Rapid and Reliable HLA-B*59:01 Genotyping to Prevent Carbonic Anhydrase Inhibitor-Induced Severe Cutaneous Adverse Reactions.

OBJECTIVE: Carbonic anhydrase inhibitors (CAIs) are intraocular pressure-reducing medications used in ophthalmology. Human leukocyte antigen-B*59:01 (HLA-B*59:01) is strongly associated with CAI-induced severe cutaneous adverse reactions (SCARs). This study aimed to develop and validate a rapid and economical screening method for HLA-B*59:01 to prevent carbonic anhydrase inhibitor-induced SCARs. METHODS: Duplex allele-specific polymerase chain reaction (PCR) with an internal control was performed for HLA-B*59:01 genotyping. The accuracy of duplex allele-specific PCR for HLA-B*59:01 genotyping was evaluated in 200 blood samples, using sequence-based typing (SBT) as the reference method. RESULTS: In total, 50 HLA-B*59:01-positive and 150 HLA-B*59:01-negative results obtained using duplex allele-specific PCR were in complete agreement with the SBT results. CONCLUSION: Duplex allele-specific PCR is a rapid, reliable, and economical assay for screening the HLA-B*59:01 allele.

Bowel obstruction secondary to internal hernia in a hereditary angioedema patient: a case report.

BACKGROUND: Gastrointestinal attacks are frequent symptoms in patients diagnosed with hereditary angioedema (HAE). Cases of self-limited bowel intussusception and unneeded exploratory laparotomy due to lack of knowledge about HAE have been reported. Furthermore, after the introduction of C1-esterase inhibitor (C1-INH) concentrate, the recommended medication for HAE attacks, treatment has become typically medical in nature. We share a rare case where operative exploration was indicated to resolve a mechanical small bowel obstruction secondary to an HAE attack. CASE REPORT: An 80-year-old woman with HAE presented with lower left abdominal pain, vomiting, and nausea. Computed tomography (CT) showed edema of the small bowel and stomach as well as possible signs of mechanical small bowel obstruction. The patient was treated with C1-INH concentrate but showed only mild signs of relief, warranting diagnostic laparoscopy. Intraoperative findings showed internal herniation and strangulation of the small bowel caused by adhesions forming a band. After surgical intervention, no bowel resection was needed. CONCLUSION: Although C1-INH concentrate remains the principal treatment for HAE, gastrointestinal attacks may potentially cause surgical emergencies.

The necessity of lumbar puncture in adult emergency patients with fever-associated seizures.

OBJECTIVE: Central nervous system (CNS) infections are often suspected in adult patients with fever-associated seizures. However, it is unclear whether lumbar puncture (LP) is routinely required in patients with fever-associated seizures. This study aimed to examine the prevalence of meningitis and encephalitis in adult patients with fever-associated seizures and to evaluate whether LP is routinely required. METHODS: We retrospectively studied patients aged ≥16 years who presented to the emergency department with complaints of seizures and fever above 37.5 °C who were admitted to the hospital between January 2017 and December 2019. LP was performed when the emergency physician suspected meningitis or encephalitis. Neurologists assessed patients with normal cerebrospinal fluid (CSF) findings and those admitted without LP after hospitalization. A neurologist confirmed the diagnoses of meningitis and encephalitis. RESULTS: The study included 148 patients. Ninety-seven patients (65.5%) were male, and the median age was 60 years. LP was performed in 105 patients (70.9%), and 14 (13.4%) had CSF pleocytosis. Meningitis and encephalitis were diagnosed in nine patients (6.1%), of whom four (2.8%) had CNS infections. Patients diagnosed with meningitis and encephalitis were more likely to have Glasgow Coma Scale <13 (P = 0.03) and less likely to have a history of seizures or epilepsy (P = 0.04) and had higher C-reactive protein levels than the other patients (P = 0.02). CONCLUSION: The prevalence of meningitis or encephalitis is relatively low in adult patients with fever-associated seizures. Lumbar puncture is considered unnecessary to be performed routinely, but its indication should be carefully considered with reference to the clinical course, comorbidities, and blood tests. Further validation studies with larger sample sizes are needed to confirm the findings of this study.

Premorbid Clinical Frailty Score and 30-day mortality among older adults in the emergency department.

OBJECTIVES: The association between frailty and short-term prognosis has not been established in critically ill older adults presenting to the emergency department. We sought to examine the association between premorbid frailty and 30-day mortality in this patient population. METHODS: This is a retrospective observational study on older adults aged over 75 who were triaged as Level 1 resuscitation with subsequent admissions to intermediate units or intensive care units (ICUs) in a single critical care center, from January to December 2019. We excluded patients with out-of-hospital cardiac arrest or those transferred from other hospitals. Frailty was evaluated by the Clinical Frailty Scale (CFS) from the patients' chart reviews. The primary outcome was 30-day mortality, and we examined the association between frailty scored on the CFS and 30-day mortality using a multivariable logistic regression model with CFS 1-4 as a reference. RESULTS: A total of 544 patients, median age: 82 years (interquartile rang 78 to 87), were included in the study. Of these, 29% were in shock and 33% were in respiratory failure. The overall 30-day mortality was 15.1%. The adjusted risk difference (95% confidence interval [CI]) in mortality for CFS 5, CFS 6, and CFS 7-9 was 6.3% (-3.4 to 15.9), 11.2% (0.4 to 22.0), and 17.7% (5.3 to 30.1), respectively; and the adjusted risk ratio (95% CI) was 1.45 (0.87 to 2.41), 1.85 (1.13 to 3.03), and 2.44 (1.50 to 3.96), respectively. CONCLUSION: The risk of 30-day mortality increased as frailty advanced in critically ill older adults. Given this high risk of short-term outcomes, ED clinicians should consider goals of care conversations carefully to avoid unwanted medical care for these patients.

Head CT findings and deterioration risk in children with head injuries and Glasgow Coma Scales of 15.

OBJECTIVE: Head injuries are an important problem in pediatric emergency care. The majority of head injuries are mild. Even when abnormalities are noted on computed tomography (CT), most patients have good outcomes. We aimed to evaluate the clinical course of pediatric patients who had head injuries and Glasgow Coma Scale (GCS) scores of 15, in whom abnormal findings were noted on head CT, to determine the impact of radiographic features on the need for hospitalization and clinical progression. METHODS: We retrospectively examined patients under 15 years of age with isolated mild head injuries, GCS scores of 15, and abnormal CT findings, and visited the emergency department between September 2011 and March 2019. RESULTS: Ninety-nine patients were included in the study. The median age was 2 years (0-15 years), and 61 (62%) patients were male. Eighty-six (87%) patients were hospitalized, and the median hospital stay was 1 day (1-10 days). Sixty-eight (69%) patients underwent repeat CT, and 12 (18%) patients showed signs of radiographic progression. These 12 patients had subdural or epidural hematomas, and surgical intervention was required for two patients (2%). In patients with isolated skull fracture or subarachnoid hemorrhage alone, no deterioration was noted radiographically or clinically. CONCLUSION: Pediatric head injuries with GCS scores of 15 may rarely require surgical intervention, even when CT shows abnormalities. In particular, patients diagnosed with isolated skull fracture or subarachnoid hemorrhage on CT may not require routine hospitalization. A validation study is needed to confirm the findings of this study.

Human iPSC-derived neural precursor cells differentiate into multiple cell types to delay disease progression following transplantation into YAC128 Huntington's disease mouse model.

OBJECTIVES: To investigate whether human HLA-homozygous induced pluripotent stem cell (iPSC)-derived neural precursor cells (iPSC-NPCs) can provide functional benefits in Huntington's disease (HD), we transplanted them into the YAC128 transgenic HD mouse model. MATERIALS AND METHODS: CHAi001-A, an HLA-homozygous iPSC line (A*33:03-B*44:03-DRB1*13:02), was differentiated into neural precursor cells, and then, they were transplanted into 6 months-old YAC128 mice. Various behavioural and histological analyses were performed for five months after transplantation. RESULTS: Motor and cognitive functions were significantly improved in transplanted animals. Cells transplanted in the striatum showed multipotential differentiation. Five months after transplantation, the donor cells had differentiated into neurons, oligodendrocytes and astrocytes. Transplantation restored DARPP-32 expression, synaptophysin density, myelin basic protein expression in the corpus callosum and astrocyte function. CONCLUSION: Altogether, these results strongly suggest that iPSC-NPCs transplantation induces neuroprotection and functional recovery in a mouse model of HD and should be taken forward for clinical trials in HD patients.

Development of a Scoring System to Predict Prolonged Post-Stroke Dysphagia Remaining at Discharge from a Subacute Care Hospital to the Home.

BACKGROUND: Prolonged dysphagia is an important stroke-related complication that imposes a substantial burden on patients and families. However, simple scoring tool to predict prolonged dysphagia is not existing. MATERIALS AND METHODS: This retrospective cohort study used data from April 2010 to March 2016. Adult patients with first-ever stroke were included. The outcome was swallowing function at discharge from the subacute care hospital to the patient's home. We collected the following factors obtained at discharge from the University of Fukui Hospital: age, sex, type of stroke, comorbidities, smoking status, alcohol use, denture use, functional dependency in daily living before admission, National Institutes of Health Stroke Scale score (NIHSS) at admission, and Functional Independence Measure(FIM). Data were divided into a training set (70%) and test set (30%). Lasso and logistic regression were used for feature selection, a scoring system was then developed, and its prediction performance evaluated. RESULTS: This study enrolled 462 patients with acute stroke. Using lasso and logistic regression, three variables (functional dependency before admission, Functional Independence Measure [FIM]-cognitive and FIM-motor scores at transfer) remained statistically significant predictors of prolonged dysphagia. Risk scores were categorized as low risk (0-2), moderate risk (3-4), and high risk (5-7), with dysphagia rates of 0%-1%, 13%-29%, and 50%-100%, respectively. A newly developed score ≥3 was the optimal cutoff for identifying patients with the potential risk of prolonged dysphagia (C-statistics, 0.92 in the test set). CONCLUSION: The developed scoring system is simple and has a high performance in predicting prolonged dysphagia after acute stroke.

High fever or hypotension predicts non-hypoglycemia in patients with impaired consciousness in prehospital settings.

AIM: To evaluate whether vital signs can predict whether hypoglycemia can be eliminated as the cause of impaired consciousness in prehospital settings. METHODS: We extracted the data of patients who underwent blood glucose measurements by paramedics in Kobe City, Japan from April 2015 to March 2019. We used receiver operating characteristic curves and calculated the area under the curve (AUC) to evaluate the validity of the vital signs in distinguishing hypoglycemia. We also calculated stratum-specific likelihood ratios to examine the threshold at which hypoglycemia becomes less likely for each vital sign. RESULTS: Of the 1,791 patients, 1,242 were eligible for analysis. Hypoglycemia was observed in 324 patients (26.1%). Significant differences in each vital sign were noted between the hypoglycemic and non-hypoglycemic groups. Body temperature was moderately accurate in differentiating between the two groups (AUC, 0.71; 95% confidence interval, 0.68-0.74). Furthermore, in patients with systolic blood pressure <100 mmHg and body temperature ≥38°C, it was unlikely that hypoglycemia caused impaired consciousness (stratum-specific likelihood ratios 0.12 and 0.15; 95% confidence intervals, 0.05-0.25 and 0.06-0.35, respectively). CONCLUSION: In the prehospital assessment of patients with impaired consciousness, high fever or hypotension was helpful in differentiating between hypoglycemia and non-hypoglycemia. In particular, body temperature ≥38°C or systolic blood pressure <100 mmHg indicated a low likelihood of hypoglycemia. A validation study is needed to confirm the findings in this study.

Calibration-Free Cuffless Blood Pressure Estimation Based on a Population With a Diverse Range of Age and Blood Pressure.

This study presents a new blood pressure (BP) estimation algorithm utilizing machine learning (ML). A cuffless device that can measure BP without calibration would be precious for portability, continuous measurement, and comfortability, but unfortunately, it does not currently exist. Conventional BP measurement with a cuff is standard, but this method has various problems like inaccurate BP measurement, poor portability, and painful cuff pressure. To overcome these disadvantages, many researchers have developed cuffless BP estimation devices. However, these devices are not clinically applicable because they require advanced preparation before use, such as calibration, do not follow international standards (81060-1:2007), or have been designed using insufficient data sets. The present study was conducted to combat these issues. We recruited 127 participants and obtained 878 raw datasets. According to international standards, our diverse data set included participants from different age groups with a wide variety of blood pressures. We utilized ML to formulate a BP estimation method that did not require calibration. The present study also conformed to the method required by international standards while calculating the level of error in BP estimation. Two essential methods were applied in this study: (a) grouping the participants into five subsets based on the relationship between the pulse transit time and systolic BP by a support vector machine ensemble with bagging (b) applying the information from the wavelet transformation of the pulse wave and the electrocardiogram to the linear regression BP estimation model for each group. For systolic BP, the standard deviation of error for the proposed BP estimation results with cross-validation was 7.74 mmHg, which was an improvement from 17.05 mmHg, as estimated by the conventional pulse-transit-time-based methods. For diastolic BP, the standard deviation of error was 6.42 mmHg for the proposed BP estimation, which was an improvement from 14.05mmHg. The purpose of the present study was to demonstrate and evaluate the performance of the newly developed BP estimation ML method that meets the international standard for non-invasive sphygmomanometers in a population with a diverse range of age and BP.

Effectiveness and safety of small-bore tube thoracostomy (≤20 Fr) for chest trauma patients: A retrospective observational study.

INTRODUCTION: Tube thoracostomy is an important treatment for traumatic hemothorax and pneumothorax. The optimal tube diameter remains unclear. To reduce invasiveness, we use small-bore chest tubes (≤20 Fr) for all trauma patients for whom tube thoracostomy is indicated in our emergency department (ED). The aim of this study was to investigate the effectiveness and safety of small-bore tube thoracostomy for traumatic hemothorax or pneumothorax. METHOD: We conducted a retrospective observational study at a single emergency medical center. This study included adult patients (≥18 years old) who had undergone tube thoracostomy for chest trauma in the ED during the 5 years from October 2013 to September 2018. We used 20 Fr chest tubes or 8 Fr pigtail catheters. The examined outcome was tube-related complications, such as tube obstruction, retained hemothorax, and unresolved pneumothorax. RESULTS: A total of 107 tube thoracostomies were performed in 102 patients. The mean Injury Severity Score of these patients was 17.8 (±9.6), and the mean duration of the tube placement period was 3.9 days (±1.8). Eight patients developed tube-related complications (7.8%) (retained hemothorax: 4 patients (3.9%), unresolved pneumothorax: 4 patients (3.9%)). None of these cases were caused by tube obstruction. Although the drainage itself was effective, they underwent definitive invasive interventions to stop bleeding or air leak. CONCLUSION: Our study showed that the use of small-bore (≤20 Fr) chest tubes to treat traumatic hemothorax/pneumothorax achieved the purposes of tube thoracostomy. It might be possible to safely manage chest trauma with small-bore chest tubes.

Intracerebral transplantation of HLA-homozygous human iPSC-derived neural precursors ameliorates the behavioural and pathological deficits in a rodent model of ischaemic stroke.

OBJECTIVES: Human-induced pluripotent stem cells (hiPSCs) are a promising cell source for treating ischaemic stroke. Although autologous hiPSCs provide the advantage of avoiding immune rejection, their practical limitations, such as substantial amount of time and costs to generate individual iPSC lines, have hampered their widespread application in clinical settings. In this study, we investigated the therapeutic potential of neural precursor cells derived from human HLA-homozygous induced pluripotent stem cells (hiPSC-NPCs) following intracerebral transplantation into a rodent model of middle cerebral artery occlusion (MCAo). MATERIALS AND METHODS: We differentiated a GMP-grade HLA-homozygous hiPSC line (CMC-hiPSC-004) into neural precursor cells for transplantation into rats at the subacute stage of ischaemic stroke (ie at 7 days after the induction of MCAo). To investigate functional recovery, the transplanted animals were subjected to five behavioural tests, namely the rotarod, stepping, mNSS, staircase and apomorphine-induced rotation tests, for up to 12 weeks, followed by histological analyses. RESULTS: We observed that the hiPSC-NPC transplantation produced significant behavioural improvements. At 12 weeks post-transplantation, a high proportion of transplanted cells survived and had differentiated into MAP2+ mature neurons, GABAergic neurons and DARPP32+ medium spiny neurons. The transplanted cells formed neuronal connections with striatal neurons in the host brain. In addition, hiPSC-NPC transplantation gave rise to enhanced endogenous repair processes, including decreases of post-stroke neuroinflammation and glial scar formation and an increase of proliferating endogenous neural stem cells in the subventricular zone as well as the perilesional capillary networks. CONCLUSIONS: These results strongly suggest that HLA-homozygous hiPSC-NPCs may be useful for treating ischaemic stroke patients.

Prognostic utility of ADAMTS13 activity for the atypical hemolytic uremic syndrome (aHUS) and comparison of complement serology between aHUS and thrombotic thrombocytopenic purpura.

BACKGROUND: Atypical hemolytic uremic syndrome (aHUS) involves dysregulation of the complement system, but whether this also occurs in thrombotic thrombocytopenic purpura (TTP) remains unclear. Although these conditions are difficult to differentiate clinically, TTP can be distinguished by low (<10%) ADAMTS13 activity. The aim was to identify the differences in complement activation products between TTP and aHUS and investigate ADAMTS13 activity as a prognostic factor in aHUS. METHODS: We analyzed patients with thrombotic microangiopathy diagnosed as TTP (N=48) or aHUS (N=50), selected from a Korean registry (N=551). Complement activation products in the plasma samples collected from the patients prior to treatment and in 40 healthy controls were measured by ELISA. RESULTS: The levels of generalized (C3a), alternate (factor Bb), and terminal (C5a and C5b-9) markers were significantly higher (all P<0.01) in the patients than in the healthy controls. Only the factor Bb levels significantly differed (P=0.008) between the two disease groups. In aHUS patients, high normal ADAMTS13 activity (≥77%) was associated with improved treatment response (OR, 6.769; 95% CI, 1.605-28.542; P=0.005), remission (OR, 6.000; 95% CI, 1.693-21.262; P=0.004), exacerbation (OR, 0.242; 95% CI, 0.064-0.916; P=0.031), and disease-associated mortality rates (OR, 0.155; 95% CI, 0.029-0.813; P=0.017). CONCLUSION: These data suggest that complement biomarkers, except factor Bb, are similarly activated in TTP and aHUS patients, and ADAMTS13 activity can predict the treatment response and outcome in aHUS patients.

Rapid, Reliable, and Inexpensive HLA-B*58:01 Detection Method Using DNA Binding Dye-based Duplex Allele-specific Melting Curve Analysis.

BACKGROUND: Allopurinol is the most commonly used drug for the treatment of gout and also one of the most common causes of severe cutaneous adverse reactions (SCARs). Human leukocyte antigen-B*58:01 (HLA-B*58:01) is strongly associated with allopurinol-induced SCARs. The aim of the present study was to develop and validate a rapid and economic screening method for HLA-B*58:01. METHODS: The accuracy of duplex allele-specific melting curve analysis using DNA-binding dye for HLA-B*58:01 was evaluated in 150 blood samples with sequence-based typing (SBT) as the reference method. RESULTS: Fifty HLA-B*58:01-positive and 100 negative results obtained by duplex allele-specific melting curve analysis were completely in agreement with the SBT results. CONCLUSION: Duplex allele-specific melting curve analysis is a rapid, reliable and inexpensive assay that is appropriate for screening for the HLA-B*58:01 allele.

Repurposing the Cord Blood Bank for Haplobanking of HLA-Homozygous iPSCs and Their Usefulness to Multiple Populations.

Although autologous induced pluripotent stem cells (iPSCs) can potentially be useful for treating patients without immune rejection, in reality it will be extremely expensive and labor-intensive to make iPSCs to realize personalized medicine. An alternative approach is to make use of human leukocyte antigen (HLA) haplotype homozygous donors to provide HLA matched iPSC products to significant numbers of patients. To establish a haplobank of iPSCs, we repurposed the cord blood bank by screening ∼4,200 high resolution HLA typed cord blood samples, and selected those homozygous for the 10 most frequent HLA-A,-B,-DRB1 haplotypes in the Korean population. Following the generation of 10 iPSC lines, we conducted a comprehensive characterization, including morphology, expression of pluripotent markers and cell surface antigens, three-germ layer formation, vector clearance, mycoplasma/microbiological/viral contamination, endotoxin, and short tandem repeat (STR) assays. Various genomic analyses using microarray and comparative genomic hybridization (aCGH)-based single nucleotide polymorphism (SNP) and copy number variation (CNV) were also conducted. These 10 HLA-homozygous iPSC lines match 41.07% of the Korean population. Comparative analysis of HLA population data shows that they are also of use in other Asian populations, such as Japan, with some limited utility in ethnically diverse populations, such as the UK. Taken together, the generation of the 10 most frequent Korean HLA-homozygous iPSC lines serves as a useful pointer for the development of optimal methods for iPSC generation and quality control and indicates the benefits and limitations of collaborative HLA driven selection of donors for future stocking of worldwide iPSC haplobanks. Stem Cells 2018;36:1552-1566.

The 2016 WHO versus 2008 WHO Criteria for the Diagnosis of Chronic Myelomonocytic Leukemia.

The 2016 WHO diagnostic criteria for chronic myelomonocytic leukemia (CMML) require both absolute and relative monocytosis (≥1×109/L and ≥10% of white blood cell counts) in peripheral blood. Moreover, myeloproliferative neoplasm (MPN) features in bone marrow and/or MPN-associated mutations tend to support MPN with monocytosis rather than CMML. We assessed the impact of the 2016 WHO criteria on CMML diagnosis, compared with the 2008 WHO criteria, through a retrospective review of the medical records of 38 CMML patients diagnosed according to the 2008 WHO classification. Application of the 2016 WHO criteria resulted in the exclusion of three (8%) patients who did not fulfill the relative monocytosis criterion and eight (21%) patients with an MPN-associated mutation. These 11 patients formed the 2016 WHO others group; the remaining 27 formed the 2016 WHO CMML group. The significant difference in the platelet count and monocyte percentage between the two groups indicated that the 2016 WHO criteria lead to a more homogenous and improved definition of CMML compared with the 2008 WHO criteria, which may have led to over-diagnosis of CMML. More widespread use of molecular tests and more sophisticated clinical and morphological evaluations are necessary to diagnose CMML accurately.

The optimal number of personnel for good quality of chest compressions: A prospective randomized parallel manikin trial.

BACKGROUND: Long durational chest compression (CC) deteriorates cardiopulmonary resuscitation (CPR) quality. The appropriate number of CC personnel for minimizing rescuer's fatigue is mostly unknown. OBJECTIVE: We determined the optimal number of personnel needed for 30-min CPR in a rescue-team. METHODS: We conducted a randomized, manikin trial on healthcare providers. We divided them into Groups A to D according to the assigned different rest period to each group between the 2 min CCs. Groups A, B, C, and D performed CCs at 2, 4, 6, and 8 min rest period. All participants performed CCs for 30 min with a different rest period; participants allocated to Groups A, B, C, and D performed, eight, five, four, and three cycles, respectively. We compared a quality change of CCs among these groups to investigate how the assigned rest period affects the maintenance of CC quality during the 30-min CPR. RESULTS: This study involved 143 participants (male 58 [41%]; mean age, 24 years,) for the evaluation. As participants had less rest periods, the quality of their CCs such as sufficient depth ratio declined over 30-min CPR. A significant decrease in the sufficient CC depth ratio was observed in the second to the last cycle as compared to the first cycle. (median changes; A: -4%, B: -3%, C: 0%, and D: 0% p < 0.01). CONCLUSIONS: A 6 min rest period after 2 min CC is vital in order to sustain the quality of CC during a 30-min CPR cycle. At least four personnel may be needed to reduce rescuer's fatigue for a 30-min CPR cycle when the team consists of men and women.

Can we truly rely on the urinary antigen test for the diagnosis? Legionella case report.

It is critical to diagnose and treat Legionella pneumonia (LP) immediately after infection because of the associated high mortality. The urine antigen test (UAT) is often used for the diagnosis of LP; however, it cannot detect the serogroups of all Legionella species. A detained medical history and several clinical findings such as liver enzyme elevation and hyponatremia are useful in diagnosis. Some specific types of Legionella are found in compost. Herein, we report a case of LP in which the patient's medical history and several clinical findings were useful for diagnosis.

Quality comparison of umbilical cord blood cryopreserved with conventional versus automated systems.

Umbilical cord blood (CB) banks usually freeze and store CB for clinical transplantation using conventional controlled-rate freezer or the automated BioArchive system. The aim of this study is to compare the quality of CB cryopreserved with conventional and automated methods and to make clear the cause of the quality difference between the two methods. The experiment used 80 CB units: 40 were conventionally cryopreserved and the remainder were cryopreserved with a BioArchive. After thawing, the following measures of CB quality were compared: recovery rates of cell count, cell viability of total nucleated cells (TNCs), mononuclear cells (MNCs), and CD34+ cells, as well as colony-forming unit-granulocyte/macrophage (CFU-GM) content. Additionally, processing and storage records were reviewed to quantify the number of exposures of CB units at room temperature (transient warming event, TWE), which was analyzed in relation to CB quality. MNC and CD34+ cell viability were as follows: MNC, 78.2% ± 6.8% (conventional), 81.7% ± 7.2% (automated); CD34+ cell, 90.6% ± 6.9% (conventional), 94.7% ± 3.5% (automated). The absolute CFU-GM content per CB unit was 7.1 × 105 ± 5.9 × 105 with conventional cryopreservation and 12.3 × 105 ± 12.0 × 105 with automated cryopreservation. CBs cryopreserved with BioArchive showed significantly higher MNC and CD34+ cell viability, and CFU-GM content than those conventionally cryopreserved. The CB quality comparison depending on the amount of TWEs showed no significant quality difference between groups that were more exposed to TWEs and groups that were less exposed. CBs cryopreserved with BioArchive were of higher quality than conventionally cryopreserved CBs, and the cause of quality difference might be due to the difference of freezing conditions rather than the TWE effect.

The first human clinical case of chronic osteomyelitis caused by Clostridium hydrogeniformans.

We present the first case report of osteomyelitis due to Clostridium hydrogeniformans in a previously healthy 18-year-old male. He was admitted to our hospital because of an open contaminated fracture of the right arm after being blown into a drain in a motorbike accident. He underwent surgical debridement and treatment course of cefazolin. Although he responded well to these initial treatments, subcutaneous abscess and ulnar osteomyelitis developed 1 month after discharge. Second debridement was performed and specimens were collected from both the abscess and bone tissues. Only anaerobic culture showed a gas-producing Gam-positive rod. Conventional methods and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry could not accurately identify this organism. However, 16S rRNA gene sequence analysis determined the isolate as C. hydrogeniformans with 99.79% homology. The patient recovered after 90 days of antibiotic treatment, and had no evidence of recurrence. Anaerobic bacteria are more common as causative pathogens in osteomyelitis related to traumatic wounds and Clostridium spp. are particularly associated with open fractures, which is consistent with our case. Although the epidemiology and clinical characteristics of C. hydrogeniformans infection is poorly understood because of the limitations of currently available conventional methods of identification, clinicians need to consider this organism as a causative pathogen in a patient with osteomyelitis in traumatic wounds, especially contaminated by sewer water.