RUFY4 deletion prevents pathological bone loss by blocking endo-lysosomal trafficking of osteoclasts.

Mature osteoclasts degrade bone matrix by exocytosis of active proteases from secretory lysosomes through a ruffled border. However, the molecular mechanisms underlying lysosomal trafficking and secretion in osteoclasts remain largely unknown. Here, we show with GeneChip analysis that RUN and FYVE domain-containing protein 4 (RUFY4) is strongly upregulated during osteoclastogenesis. Mice lacking Rufy4 exhibited a high trabecular bone mass phenotype with abnormalities in osteoclast function in vivo. Furthermore, deleting Rufy4 did not affect osteoclast differentiation, but inhibited bone-resorbing activity due to disruption in the acidic maturation of secondary lysosomes, their trafficking to the membrane, and their secretion of cathepsin K into the extracellular space. Mechanistically, RUFY4 promotes late endosome-lysosome fusion by acting as an adaptor protein between Rab7 on late endosomes and LAMP2 on primary lysosomes. Consequently, Rufy4-deficient mice were highly protected from lipopolysaccharide- and ovariectomy-induced bone loss. Thus, RUFY4 plays as a new regulator in osteoclast activity by mediating endo-lysosomal trafficking and have a potential to be specific target for therapies against bone-loss diseases such as osteoporosis.

Negative pressure pulmonary edema after general anesthesia using the i-gel.

Negative pressure pulmonary edema (NPPE) is a rare complication that occurs mainly after tracheal extubation. We report a case of postoperative NPPE associated with the use of the i-gel. A 28-year-old woman was scheduled for an emergency right axillary sentinel lymph node excision. During emergence, the patient experienced a sudden onset of airway obstruction, and spontaneous ventilation through the i-gel was impossible. Pink and frothy secretions were noted in the i-gel and the patient's oral cavity. Positive airway pressure with 100% oxygen was applied using a facemask, and the patient was subsequently treated with high-flow oxygen therapy. In this case, laryngospasm or displacement of the i-gel was believed to be the cause of airway obstruction. We recognized that NPPE is likely to occur regardless of the airway device, and the use of the i-gel cannot completely eliminate the possibility of NPPE occurrence.

Differential Expression Patterns of Toll-like Receptors in COVID-19 Patients.

Since Toll-like receptors (TLRs) recognize the earliest signs of infection or cell damage, they play fundamental roles in innate immunity. This review summarizes the numerous studies on the expression of TLRs in patients with Coronavirus disease 2019 (COVID-19). We show that infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can stimulate at least six of the ten TLRs in humans and that this can shape the severity of COVID-19. Specifically, TLR2, TLR4, and TLR9 appear to play pathogenic roles while TLR3, TLR7, and TLR8 may be protective. Most have mutations that could partly explain the susceptibility phenotypes of COVID-19. Further understanding the roles of TLRs in COVID-19 immunopathogenesis could reveal prognostic biomarkers and help drive the development of novel and effective therapeutics for COVID-19.

Severe bradycardia and hypotension induced by autonomic neuropathy in a diabetic patient during general anesthesia - A case report.

Cardiovascular autonomic neuropathy (CAN) is characterized by dysregulation of sympathetic and parasympathetic nervous systems that causes cardiovascular and respiratory disorders. The number of diabetic patients undergoing surgery is increasing in line with the prevalence of DM. Anesthesiologists should pay attention to diabetic patients with CAN because it is related to serious cardiovascular morbidity and mortality. We report an 80-year-old male who underwent cervical laminoplasty. He had severe bradycardia and hypotension from induction to the suspension of surgery. His blood pressure dropped to 70/40 mmHg and his heart rate to 20 bpm. Ephedrine, phenylephedrine, and atropine administration had minimal effect, but after epinephrine administrations, his heart rate and blood pressure increased to 70 bpm and 170/90 mmHg. The operation was discontinued because of the patient's unstable, fluctuating vital signs. The results of autonomic nervous system function examination indicated postganglionic cholinergic sympathetic dysfunction, sympathetic adrenergic dysfunction, and parasympathetic cholinergic dysfunction.

Prophylactic antibiotics induce early postcraniotomy seizures in neurosurgery patients: A case series.

RATIONALE: Antibiotics can cause central nervous system disturbances, manifesting as dizziness, confusion, headache, and seizures. Seizures due to antibiotic administration are related to increased excitatory neurotransmission because antibiotics act as competitive antagonists of the γ-aminobutyric acid type A receptor. PATIENT CONCERNS AND CLINICAL FINDINGS: All 5 patients, comprising 4 females and one male and aged 45 to 72 years, underwent open craniotomy with additional surgical maneuvers according to their specific disease. All patients presented American Society of Anesthesiologists Physical Status grades 1 to 2. There were no specific underlying diseases, except hepatitis C and hypertension. However, seizures developed sequentially in the 5 patients after neurosurgery. DIAGNOSES, INTERVENTIONS, AND OUTCOMES: Early postcraniotomy seizures (PCS) developed in the patients after neurosurgery. Prophylactic antibiotics were administered in all cases to prevent infection due to open craniotomy. This included the administration of 10 g and 2 g of an antibiotic (cefotiam HCL; Jetiam Intravenous Injection 1g®) an hour before the surgery in the ward and half an hour before the surgery in the operating room, respectively. After surgery, cefotiam HCL 2 g was administered in all patients on the day of surgery. Five patients had myoclonic seizure or generalized tonic-clonic seizure several times at emergence or in the intensive care unit. LESSONS: Early PCS occurred in every patient when an overdose of the prophylactic antibiotic was administered. This report showed that the preoperative prophylactic antibiotic cefotiam administered in double doses evoked early PCS within a few hours of drug administration. Furthermore, such experiences caution that preoperative intravenous cephalosporins, including cefotiam, should be administered carefully in open craniotomy.

Pim1 promotes IFN-ß production by interacting with IRF3.

The Pim (proviral integration site for Moloney murine leukemia virus) proteins compose a serine threonine kinase family whose members regulate cell proliferation, migration and cell survival. However, whether Pim kinases participate in innate immune responses is unclear. Here, we show for the first time that Pim1 plays an essential role in the production of interferon (IFN)-ß by macrophages after their Toll-like receptor (TLR) pathway is activated by pathogen-associated molecular patterns (PAMPs). Specifically, Pim1 was quickly upregulated in an NF-κB-dependent manner after TLR stimulation with PAMPs. Pim1 deficiency reduced TLR3- or TLR4-stimulated IFN-ß and IFN-stimulated gene (ISG) expression but not proinflammatory cytokine expression in macrophages. Mechanistically, Pim1 specifically upregulates IRF3 phosphorylation and nuclear translocation. However, this role is not dependent on Pim1 kinase activity. Rather, Pim1 appears to promote IRF3 phosphorylation by enhancing the formation of IFN-ß signaling complexes composed of TRIF, TRAF3, TBK1, and IRF3. Poly (I:C)-treated Pim1-/- mice produced less serum IFN-ß and were less likely to survive than wild-type mice. These findings show for the first time that Pim1 participates in TLR-mediated IFN-ß production, thus revealing a novel target for controlling antiviral innate immune responses.

Glycogen Synthase Kinase 3ß Regulates Antiviral Responses of TLR3 via TRAF2-Src Axis.

The protein tyrosine kinase Src regulates the synthesis of TLR3-mediated IFN-ß via the TBK1-IFN regulatory factor 3 axis. However, the molecular mechanisms regulating Src activity in TLR3 signaling remain unclear. In this study, we report that GSK3ß regulates Src phosphorylation via TNFR-associated factor 2 (TRAF2)-mediated Src ubiquitination. GSK3ß deficiency in mouse embryonic fibroblasts significantly reduces polyinosinic:polycytidylic acid-induced IFN-ß and IFN-stimulated gene expression, which is caused by diminished phosphorylation of Src at tyrosine 416. Src undergoes polyinosinic:polycytidylic acid-dependent lysine 63 chain ubiquitination, and TRAF2 is a direct E3 ligase for Src. Our study reveals novel mechanisms underlying TLR3-mediated antiviral responses mediated via the GSK3ß-TRAF2-Src axis.

Optimization of the enzymatic production of 20(S)-ginsenoside Rg(3) from white ginseng extract using response surface methodology.

The optimization of the conversion of ginseng saponin glycosides to 20(S)-ginsenoside Rg(3) by enzymatic transformation was carried out using response surface methodology (RSM) based on a 2(3) factorial central composite design. The production of 20(S)-ginsenoside Rg(3) using several commercial enzymes indicated that the enzyme Cellulase-12T was the most efficient at producing 20(S)-ginsenoside Rg(3). To optimize the enzymatic production of 20(S)-ginsenoside Rg(3), response surface methodology was applied to determine the ideal amount of white ginseng extract, Cellulase-12T and reaction time. These results indicate that white ginseng extract (1.67%) treated with Celluase-12T (3.67%) for 72 hours had 4 times the quantity of 20(S)-ginsenoside Rg(3) compared to commercial white ginseng extract.