preoperative: 5.96 (SD=3.81), postoperative: 9.42 (SD=9.6) (P=0.097); 2) O-C2 preoperative: 13.56 (SD=10.58), postoperative: 14.67 (SD=10.66) (P=0.32); 3) C2-C7 preoperative: 23.71 (SD=12.10), postoperative: 18.29 (SD=13.68) (0.128); 4) O-C7 preoperative: 20.04 (SD=8.85), postoperative: 25.33 (SD=11.08) (P=0.069); and 5) T1 slope preoperative: 14.42 (SD=10.68), postoperative: 16.58 (SD=8.78) (P=0.291). There was significant positive correlation (r=0.384, P=0.046) between cervical lordosis and T1 slope and a significant negative correlation between O-C2 and C2-C7 (r=-0.415, P=0.044). Subgrouping of fixation angles at O-C2 beyond 20 degrees had major changes in the subaxial spine which also percolated to the thoraco-lumbar spine. CONCLUSIONS: Fixation angles of occipito-cervical spine do impact the sagittal alignment of the subaxial spine.

Procalcitonin Assay Has No Role in the Routine Assessment of Severe Trauma Patients at Admission to the Emergency Department.

Introduction A key challenge in emergency departments (ED) is the early recognition of sepsis or the potential for sepsis in patients. Appropriate and accurate ED triage will ensure improved case management. This study analysed the association between ED findings at admission and outcomes in patients presenting with severe trauma. Methods This was a prospective study conducted at a tertiary level ED and included severely injured adult patients who presented to the ED within 24 hours of injury. Data collected included clinical findings and imaging reports at initial assessment, serum procalcitonin (PCT), length of ICU and hospital stay, the incidence of bloodstream and other infections, and patient outcome as discharge from care or death. Multiple logistic regression was used to assess the association between outcome variables and independent variables. Results A total of 155 patients were included in the study. Head and neck (61.9%), extremity (58%), and chest (45%) were more commonly injured. Injury Severity Score (ISS) >25, Glasgow Coma Scale (GCS) score <8, head and neck injuries, and extremity injuries were found to be significantly associated with mortality. Bloodstream infections were more common in the presence of lung contusions, abdominal injury, operative management, and blood transfusions. PCT levels at admission did not have a significant predictive value for mortality, bloodstream infections, other infectious complications, or length of ICU stay. Conclusions Head injuries were the most common cause of mortality in our study. In addition to the anatomical region involved, ISS and GCS have a significant association with mortality. PCT levels at ED admission do not have any prognostic value and need not be routinely analysed.

Separation Anxiety-An Unseen Cause for Development of Abnormal Oral and Paraoral Habits and Malocclusion: A Review of Literature and Report of Two Cases.

A child's overall well-being depends on its mutual relationship with the caregiver and social environment. Its attachment with the caregiver forms an emotional bond at six months to two years of age. Separation anxiety refers to fear of separation from the attachment figure. In some neglected children in early age of attachment formation, symptoms of separation anxiety persist and they may show debilitating nervous behaviors during growth. It thus becomes an unseen cause of psychological problem and development of abnormal oral habits occlusal dysfunctions and further psychological problems. Oral habits being learnt patterns of muscle contraction are displayed for release of mental tension due to fear and anxiety often associated with anger, hunger, sleep, tooth eruption and fear. Pediatric dentist plays an important role in patient education through anticipatory guidance. Proper diagnosis and early interception of oral habits as a result of separation anxiety disorder can prevent occlusal disturbances in children. The present paper includes review of literature along with the management of two cases of separation anxiety as an unseen cause of development of unique socially unacceptable abnormal oral and paraoral habits and resultant malocclusion. How to cite this article: Dhote VS, Dharmadhikari PM, Bahadure RN, et al. Separation Anxiety-An Unseen Cause for Development of Abnormal Oral and Paraoral Habits and Malocclusion: A Review of Literature and Report of Two Cases. Int J Clin Pediatr Dent 2021;14(S-2):S199-S205.

Modulation of protein S and growth arrest specific 6 protein signaling inhibits pancreatic cancer cell survival and proliferation.

Thrombotic complications and hypercoagulopathies are commonly associated with the progression of pancreatic ductal adenocarcinoma (PDAC). Although the mechanistic link between the two phenomena is uncertain, there is evidently an increase in procoagulant proteins and a decrease in anticoagulants in PDAC patients. For example, the anticoagulant protein S (PS) is decreased during the progression of PDAC, a condition that possibly contributes to the hypercoagulopathies. PS is also an important signaling molecule that binds a family of tyrosine kinase receptors known as TAM (Tyro3, Axl and Mer) receptors; TAM receptors are often upregulated in different cancers. Growth Arrest Specific 6 or GAS6 protein, a homolog of PS, is also a TAM receptor family ligand. The downstream signaling pathways triggered by this ligand­receptor interaction perform diverse functions, such as cell survival, proliferation, efferocytosis, and apoptosis. Targeting the TAM receptors to treat cancer has had limited success; side effects are a significant obstacle due to the widespread numerous functions of TAM receptors. In the present study, it was revealed that PS­TAM interaction was pro­apoptotic, whereas GAS6­mediated TAM signaling promoted proliferation and survival in select PDAC cell lines. Furthermore, by regulating the balance between these two signaling pathways (by overexpressing PS or knocking down GAS6), the proliferative potential of the cells was decreased. Both long­term and short­term effects of natural PS overexpression were comparable to the treatment of the cells with the drug UNC2025, which inhibits the Mer­receptor. The present study lays the foundation for investigation of PS as a therapeutic agent to control cancer progression and to concurrently arrest thrombotic events.

Padua FIXa resistance to Protein S and a potential therapy for hyperactive FIXa.

INTRODUCTION: Abnormalities in the levels and functions of proteins that maintain hemostasis can cause thrombosis. Factor IX (FIX) R338L, i.e., Factor IX Padua, is a hyperactive clotting factor that promotes thrombosis. The R338L mutation increases the clotting rate by 8-fold despite increasing the Factor IXa enzymatic activity by only 2-fold. Protein S (PS) is a natural anticoagulant that directly inhibits FIXa. Because individuals affected by the R338L mutation have normal concentrations of PS, we speculated that the Padua hypercoagulation phenotype is due to decreased inhibition of FIXa R338L by PS. METHODS: We measured the ability of PS to inhibit FIX R338L, and we assessed the ability of PS to mitigate the prothrombotic effect FIX R338L. RESULTS: Plasma clotting assays demonstrated that 3-fold more PS was required to inhibit FIXa R338L compared with inhibition of wild type FIXa. Thrombin generation assays with Padua patient plasma recapitulated this biochemical consequence of the R338L mutation. Importantly, the less efficient inhibition of FIXa R338L was reversed by increasing PS concentration. Binding and co-immunoprecipitation studies revealed that the decrease in the inhibition of FIXa R338L by PS was caused by a 3- to 4-fold reduction in FIXa R338L affinity for PS. CONCLUSION: In summary, the resistance of FIXa R338L to inhibition by PS likely contributes to the unexpectedly high clotting rate in Padua individuals. Moreover, PS-mediated reversal of the pathological properties of FIXa R338L suggests that PS administration may be a novel and effective means to mitigate thrombophilia caused by any source of elevated FIXa activity.

Surgical Management of Large Radicular Cyst Associated with Mandibular Deciduous Molar Using Platelet-rich Fibrin Augmentation: A Rare Case Report.

Radicular cysts arising from deciduous teeth are rare and usually cause a large bony defect. Autologous platelet-rich fibrin (PRF) is an easily available healing biomaterial in oral surgical defect with the new perspective of accelerated healing of a large bony defect. The present case is of unusually large radicular associated with neglected carious mandibular deciduous second molar in 10-year-old girl and its surgical management with PRF augmentation as a healing biomaterial in the bony defect. One-year follow-up showed uneventful healing and eruption of succedaneous tooth. Healing was relatively faster and facilitated by PRF placement. Furthermore, the importance of anticipatory guidance about the treatment of diseased primary teeth and their preservation gets highlighted.

Physicochemical characterization of black seed oil-milk emulsions through ultrasonication.

The ultrasonic formation of stable emulsions of a bioactive material, black seed oil, in skim milk was investigated. The incorporation of 7% of black seed oil in pasteurised homogenized skim milk (PHSM) using 20kHz high intensity ultrasound was successfully achieved. The effect of sonication time and acoustic power on the emulsion stability was studied. A minimum process time of 8min at an applied acoustic power of 100W was sufficient to produce emulsion droplets stable for at least 8days upon storage at 4±2°C, which was confirmed through creaming stability, particle size, rheology and color analysis. Partially denatured whey proteins may provide stability to the emulsion droplets and in addition to the cavitation effects of ultrasound are responsible for the production of smaller sized emulsion droplets.

Brainstem epidermoid cyst: An update.

The incidence of epidermoid tumors is between 1% and 2% of all intracranial tumors. The usual locations of epidermoid tumor are the parasellar region and cerebellopontine angle, and it is less commonly located in sylvian fissure, suprasellar region, cerebral and cerebellar hemispheres, and lateral and fourth ventricles. Epidermoid cysts located in the posterior fossa usually arise in the lateral subarachnoid cisterns, and those located in the brain stem are rare. These epidermoids contain cheesy and flaky white soft putty like contents. Epidermoid cysts are very slow growing tumors having a similar growth pattern of the epidermal cells of the skin and develop from remnants of epidermal elements during closure of the neural groove and disjunction of the surface ectoderm with neural ectoderm between the third and fifth weeks of embryonic life. We are presenting an interesting case of intrinsic brainstem epidermoid cyst containing milky white liquefied material with flakes in a 5-year-old girl. Diffusion-weighted imaging is definitive for the diagnosis. Ideal treatment of choice is removal of cystic components with complete resection of capsule. Although radical resection will prevent recurrence, in view of very thin firmly adherent capsule to brainstem, it is not always possible to do complete resection of capsule without any neurological deficits.

Respiratory Complication in Liver Disease.

Cirrhosis, the twelfth leading cause of death, accounts for 1.1% of all deaths in the United States. Although there are multiple pulmonary complications associated with liver disease, the most important complications that cause significant morbidity and mortality are hepatopulmonary syndrome, hepatic hydrothorax, and portopulmonary hypertension. Patients with cirrhosis who complain of dyspnea should be evaluated for these complications. This article reviews these complications.