Diphenyl-tetrazol-propanamide Derivatives Act as Dual-Specific Antagonists of Platelet CLEC-2 and Glycoprotein VI.

BACKGROUND: Platelet C-type lectin-like receptor 2 (CLEC-2) induces platelet activation and aggregation after clustering by its ligand podoplanin (PDPN). PDPN, which is not normally expressed in cells in contact with blood flow, is induced in inflammatory immune cells and some malignant tumor cells, thereby increasing the risk of venous thromboembolism (VTE) and tumor metastasis. Therefore, small-molecule compounds that can interfere with the PDPN-CLEC-2 axis have the potential to become selective antiplatelet agents. METHODS AND RESULTS: Using molecular docking analysis of CLEC-2 and a PDPN-CLEC-2 binding-inhibition assay, we identified a group of diphenyl-tetrazol-propanamide derivatives as novel CLEC-2 inhibitors. A total of 12 hit compounds also inhibited PDPN-induced platelet aggregation in humans and mice. Unexpectedly, these compounds also fit the collagen-binding pocket of the glycoprotein VI molecule, thereby inhibiting collagen interaction. These compounds also inhibited collagen-induced platelet aggregation, and one compound ameliorated collagen-induced thrombocytopenia in mice. For clinical use, these compounds will require a degree of chemical modification to decrease albumin binding. CONCLUSION: Nonetheless, as dual activation of platelets by collagen and PDPN-positive cells is expected to occur after the rupture of atherosclerotic plaques, these dual antagonists could represent a promising pharmacophore, particularly for arterial thrombosis, in addition to VTE and metastasis.

Comparison of serum and whole blood concentrations in quetiapine overdose cases.

This study aimed to compare whole blood and serum concentrations of quetiapine in acute poisoning cases. Authentic whole blood and respective serum samples were routinely collected from patients diagnosed with blood poisoning at our University Hospital. Accordingly, whole blood and serum paired samples from nine patients (one male and eight female patients) were analyzed for quetiapine using liquid chromatography-mass spectrometry (LC-MS). Quetiapine concentrations in whole blood and serum samples ranged widely from 5.4 to 2780 ng/mL and 9.9 to 2500 ng/mL, respectively. The whole blood/serum concentration ratio was 0.5-1.1 and increased together with an increase in whole blood and serum quetiapine concentrations. The ratio was reversed at around 2500 ng/mL to > 1. Our findings suggest that whole blood concentrations are more useful than serum concentrations in diagnosing quetiapine poisonings.

Analysis of Grade Classification for Prehospital Injury Severity Scores Applied in Shonan-area Medical Control Council.

OBJECTIVE: To clarify the usefulness of grade classification for injury severity scores applied in Shonan-area Medical Control Council. METHODS: The participants included 11,668 injury cases that occurred in this jurisdiction from April to September 2016. Multivariate analysis was performed using "severity at the time of the disease" a s the response variable. The AUC-ROC was also compared with and without Grade classification, and potential improvements in discrimination ability were examined. RESULTS: There were 11,271 subjects in the "mild/moderate" group and 397 subjects in the "severe/dead" group. Almost all explanatory variables were significant and independent risk factors in the multivariate analysis, and the "Load & Go adaptation" had a particularly high odds ratio of 20.2. Discrimination ability improved (AUC-ROC: 0.773 VS. 0.787) when Grade classification was added to the conventional pre-hospitalization evaluation items. CONCLUSION: Load & Go adaptation has a great influence on severity, and discrimination ability is improved through Grade classification.

Serum potassium level as a biomarker for acute caffeine poisoning.

AIM: Acute caffeine poisoning presents with hypokalemia, although a relationship between potassium levels and blood concentrations of caffeine has not been established. A correlation between serum potassium level and blood caffeine concentration could establish serum potassium as a simple marker to assess caffeine toxicity in patients with acute toxicity. We investigated whether serum potassium, a symptom of acute caffeine poisoning, could be a parameter correlated with blood caffeine levels. METHODS: We enrolled 85 patients treated for acute caffeine poisoning between January 2012 and March 2019 with blood caffeine levels measured after an overdose of a caffeine-containing over-the-counter drug and for whom serum potassium levels were available. We examined the correlation between serum potassium and blood caffeine concentration. A receiver operating characteristic curve was created with serum potassium values to stratify participants into two groups by blood caffeine concentrations: <20 or ≥20 mg/L (toxic dose) and <80 or ≥80 mg/L (lethal dose). The lethal cut-off value was calculated. RESULTS: The correlation coefficient between serum potassium level and blood caffeine concentration was -0.612 (R 2 = 0.374), indicating a negative correlation. The areas under the curve at blood caffeine concentrations of 20 mg/L (toxic dose) and 80 mg/L (lethal dose) and serum potassium levels were 0.716 and 0.888 (sensitivity, 0.829 and 0.919; specificity, 0.568 and 0.818; cut-off, 3.3 mEq/L and 2.9 mEq/L), respectively. CONCLUSION: Serum potassium levels are associated with blood caffeine concentrations; K+ of 3.3 mEq/L and 2.9 mEq/L indicate acute caffeine poisoning in the toxic and lethal dose, respectively.

Podoplanin is indispensable for cell motility and platelet-induced epithelial-to-mesenchymal transition-related gene expression in esophagus squamous carcinoma TE11A cells.

BACKGROUND: The transmembrane glycoprotein podoplanin (PDPN) is upregulated in some tumors and has gained attention as a malignant tumor biomarker. PDPN molecules have platelet aggregation-stimulating domains and, are therefore, suggested to play a role in tumor-induced platelet activation, which in turn triggers epithelial-to-mesenchymal transition (EMT) and enhances the invasive and metastatic activities of tumor cells. In addition, as forced PDPN expression itself can alter the propensity of certain tumor cells in favor of EMT and enhance their invasive ability, it is also considered to be involved in the cell signaling system. Nevertheless, underlying mechanisms of PDPN in tumor cell invasive ability as well as EMT induction, especially by platelets, are still not fully understood. METHODS: Subclonal TE11A cells were isolated from the human esophageal squamous carcinoma cell line TE11 and the effects of anti-PDPN neutralizing antibody as well as PDPN gene knockout on platelet-induced EMT-related gene expression were measured. Also, the effects of PDPN deficiency on cellular invasive ability and motility were assessed. RESULTS: PDPN-null cells were able to provoke platelet aggregation, suggesting that PDPN contribution to platelet activation in these cells is marginal. Nevertheless, expression of platelet-induced EMT-related genes, including vimentin, was impaired by PDPN-neutralizing antibody as well as PDPN deficiency, while their effects on TGF-ß-induced gene expression were marginal. Unexpectedly, PDPN gene ablation, at least in either allele, engendered spontaneous N-cadherin upregulation and claudin-1 downregulation. Despite these seemingly EMT-like alterations, PDPN deficiency impaired cellular motility and invasive ability even after TGF-ß-induced EMT induction. CONCLUSIONS: These results suggested that, while PDPN seems to function in favor of maintaining the epithelial state of this cell line, it is indispensable for platelet-mediated induction of particular mesenchymal marker genes as well as the potentiation of motility and invasion capacity.

Urinary glucose and ketone bodies as indicators of acute caffeine poisoning.

AIM: In various countries, many fatal health problems have been reported due to high intake of caffeine-rich energy drinks, tablets, and powders. In patients with acute caffeine poisoning, determination of blood caffeine concentration is an important yet difficult task. We aimed to assess whether the presence of glucose and ketone bodies in urine reflected the blood caffeine concentration in patients with acute caffeine poisoning. METHODS: From April 2010 to March 2018, 25 patients with an overdose of only caffeine-rich tablets were admitted to our hospital. Their clinical features were investigated. In addition, we investigated whether the glucose and ketone bodies in the urine reflected blood caffeine concentration in 23 patients who underwent the urine qualitative test at admission. RESULTS: The majority of the patients were young healthy women, whose average caffeine ingestion was 15.6 ± 8.1 g. Initial urine examinations showed glucose in 60% (14/23) of patients and ketone bodies in 57% (13/23) of patients. Ketone bodies or glucose were found in 78% (18/23) of the patients. The correlation between blood caffeine concentration and urinary glucose was R = 0.625, blood caffeine concentration and ketone bodies was R = 0.596, and blood caffeine and both was R = 0.76. CONCLUSION: Urine qualitative test is effective for differential diagnosis and severity assessment of acute caffeine poisoning in patients.

Usefulness of next-generation DNA sequencing for the diagnosis of urinary tract infection.

Acute urinary tract infection (UTI) is a highly common clinical condition. Although bacterial culture is the gold standard diagnostic test, false negative results may be possible, leading to the pathogen being unidentified. In recent years, bacterial DNA sequencing analysis has garnered much attention, but clinical studies are rare in Japan. In this study, we assessed the usefulness of next-generation DNA sequencing (NGS) analysis for acute UTI patients. We thus performed an observational, retrospective case series study. Urine and blood samples were collected from ten acute UTI patients, of whom four had also been diagnosed with urosepsis. Seven variable regions of bacterial 16S rRNA genes were amplified by PCR and then sequenced by IonPGM. The identified bacterial species were compared with those identified using the culture tests and the clinical parameters were analyzed. As a result, the NGS method effectively identified predominant culture-positive bacteria in urine samples. The urine NGS also detected several culture-negative species, which have been reported to be potentially pathogenic. Out of four urosepsis cases, three were pathogen-positive in blood NGS results, while two were pathogen-negative in blood culture. In one sepsis case, although blood culture was negative for Escherichia coli, this species was detected by blood NGS. For non-sepsis cases, however, blood NGS, as well as blood culture, was less effective in detecting bacterial signals. In conclusion, NGS is potentially useful for identifying pathogenic bacteria in urine from acute UTI patients but is less applicable in patients who do not meet clinical criteria for sepsis.

Assessment of Concentrations of Four Phenothiazine Antipsychotics in Serum and Whole Blood Using Different Diatomaceous Earth-based Solid-phase Columns: A Comparative Analysis.

This study attempted to determine the phenothiazine antipsychotics concentration in serum and whole blood samples using various diatomaceous earth-based solid-phase columns and elution solvents and subsequently evaluate their efficiency. Phenothiazine antipsychotics concentrations of 5 - 2000 ng/mL were extracted from serum and whole blood using each column. All compounds were analyzed using liquid chromatography-tandem mass spectrometry. Phenothiazine antipsychotics extraction in serum and whole blood using diatomaceous earth-based solid-phase columns seemed to have an affinity with the elution solvent.

A case of crush syndrome induced by the kneeling seiza position.

Crush syndrome results in a characteristic syndrome of rhabdomyolysis with myoglobinuric acute renal failure. The most commonly described crush injury is that which affects victims of natural disasters such as earthquakes. Here, we report a rare case of crush syndrome that was induced by the kneeling seiza position.

Reduction of immunocompetent T cells followed by prolonged lymphopenia in severe sepsis in the elderly.

OBJECTIVE: To investigate the immunological changes caused by severe sepsis in elderly patients. DESIGN: One-year, prospective observational study. SETTING: Emergency department and intensive care unit of a single university hospital. PATIENTS: Seventy-three patients with severe sepsis and 72 healthy donors. MEASUREMENTS AND MAIN RESULTS: In elderly septic patients (aged 65 yr and over), 3-month survival was significantly reduced compared with that for adult patients (18-64 yr) (60% vs. 89%, p < 0.01). We found that lymphopenia was prolonged for at least 21 days in elderly nonsurvivors of sepsis, while the number of lymphocytes recovered in both adult and elderly survivors of sepsis. In order to examine the immunological status of septic patients, blood samples were collected within 48 hrs of diagnosis of severe sepsis, and peripheral blood mononuclear cells were purified for flow cytometric analysis. T cell levels were significantly reduced in both adult and elderly septic patients, compared with those in healthy donors (56% and 57% reduction, respectively). Interestingly, the immunocompetent CD28+ subset of CD4+ T cells decreased, whereas the immunosuppressive PD-1+ T cells and the percentage of regulatory T cells (CD4+ T cells that are both Foxp3+ and CD25+) increased in elderly patients, especially nonsurvivors, presumably reflecting the initial signs of immunosuppression. CONCLUSION: Reduction of immunocompetent T cells followed by prolonged lymphopenia may be associated with poor prognosis in elderly septic patients.

Partial splenic embolisation using n-butyl cyanoacrylate: intraprocedural evaluation by magnetic resonance imaging.

OBJECTIVES: To evaluate the use of diffusion-weighted imaging (DWI) for estimating infarcted splenic volume during partial splenic embolisation (PSE) using n-butyl cyanoacrylate (NBCA). METHODS: Twenty consecutive patients (57.2 ± 11.7 years) with hypersplenism underwent PSE. Intrasplenic branches were embolised using NBCA via a 2.1-French microcatheter aiming at infarction of 50 to 80 % of total splenic volume. Immediately after PSE, signal intensities (SI) of embolised and non-embolised splenic parenchyma were measured on DWI. Semi-automated volumetry (SAV) on DWI was compared with conventional manual volumetry (MV) on contrast-enhanced CT 1 week after PSE. Platelet counts were recorded before and after PSE. RESULTS: The SI on DWI in the embolised parenchyma decreased significantly (P < 0.01) to 24.7 ± 8.1 % as compared to non-embolised parenchyma. SAV and MV showed a strong correlation (r = 0.913 before PSE, r = 0.935 after PSE, P < 0.01) and significant (P < 0.01) reduction of normal splenic volume was demonstrated on both SAV (71.9 ± 12.4 %) and MV (73.6 ± 9.3 %) after PSE. Based on the initial SAV, three patients (15 %) underwent additional branch embolisation to reach sufficient infarction volume. Platelet counts elevated significantly (522.8 ± 209.1 %, P < 0.01) by 2 weeks after PSE. No serious complication was observed. CONCLUSION: Immediate SI changes on DWI after PSE allowed semi-automated splenic volumetry on site. KEY POINTS: • Partial splenic embolisation (PSE) is an important interventional technique for hypersplenism • Diffusion-weighted MR reveals an immediate decrease in signal in the embolised parenchyma • Such signal reduction permits semi-automated splenic volumetry on site. • This allows precise quantification of the amount of parenchyma infarcted, avoiding additional PSE.

Efficacy of early anticoagulant therapy for venous thromboembolism in polytrauma patients in the acute phase.

OBJECTIVE: To determine whether antithrombotic therapy with warfarin is effective and safe in patients who developed venous thromboembolism in the acute stage of polytrauma, which is associated with bleeding risk. METHOD: A retrospective study of 11 patients (8 males, 3 females; mean age, 39.8 years; injury severity score, 30.1; no fatalities) with deep venous thromboembolism and/or pulmonary embolism who were medicated with heparin and warfarin during their iCU stay. RESULTS: Thrombosis was diagnosed at an average of 11.8 days after admission. Thrombus formation was confirmed in pulmonary arteries in 5 cases and in deep veins in 9 cases. Diagnosis was based on Doppler ultrasound findings in 6 cases and on computed tomography findings in 5 cases. anticoagulant therapy was used in 10 cases, but not in 1 case with cerebral contusion. approximately 33 days after starting anticoagulant therapy, thrombi had disappeared or were reduced in size in 9 of 10 patients with no complications observed. CONCLUSIONS: Heparin and warfarin therapy cleared deep vein and pulmonary artery thrombosis after polytrauma without any bleeding complications. Further studies are necessary to determine the safe anticoagulant dosage and duration for rapid thrombus removal.

Early diagnosis of anti-N-methyl-D-aspartate receptor encephalitis in a young woman with psychiatric symptoms.

A previously healthy 21-year-old woman, transported to our medical emergency center for excluding organic brain disease, had undergone medical examination 9 days before for trembling in her left hand, which was caused by stress. The patient exhibited fever and strange behaviors, e.g., wandering around, babbling, and making smoking gestures; hence, psychiatric examination was performed. The patient's Glasgow Coma Scale score was 4-3-5, and involuntary movement was observed. Cerebrospinal fluid examination revealed increased cell count; hence, we suspected anti-N-methyl-d-aspartate (NMDA) receptor encephalitis. We conducted an abdominal CT scan, which revealed a neoplastic lesion with calcification in the right ovary. Early steroid pulse therapy was started. On hospital day 25, she tested positive for anti-NMDA receptor antibodies; hence, anti-NMDA receptor encephalitis and concomitant ovarian teratoma was diagnosid. She underwent right adnexectomy; subsequently, immunotherapy was performed. The patient recovered and was discharged on hospital day 105. Anti-NMDA receptor encephalitis is not uncommon; however, this disease must be considered for young encephalitis patients exhibiting psychiatric symptoms. If patients (aged ≤ 30 years) presents with encephalitis of uncertain etiology, psychiatric symptoms, seizures, movement disorders, or psychosis, clinicians should consider anti-NMDA encephalitis as a possible diagnosis. Clinical diagnosis should be waged early to ensure timely treatment.

Arterial embolization for iatrogenic life-threatening bleeding from subcutaneous hypervascular tumor in prone position.

Performing angiography in the prone position is a difficult technique; however it is useful in some emergency situation. We experienced a 60 years old male who was performed lipoma excision on his back in his family doctor's clinic. Since massive arterial bleeding could not be controlled with manual astriction, he transferred to our hospital in prone position with hemodynamic instability. Operating field was not kept because of massive bleeding; therefore surgical treatment was impossible. We planed emergency arterial embolization (AE) in prone position. Hence we chose the left radial artery for vascular access. The left subclavicle arteriography showed many major and minor feeding arteries from left subclavicular and axillary arteries and a massive extravasation of the contrast medium. Three major feeding arteries were performed AE with gelatin sponge and steel coils. After AE, massive bleeding was controlled. He could discharge from our hospital on the 5th hospital day without any complication. Arterial embolization for life-threatening bleeding from subcutaneous hypervascular tumor in the prone position is first report to our knowledge, and it is extremely rare. However we thought that this technique is useful for patients who could not turn in the supine position, e.g. massive bleeding during renal biopsy and penetrating trauma from back.

Arterial embolization in patients with grade-4 blunt renal trauma: evaluation of the glomerular filtration rates by dynamic scintigraphy with 99mTechnetium-diethylene triamine pentacetic acid.

BACKGROUND: High-grade blunt renal trauma has been treated by arterial embolization (AE). However, it is unknown whether AE preserves renal function, because conventional renal function tests reflect total renal function and not the function of the injured kidney alone. Dynamic scintigraphy can assess differential renal function. METHODS: We performed AE in 17 patients with grade-4 blunt renal trauma and determined their serum creatinine (sCr) level and glomerular filtration rate (GFR; estimated by dynamic scintigraphy) after 3 months. In 4 patients with low GFR of the injured kidney (<20 ml.min-1.1.73 m-2), the GFR and sCr were measured again at 6 months. Data are presented as median and interquartile range (25th, 75th percentile). RESULTS: The median GFR of the injured kidney, total GFR, and median sCr at 3 months were 29.3 (23.7, 35.3) and 96.8 (79.1, 102.6) ml.min-1.1.73 m-2 and 0.6 (0.5, 0.7) mg/dl, respectively. In the patients with low GFR (ml.min-1.1.73 m-2), the median GFR of the injured kidney, total GFR, and median sCr (mg/dl) were 16.2 (15.7, 16.3), 68.7 (61.1, 71.6), and 0.7 (0.7, 0.9), respectively, at 3 months and 34.5 (29.2, 37.0), 90.9 (79.1, 98.8), and 0.7 (0.7, 0.8), respectively, at 6 months. CONCLUSIONS: The function of the injured kidney was preserved in all patients, indicating the efficacy of AE for the treatment of grade-4 blunt renal trauma.

Arterial embolization of an extrapleural hematoma from a dislocated fracture of the lumbar spine: a case report.

BACKGROUND: We present a report of a blunt-trauma patient who developed an atypical extrapleural hematoma with hemodynamic instability following a dislocation fracture of the first lumbar vertebra. We successfully treated her with arterial embolization (AE) of the lumbar and intercostal arteries. CASE REPORT: The patient, a 74-year-old woman, was injured in a traffic accident. At the scene of the accident, she was found to be alert, and her hemodynamic condition was stable. She arrived at our hospital complaining of lumbago. A thoracoabdominal computed tomography (CT) scan with contrast enhancement showed a dislocation fracture of the first lumbar vertebra along with paravertebral and retroperitoneal hematomas. Therefore, we managed the patient conservatively with bed rest. However, 3 h after admission, her blood pressure suddenly decreased. A repeated thoracoabdominal CT scan showed enlargement of the right retroperitoneal hematoma with extravasation of the contrast medium into the right extrapleural space. Angiography was immediately performed, showing extravasation of the contrast media from the right intercostal (Th12) and lumbar arteries (L1). After arterial embolization (AE) with gelatin-sponge particles, extravasation of the contrast medium ceased, and the patient's hemodynamic condition stabilized without massive fluid resuscitation. CONCLUSION: The extrapleural hematoma reduced in size after AE, and almost disappeared on the 14th day of hospitalization. The lumbar spinal fracture was successfully repaired on day 16, and the patient was kept in the hospital to recuperate. We believe that AE is effective for the management of intractable bleeding following fractures of the spine.

Lactate as a prognostic factor in carbon monoxide poisoning: a case report.

Carbon monoxide (CO) poisoning results in various neuropsychological impairments, including delayed encephalopathy (DE) and death. However, factors related to these outcomes are unknown. A group suicide was attempted by 3 young people--a 31-year-old man (patient 1), a 21-year-old woman (patient 2), and a 20-year-old man (patient 3)--by burning charcoal in a closed car. At the emergency department, hypotension and hyperthermia were severe in patient 1, moderate in patient 2, and absent in patient 3, although all the patients were comatose. The initial serum lactate levels were 75.1 mg/dL in patient 1, 41.9 mg/dL in patient 2, and 26.3 mg/dL in patient 3, although the carboxyhemoglobin levels were approximately equal in all the patients. Hyperbaric oxygen therapy (HBOT) was immediately initiated and continued for 10 days in all the cases; however, the outcomes of these patients varied considerably. Patient 1 remained comatose and died on day 31 because of central diabetes insipidus after shock. Patient 2 recovered from coma and was discharged; however, she was rehospitalized for DE on day 45 and recovered completely after another 10-day HBOT. Patient 3 gained consciousness and recovered completely with no sequelae during the 1-year follow-up. From these cases, we can consider that the initial blood lactate may correlate with the patient outcomes and prove to be a useful prognostic factor. Thus, we should particularly consider elevated lactate levels in CO poisoning.