Construction of an Automatic Quantification Method for Bone Marrow Cellularity Using Image Analysis Software.

Although rapid, the evaluation of bone marrow (BM) cellularity is semi-quantitative and largely dependent upon visual estimates. We aimed to construct an automatic quantification method using image analysis software. We used hematoxylin and eosin (HE)-stained specimens of BM biopsies and clots from patients who underwent BM examination at Tottori University Hospital from 2020 to 2022. We compared image analysis (Methods A, B, and C) with visual estimates in pathology reports of 91 HE specimens in 54 cases (29 males, 25 females), including 38 biopsy and 53 clot specimens. Cellularity was visually scored as hypocellular (n = 17), normocellular (n = 44), or hypercellular (n = 30). Compared with the visual estimates, intraclass correlation coefficients for Methods A, B, and C were 0.80, 0.85, and 0.88, respectively. The most appropriate values were obtained with Method C which detected both non-fatty and cell nuclear areas.

Fibrin monomer complex as a potential thrombosis marker related to venous thromboembolism risk in pregnant women.

BACKGROUND: Pregnancy is a risk factor for venous thromboembolism (VTE) due to increased coagulation factor activity and decreased protein S activity. However, thrombosis markers for predicting VTE in pregnancy remain controversial. This study aimed to investigate the relationship between VTE risk and thrombosis markers in pregnant women and to identify markers related to VTE risk. METHODS: Archived plasma samples from 107 pregnant women were used in this study, and the concentrations of D-dimer, fibrin monomer complex (FMC), plasmin-plasmin inhibitor complex, prothrombin time, activated partial thromboplastin time, and fibrinogen were measured. VTE risk was scored according to the Royal College of Obstetricians and Gynaecologists green-top guidelines and the patients were divided into low- or high-risk groups. RESULTS: The median (range) of risk score for deep vein thrombosis was 2 (0-8), and we defined the high-risk group included those with a score of ≧3. D-dimer and FMC concentrations were significantly higher in the high-risk group than in the low-risk group (D-dimer 4.5 vs 2.6 µg/mL, p = 0.008; FMC 14.6 vs 3.4 µg/mL, p < 0.001). Although D-dimer concentration significantly increased with gestational age (Spearman's correlation coefficient [rs] = 0.317, p < 0.001), FMC concentration did not (rs = -0.081, p = 0.409). The area under the receiver operating characteristic curve values of D-dimer, FMC, and both D-dimer and FMC for the high-risk group were 0.656, 0.713, and 0.738, respectively. CONCLUSIONS: FMC may be a thrombosis marker related to VTE risk in pregnancy and is potentially preferable over D-dimer concentrations.

Regression Analysis to Estimate the Factor VIII Activity of Patients with Hemophilia A Without Inhibitor who Received Emicizumab Therapy.

BACKGROUND: Emicizumab, a bispecific monoclonal antibody for hemophilia A (HA), has strong pharmacodynamic effects in several coagulation assays resulting in dosing difficulties with Factor VIII (FVIII) concentrates during bleeding emergencies. MATERIALS AND METHODS: Single and multiple regression models were studied to estimate FVIII activity using 27 archived plasma samples from three patients with HA without inhibitor under emicizumab treatment. Explanatory variables were FVIII chromogenic assay (CSA), Ad|min1|, Ad|min2|, the number of seconds of APTT, and the FVIII one-stage assay (OSA), which were measured without idiotype antibodies. The response variable was FVIII OSA measured with idiotype antibodies. RESULTS: In the simple linear model, the FVIII CSA regression coefficient was 1.04 and the intercept was -14.55 (r2 = 0.95; p < 0.001). In the multiple regression model, FVIII OSA and FVIII CSA were selected based on the Akaike Information Criterion, with regression coefficients of 1.74 and 1.15, respectively, and an intercept of -92.03 (r2 = 0.96, p < 0.001). CONCLUSIONS: The regression models can estimate the FVIII:C levels in patients with HA receiving emicizumab and would be useful in a bleeding emergency and/or surgery.

Analysis of family history of palmoplantar hyperhidrosis in Japan.

Palmoplantar hyperhidrosis (PPH) is a disorder characterized by excessive sweating of the palmar surfaces of the hands and feet due to emotional sweating. There have been reports based on family histories, and the involvement of genetic factors has been suggested. Among 410 PPH patients who visited our hospital from August 2006 to October 2008, onset age and family history were investigated in order to prepare pedigree charts, and family histories were confirmed in 147 patients (36%). Between the family history and negative family history (sporadic) groups, no significant differences were seen in onset age, sex or sweat volume. With regard to the patterns of incidence within families, parent-child was the most common at 58%, followed by sibling cases at 18%. The incidence of PPH in three generations was 13%. Pedigree charts prepared based on data obtained by patient interviews suggested autosomal dominant inheritance.

Reduction in QSART and vasoactive intestinal polypeptide expression in the skin of Parkinson's disease patients and its relation to dyshidrosis.

BACKGROUND: With regards to dyshidrosis in Parkinson's disease (PD), there is no established and consistent view on the occurrence sites, frequency and etiology, although there have been several reports on hypohidrosis of the limbs and sudoresis on the face/cervical region. METHODS: Hydrosis in the forearms of PD patients and healthy individuals were compared by quantitative sudomotor axon reflex test (QSART). The expression of various neuropeptides and alpha-synuclein was examined with immunohistochemical staining. RESULTS: There was a significant reduction in QSART of PD patients but not of healthy controls. Reduced expression of vasoactive intestinal polypeptide (VIP) was also detected in the sweat glands of PD patients. CONCLUSION: Reduction in QSART and VIP expression in the sweat glands might be involved in the dyshidrosis of PD patients.

Generational differences in factors influencing job turnover among Japanese nurses: an exploratory comparative design.

BACKGROUND: Although nurse turnover is a serious problem, the fact that each nurse has different work-related needs/values, and leaves their job for different reasons makes it difficult for organisations to develop effective countermeasures against it. Understanding nurses' needs and the reasons for job turnover by the generation in which they were born may provide some feasible solutions. OBJECTIVES: The purpose of the study was to identify specific work-related needs and values of nurses in three generations (born in 1946-1959, 1960-1974, 1975-present). The study also aimed to explore generation-specific reasons that might make nurses consider leaving the jobs. SETTINGS: The study was conducted in three public hospitals in Japan. PARTICIPANTS: A convenience sample of 315 registered nurses participated in the study. METHODS: A survey method was used to collect quantitative and qualitative data. Quantitative data were analysed by ANOVA, and qualitative data were analysed by content analysis. RESULTS: Nurses born between 1960 and 1974 embraced high needs and values in professional privileges such as autonomy and recognition, while those born after 1975 expressed low needs and values in the opportunities for clinical challenge. For nurses born between 1960 and 1974, the imbalance between their jobs and personal life made them consider leaving their jobs. For those born after 1975, losing the confidence to care made them consider turning over. Nurses born after 1960 tended to value economic return and job security more highly compared to those born between 1946 and 1959. CONCLUSIONS: Nurses in different generations have different sets of needs/values and reasons for job turnover. Understanding generation-specific needs and values of nurses may enable organisations and Nurse Managers to develop feasible and effective countermeasures to reduce nurse turnover.

Local injection of botulinum toxin A for palmar hyperhidrosis: usefulness and efficacy in relation to severity.

Botulinum toxin A is widely used in Europe and the USA for the treatment of localized hyperhidrosis, and its efficacy has been recognized. In this study, botulinum toxin A (Botox) was locally injected at 30 sites (2 U/injection) on the right palm in 27 patients with palmar hyperhidrosis (14 severe patients, 13 mild patients), and the results confirmed the efficacy of injection. The amount of sweat was then quantified for the left and right hands every month after local injection. The quantity of sweat on the treated hand was approximately one-fifth that on the untreated hand. In addition, the quantity of sweat on the untreated hand decreased slightly. Over time, the quantity of sweat on the treated hand increased slightly, but the quantity of sweat on the treated hand at 6 months after injection was less than half that before injection, and there were significant differences before and after injection. In the present study, severe sweating was defined as 1 mg/cm2/min or more and mild sweating as less than 1 mg/cm2/min, and the therapeutic effects of botulinum toxin A were analyzed in relation to severity. When compared to the mild cases, the quantity of sweat remained higher in the severe cases after botulinum toxin A therapy. Therefore, to achieve satisfactory effects in severe cases, it would be necessary to increase the number of injection sites, as well as injection dose.

Thioredoxin upregulation by 5-aminolaevulinic acid-based photodynamic therapy in human skin squamous cell carcinoma cell line.

BACKGROUND/PURPOSE: 5-aminolaevulinic acid-based photodynamic therapy (ALA-PDT) is widely performed in the clinical setting for superficial skin cancers, giving favorable results, but residual tumor and recurrence occur occasionally. Thioredoxin is a common antioxidant that suppresses apoptosis and facilitates cell growth. We investigated the expression of thioredoxin following ALA-PDT in human skin squamous cell carcinoma cell line, HSC-5. METHODS: ALA-PDT was performed in HSC-5 cells using low-dose (5 J/cm(2), 100 mW/cm(2)) or high-dose (30 J/cm(2), 100 mW/cm(2)) irradiation, and the expression of thioredoxin was measured by Western blotting. An MTT assay was used to assess cell growth following a low dose of multiple irradiations. Cell death was examined by Western blotting for caspase-3 and PARP. Immunofluorescence double staining using annexin V and propidium iodine was also performed. RESULTS: Expression of thioredoxin was only observed following low-dose exposure ALA-PDT. Multiple low-dose exposure ALA-PDT significantly proliferated cell growth. With high-dose exposure ALA-PDT, caspase-3 and PARP expression were seen, and cell death due to apoptosis and/or necrosis was observed, but thioredoxin was barely detected. CONCLUSION: Low-dose exposure ALA-PDT increased the expression of thioredoxin and facilitated the growth of HSC-5 cells.

Nurses' leaving intentions: antecedents and mediating factors.

AIM: This paper is a report of a study to investigate how nurses' work values, perceptions of environmental characteristics, and organizational commitment are related to their leaving intentions. BACKGROUND: Nurse leaving is a serious international problem as it contributes to the nursing shortage that threatens the welfare of society. The characteristics of nurses, the work environment and nurses' feelings towards their jobs (or organizations) have an impact on their leaving intentions. METHOD: A convenience sample of 849 Registered Nurses was recruited from three public hospitals in the central-west region of Japan during 2006 and 319 completed questionnaires were returned (response rate 39%). Data were analysed using regression analysis. FINDINGS: Nurses' work values and their perceptions of their workplace environment interacted to influence leaving intentions. When there was a match between the importance nurses placed on being able to challenge current clinical practices and the number of the actual opportunities to do so, leaving intentions were low. When there was a mismatch, intention to quit the job became stronger. In addition, organizational commitment intervened between nurses' perceptions of the match in clinical challenges and leaving intention. CONCLUSION: Nurses' leaving intentions, deserve extensive exploration of their causes. Such exploration should include attending to both nurses' needs and organizational characteristics, investigating how the match between them could affect nurses' leaving intention, and exploring factors that intervene between nurses' perceptions of the match and leaving intention.

Rhombomere formation and hind-brain crest cell migration from prorhombomeric origins in mouse embryos.

Prior to rhombomere development, structures called prorhombomeres appear in the mammalian hindbrain. This study clarifies the developmental relationship between prorhombomeres and their descendent rhombomeres and hindbrain crest cells in mouse embryos by focal dye injections at various levels of prorhombomere A (proRhA), proRhB, and proRhC, as well as at their boundaries. ProRhA gives rise to two rhombomeres, rhombomeres 1 and 2 (r1 and r2), as well as to crest cells that migrate into the first pharyngeal arch, including the trigeminal ganglion. ProRhB develops into r3 and r4 and produces crest cells populating the second arch and acousticofacial ganglion. The anterior portion of proRhC gives rise to r5 and r6 and to crest cells migrating into the third pharyngeal arch and the IXth ganglion; its posterior portion develops into r7 and releases crest cells into the fourth pharyngeal arch region as well as the Xth ganglion. These results suggest that the boundaries between prorhombomeres serve as lineage restrictions for both hind-brain neuroepithelial cells and for segmental origins of crest cell populations in mouse embryos. The Hox code of the mouse head can be schematized in a much simpler way based on this prorhombomeric organization of the hind-brain, suggesting that prorhombomeres primarily underlie mammalian hind-brain segmentation.

Whole embryo culture and the study of postimplantation mammalian development.

In the past decade, striking advances have been made in the field of gene introducing/disrupting technology including generation of transgenic and knockout mice, which have enabled us to elucidate roles of specific genes in development. In this technology, embryos introduced with exogenous genes or chimeric embryos aggregated/injected with embryonic stem (ES) cells carrying targeted genes are allowed to develop in the uterus of foster mothers. The uterus, however, is like a black box for researchers investigating postimplantation development of mammalian embryos. Embryo culture is one of the powerful techniques that can open this black box. In this review, we focus on the applicable aspects of the whole embryo culture in the study of mammalian development and discuss the future possibilities of this technique.

Retinoic Acid Treatment Induces Cell Death and the Protein Expression of Retinoic Acid Receptor ß in the Mesenchymal Cells of Mouse Facial Primordia in Vitro: (RA/facial mesenchymal cells/chondrogenesis/cell death/RAR b and g protein expression/ micromass culture).

We isolated mesenchymal cells from individual facial primordia of mouse embryos on 11 days post coitum and examined the effects of retinoic acid (RA) on chondrogenesis, induction of cell death, and the protein expression of retinoic acid receptor (RAR) ß and γ in micromass culture. Under the control condition, cells of both medial and lateral nasal prominences (MNP and LNP) displayed high chondrogenic potential, while those of maxillary and mandibular prominences (Mx and Md) had constant growth activity and low chondrogenic potential. Though none of the cells expressed detectable levels of the RAR ß protein, RAR γ was expressed in the cells of all the facial primordia. One µM RA inhibited the chondrogenesis, and induced cell death accompanied with the induction of the RAR ß protein in LNP, MX and Md cells within 6 hr. On the contrary, both cell death and RAR ß protein induction were detected in the MNP cells treated with RA for 24 hr. These results suggest that the RAR ß is involved in the process of the cell death induced by the RA treatment in the mesenchymal cells of the mouse facial primordia.

Retinoic Acid Treatment Induces the Ectopic Exporession of Retinoic Acid Receptor ß Gene and Excessive Cell Death in the Embryonic Mouse Face.

Maternal treatment with 100 mg/kg of retinoic acid (RA) on day 9 of gestation in mice caused craniofacial abnormalities of the mandibulofacial dysostosis type. The abnormal morphology was attributed to the excessive cell death in the dorsal aspects of the maxillary and mandibular prominences of the first pharyngeal arch and in the proximal region of the mandibular prominence. To investigate the expression of the RA receptor (RAR) genes in abnormal face morphogenesis, in situ hybridization was performed. The distribution patterns of RAR α and γ transcripts were not altered in the treated embryos. By contrast, the teratogenic dose of RA increased the level of RAR ß transcripts, as early as 3 hr after RA-treatment, in the regions where the RAR ß expression is at a low level in normal development. The increase of RAR ß transcripts was detected by 12 hr, and declined to the low level within 24 hr after the treatment. The regions where ectopic expression of RAR ß gene was observed included the areas where the excessive cell death occurred 9-12 hr after RA-treatment. These results suggest that ectopic induction of RAR ß by RA may lead to the excessive cell death, threfore may cause abnormal morphogenesis.