Coproduction of qnrB and armA from Extended-Spectrum beta-lactamase-producing Klebsiella pneumoniae / 대한진단검사의학회지

BACKGROUND: Multidrug-resistant Enterobacteriaceae is a worldwide problem. Although various resistance mechanisms have been recognized with increasing frequency, only a few cases of triple resistance of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae have been reported. This study was designed to evaluate the coexistence of qnr (qnrA, qnrB, and qnrS) and 16S rRNA methylase (armA, rmtA, rmtB, and rmtC) in ESBL-producing K. pneumoniae. METHODS: We tested 44 isolates of ESBL-producing K. pneumoniae at Chungnam National University Hospital from March to September 2006. Antimicrobial susceptibilities were tested by broth microdilution method, and transconjugation test was performed using E. coli J53 with azide resistance. Search for qnr (qnrA, qnrB, and qnrS) and 16S rRNA methylase (armA, rmtA, rmtB, and rmtC) genes was conducted by PCR amplification, and the genotypes were determined by direct nucleotide sequence analysis of the amplified products. Epidemiologic study was performed by Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). RESULTS: All ESBL-positive strains produced qnrB; however, armA was detected in 68.2%. The coproduction rate of qnrB and armA in ESBL-producing K. pneumoniae was 68.2%. Two types (A and B) were dominant in ERIC-PCR results. CONCLUSIONS: K. pneumoniae producing qnrB, armA, and ESBL are spreading widely.

A case of prenatally diagnosed Non-15, Non-22 marker chromosome / 대한산부인과학회지

Midtrimester genetic amniocentesis is an important diagnostic tool in prenatal genetic diagnosis and counseling. We can identify karyotypes with metaphase chromosome analysis of cultured amniocytes. Marker chromosomes are defined as unidentified structurally abnormal chromosomes. Incidence of marker chromosomes in the previous reported studies was 0.6-1.5/1,000. They occurred more frequently with advanced maternal age. Ascertainment of chromosomal origin is important because it may be associated with malformation and developmental abnormalities. Recently, identification of the origin and composition of marker chromosomes has been made possible by the use of fluorescent in situ hybridization (FISH). Most marker chromosomes are known to be originated from chromosome 15 or 22, X, Y. We have experienced a case of non-15, non-22 marker chromosome prenatally detected in amniocentesis and FISH, so we reported it with a brief review of literature.

Changes in the Outcomes of Very Low Birth Weight Infants

PURPOSE: The outcomes of infants weighing less than 1,500 gm(very low birth weight infant : VLBWI) reflect recent progress in neonatal intensive care. In this study, we analyzed changes over time in survival rate and morbidity of VLBWIs during the past seven years. METHODS: A retrospective review of medical records was analyzed for VLBWIs admitted to the neonatal intensive care unit of Samsung Medical Center within three days from birth. We compared the outcomes of previous corresponding data(period I : Oct. 1994 to Sept. 1996), with the outcomes of period II(Oct. 1996 to Dec. 1998) and period III(Jan. 1999 to Dec. 2000). RESULTS: As shown in Tables 1 and 3, the distribution of birth weight, gestational age(GA), gender, and inborn admissions did not change during the 7-year study. The overall survival rate of VLBWI increased significantly over time(period I : 72% vs period III : 88.3%, P<0.05). Between period I and period II, the birth weight-specific survival rate increased by 23.6%(75% vs 92.7%, P<0.05) for infants 1,000 to 1,249 gm. Between period II and period III, the birth weight-specific survival rate increased three times(20% vs 66.7%, P<0.05) for infants <750 gm. The survivors of lowest birth weight included infants at 624 gm(GA : 26(+5) weeks), 667 gm(GA : 25(+6) weeks) and 480 gm(GA : 26(+2) weeks) in each period. The gestational age-specific survival rate in period III increased significantly in GA 25-26 weeks and 29-30 weeks(vs period I and period II, P<0.05). The survivors of lowest gestational age included infants at GA 26 weeks(970 gm), GA 23(+5) weeks(791 gm) and GA 24(+1) weeks(740 gm) in each period. The incidence of severe IVH(grade III, IV) and the early death rate(

A Case of Identification of Marker Chromosome by comparative genomic hybridization and fluorescence in situ hybridization / 대한임상병리학회지

Comparative genomic hybridization (CGH) has been used to identify deletions and amplifications, particularly in neoplastic samples. CGH provides a new possibility searching genomes for imbalances of genetic material. We described the combined use of CGH and fluorescence in situ hybridization (FISH) to identify the origin of a marker chromosome in a child with mental retardation. Giemsa banding of metaphases from cultured lymphocytes showed a marker chromosome. The Karyotype was 47,XX,+mar. CGH revealed that the additional material originated from 15q. FISH confirmed this finding with whole chromosome paint for chromosome 15 and with a D15S10 (15q11-13) probe. This case demonstrates the efficient use of CGH and confirmatory FISH for the identification of chromosomal material of unknown origin.

Effects of Sample Pretreatment in Amino Acid Analysis / 대한임상병리학회지

BACKGROUND: Measurements of the concentrations of free amino acids in the blood are used as useful biochemical indicators. The sample pretreatments, including anticoagulant selection and deproteinization, are important steps in plasma-free amino acid analysis for accurate and stable results. Heparin and EDTA venous plasma in a frozen state are most commonly applied sample sources in our laboratory. Therefore, we investigated the effects of the anticoagulant and delayed deproteinization in amino acid measurement using ion-exchange chromatography. METHODS: We used Biochrom 20 amino acid analyzer (Biochrom, U.K). Blood samples were taken from 3 healthy adults after a minimum of 8 hours fasting. Two different types of vacutainer tubes, including sodium heparin and EDTA were used. To investigate variations by heparin volume, 3 mL and 6 mL of blood were drawn in 10 mL heparin tubes. We used an aqueous solution of SSA for deproteinization. To investigate variations through delayed deproteinization, we deproteinized the samples immediately and 24 hours later after plasma separation. RESULTS: There were no significant differences in concentrations except for cystine, glutamic acid and taurine, and the retention time between the 6 sample groups. The concentration of taurine was higher in the groups of late deproteinized plasma. In the groups of the same deproteinization time, there were no significant differences in concentration by different heparin concentrations. When we compared the results of 3 mL EDTA plasma with that of heparin-treated 6 mL of blood, the most widely used sample type, there was a significant difference in cystine concentration in the delayed deproteinized group but there were no differences in the immediately deproteinized group. CONCLUSIONS: Both 3 mL EDTA blood and 6 mL heparin-treated blood can be used commonly in case of using high-resolution ion-exchange chromatography and an immediately deproteinized sample. But, the results in amino acids can be affected in delayed pretreatment samples. Their effects should always be considered when interpreting laboratory results. The laboratories should standardize adequate sample preparation for the accurate analysis of amino acids.

Measurement of Pancreatic Islet Cell Autoantibodies by ELISA and RIA in Patients with Diabetes Mellitus / 대한임상병리학회지

BACKGROUND: The isoenzyme of glutamate decarboxylase (GAD), islet associated antigen (IA2, IAA) and insulin are known to be the major target antigens of pancreatic islet cell autoantibody as a predictor of type 1 diabetes mellitus (DM). Generally radioimmunoassay (RIA) methods are used for these autoantibodies but inconvenience of dealing with radioisotope have made enzyme-linked immunosorbent assay (ELISA) developed for clinical utilization. But, lack of evaluation or comparison studies of these two methods for autoantibodies make laboratories hesitate to adopt. METHODS: We measured the glutamate decarboxylase autoantibody (GADA), insulin autoantibody (IAA) and pancreatic islet cell autoantibodies (ICA) by a commercial ELISA method in 34 patients with type 1 DM, and 31 patients with type 2 DM, and 32 healthy control group. Conventional RIA was performed concurrently and compared for GADA and IAA. ICA was measured by conventional indirect immunofluorescent assay (IFA). The obtained results were compared and also C-peptide level was measured as a marker for residual function of islet cell of pancreas. RESULTS: Each autoantibody measured by ELISA in type 1 DM showed positive rate of 11.8% and for ICA, 26.5% for GADA, and 35.3% for IAA. The positive rate of the same group of type 1 DM when using RIA were 76.5% for GADA far exceeding that of ELISA method, and 29.4% for IAA. The percentage of positivity in combination of the ELISA methods for ICA and GAD yielded 29.4%, ICA plus IAA showed 38.2%, and GAD plus IAA was 52.9%, respectively. IAA positive rates in two groups divided by the age of 10 showed no significant difference. The presence of the autoantibodies did not influenced the C-peptide level. CONCLUSIONS: Further large scale studies including prediabetic state and autoimmune diabetes are required to establish the accurate diagnostic method of islet cell autoantibodies. But, presently ELISA method was considered that more improvement was needed for reliable and comparable results especially GADA.

Evaluation of Theophylline, Valproic Acid and Phenytoin by the CEDIA Method / 대한임상병리학회지

BACKGROUND: CEDIA is a newly developed method for therapeutic drug monitoring (TDM) and has some merits such as easy application to routine chemical analyzers, rapid and precise quantitation even in low concentrations and less cross reactivity. We evaluated the CEDIA(epsilon) (Microgenics Co., CA, USA) in measurement of theophyllin, valproic acid and phenytoin levels using 502X(epsilon) (A & T, Tokyo, Japan) and compared the results to those of the TDx(epsilon) (Abbott Laboratories, IL, USA) in order to assess the utility of the CEDIA(epsilon). METHODS: We evaluated the performance of 502X(epsilon) in the aspects of the within-runs and the between-runs precision, linearity, and carry-over. We compared the results of the CEDIA(epsilon) reagent with those of TDx(epsilon). The control materials (Bio-Rad TDM control level 1 and level 3; Bio-Rad laboratories, CA, USA) and clinical specimens were used for these studies. RESULTS: The coefficients of variation (CV) for the within-run and the between-run imprecision of 502X(epsilon) were 2.0-7.6% and 4.0-6.5%, respectively. The carry-over rate for theophyllin, valproic acid and phenytoin was 1.33%, 0.45% and 0.53%, respectively. The linearity (r(2)) of theophyllin, valproic acid and phenytoin was 0.9941, 0.9983 and 0.9947, respectively. The correlation coefficients (r) of theophyllin, valproic acid and phenytoin levels of CEDIA(epsilon), with those determined by the TDx(epsilon), were 0.9730, 0.9703 and 0.9695, respectively (P<0.001). CONCLUSIONS: The recentlydeveloped CEDIA(epsilon) proved to be highly precise and linear for quantitative analysis of theophyllin, phenytoin and valproic acid. Correlations with TDx(epsilon) were significantly high. CEDIA(epsilon) was thought to be clinically useful for TDM.

The Role of Aldosterone on the Development of Renal Tubular Reabsorption in Low Birth Weight Infants

PURPOSE: To determine the postnatal changes in aldosterone action on the renal tubular reabsorption in low birth weight(LBW) infants, we assessed the relation of the aldosterone concentrations to renal parameters during the first 10 days of life. METHODS: Twenty LBW infants were evaluated and their gestational ages ranged from 32.4 to 39.3 weeks and their birth weights ranged from 1,440 to 2,500 g. Estimated glomerular filtration rate, fractional excretion of sodium(FENa) and potassium(FEK), and plasma aldosterone concentrations were analyzed according to the postnatal age and the conceptional age(CA). RESULTS: Glomerular functions were improved after birth and were correlated with CA. FENa and FEK decreased after birth and correlated with CA. Plasma aldosterone concentrations increased to 318.6 +/- 147.2 ng/dL at 48 hours and then decreased to 162.0 +/- 72.2 ng/dL at 10 days after birth. Plasma aldosterone concentrations of infants less than 38th week of CA were higher than that of infants more than 38th week. There was a significant negative correlation coefficient between plasma aldosterone concentrations and FENa in infants more than 34th week of CA, but not in that of less than 34th week. CONCLUSIONS: LBW infants have higher plasma aldosterone concentrations, but a poor correlation between plasma aldosterone concentration and urinary sodium excretion for the first few days of life and in lower chronologic aged infants. These results show that the renal tubule reabsorption of sodium is less responsive to plasma aldosterone in these infants and, therefore, the careful management of fluid and electrolyte balance is mandatory.

Detailed Characterization of Chromosomal Breakpoints in an ALL, L3 using Cross Species Color Banding / 대한임상병리학회지

Karyotype analysis by G-anding is the standard method for identifying numerical and structural chromosomal aberrations in cytogenetic laboratories. However, the origins of marker chromosomes, subtle translocations, or complex chromosomal rearrangements are often difficult to identify with certainty. Cross-pecies color banding is a new FISH-ased screening technique that enables the generation of a specific color banding pattern for each human chromosome based on the genomic homologies between man and various species of apes. We report application of Cross-pecies color banding (RxFISH) to characterize the chromosomal rearrangements of one leukemia sample the G-and karyotype of which were incomplete. The combination of G-anding and RxFISH in this case study yielded additional information beyond that obtained by either technique used alone in determining the precise breakpoints in complex chromosomal rearrangements.

Detailed Characterization of Chromosomal Breakpoints in an ALL, L3 using Cross Species Color Banding / 대한임상병리학회지

Karyotype analysis by G-anding is the standard method for identifying numerical and structural chromosomal aberrations in cytogenetic laboratories. However, the origins of marker chromosomes, subtle translocations, or complex chromosomal rearrangements are often difficult to identify with certainty. Cross-pecies color banding is a new FISH-ased screening technique that enables the generation of a specific color banding pattern for each human chromosome based on the genomic homologies between man and various species of apes. We report application of Cross-pecies color banding (RxFISH) to characterize the chromosomal rearrangements of one leukemia sample the G-and karyotype of which were incomplete. The combination of G-anding and RxFISH in this case study yielded additional information beyond that obtained by either technique used alone in determining the precise breakpoints in complex chromosomal rearrangements.

Effect of Fluid Restriction Through High Humidification on the Management of Very Low Birth Weight Infant

PURPOSE: The aim of this study is to investigate the effects of high humidification of nearly 100% on insensible water loss and total fluid requirement in very low birth weight infants during their first week of life. METHODS: We retrospectively compared twenty-five infants (GA 28.1+/-1.7 wk, and BW 970+/-186 g) who were cared for in a double walled incubator with high humidification with 24 infants (GA 27.6+/-2.2 wk, and BW 972+/-186 g) who were in a double walled incubator without raised humidity during the first week of their life. We analyzed the changes in body weight, total fluid intake, insensible water loss, urine output, input of sodium and potassium, serum levels of sodium and potassium, and the incidence of complications during the hospitalization in the high humidified group and the control group. RESULTS: High humidification during the first week of life in very low birth weight infants reduced insensible water loss and total fluid requirement. However the imbalance of serum electrolyte or oliguria did not increase. The high humidification group (n=25) acquired more physiologic weight loss during the first two weeks of life than the control group (n=24) but gained more weight than the control group (n=24) on the third month of life. Infants nursed with high humidification showed an increased tendency toward spontaneous closure of the patent ductus arteriosus and a decreased tendency toward surgical ligation of the patent ductus arteriosus and intraventricular hemorrhage without the evidence of an increase in infection. No significant differences were noted in the incidence of patent ductus arteriosus, bronchopulmonary dysplasia and retinopathy of prematurity. CONCLUSION: The high humidification of nearly 100% and fluid restriction therapy during the first week of life were effective in the fluid and electrolyte management of very low birth weight infants.

A case of multiple myeloma with ascites / 대한내과학회지

Ascites is a rare complication of multiple myeloma. When it develops, it is usually associated with extensive liver infiltration with plasma cells, infectious peritonitis or myelomatous peritoneal infiltration. Ascites caused by peritoneal infiltration is even less frequent than others. The majority of previously reported cases were characterized by an IgA paraprotein and lack of skeletal lesions. This rare extramedullary complication of myeloma has been unresponsive to therapy and rapidly fatal. Therefore, it is important to recognize myeloma as a cause of ascites and the presence of ascites heralds a poor prognosis of myeloma. We recently experienced a case of myeloma with ascites and reviewed the relevant literature of human myeloma presenting with the triad of ascites, relative or absolute sparing of the skeleton, and an IgA paraprotein. A 76-year-old man was presented with ascites early in the course of myeloma. He had no evidence of intra-abdominal plasmacytoma and skeletal lesions. Myelomatous ascites was demonstrated by the monoclonal immunoglobulin of IgA type in ascitic fluid. He was treated by plasmapheresis due to hyperviscosity syndrome and VAD combination chemotherapy. He was discharged with the improved clinical condition.

A Case of mos 45,X/46,X, +mar. ish der(X)(wcpX+) Turner Syndrome / 대한임상병리학회지

Turner syndrome is a genetic disorder that affects about 1/2,000-1/5,000 females born. The typical female with Turner syndrome has only one X chromosome in each of her cells. There are several variations on this theme as other similar chromosome anomalies occur in females with Turner syndrome. We observed a patient with short stature, abscent vagina and chromosomal abnormality. Chromosomal analysis of the patient showed 45,X/46,X, +mar. The marker chromosome was revealed as X chromosome in fluorescent in situ hybridization (FISH). We report a case of mos 45,X/46,X,+mar.ish der(X)(wcp X+) in Turner syndrome with a brief review of literature.