Assessment of myopic rebound effect after discontinuation of treatment with 0.01% atropine eye drops in Japanese school-age children.

PURPOSE: Having previously demonstrated the efficacy of 0.01% atropine eye drops for inhibiting progression of childhood myopia, we conducted additional analyses to assess post-treatment changes in myopia progression. STUDY DESIGN: Analysis of follow-up data from a previously reported randomized controlled trial METHODS: A mixed-effects model was used to compare intergroup changes in spherical equivalent (SE) and axial length (AL) at 1 month and 12 months after discontinuation of 2-year treatment with atropine or placebo in 167 school-age children. RESULTS: Follow-up measurements were available for 149 participants at 1 month after discontinuation of treatment and for 51 participants at 12 months after discontinuation. At 1 month post-treatment, differences between the atropine and placebo groups in least squares (LS) mean changes in SE and AL, respectively, from 24 months were -0.06 diopters (D) (95% CI: -0.21, 0.08; P = .39) and 0.02 mm (95% CI: -0.05, 0.08; P = .60). At 12 months post-treatment, intergroup differences (atropine vs placebo) in LS mean changes in SE and AL, respectively, were -0.13 D (95% CI: -0.35, 0.10; P = .26) and -0.02 mm (95% CI: -0.12, 0.09; P = .75). LS mean changes in SE and AL from treatment discontinuation did not differ between the groups at 1 or 12 months post-treatment. CONCLUSION: Axial elongation was significantly less in the atropine group than in the placebo group. The suppression effect obtained at 2 years was maintained after 12 months. The absence of intergroup differences in myopia progression since treatment cessation suggests that myopic rebound did not occur.

Occurrence and disease burden of respiratory syncytial virus and other respiratory pathogens in adults aged ≥65 years in community: A prospective cohort study in Japan.

BACKGROUND: The frequency and clinical profile of respiratory syncytial virus (RSV)-acute respiratory disease (ARD) in older adults in Japan has not been well-characterized. METHODS: This was a multicenter prospective observational cohort study to evaluate the occurrence rate of ARD in 1000 older adult participants (≥65 years) for 52 weeks during the 2019 to 2020 season. A multiplex polymerase chain reaction panel was used for pathogen detection in nasopharyngeal swab from participants diagnosed with ARD. Symptoms and impact of ARD was assessed using the Respiratory Infection Intensity and Impact Questionnaire (RiiQ™). The study was registered at UMIN (https://www.umin.ac.jp/ctr/): UMIN000037891. RESULTS: RSV-ARD was detected in 24/1000 (2.4%) participants and RSV-lower respiratory tract disease in 8/1000 (0.8%) participants. The median duration of RSV-ARD was 18 days. All 24 participants had utilized the medical services of outpatient visits and only 1 (4.2%) participant was hospitalized for RSV-ARD. The most common viruses other than RSV that caused ARD (detected in >10 participants) were human rhinovirus/enterovirus, parainfluenza 3, coronavirus OC43, human metapneumovirus, and influenza A/H1. The most frequent symptoms of RSV-ARD were cough, sore throat, nasal congestion, and expectoration. CONCLUSIONS: RSV was reported as a major pathogen for respiratory infections in older adults in Japan.

Computed Tomographic Assessment of Age- and Gender-Specific Sagittal Lumbopelvic Alignment in a Japanese Population.

INTRODUCTION: Previous studies have reported differences in lumbopelvic alignment between standing and supine positions. Computed tomography (CT) images taken in the supine position are often used for clinical studies in addition to standing radiographs, although not frequently. Our study aims to clarify normative values of CT-evaluated lumbopelvic parameters and the characteristics of age- and gender-related lumbopelvic alignment in the supine position. METHODS: Patients undergoing CT scans of abdominal or lumbar regions for reasons other than low back disorders were included (n=581). Sagittal multiplanar reconstruction CT images were obtained, and lumbar lordosis (LL), L5-S1 angle, and sacral slope (SS) were measured. Mean values of the parameters in patients aged 59 years and under, 60-69, 70-79, and 80 and over, and in males and females, were calculated. Age- and gender-related differences in these parameters were statistically analyzed. RESULTS: Among the four age groups, patients 80 years and over showed significantly lower LL and SS than patients aged 70-79. Females 80 years and over showed significantly lower LL and SS than all other age groups, but those in males did not. The comparison between males and females showed no significant differences in LL and SS; however, the L5-S1 angle was significantly higher in males than in females. In patients 80 years and over, females showed significantly lower LL and SS than in males. CONCLUSIONS: This study provides normative CT-evaluated lumbopelvic parameters, such as LL, L5-S1 angle, and SS, which will be utilized for comparisons in future studies. The present study first revealed that pelvic retroversion and lumbar kyphosis occur in elderly females in the supine position, which raised a possibility that age-related decrease of LL and SS in females occurs at an older age in the supine position than in the standing position.

Combined Assessment of D-Dimer with the Get with the Guidelines-Heart Failure Risk Score and N-Terminal Pro-B-Type Natriuretic Peptide in Patients with Acute Decompensated Heart Failure with Preserved and Reduced Ejection Fraction.

The prognostic role of D-dimer in different types of heart failure (HF) is poorly understood. We investigated the prognostic value of D-dimer on admission, both independently and in combination with the Get With The Guidelines-Heart Failure (GWTG-HF) risk score and N-terminal pro-B-type natriuretic peptide (NT-proBNP), in patients with preserved left ventricular ejection fraction (LVEF) and acute decompensated HF (HFpEF) or reduced LVEF (HFrEF). Baseline D-dimer levels were measured on admission in 1670 patients (mean age: 75 years) who were hospitalized for worsening HF. Of those patients, 586 (35%) were categorized as HFpEF (LVEF ≥ 50%) and 1084 as HFrEF (LVEF < 50%). During the 12-month follow-up period after admission, 360 patients died. Elevated levels (at least the highest tertile value) of D-dimer, GWTG-HF risk score, and NT-proBNP were all independently associated with mortality in all HFpEF and HFrEF patients (all p < 0.05). Adding D-dimer to a baseline model with a GWTG-HF risk score and NT-proBNP improved the net reclassification and integrated discrimination improvement for mortality greater than the baseline model alone in all populations (all p < 0.001). The number of elevations in D-dimer, GWTG-HF risk score, and NT-proBNP were independently associated with a higher risk of mortality in all study populations (HFpEF and HFrEF patients; all p < 0.001). The combination of D-dimer, which is independently predictive of mortality, with the GWTG-HF risk score and NT-proBNP could improve early prediction of 12-month mortality in patients with acute decompensated HF, regardless of the HF phenotype.

On-site assessment of computed tomography-derived fractional flow reserve in comparison with myocardial perfusion imaging and invasive fractional flow reserve.

Myocardial perfusion imaging (MPI) using Single Photon Emission Computed Tomography has been established as a standard noninvasive tool for risk stratification of coronary artery disease (CAD). We evaluated the diagnostic performance of on-site workstation-based computed tomography-derived fractional flow reserve (CT-FFR) in comparison with MPI using invasive fractional flow reserve (invasive FFR) as a gold standard. We enrolled 97 patients with suspected CAD. Diagnostic performance of CT angiography (CTA), and CT-FFR was compared in 105 lesions of 97 patients. Invasive FFR ≤ 0.8 was detected in 38 (36%) lesions. Diagnostic performance of CT-FFR was improved compared with CTA (AUC 0.83 vs. 0.60, p < 0.0001). The lesions with both CTA and MPI findings (n = 47), invasive FFR ≤ 0.8 was detected in 19 (40.4) lesions. CT-FFR (AUC 0.81, 95% CI 0.72-0.94) significantly improved diagnostic performance compared with CTA-50% (AUC 0.59, p = 0.00019) and MPI (AUC 0.64, p = 0.0082). In lesions with ≥ 50% on CTA (n = 42), diagnostic accuracy of CT-FFR (AUC 0.81) was significantly superior to MPI (AUC 0.64, p = 0.0239). In conclusions, CT-FFR improved diagnostic accuracy to detect invasive FFR ≤ 0.8 compared with luminal stenosis on CTA and ischemia on MPI. Patients with ≥ 50% stenosis on CTA would be the candidates for CT-FFR.

Pilot study of oblique lumbar interbody fusion using mobile percutaneous pedicle screw and validation by a three-dimensional finite element assessment.

The purpose of this study was to try oblique lateral interbody fusion (OLIF) using percutaneous pedicle screws (PPS) with mobility. Twelve patients who underwent single-level OLIF were observed for at least one year. These included 6 patients with conventional PPS (rigid group), and 6 with movable PPS (semi-rigid group). Mobile PPS used cosmicMIA, which is a load sharing system. The anterior and posterior disc height, screw loosening and bone healing period, and implant failure were evaluated at final observation by CT. Moreover, the stress on the vertebral body-cage, on the vertebral body-screw/rod and on the bone around the screw was estimated using a three-dimensional finite element assessment in both groups. There was no significant difference in surgical time, amount of bleeding, JOA score, or low back pain VAS between groups. There were no differences between groups in anterior and posterior disc height, screw loosening, and implant failure at final observation. The bone healing period was significantly shorter in the semi-rigid screw group (18.3 months vs 4.8 months, p = 0.01). The finite element analysis showed that the lower stress on the rod/screw would contribute to fewer implant fractures and that lower stress on the bone around the screw would reduce screw loosening, and that higher compressive force on the cage would promotes bone healing. OLIF combined with a movable screw accelerated bone healing by nearly 75%. We conclude that mobile PPS in combination with OLIF promotes bone healing and can be a better vertebral fusion technique.

Ultra-High-Resolution Computed Tomography Angiography for Assessment of Coronary Artery Stenosis.

BACKGROUND: Limitations of coronary computed tomography (CTA) include false-positive stenosis at calcified lesions and assessment of in-stent patency. A prototype of ultra-high resolution computed tomography (U-HRCT: 1,792 channels and 0.25-mm slice thickness×128 rows) with improved spatial resolution was developed. We assessed the diagnostic accuracy of coronary artery stenosis using U-HRCT.Methods and Results:Seventy-nine consecutive patients who underwent CTA using U-HRCT were prospectively included. Coronary artery stenosis was graded from 0 (no plaque) to 5 (occlusion). Stenosis grading at 102 calcified lesions was compared between U-HRCT and conventional-resolution CT (CRCT: 896 channels and 0.5-mm slice thickness×320 rows). Median stenosis grading at calcified plaque was significantly improved on U-HRCT compared with CRCT (1; IQR, 1-2 vs. 2; IQR, 1-3, P<0.0001). Assessability of in-stent lumen was evaluated on U-HRCT in 79 stents. Stent strut thickness and luminal diameter were quantitatively compared between U-HRCT and CRCT. Of 79 stents, 83.5% were assessable on U-HRCT; 80% of stents with diameter 2.5 mm were regarded as assessable. On U-HRCT, stent struts were significantly thinner (median, 0.78 mm; IQR, 0.7-0.83 mm vs. 0.83 mm; IQR, 0.75-0.92 mm, P=0.0036), and in-stent lumens were significantly larger (median, 2.08 mm; IQR, 1.55-2.51 mm vs. 1.74 mm; IQR, 1.31-2.06 mm, P<0.0001) than on CRCT. CONCLUSIONS: U-HRCT with improved spatial resolution visualized calcified lesions with fewer artifacts. The in-stent lumen of stents with diameter ≥2.5 mm was assessable on U-HRCT.

Noninvasive Assessment of Stenotic Severity and Plaque Characteristics by Coronary CT Angiography in Patients Scheduled for Carotid Artery Revascularization.

AIMS: Coronary artery atherosclerosis in patients needing carotid revascularization has not been fully clarified. The aim of this study was to evaluate the stenotic severity and plaque characteristics of coronary arteries by coronary computed tomography angiography (CTA) in patients scheduled for carotid-artery stenting (CAS) or carotid endarterectomy (CEA). METHODS: We performed coronary CTA after carotid ultrasound (US) in 164 patients (81.7% male, aged 68.1± 12.2 years) from 2014 to 2016. Of all, 70 were scheduled for CAS or CEA (CAS/CEA group) and 94 were not (non-CAS/CEA group). Carotid US and coronary CTA were compared for the evaluation of stenotic severity and plaque characteristics of each vessel between CAS/CEA and non-CAS/CEA groups. RESULTS: Between the two groups, there were significant differences in the presence of significant stenosis (SS: ≥70% stenosis of coronary artery) (55.7% vs. 39.4%, P=0.038), triple-vessel disease (TVD)/left main trunk (LMT) (SS in each of three epicardial vessels and/or LMT) (24.3% vs. 7.5%, P= 0.0025), and high-risk plaque (HRP: positive remodeling and/or low attenuation) (55.7% vs. 24.5%, P＜0.0001). CAS/CEA was independently associated with TVD/LMT (OR=2.30, 95%CI: 1.14-8.59, P=0.026) and HRP (OR=3.17, 95%CI: 1.57-6.54, P=0.0012) in multivariable logistic regression analysis. Similarly, vulnerable plaque (78.6% vs. 2.1%, P＜0.0001) as well as severe stenosis of carotid artery (98.6% vs. 0%, P＜0.0001) was seen more often in CAS/CEA than in non-CAS/CEA group. CONCLUSIONS: The prevalence of TVD/LMT and HRP determined by coronary CTA is higher in patients needing CAS/CEA than in those without. Management of systemic atherosclerosis is required in the perioperative period of CAS/CEA.

Health assessment questionnaire-disability index (HAQ-DI) score at the start of biological disease-modifying antirheumatic drug (bDMARD) therapy is associated with radiographic progression of large joint damage in patients with rheumatoid arthritis.

OBJECTIVES: Radiographic progression of damage (RPD) to large joints in patients with rheumatoid arthritis (RA) has not been fully studied. We previously demonstrated that Larsen grade of the large joints was associated with RPD of large joints in patients treated with biological disease-modifying anti-rheumatic drugs (bDMARDs); however, no factors associated with background characteristics of patients were identified. METHODS: A total of 400 large joints in the upper and lower extremities, including the shoulder, elbow, knee, and ankle, of 88 patients with RA treated with bDMARDs for 1-3 years were investigated. Radiographs of tender and/or swollen large joints were acquired at least twice during the study period (mean, 16.4 months), and the RPD was evaluated. RESULTS: A multivariate analysis revealed that health assessment questionnaire-disability index (HAQ-DI) score at the start of bDMARD treatment was associated with RPD. The cutoff value that discriminated progression from non-progression, determined by a receiver operating characteristic (ROC) curve, was 1.4375 (sensitivity: 0.778, specificity: 0.894). CONCLUSIONS: HAQ-DI score at the start of bDMARD treatment was associated with RPD to large joints during a therapeutic period of 1-3 years. Progressive damage is expected to increase when functional disability exceeds an HAQ-DI score of 1.5.

Assessment of trough rivaroxaban concentrations on markers of coagulation activation in nonvalvular atrial fibrillation population.

Whether trough-phase rivaroxaban concentrations provide sufficient anticoagulation needs more study. We evaluated levels of coagulation activation markers in the trough concentration phase in nonvalvular atrial fibrillation (NVAF) patients, and the correlation between these markers and rivaroxaban concentration. Fifty-five Japanese NVAF patients received 24-week rivaroxaban treatment of either 15 or 10 mg once-daily in the morning. Of these, 26 patients had no history of anticoagulant therapy (naive group) and 29 had switched from warfarin (warfarin group). D-dimer and prothrombin fragment 1 + 2 (F1 + 2) levels, and protein C activities were measured at 0 (baseline), 12 and 24 weeks of rivaroxaban treatment just before the patient's regular dosing time (trough phase). For 49 patients, D-dimer, F1 + 2, and rivaroxaban concentrations were also measured twice between 28 and 32 weeks of rivaroxaban treatment at non-trough times to achieve a range of drug concentrations for correlation analysis. For the naive group, D-dimer and F1 + 2 levels were significantly reduced (p < 0.01) from baseline at 12 and 24 weeks. For the warfarin group, these values were unchanged for D-dimer but significantly increased (p < 0.01) for F1 + 2. Protein C activity was unchanged in the naive group and was increased (p < 0.01) in the warfarin group. Prothrombin time (r = 0.92, p < 0.0001) and activated partial thromboplastin time (r = 0.54, p < 0.0001) correlated with rivaroxaban concentration, but not D-dimer and F1 + 2 levels. In conclusion, rivaroxaban in the trough phase is comparable to warfarin in reducing D-dimer levels. Although trough level rivaroxaban suppresses F1 + 2 less than warfarin, the higher activities of protein C with rivaroxaban treatment compared to warfarin treatment may counterbalance this. Lack of correlation between rivaroxaban concentration and D-dimer and F1 + 2 levels suggests that trough concentrations of rivaroxaban reduce their concentrations as effectively as higher levels do.

Prediction of thrombolytic therapy after stroke-bypass transportation: the Maria Prehospital Stroke Scale score.

There is no prehospital stratification tool specifically for predicting thrombolytic therapy after transportation. We developed a new prehospital scale named the Maria Prehospital Stroke Scale (MPSS) by modifying the Cincinnati Prehospital Stroke Scale. Our objective is to evaluate its utility in a citywide bypass transportation protocol for intravenous (IV) tissue plasminogen activator (tPA). In the MPSS, facial droop, arm drift, and speech disturbance are tested by emergency medical technicians (EMTs). Facial droop is graded as normal (0) or abnormal (1), and the other 2 items are graded in 3 levels as normal (0), not severe (1), and severe (2). Thus, the total MPSS score ranges from 0 to 5. The predictive value of MPSS for thrombolytic therapy after bypass transportation was evaluated in 1057 patients. The MPSS scored by EMTs was significantly correlated with the National Institutes of Health Stroke Scale score in the emergency room (Spearman rho = .67, P = .000). The onset-to-door time was significantly longer with a low MPSS score (analysis of variance, F5,4.21 = .001). The rate of thrombolytic therapy was increased when the MPSS score increased from 0 to 5: 0%, 4.1%, 8.8%, 13.0%, 20.3%, and 31.5%, respectively. The areas under the receiver operating characteristic curve for the correct diagnosis of stroke and prediction of IV tPA therapy were calculated as .737 (95% confidence interval [CI]: .688-.786) and .689 (95% CI: .645-.732), respectively. Multivariate logistic regression analysis showed that the MPSS score and the detection-to-door time were independent predictors of tPA use after transportation. The MPSS is a novel prehospital stratification tool for the prediction of thrombolytic therapy after transportation.

[Condition of patients who require heart surgery during treatment for advanced digestive cancer and early recurrence after surgery- an assessment from the viewpoint of digestive surgeons].

The need for cardiac surgery among patients undergoing treatment for advanced digestive cancer is limited to the following situations:(i) heart diseases that can be life threatening if left untreated and that cannot be cured by medicinal treatment alone (e.g., cardiac tumors) and (ii) heart diseases (e.g., infectious endocarditis and pulmonary thromboembolism) occurring after digestive cancer surgery that need emergency treatment and that are resistant to medicinal treatment. We encountered 2 cases that required cardiac surgery.( Case 1) A 68-year-old woman with advanced gastric carcinoma accompanied by pyloric stenosis and left atrial myxoma underwent radical surgery for gastric cancer( Stage IIIA). Subsequently, the left atrial myxoma was resected before adjuvant chemotherapy for the treatment of gastric cancer was administered. One month after the surgery, multiple liver metastases appeared. However, they disappeared after chemotherapy was completed, and the patient survived for more than 3 years with complete response. (Case 2) A 67-year-old woman who underwent a Hartmann operation for obstructive rectal cancer (Stage II) experienced infectious endocarditis after the surgery. Because the endocarditis was resistant to medicinal treatment and acute heart failure was anticipated, cardiac surgery was performed. Approximately 2 months after the surgery, the bacilli( methicillin-resistant Staphylococcus aureus [MRSA]) were not found in blood culture. However, multiple liver metastases appeared immediately after the disappearance of the bacilli, and the patient died 3 months after the surgery. In both cases, cancer recurrence occurred early after cardiac surgery. Excessive surgical stress due to cardiac surgery may have promoted cancer recurrence. A decision pertaining to the timing of cardiac surgery is difficult in cases of patients with advanced digestive cancer and co-existing heart disease, which cannot be cured by medicinal treatment.

Coronary plaque regression and lifestyle modification in patients treated with pravastatin. - Assessment mainly by daily aerobic exercise and an increase in the serum level of high-density lipoprotein cholesterol -.

BACKGROUND: The purpose of this study was to explore the effect of lifestyle modification, mainly daily aerobic exercise, on coronary atherosclerosis in patients with coronary artery disease (CAD). METHODS AND RESULTS: A 6-month prospective observational study was conducted with 84 CAD patients receiving pravastatin treatment in order to evaluate the relationship between lifestyle modification, in particular aerobic exercise, and plaque volume as assessed by intravascular ultrasound (IVUS). Lifestyle during the study period was assessed by the-lifestyle modification score. A significant decrease in plaque volume by 12.9% was observed after 6 months of pravastatin therapy (P<0.0001 vs baseline). The change in plaque volume correlated with the change in the serum level of high-density lipoprotein cholesterol (HDL-C) (r=-0.549, P<0.0001), non-HDL-C (r=0.248, P=0.03), low-density lipoprotein cholesterol/HDL-C (r=0.505, P<0.0001), apolipoprotein (apo) A-1 (r=-0.335, P=0.007) and apoB/apoA-1 (r=0.335, P=0.007), and lifestyle modification score (r=-0.616, P<0.0001). There was a clear positive correlation between a change in the serum HDL-C level and lifestyle modification score. Multivariate regression analysis revealed that the increase in serum HDL-C level and lifestyle modification score were independent predictors of coronary plaque regression. CONCLUSIONS: An appropriate combination of statin therapy and lifestyle modification, in particular, physical activity, may result in coronary plaque regression. This combined treatment strategy, inducing an increase of the serum HDL-C, may contribute to coronary plaque regression.

Developing a uniformed assessment tool to evaluate care service needs for disabled persons in Japan.

Until recently, the care services for disabled persons have been under rigid control by public sectors in terms of provision and funding in Japan. A reform was introduced in 2003 that brought a rapid increase of utilization of services and serious shortage of financial resources. Under these circumstances, the "Services and Supports for Persons with Disabilities Act" was enacted in 2005, requiring that the care service provision process should be transparent, fair and standardized. The purpose of this study is to develop an objective tool for assessing the need for disability care. In the present study we evaluate 1423 cases of patients receiving care services in 60 municipalities, including all three categories of disabilities (physical, intellectual and mental). Using the data of the total 106 items, we conducted factor analysis and regression analysis to develop an assessment tool for people with disabilities. The data revealed that instrumental activities of daily living (IADL) played an essential role in assessing disability levels. We have developed the uniformed assessment tool that has been utilized to guide the types and quantity of care services throughout Japan.

Assessment of lower limb ischemia with measurement of skin perfusion pressure in patients on hemodialysis.

Measurement of skin perfusion pressure (SPP) using laser Doppler has become available for the assessment of peripheral arterial disease. We studied whether measurements of SPP can be used to identify hemodialyzed patients with peripheral arterial disease by comparing it with measurements of the ankle brachial pressure index (ABI). The ABI at rest and the SPP in the foot were measured in 59 Japanese hemodialyzed patients (118 limbs). Twenty-one patients had diabetes mellitus. Five had intermittent claudication; however, 20 patients were accompanied by other exertional leg symptoms. The SPP could not be measured in three limbs because of involuntary movement due to previous stroke or restless leg syndrome. The SPP was correlated with the ABI. Depending upon these results of the ABI, the 114 limbs from which both the ABI and the SPP could measured were divided into three groups: (A) ABI > or = 1.3, (B) 0.9 < or = ABI < 1.3, and (C) ABI < 0.9. The average SPP of group C was significantly decreased among the three groups. All subjects of the three groups were divided into an extra two groups according to the presence of diabetes (non-diabetes groups, -I; diabetes groups, -II). The average SPP of group B-II was significantly decreased compared with those of group B-I. The SPP measurement is a noninvasive, useful screening method for limb ischemia that can be applied to exercise tolerance limited patients. The SPP measurements are expected to be useful for the evaluation of limb ischemia in hemodialyzed patients at risk.

Measuring mouse sperm parameters using a particle counter and sperm quality analyzer: a simple and inexpensive method.

This study examined a method for analyzing the count, motility, and morphology of mouse epididymal sperm, optimizing the diluent, incubation time, sample concentration, and temperature, using a particle counter (CDA-500) to count and size sperm and a sperm quality analyzer (SQA-IIC) to measure sperm motility, quantified as the sperm motility index (SMI). The optimal conditions consisted of a 30-min incubation in D-MEM (Dulbecco's modified Eagle's medium; considering cost and availability) at 37 degrees C, with 5 x 10(6)cells mL(-1) in the original solution. Furthermore, the influence of formalin fixation, and the correlation between the automated counter and a manual method were investigated. The sample fixation had no marked effect on the sperm count or morphology assessment. A linear correlation was observed between the manual and automated methods (y=0.920x +0.276; r(2)=0.571; p<0.001; range: (3-6) x 10(6)). The suitability of the proposed method was confirmed using spermatozoa prepared from mice treated with the reproductive toxin diethylstilbestrol (DES). Using sperm from the cauda epididymidis on one side per mouse, we confirmed that measurement of these sperm parameters using the two devices was simple, rapid, inexpensive, and reproducible.

Application of the crypt isolation technique to the assessment of genetic alterations of colorectal carcinomas.

The crypt isolation technique (CIT) allows for the isolation of pure tumor crypts from colon tumor tissue. In a previous study we reported on the genetic alterations found in colorectal tumor crypts using the CIT; however, a direct comparison of the genetic alterations found in colorectal carcinomas using either conventional methods (CM) or the CIT has not previously been performed. Here, we analyzed the impact of this method on the genetic analysis of colon tumor cells by comparing the observed frequency of genetic alterations in colon tumors isolated using CM or the CIT. We used a combination of the CIT and the fluorescent polymerase chain reaction assay to accurately assess the incidence of allelic imbalances (AI) at a number of chromosomal loci (17p, 5q, 18q, 1p, 8p, 22q), microsatellite instability (MSI), and mutations of cancer-related genes (p53 and APC genes) in 48 sporadic colorectal carcinomas. In addition, genetic alterations seen in multiploid tumors (defined as tumors with both diploid and aneuploid cell populations) identified by the CIT were also examined. The incidence of AI at the chromosomal loci tested was more frequently detected in samples isolated from tumors using the CIT than in those isolated from the same tumors using CM. In contrast, we observed no differences in the frequency of MSI or cancer-related gene mutation between the two groups. Although there was no difference in the frequency of genetic alterations between tumors with evidence of multiploidy, sorting of diploid and aneuploid populations allowed detection of distinct genetic changes. The crypt isolation method thus appears to be useful in that it allows purification of tumor cells and the accurate assessment of their genetic alterations. In addition, it may also be of benefit in clarifying the genetic profile of multiploid tumor cell populations.