Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Zl, DXZI, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.
Blastomeres ; Chromosome Aberrations ; DNA Probes ; Fluorescence* ; Germ Cells ; Humans ; In Situ Hybridization* ; Interphase ; Lymphocytes ; Metaphase ; Spermatozoa

BACKGROUND: The purposes of this study were to assess the resting volume of the modified bronchial cuff of left-sided double-lumen bronchial tubes (DLT) and to determine the maximum range of the mainstem bronchial diameter indicated for DLT. METHODS: Left-sided DLTs (Broncho-Cath(R)) of 35-41 Fr (n = 5 each) were used for the study. The cuff was inflated with air in 0.5-ml increments to a volume of 5 ml and the corresponding cuff pressure was recorded. The smallest cuff volume, beyond which a 0.5 ml increase resulted in more than 10 mm Hg increase in cuff pressure, was considered to be the resting volume of that cuff. The resting volume was also calculated by differentiation on the fitted curve. The minimum required bronchial diameter was considered to be the reported OD of the bronchial tube and the maximum diameter was the measured OD of the bronchial cuff at a cuff pressure of 30 mmHg, which was measured with a precision caliper (0.1 mm intervals) at the midcuff level. RESULTS: The resting volume of the bronchial cuff, measured both traditionally and using the curve fitting analysis, were comparable. A DLT of any size > or = 35 Fr can be used for a bronchus with a diameter of 10.7-20.6 mm. CONCLUSIONS: There were extensive overlaps in the range of bronchial diameters indicated for each DLT size. Contrary to a common belief, the upper limits of the bronchial diameters indicated for all the DLTs > or = 35 Fr seemed to be the same regardless of the DLT size.
Bronchi*

OBJECTIVE: The purpose of this study was to determine whether intracytoplasmic sperm injection(ICSI) could overcome the defects of oocytes in IVF-ET patients with previous fertilization failure by conventional fertilization technique. Design: Retrospective study Materials and METHODS: A total of 119 ICSI cycles in 57 IVF-ET patients performed from May, 1995 to December, 1997 was enrolled. Subjects were divided into two groups: FR group included 66 ICSI cycles in 35 patients with normal sperm who underwent ICSI due to past history of failed or poor fertilization in the previous IVF-ET cycles, and OAT group included 53 ICSI cycles in 22 patients with severe oligoasthenoterato- zoospermia(OAT) which was defined as sperm concentration < 20 million/ml, mo#dlity < 30% and normal morphology < 4% by strict morphologic criteria. The outcomes of ICSI were analyzed and compared in both groups. RESULTS: The age of female patients, basal serum FSH level, and the numbers of oocytes retrieved and metaphase II oocytes were all comparable in both groups. The fertilization rate after ICSI was similar in both groups(68.7+/-25.3% vs. 67.7+/-24.5%), as were the cleavage rate of normally fertilized oocytes(93.1+/-21.4% vs. 89.3+/-21.6%), the number of embryos transferred(4,00+/-1.98 vs. 4.64+/-2.10), and cumulative embryo score(CES) indicating the quality of embryos(47.3+/-33.2 vs. 54.1+/-33.2). The implantation rate(4.3+/-10.5% vs. 3.8+/-11.0%) and the clinical pregnancy rate per cycle(15.2% vs. 13.2%) were also comparable in both groups. CONCLUSIONS: Although it has been shown that there is a higher risk of chromosomal abnormalities in oocytes from IVF-ET patients with pevious failed or poor fertilization, higher implantation and clinical pregnancy rates wer#e not observed in patients with OAT following ICSL Therefore, the functional defect of sperm such as loss of capacitation, defect of aaasome reaction, and abnormality of nucleus decondensation should be also considered in patients with previous failed or poor fertilization.
Avena ; Chromosome Aberrations ; Embryonic Structures ; Female ; Fertilization* ; Humans ; Male* ; Metaphase ; Oocytes ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; Spermatozoa*

Interleukin (IL)-27 is a novel cytokine of the IL-6/IL-12 family that has been reported to be involved in the pathogenesis of autoimmune diseases and has a pivotal role as both a pro- and anti-inflammatory cytokine. We investigated the in vivo effects of IL-27 on arthritis severity in a murine collagen-induced arthritis (CIA) model and its mechanism of action regarding control of regulatory T (Tregs) and IL-17-producing T helper 17 (Th17) cells. IL-27-Fc-treated CIA mice showed a lower severity of arthritis. IL-17 expression in the spleens was significantly decreased in IL-27-Fc-treated CIA mice compared with that in the CIA model. The Th17 population was decreased in the spleens of IL-27-Fc-treated CIA mice, whereas the CD4+CD25+Foxp3+ Treg population increased. In vitro studies revealed that IL-27 inhibited IL-17 production in murine CD4+ T cells, and the effect was associated with retinoic acid-related orphan receptor gammaT and signal transducer and activator of transcription 3 inhibition. In contrast, fluorescein isothiocyanate-labeled forkhead box P3 (Foxp3) and IL-10 were profoundly augmented by IL-27 treatment. Regarding the suppressive capacity of Treg cells, the proportions of CTLA-4+ (cytotoxic T-lymphocyte antigen 4), PD-1+ (programmed cell death protein 1) and GITR+ (glucocorticoid-induced tumor necrosis factor receptor) Tregs increased in the spleens of IL-27-Fc-treated CIA mice. Furthermore, in vitro differentiated Treg cells with IL-27 exerted a more suppressive capacity on T-cell proliferation. We found that IL-27 acts as a reciprocal regulator of the Th17 and Treg populations in CD4+ cells isolated from healthy human peripheral blood mononuclear cells (PBMCs), as well as from humans with rheumatoid arthritis (RA) PBMCs. Our study suggests that IL-27 has the potential to ameliorate overwhelming inflammation in patients with RA through a reciprocal regulation of Th17 and Treg cells.
Animals ; Arthritis, Experimental/*drug therapy/immunology ; Cells, Cultured ; Humans ; Interleukins/immunology/*therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; T-Lymphocytes, Regulatory/*immunology ; Th17 Cells/*immunology

The proportion of male factor infertility due to quantitative and qualitative sperm disorders is approximately 50-60% in infertile couples. In IVF-ET, lower or failed fertilization of oocytes usually results from subnormal count of total motile sperms, but this may occur in infertile couples even with normal sperm count. It has been suggested that some functional defects in sperms are responsible for lower or failed fertilization. Routine semen analysis based on numerical background has limits for the assessment of fertilization capacity of sperm in infertile males, and the andrologic test for the prediction of fertilization capacity must be objective, repeatable, quick, economic, and easily applicable for the clinical settings. The purposes of this study were to develop the analysis method of strict morphology of sperm using the strict criteria as a simple, inexpensive and useful test of sperm fertilization capacity, to establish the normal fertile range and the cut-off value of strict morphology, and to evaluate the validity of strict morphology as a prognostic indicator of fertilization capacity in IVF-ET. In establishing the effectiveness of strict morphology of sperm, ROC curve was used. Among the various thresholds for the prediction of fertilizing ability, normal morphologic value 10.0 corresponding to the value with higher sensitivity and lesser false positive rates was determined as a cut-off value. Using this cut-off point, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of strict morphology for the prediction of fertilization capacity was 73.9%, 81.0%, 80.6%, and 72.7%, respectively. To evaluate the clinical validity of strict morphology as a prognostic indicator of fertilization capacity, this cut-off point was applied to 133 patients undergoing IVF-ET. For the prediction of fertilization rate >30% in IVF-ET, the sensitivity, specificity, PPV, and NPV was 77.3%, 77.8%, 87.2%, and 63.6%, respectively. In conclusion, the strict morphology of sperm is one of the most simple and useful test for the assessment of fertilization capacity of sperm and the prediction of IVF-ET outcomes in infertile couples.
Family Characteristics ; Fertilization ; Humans* ; Infertility ; Male ; Oocytes ; ROC Curve ; Semen Analysis ; Sensitivity and Specificity ; Sperm Count ; Spermatozoa*

Background: Current international guidelines recommend against deep sedation as it is associated with worse outcomes in the intensive care unit (ICU). However, in Korea the prevalence of deep sedation and its impact on patients in the ICU are not well known. Methods: From April 2020 to July 2021, a multicenter, prospective, longitudinal, noninterventional cohort study was performed in 20 Korean ICUs. Sedation depth extent was divided into light and deep using a mean Richmond Agitation–Sedation Scale value within the first 48 hours. Propensity score matching was used to balance covariables; the outcomes were compared between the two groups. Results: Overall, 631 patients (418 [66.2%] and 213 [33.8%] in the deep and light sedation groups, respectively) were included. Mortality rates were 14.1% and 8.4% in the deep and light sedation groups (P = 0.039), respectively. Kaplan-Meier estimates showed that time to extubation (P < 0.001), ICU length of stay (P = 0.005), and death P = 0.041) differed between the groups. After adjusting for confounders, early deep sedation was only associated with delayed time to extubation (hazard ratio [HR], 0.66; 95% confidence inter val [CI], 0.55– 0.80; P < 0.001). In the matched cohort, deep sedation remained significantly associated with delayed time to extubation (HR, 0.68; 95% 0.56–0.83; P < 0.001) but was not associated with ICU length of stay (HR, 0.94; 95% CI, 0.79–1.13; P = 0.500) and in-hospital mortality (HR, 1.19; 95% CI, 0.65–2.17; P = 0.582). Conclusion In many Korean ICUs, early deep sedation was highly prevalent in mechanically ventilated patients and was associated with delayed extubation, but not prolonged ICU stay or in-hospital death.

Objective: The incidence of hip fractures is increasing worldwide with the aging population, causing a challenge to healthcare systems due to the associated morbidities and high risk of mortality. After hip fractures in frail geriatric patients, existing comorbidities worsen and new complications are prone to occur. Comprehensive rehabilitation is essential for promoting physical function recovery and minimizing complications, which can be achieved through a multidisciplinary approach. Recommendations are required to assist healthcare providers in making decisions on rehabilitation post-surgery. Clinical practice guidelines regarding rehabilitation (physical and occupational therapies) and management of comorbidities/complications in the postoperative phase of hip fractures have not been developed. This guideline aimed to provide evidence-based recommendations for various treatment items required for proper recovery after hip fracture surgeries. Methods Reflecting the complex perspectives associated with rehabilitation post-hip surgeries, 15 key questions (KQs) reflecting the complex perspectives associated with post-hip surgery rehabilitation were categorized into four areas: multidisciplinary, rehabilitation, community-care, and comorbidities/complications. Relevant literature from four databases (PubMed, EMBASE, Cochrane Library, and KoreaMed) was searched for articles published up to February 2020. The evidence level and recommended grade were determined according to the grade of recommendation assessment, development, and evaluation method. Results A multidisciplinary approach, progressive resistance exercises, and balance training are strongly recommended. Early ambulation, weigh-bearing exercises, activities of daily living training, community-level rehabilitation, management of comorbidities/complication prevention, and nutritional support were also suggested. This multidisciplinary approach reduced the total healthcare cost. Conclusion This guideline presents comprehensive recommendations for the rehabilitation of adult patients after hip fracture surgery.

Objective: The incidence of hip fractures is increasing worldwide with the aging population, causing a challenge to healthcare systems due to the associated morbidities and high risk of mortality. After hip fractures in frail geriatric patients, existing comorbidities worsen and new complications are prone to occur. Comprehensive rehabilitation is essential for promoting physical function recovery and minimizing complications, which can be achieved through a multidisciplinary approach. Recommendations are required to assist healthcare providers in making decisions on rehabilitation post-surgery. Clinical practice guidelines regarding rehabilitation (physical and occupational therapies) and management of comorbidities/complications in the postoperative phase of hip fractures have not been developed. This guideline aimed to provide evidence-based recommendations for various treatment items required for proper recovery after hip fracture surgeries. Methods Reflecting the complex perspectives associated with rehabilitation post-hip surgeries, 15 key questions (KQs) reflecting the complex perspectives associated with post-hip surgery rehabilitation were categorized into four areas: multidisciplinary, rehabilitation, community-care, and comorbidities/complications. Relevant literature from four databases (PubMed, EMBASE, Cochrane Library, and KoreaMed) was searched for articles published up to February 2020. The evidence level and recommended grade were determined according to the grade of recommendation assessment, development, and evaluation method. Results A multidisciplinary approach, progressive resistance exercises, and balance training are strongly recommended. Early ambulation, weigh-bearing exercises, activities of daily living training, community-level rehabilitation, management of comorbidities/complication prevention, and nutritional support were also suggested. This multidisciplinary approach reduced the total healthcare cost. Conclusion This guideline presents comprehensive recommendations for the rehabilitation of adult patients after hip fracture surgery.