Lower peripheral circulation in eumenorrheic young women with premenstrual symptoms.

BACKGROUND: A majority of women from all cultures and socioeconomic levels experience diverse psychosomatic and behavioral symptoms premenstrually, a phenomenon commonly termed premenstrual syndrome, although symptoms and discomfort levels vary from woman to woman. The underlying pathological mechanisms of premenstrual syndrome remain unknown; however, altered function or even slight disorder of the blood circulation system, which contributes to the orchestrations of the human internal environment, could cause bio-psychological changes leading to complaints and ultimately compromising a woman's overall health. The present study, therefore, investigates to what extent and how the menstrual cyclicity of peripheral circulation is associated with premenstrual symptomatology. METHODS: Twenty-one eumenorrheic young women participated in this study. All subjects were investigated during the follicular and late luteal phases. Cycle phase was determined by the onset of menstruation and oral temperature and was verified by concentrations of ovarian hormones, estrone, and pregnanediol in a urine sample taken early in the morning. Peripheral circulation was evaluated with the Astrim (Sysmex, Kobe), a portable non-invasive monitoring device using the principle of near-infrared spectroscopy, which calculates the venous oxygenation index (VOI) based on the ratio of light absorption of oxyhemoglobin and deoxyhemoglobin, a proven reliable indicator of peripheral blood circulation. The Menstrual Distress Questionnaire was applied to measure physical, emotional, and behavioral symptoms accompanying the menstrual cycle of the subjects. RESULTS: The oral temperature and urinary ovarian hormones adjusted for creatinine significantly increased in the late luteal phase in all subjects. While 10 subjects experienced no symptoms during the menstrual cycle, 11 subjects had apparent physical and psychological discomfort in the late luteal phase. We found that VOI decreased more significantly in the late luteal phase than in the follicular phase only in women with premenstrual discomfort although the symptoms were not unbearable enough to cause the disruption of daily activities. CONCLUSION: Several models have tried to explain the etiopathogenesis of premenstrual syndrome. Although causes and consequences remain enigmatic, our data suggest that the peripheral circulation could alter in the luteal phase, which might be partly associated with premenstrual psychosomatic symptoms in eumenorrheic young women.

Detection of Coxiella burnetii in market chicken eggs and mayonnaise.

We tried to detect C. burnetii in market chicken eggs and mayonnaise by nested PCR assay. The PCR target was the com 1 gene of C. burnetii. The positive rate for egg and mayonnaise samples was 4.2% and 17.6%, respectively. Direct sequence of some of the positive egg samples shows mutations whereas no mutation was found in the positive mayonnaise samples. The number of molecules of the Q fever agent is estimated at 10(4) to 10(6) per egg, according to our quantitative PCR test.

Autonomic nervous system activity in the late luteal phase of eumenorrheic women with premenstrual symptomatology.

The majority of women of reproductive age experience a regular recurrence of various symptoms in the premenstrual phase. The etiopathogenesis of premenstrual symptomatology, however, remains inconclusive. The present study was proposed to evaluate whether the activity of the autonomic nervous system (ANS), which largely contributes to the relative stability of a human's internal environment, is altered during the menstrual cycle of women with premenstrual symptomatology. Thirty eumenorrheic young women participated in this study. All subjects were investigated during the follicular and late luteal phases. The ANS activity was assessed by means of heart rate variability power spectral analysis during supine rest. No intramenstrual cycle differences in the ANS activity were found in women experiencing no or small increases in premenstrual symptoms. In contrast, the sympathetic nervous system (SNS) activity significantly increased and the parasympathetic nervous system (PNS) activity apparently decreased in the late luteal phase in subjects whose premenstrual symptomatology was not unbearable, but substantially increased (> 20%) compared to the symptom-free follicular phase. The women with greater degrees of premenstrual distress possessed higher SNS activity and lower PNS activity in the late luteal phase than the women with less symptomatology. The ANS activity in the follicular phase did not differ among the subjects regardless of their premenstrual symptoms. Although causes and consequences continue to elude, the present study provides additional intriguing evidence that the altered functioning of ANS in the late luteal phase could be associated with diverse psychosomatic or behavioral symptoms appearing premenstrually.

Validation of platelet counting accuracy with the celltac f automated hematology analyzer.

Rapid and accurate analysis of platelet count plays an important role in evaluating hemorrhagic status. Therefore, we evaluated platelet counting performance of a hematology analyzer, Celltac F (MEK-8222, Nihon Kohden Corporation, Tokyo, Japan), that features easy use with low reagent consumption and high throughput while occupying minimal space in the clinical laboratory. All blood samples were anticoagulated with dipotassium ethylenediaminetetraacetic acid (EDTA-2K). The samples were stored at room temperature (18(;)C-22(;)C) and tested within 4 hours of phlebotomy. We evaluated the counting ability of the Celltac F hematology analyzer by comparing it with the platelet counts obtained by the flow cytometry method that ISLH and ICSH recommended, and also the manual visual method by Unopette (Becton Dickinson Vacutainer Systems). The ICSH/ISLH reference method is based on the fact that platelets can be stained with monoclonal antibodies to CD41 and/or CD61. The dilution ratio was optimized after the precision, coincidence events, and debris counts were confirmed by the reference method. Good correlation of platelet count between the Celltac F and the ICSH/ISLH reference method (r = 0.99, and the manual visual method (r= 0.93) were obtained. The regressions were y = 0.90 x+9.0 and y=1.11x+8.4, respectively. We conclude that the Celltac F hematology analyzer for platelet counting was well suited to the ICSH/ISLH reference method for rapidness and reliability.

Performance evaluation of the complete blood count and white blood cell differential parameters obtained using a Celltac F automated hematology analyzer.

Performance of a new hematology analyzer, the Celltac F, MEK-8222 (Nihon Kohden, Tokyo, Japan), was evaluated. This analyzer simultaneously measures 22 parameters including leukocyte differentials. Within- and between-batch precision, linearity, and absence of carryover were all excellent. Accurate measurements of complete blood count parameters were possible during the 48-hour test periods for stability at 4 degrees C and room temperature. Automated differential parameters showed acceptable stability during the 24-hour test period. The performance was evaluated in comparison with the CD-4000 (Abbott, Abbott Park, IL, USA). There was a good correlation between findings with the Celltac F and those with the CD-4000 hematology analyzer for complete blood count parameters and leukocyte differential parameters other than percentages of basophils. Comparisons of differential leukocyte counts to microscopic differentials were excellent for neutrophils, lymphocytes, and eosinophils and were acceptable for monocytes. The clinical utility and flagging performance were excellent. The Celltac F is compact and it is able to process 80 samples per hour. The volumes of sample and reagent consumed were slight. The instrument has good potential to contribute to effective total cost management and to be a useful backup system with high throughput, while taking up minimal space in the clinical laboratory.

A new method of purifying fibrinogen with both biological and immunological activity from human plasma.

Using a combination of Cohn ethanol fractionation, virus inactivation, glycine and sodium chloride precipitation, and lysine-Sepharose affinity chromatography, a unique and rapid simplified method was developed to obtain highly purified fibrinogen for diagnostic use with both biological (Clauss method) and immunological (Jacobsson method) activity. Yield was 0.66 g of fibrinogen per liter of starting pooled plasma, and the purified product showed good agreement in activity with the starting material. The purified fibrinogen solution contained over 95% clottable protein and had a clear appearance. No degradation was observed after urokinase treatment and the preparation provided good precision in fibrinogen measurement compared to pooled plasma. The simplified method was, thus, shown to result in a high-purity fibrinogen preparation, suitable for in vitro diagnostic use, as well as for use to prepare a fibrinogen reference material and to perform fibrinogen quality control using an automated coagulation analyzer.

[Past, present and future of external quality assessment of medical laboratory testing in Japan].

In Japan, three major bodies have conducted external quality surveillance of medical laboratory performance: the Japan Medical Association, the Japan Association of Medical Technologists, and the Japan Society of Registered Medical Laboratories. Although these systems are working very well to elevate the quality of laboratory tests, each has adopted its own surveillance system with little communication. All of these surveillance systems have problems in the evaluation of laboratory proficiency. Thus, the 3 bodies should be unified first to establish an efficient National External Quality Assurance Scheme. New IT technology should also be introduced into the system in the future.

[Education of medical technologists in North America].

The increasing complexity of diseases and advancing medical technologies have recently raised the number of test items and their use. This has caused each department to ask for clinical support by medical technologists, including consultation services on clinical laboratory tests in Japan. Under these circumstances, we think it is necessary to consider a Japanese education system for medical technologists to foster people with advanced ability capable in providing adequate clinical support. It is important that we study medical technologist systems and education systems superior to than ours. Therefore, to investigate the state of the Japanese education system for medical technologists, we conducted a comparison between the medical technologist systems and education systems in foreign countries and those of Japan. The social background in which medical technologists are positioned, their scope of work, and the education systems overseas are different from Japan on many points and we were given many suggestions for what a system in Japan should be like.

Expression of the inhibitor of apoptosis (IAP) family members in human neutrophils: up-regulation of cIAP2 by granulocyte colony-stimulating factor and overexpression of cIAP2 in chronic neutrophilic leukemia.

Human neutrophils were found to express members of the inhibitor of apoptosis (IAP) family, namely cellular IAP1 (cIAP1), cIAP2, and X-linked IAP. Among these members, cIAP2 expression was selectively up-regulated by stimulation with granulocyte colony-stimulating factor (G-CSF), but not with granulocyte-macrophage CSF. The increased expression of cIAP2 mRNA was detected as early as 30 minutes after in vitro stimulation with G-CSF, and the elevated level of cIAP2 protein was detected at 1 hour. The elevated level of cIAP2 protein was also detected in peripheral blood neutrophils obtained from healthy donors receiving G-CSF administration. G-CSF-induced up-regulation of cIAP2 mRNA and protein, phosphorylation of signal transducer and activator of transcription 3 (STAT3), and the antiapoptotic effects were inhibited by pretreatment of cells with AG490, a specific inhibitor of Janus kinase 2 (JAK2). Mature neutrophils from a patient with chronic neutrophilic leukemia exhibited remarkable overexpression of cIAP2 mRNA and prolongation of survival, whereas cIAP2 mRNA expression and survival in mature neutrophils from patients with chronic myelogenous leukemia were essentially similar to those in normal neutrophils. These findings suggest that cIAP2 expression is up-regulated by G-CSF through activation of the JAK2-STAT3 pathway, and increased expression of cIAP2 protein may contribute to G-CSF-mediated antiapoptosis. In addition, overexpression of cIAP2 may be partly responsible for sustained neutrophilia at least in some cases of chronic neutrophilic leukemia.

Proposed fibrinogen standard material with purified fibrinogen for plasma fibrinogen measurement on coagulation analyser.

A commercially available fibrinogen standard calibrated by a World Health Organization (WHO) reference material is widely used in Japan, and most clinical laboratories use the Clauss method for plasma fibrinogen measurement. However, a current issue in fibrinogen measurement is poor laboratory-to-laboratory variability. To improve the reliability of fibrinogen values and thereby solve the poor precision and accuracy of plasma fibrinogen testing, the present paper develops a simple and large preparation procedure for a suitable fibrinogen standard and quality control material and evaluates their basic performance. With a new procedure getting high purified fibrinogen by glycine precipitation, the calibrator determined by both the Clauss and Jacobson methods produced a fibrinogen concentration of 2.20 g l(-1). The total precision of the calibrator was excellent (coefficient of variation 1.4-2.1%) in comparison with current plasma fibrinogen materials from the WHO (#98/612) and with a commercial standard (CV 1.9-3.9%). The within-run precision of the calibrator on the coagulation analysers was 1.7-2.8%. Within-analyser variability among the five instruments had good consistency (mean 2.20 +/- 0.022 g l(-1); CV 1.0%). The degradation study of the calibrator suggested that storage at 9 degrees C for two years was as predicted. In conclusion, the results show that the calibrator prepared herein can be useful as a candidate Japanese fibrinogen standard and is applicable to automated and semi-automated coagulation analysers. Additionally, it is expected that it will be widely used in Japan by diagnostic manufacturers and clinical laboratories as a recommended secondary standard to estimate a fibrinogen value according to the WHO primary standard.

Quality control material for plasma fibrinogen test produced from purified human fibrinogen.

Plasma fibrinogen measurement is a routine laboratory procedure commonly performed on automated coagulation analysers. Its determination is quantitative, not quantitative. Yet, a lack of precision has been an issue for fibrinogen measurement. A control material derived from plasma comprises many proteins, inhibitors and fatty acids, any or all of which can interfere in the fibrinogen assay. This study has attempted to develop a quality control material using purified human fibrinogen and has compared measurement precision between both purified and plasma materials. Purified fibrinogen was prepared using Cohn fraction 1 and glycine precipitation. Purified fibrinogen clottability was greater than 95%, with no main plasma proteins, lipids or fibrinogen degradation products observed. Two purified control materials were lyophilized at normal (2.30 g l(-1)) and abnormal (1.20 g l(-1)) levels of fibrinogen concentration. Precision was evaluated using a liquid-type reagent, Thrombocheck Fib(L), on automated coagulation analysers. Coefficient of variation for within-run, intraday and between-day precision of the purified materials was 0.7-3.5%. In comparison, the coefficient of variation for plasma materials ranged from 1.2 to 5.3%. These results suggest that materials prepared from purified fibrinogen can be useful to laboratory quality control by improving overall precision of fibrinogen measurement and are applicable to automated coagulation analysers.

[Chronic myelogenous leukemia treated with non-myeloablative stem cell transplantation after discontinuing myeloablative stem cell transplantation due to mental aberrations].

A 54-year-old woman with chronic myelogeneous leukemia was admitted to our hospital on February 15th, 2001 to undergo allogeneic bone marrow transplantation (BMT). We started the transplantation preconditioning with busulfan and high-dose cyclophosphamide on February 22nd, 2001. However, symptoms of a psychiatric nature, such as hallucination, persecution complex, auditory hallucination and sleeplessness, occurred by the third day of treatment with busulfan. Thus, we decided to discontinue conditioning and stopped the administration of BMT at that point. However, pancytopenia persisted for more than 20 days. She finally underwent BMT followed by reduced-intensity conditioning with fludarabine and ATG from a sex-mismatched, HLA-identical sibling donor on April 19th, 2001. To prevent any exacerbation of the psychotic symptoms, the patient was hospitalized in a laminar flow instead of a bio-free room. Graft-versus-host disease occurred on the 32nd hospital day, and was brought under control by steroid treatment. Achievement of complete chimeras was confirmed on the 54th hospital day. Her mental condition was kept stable with antidepressant drugs and tranquilizers, although minor changes in the combination of drugs were required to treat transient exacerbation of psychosis after a short period at home. She was discharged on September 1st, 2001. We think that non-myeloablative stem cell transplantation is a useful treatment for patients with hematological malignancy complicated with psychiatric disorders.

[A case of amelanotic melanoma of the nasal cavity].

A 68-year-old male visited Hospital A for treatment of epistaxis, his chief complaint. He was told that he had an easily-bleeding tumor in the nasal cavity. Based of biopsy, a diagnosis of amelanotic melanoma was made. Operation was performed for removal of the tumor. About 8 months after discharge, he visited Hospital B with complaints of lumbar pain and epistaxis. After biopsy at Hospital B, malignant lymphoma (diffuse large cell) was diagnosed, and the patient was referred to our hospital. On bone marrow puncture and biopsy, tumor cell infiltration was observed. Flow cytometric surface marker analysis revealed that these tumor cells were negative for CD45. Results of HE staining of the nasal cavity tumor were insufficient for diagnosis, and staining by immunohistochemistry was necessary to confirm the diagnosis. On immunohistochemical staining of the nasal cavity tumor tissue and bone marrow biopsy tissue, LCA, L26 and UCHL-1 were negative, and S-100 and HMB-45 positive. Recurrence of amelanotic melanoma accompanied by bone marrow infiltration was therefore diagnosed. The incidence of amelanotic melanoma with primary lesions in the nasal cavity is low. However, in making the diagnosis of a nasal cavity lesion, the possibility of such a melanoma should be kept in mind. In many cases, it is difficult to diagnose amelanotic melanoma with HE staining alone, and immunohistochemistry must be used.

Changes in hypervariable region 1 in patients with chronic hepatitis C of genotype 1b with biochemical response to interferon.

BACKGROUND/AIMS: The outcome of interferon (IFN) therapy of hepatitis C virus (HCV) infection can be classified as a complete viral response (CR), biochemical response (BR), or no response (NR). Why alanine aminotransferase (ALT) activity decreases in patients with BR despite viral persistence is unknown. METHODS: Of 158 patients infected with HCV genotype 1b, all 20 patients with BR and 20 of the 114 patients with NR, matched for viral load to the BR group, were studied. We sequenced nucleotides in the hypervariable region (HVR) of serum HCV RNA, and analyzed quasispecies of this region by the polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP). RESULTS: In HVR 1, SSCP patterns differed after therapy; the major clone before therapy disappeared with therapy in eight of the 20 BR patients, but in none of the 20 NR patients (P=0.0033; Fisher's exact test). PCR products of HVR 1 from six patients were cloned before and after therapy, and 40 clones from each patient were sequenced each time. Results of cloning and sequencing were generally consistent with those of SSCP. For the six patients, a major clone could be identified both before and after therapy. In two patients with BR, there were many changes in the amino acid sequence of the major clone after IFN; in one patient with NR, mutations were not found. CONCLUSION: Changes in the major viral clone with IFN treatment may be related to the decrease in ALT activity in some patients, in spite of the continued presence of HCV RNA.

Differential effects of fibroblast growth factor-4, epidermal growth factor and transforming growth factor-beta1 on functional development of stromal layers in acute myeloid leukemia.

The hematopoietic supporting abilities are known to be impaired in marrow stromal layers developed from patients with acute myeloid leukemia (AML). In this study, fibroblast growth factor-4 (FGF-4), epidermal growth factor (EGF) or transforming growth factor-beta1 (TGF-beta1) were studied to see whether these growth factors can modify the functional development of leukemic stromal layers. Adherent stromal layers from 13 patients with AML and from six non-leukemic controls were established with 3ng/ml of FGF-4, EGF or TGF-beta1. Established stromal layers were washed three times and irradiated, followed by recharge of allogenic peripheral CD34 positive cells as an indicator of supportive function. Progenitor-outputs into supernatant were evaluated at biweekly interval with colony-forming assay until 6 weeks. The results showed that both leukemic and non-leukemic stromal cells established with FGF-4, but not with EGF, showed significantly higher progenitor cell-outputs compared with control stromal cells. By contrast, stromal cells developed with TGF-beta1 showed significantly lower progenitor cell-outputs compared with control. These differences were significant at later than 4 weeks after the recharge of indicator cells, suggesting that the stromal layer developed with EGF or TGF-beta1 preferentially affected the primitive progenitors rather than committed ones. These results indicate that FGF-4 and TGF-beta1 differentially affect the functional development of leukemic as well as of normal stromal layers.