Liquid culture enhances diagnosis of patients with milder forms of non-tuberculous mycobacterial lung disease.

OBJECTIVE: To evaluate the proportion and clinical characteristics of patients with non-tuberculous mycobacteria (NTM) lung disease diagnosed based on positive culture results in liquid medium only. METHODS: We reviewed the medical records of 978 patients diagnosed with NTM lung disease. All clinical samples were cultured in both solid and liquid media. RESULTS: Of the 978 patients, 111 (11.3%) were culture-positive in liquid medium only (liquid culture group), and 867 (88.7%) (solid culture group) on solid medium, regardless of the culture results in liquid medium. At the time of diagnosis, the liquid culture group was less likely than the solid culture group to have haemoptysis (11.7% vs. 20.0%, P = 0.04), positive sputum smear for acid-fast bacilli (14.4% vs. 50.2%, P < 0.001) or the fibrocavitary form of NTM lung disease (3.6% vs. 14.6%, P = 0.001). During the median follow-up period of 28.9 months (interquartile range 19.1-41.6), the proportion of patients requiring antibiotic treatment was lower in the liquid culture group than in the solid culture group (44.1% vs. 61.6%, P < 0.001). CONCLUSIONS: Liquid media culture is helpful in the diagnosis of patients with less severe forms of NTM lung disease.

Coagulase-negative staphylococcal bacteremia: risk factors for mortality and impact of initial appropriate antimicrobial therapy on outcome.

It is uncertain whether an initial inappropriate empirical antibiotic treatment of coagulase-negative staphylococci (CoNS) bacteremia adversely affects the outcome. A retrospective cohort study of CoNS bacteremia was performed at the Dongguk University Ilsan Hospital during a 3-year period. During the study period, 109 patients with CoNS bacteremia were enrolled. The median age of the patients was 72 years and most (96%, 105/109) had one or more comorbid diseases. Among the participants, 29% (32/109) received an appropriate empirical antimicrobial therapy. The 30-day mortality was 24% (26/109) and CoNS bacteremia-related mortality was 14% (15/109). There was no difference in the CoNS bacteremia-related mortality between the group with an inappropriate empirical treatment (13%, 10/77) and that with an appropriate treatment (16%, 5/32) (p = 0.46). In the multivariate analysis using the Cox regression analysis method, Pitt bacteremia scores [hazard ratio (HR) 1.48; 95% confidence interval (CI) 1.09-2.01; p = 0.01] and retention of eradicable focus (HR 5.0; 95% CI 1.39-17.9; p = 0.01) were found to be associated with CoNS bacteremia-related mortality. The results suggest that inappropriate empirical therapy might not necessarily be associated with the 30-day mortality or CoNS bacteremia-related mortality. Conversely, Pitt bacteremia scores and retention of eradicable focus were associated with poor outcomes.

Clinical characteristics of Raoultella ornithinolytica bacteremia.

PURPOSE: Raoultella ornithinolytica is not well known as a clinical pathogen. We performed a retrospective review of R. ornithinolytica bacteremia to investigate its clinical features, antimicrobial susceptibility, and overall patient outcomes. METHODS: R. ornithinolytica bacteremia cases were collected from an electronic database of all cases of bacteremia over a 10-year period. Medical records were retrospectively reviewed. Demographic data, clinical information, the presence of underlying comorbidities, the results of antimicrobial susceptibility testing, and the antimicrobial regimen administered were investigated. RESULTS: R. ornithinolytica was isolated from blood culture specimens in 16 cases. The majority of these patients had an underlying malignant condition of advanced stage (15 patients, 94 %). Seven of these patients had a solid tumor with lesions or metastases that extended to the bile duct or biliary tract. Neutropenic fever following hematologic stem cell transplantation was found in three cases. No resistance to piperacillin/tazobactam or imipenem was found. Four cases showed resistance to cefoxitin, while one of these cases showed resistance to multiple cephalosporins. In overall outcomes, seven patients (44 %) did not recover from the infection and subsequently expired. CONCLUSIONS: R. ornithinolytica bacteremia occurs mainly in patients with underlying malignancies. The overall outcome was not favorable, despite favorable antimicrobial susceptibility test results. The findings of this study contradict those of other studies that demonstrated that infection from Raoultella species have good prognoses.

Genomic aberrations of myeloproliferative and myelodysplastic/myeloproliferative neoplasms in chronic phase and during disease progression.

INTRODUCTION: Myeloproliferative neoplasms (MPN) and myelodysplastic/myeloproliferative neoplasms (MDS/MPN) may transform into secondary myelofibrosis (MF) or evolve into acute myeloid leukemia (AML). The genetic mechanisms underlying disease progression in MPN and MDS/MPN patients remain unclear. The purpose of this study was to investigate sequential genomic aberrations identified by single nucleotide polymorphism array (SNP-A)-based karyotyping that can detect cryptic aberrations or copy neutral loss of heterozygosity (CN-LOH) in the chronic phase and during disease progression of MPN and MDS/MPN patients. METHODS: The study group included 13 MPN and four MDS/MPN patients (seven polycythemia vera (PV); four essential thrombocythemia (ET); two MPN-unclassifiable (MPN-U); one chronic myelomonocytic leukemia (CMML); one atypical chronic myeloid leukemia, BCR-ABL1 negative (aCML); and two MDS/MPN-unclassifiable (MDS/MPN-U)). Among them, five patients (two PV, two MPN-U, and one MDS/MPN-U) progressed to MF and three patients (one CMML, one aCML, and one MDS/MPN-U) transformed to AML. The median follow-up period was 70 months (range, 7-152). Whole-genome SNP-A (SNP 6.0; Affymetrix, Santa Clara, CA, USA)-based karyotyping and JAK2 mutation analysis were performed according to the manufacturer's instructions. RESULTS: SNP-A showed 19 kinds of genomic aberrations, including seven gains, eight deletions, and four CN-LOH. CN-LOH of 9p involving JAK2 was the most common aberration, followed by 5q deletion and 9p gain. The incidence of genomic changes identified by SNP was not different in patients with disease progression (75%), compared with those without disease progression (56%) (P = 0.4). However, when excluding 9p CN-LOH, the incidence of genomic changes was significantly higher in patients with disease progression than in patients without disease progression (63% and 0%, respectively, P = 0.01). Among eight patients with disease progression, two patients (two MPN-U) showed abnormal SNP-A results, whereas metaphase cytogenetics (MC) analysis showed normal results at diagnosis and during follow-up. In nine patients without disease progression, SNP-A did not show any genomic aberrations except for 9p CN-LOH. In three patients (one PV, one aCML, and one MDS/MPN-U), clonal evolutions were identified by both MC and SNP-A according to disease progression. One PV patient who progressed to MF at 45 months after diagnosis showed sequential genomic changes from 9p CN-LOH to 9p gain by SNP-A. Results of JAK2 mutation analysis were variable depending on the patient. Most of the patients with 9p CN-LOH or 9p gain showed more than 50% of the JAK2 mutant alleles. In one patient (MDS/MPN-U) evolving to AML, the number of JAK2 mutant alleles decreased according to disease progression. CONCLUSION: This study suggests sequential genomic changes identified by SNP-A may be associated with disease progression.

Low virulence? Clinical characteristics of Raoultella planticola bacteremia.

PURPOSE: Numerous case reports regarding Raoultella planticola infection have accumulated in the literature; however, its significance as a clinical pathogen remains unknown. We performed a retrospective review of R. planticola bacteremia to characterize its clinical features, antimicrobial susceptibility, and patient outcome. METHODS: Raoultella planticola bacteremia cases were culled from an electronic database of all bacteremia cases occurring over a 4-year-period. Medical records were retrospectively reviewed and demographic data, clinical findings, presence of underlying disease, results of antimicrobial susceptibility testing, and the antibiotic regimens administered during the treatment were evaluated. RESULTS: Raoultella planticola was isolated from blood culture specimens in 20 cases. The majority of these patients had underlying malignant conditions (17 patients, 85%). The most prevalent causes of malignancy were adenocarcinoma involving the gallbladder or bile duct (7 patients) and hematologic malignancies (6 patients). No cases with resistance to carbapenem or third generation cephalosporins were found. All 14 patients with R. planticola as the sole microbial isolate recovered with the use of empirical antibiotics. Of the six patients with polymicrobial infection, three did not recover and subsequently expired. CONCLUSIONS: Raoultella planticola bacteremia seemed to occur mainly in immunocompromised patients, and was also frequently found in patients with lesions involving the gallbladder or bile duct. The overall outcome was favorable when R. planticola was treated with administration of empirical antibiotics. Mixed outcomes were found when blood cultures yielded multiple species of microbes.

Group B streptococcal bacteremia in non-pregnant adults: results from two Korean centers.

Patients with liver cirrhosis (LC) have impaired immunity and are, thus, predisposed to infection. Few studies have attempted to evaluate group B streptococcal (GBS) bacteremia in LC patients. A retrospective study of patients with GBS bacteremia was performed at the Dongguk University Ilsan Hospital and National Health Insurance Service Ilsan Hospital over a 13-year period (October 2000 to July 2013). During the study period, 97 patients with GBS bacteremia were enrolled. The median age of the patients was 67 years and 54 % were men. Among them, 23 (24 %) patients were classified as LC patients. The 30-day mortality rate of LC patients was significantly higher than that of patients with other diseases (26 % vs. 8 %, p = 0.03). The multivariate analysis indicated that LC was associated with an increased risk of 30-day mortality [hazard ratio (HR) 5.0; 95 % confidence interval (CI) 1.53-16.3; p = 0.008], as well as age (HR 1.07; 95 % CI 1.03-1.13; p = 0.02) and high Pitt bacteremia score (HR 1.23; 95 % CI 1.02-1.46; p = 0.03). The probability of survival at day 30 was significantly different for the Child-Pugh class C and the Child-Pugh classes A or B (44 % vs. 93 %, respectively; p = 0.01 by the log-rank test). The mortality rates of LC patients with GBS bacteremia were significantly higher than those of patients with other diseases. The severity of hepatic dysfunction plays an important role in the development of adverse events. Cirrhosis-specific scores such as the Child-Pugh class might be useful for predicting the prognosis of GBS bacteremia in LC patients.

Clinical characterization and molecular classification of 12 Korean patients with pseudohypoparathyroidism and pseudopseudohypoparathyroidism.

CONTEXT: Pseudohypoparathyroidism (PHP) is defined as resistance toward parathyroid hormones. PHP and pseudopseudohypoparathyroidism (PPHP) are rare disorders resulting from genetic and epigenetic aberrations within or upstream of the GNAS locus. This study investigated the clinical characteristics and performed a molecular analysis of PHP and PPHP. METHODS: A total of 12 patients with (P)PHP from 11 unrelated families (4 with PHP-Ia, 6 with PHP-Ib, and 2 with PPHP) were characterized using both clinical and molecular methods. Clinical features included the presenting symptoms, Albright hereditary osteodystrophy features, and resistance to hormones. Comprehensive analysis of the GNAS and STX16 loci was undertaken to investigate the molecular defects underlying (P)PHP. RESULTS: All PHP-Ib patients displayed hypocalcemic symptoms. All PHP-Ia patients showed resistance toward TSH, in addition to PTH. In most patients with PHP, when the diagnosis of PHP was first established, hypocalcemia and hyperphosphatemia were associated with a significant increase in serum PTH levels. One patient with PHP-Ia was diagnosed with growth hormone deficiency and showed a good response to human recombinant growth hormone therapy. 6 patients with PHP-Ia and PPHP showed 5 different mutations in the GNAS gene. 5 patients with PHP-Ib displayed a loss of differentially methylated region (DMR) imprints of the maternal GNAS. One PHP-Ib patient showed a de novo microdeletion in STX16 and a loss of methylation of exon A/B on the maternal allele. No patients revealed paternal disomy among 4 patients with PHP-Ib. CONCLUSIONS: Identification of the molecular causes of PHP and PPHP explains their distinctive clinical features and enables confirmation of the diagnosis and exact genetic counseling.

Microscopic schistocyte determination according to International Council for Standardization in Hematology recommendations in various diseases.

INTRODUCTION: Recently, a consensus report for microscopic schistocyte determination was prepared by International Council for Standardization in Hematology (ICSH). ICSH focused on diagnosis of thrombocytopenic purpura (TTP)/hemolytic uremic syndrome (HUS). We aimed to reanalyze schistocytes according to ICSH recommendations, to study diseases other than TTP/HUS related to the schistocytes, and to compare the percentage of schistocytes among the various diseases. METHODS: We retrieved all reported cases of peripheral blood (PB) smear in a single institution during 6 years. Schistocytes on 282 PB smears showing previous peripheral schistocytes and hemoglobin ≤10 g/dL were recounted according to ICSH recommendations. RESULTS: The schistocytes were frequently observed in patients with microangiopathic hemolytic anemia (MAHA), metastatic carcinoma, sepsis, chronic renal failure, preterm infant, and infection. Only two among 34 patients categorized as MAHA were diagnosed as TTP/HUS. Schistocytes were observed with other morphological changes in 169 of 170 cases with schistocyte ≤1% and in 102 of 112 with schistocyte >1%. The median schistocyte percentages of patients with hematologic malignancy, megaloblastic anemia, acute renal failure, and preterm infant were 1.20%, 1.30%, 1.35%, and 1.70%, respectively. CONCLUSION: Schistocytes were observed above 1% in many diseases other than TTP /HUS. Therefore, it is important to understand that schistocytes could be seen in various diseases, and in these cases, schistocytes were usually detected together with other red blood cell morphologic changes. These data support ICSH recommendation that a schistocyte count should be considered clinically meaningful if schistocytes represent the main morphological abnormality in the PB smear.

Evaluation of overnight storage conditions for autologous peripheral blood stem cell products: comparison of three different conditions.

BACKGROUND: Overnight (ON) storage of peripheral blood stem cell (PBSC) occurs frequently in clinical settings. However, there are no standard guidelines for optimal storage conditions of freshly harvested PBSC. The aim of this study was to investigate the influence of storage temperatures on the quality of autologous PBSC and establish optimal storage conditions before cryopreservation. METHODS: A retrospective analysis was performed on 260 PBSC harvests according to pre-cryopreservation conditions: immediate processing or ON storage at room temperature (RT). For direct comparison, 30 autologous PBSC products were collected prospectively and prepared under three different pre-cryopreservation conditions: immediate processing, ON storage at 4°C and ON storage at RT. The recovery of CD34(+) cells, post-thaw CFU-GM count and viability were analysed. RESULTS: Retrospective analysis revealed that post-thaw CFU-GM count was significantly lower when PBSC were stored ON at RT compared to when immediately processed (136·4 vs. 409·6/µl). Prospective analysis showed a mean recovery of CD34(+) cells of 65·5 ± 25·1%, 70·5 ± 27·4% and 35·9 ± 25·1% for immediate processing, ON storage at 4°C and ON storage at RT, respectively. Similarly, mean viability and CFU-GM counts were significantly reduced when stored ON at RT compared to when immediately processed or stored ON at 4°C (60·4 ± 25·6 vs. 84·1 ± 12·9 vs. 82·7 ± 12·6%, 15·7 ± 25·7 vs. 398·5 ± 906·2 vs. 350·0 ± 847·9/µl, respectively). CONCLUSIONS: ON storage of autologous PBSC at RT significantly decreased the quality of HPCs. These data indicate that ON storage of autologous PBSC at 4°C would be the most reasonable approach for maintaining the quality of HPCs when immediate processing is not possible.

Detection of methicillin-resistant Staphylococcus aureus (MRSA) from nasal samples by multiplex real-time PCR based on dual priming AT-rich primers.

In this study, we reported on the design of a multiplex real-time PCR assay based on SYBR Green I, incorporating dual priming adenine-thymine (AT)-rich primers for direct detection of MRSA from nasal samples. The multiplex real-time polymerase chain reaction (RT-PCR) assay reported in this study is based on SYBR Green I with incorporation of six dual priming AT-rich primers designed from the SCCmec/orf junction. A string (4-6 bp) of low-melting bases, such as adenine and thymine, was incorporated into the primers, which virtually divided a single primer in two functional regions, thus decreasing non-specific PCR products. The analytical sensitivity and specificity of the RT-PCR assay was determined with genomic DNA of reference strains (MRSA, MSSA, and MRCoNS). RT-PCR assay was performed for analysis of 72 nasal swab specimens, and the results were confirmed by use of a culture method. Furthermore, the results of RT-PCR were compared with LightCycler MRSA advance test. The multiplex RT-PCR assay reproducibly detected a minimum of 1 pg genomic DNA (31.5 copy of genome) of MRSA reference strains and clinical isolates, with a specific melting peak at 83.5 ± 1.5°C, and neither fluorescence nor a melting peak was detected in non-target isolates. The concordance rate between RT-PCR assay and culture method was 87.5% with Cohen's kappa value (κ) 0.75, which showed good agreement between the two assays. The sensitivity, specificity, positive predictive value, and negative predictive value of the assay were 93.5%, 82.9%, 80.5%, and 94.4%, respectively. In a comparative study for the detection of 72 nasal samples, the sensitivity, specificity, positive predictive value, and negative predictive value of the multiplex RT-PCR assay with respect to LightCycler MRSA advance test was 84.2%, 88.2%, 89%, and, 83.3%, respectively. The results of RT-PCR assay demonstrated high specificity (88.2%) and positive predictive value (89%) for the direct detection of MRSA from nasal samples.

CD34, RAB20, PU.1 and GFI1 mRNA expression in myelodysplastic syndrome.

Myelodysplastic syndrome (MDS) with hypocellular bone marrow (BM) is often difficult to distinguish from aplastic anemia (AA). Furthermore, the diagnosis of MDS with low blast counts and normal karyotype may be problematic. These issues highlight the need for a reliable marker for the diagnosis of MDS. This study was conducted to determine if changes of mRNA expression in any of the four selected genes would be useful markers for differentiation of hypoplastic MDS from AA, and MDS from benign disease, as well as to investigate whether mRNA expressions differ between MDS risk subgroups. Thirty-five patients diagnosed with MDS, 27 patients with AA and 17 patients with benign diseases were included. The CD34, RAB20, PU.1 and GFI1 mRNA levels were measured by real-time RT-PCR. The CD34 mRNA expressions in hypoplastic MDS were higher than those found in AA. PU.1 and GFI1 mRNA expressions were significantly lower in MDS with low blast counts and normal karyotype than those of benign disease. High-risk MDS showed higher CD34 expressions than those of low-risk MDS. This study suggests that measurement of CD34 and GFI1 mRNA expressions could be useful as a diagnostic and prognostic marker for MDS.

Qi therapy as an intervention to reduce chronic pain and to enhance mood in elderly subjects: a pilot study.

Qi therapy (or external Qi) is an oriental complementary therapy preventing, curing disease and strengthens health and improving the human potentiality through regulation of body. It is increasingly being used to improve the quality of life, but there is little direct evidence of its efficacy. This study assessed the effects of Qi therapy (QT) on reducing pain and enhancing mood states in elderly subjects with chronic pain. We studied 40 elderly participants with chronic pain, who were randomly allocated to receive QT (n=20) or standard care (n=20). The experimental group receives Qi therapy twice a week for 2 weeks (total 4 times), and control group received general care at the same time and the same amount of duration. We measured pain level and Profile of Mood State (POMS) to explore participants' response to Qi therapy. There was a significant reduction in pain (p<0.0001) after QT and an improved positive mood state (p<0.0001). These findings suggest that Qi therapy may have a role in helping the elderly to cope with their pain and mood disturbances.

Effects of emitted Qi on in vitro natural killer cell cytotoxic activity.

The present study investigated the effects of Korean Qi-therapy, ChunSoo Energy Healing, on natural killer (NK) cell cytotoxicity in vitro depending on Qi-treatment time and the types of cells treated. NK cell cytotoxicity was assayed by measuring LDH release from tumor target cells (K562 cell lines). NK activity was significantly increased by emitted-Qi treatment of 30 sec duration. Three and 5 minutes of Qi projection created the greatest increase in NK cell activity when mixtures of NK cells and K562 cells were treated (1.81 and 2.12 fold for 4 hr culture; 1.54 and 1.36 for 16 hr culture, respectively). NK cell activity increased significantly in Qi-treated K562 cells alone (1.13 fold, p<0.05) compared to control. These results are consistent with in vivo Qi-therapy on humans and suggests that emitted-Qi has an acute stimulatory effect on NK cell activity. This study provides direct scientific support that Qi as such may positively affect human cellular immunity.

Effect of Qi-training on blood pressure, heart rate and respiration rate.

To examine the physiological effects of Korean traditional Qi-training, we investigated the changes in blood pressure, heart and respiratory rates before, during and after ChunDoSunBup (CDSB) Qi-training. Twelve normal healthy CDSB Qi-trainees (19-37 years old; trained for 1.3 +/- 0.2 years; 9 men and 3 women) volunteered to participate in this study. Heart rate, respiratory rate, systolic blood pressure and rate-pressure product were significantly decreased during Qi-training. From these results, we suggest that CDSB Qi-training has physiological effects that indicate stabilization of cardiovascular system.

Acute effects of chundosunbup qi-training on blood concentrations of TSH, calcitonin, PTH and thyroid hormones in elderly subjects.

The present study investigated how the systemic treatment of a programmed exercise, ChunDoSunBup (CDSB) Qi-training, affects the secretion of thyroid and parathyroid hormones in elderly subjects (10 male and 5 female). Plasma concentrations of thyroid stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), parathyroid hormone (PTH), ionized calcium, and calcitonin were determined. CDSB Qi-training induces a slight increase in TSH. Both T4 and T3 were increased at the mid-time of CDSB Qi-training (p < 0.05). There were significant correlations only between T3 and T4 at mid-training. This shows that increase in the plasma level of T3 was associated with the secretion of T4. The plasma concentrations of calcitonin and PTH were increased at mid-time and post-time of CDSB Qi-training. But ionized calcium was decreased slightly by CDSB Qi-training. These results suggest that Qi-training modulates the secretion of thyroid hormones, calcium metabolism, and parathyroid hormones in the elderly. However, whether the long-term practice of CDSB Qi-training might change bone metabolism and have longitudinal effects on the thyroid hormone of the elderly need further investigation.