Diagnostic Agreement for High-Grade Urothelial Cell Carcinoma in Atypical Urine Cytology: A Nationwide Survey Reveals a Tendency for Overestimation in Specimens with an N/C Ratio Approaching 0.5.

In the Paris System (TPS), standardized cytomorphological criteria and diagnostic categories were proposed for reporting urine cytology. To evaluate the diagnostic agreement and interobserver concordance for assessing TPS criteria, the Taiwan Society of Clinical Cytology organized an online survey with 10 atypical urine cytology cases. A total of 137 participants completed the survey. The mean agreement of diagnosis was 51.2%, ranging from 34.3% to 83.2% for each case. For 60% (6/10) of cases, the agreement was <50%. The interobserver concordance of diagnosis and cytological criteria assessment showed poor agreement. The nuclear-to-cytoplasmic (N/C) ratio had the highest kappa value of 0.386, indicating a significantly higher interobserver concordance and reproducibility than the other three TPS criteria. The correct rate of assessing the N/C ratio increased as the N/C ratio increased (correlation coefficient: 0.891, p < 0.01). Three cases with an N/C ratio near 0.5 were overestimated. Poor interobserver concordance of diagnosis and TPS criteria was revealed. Compared with other cytological features, the N/C ratio assessment was quantitative and more reproducible, but a tendency to overestimate cells was noted when the N/C ratio was approximately 0.5. Continuing education programs should emphasize the accurate assessment of N/C ratio to improve the application of TPS.

Transferred-tissue Microarray for Fluorescence In Situ Hybridization Test for Human Epidermal Growth Factor Receptor 2 in Breast Cancer.

CONTEXT: Human epidermal growth factor receptor 2 (HER2) status of breast carcinomas is usually determined by immunohistochemical (IHC) staining and, if the IHC results are equivocal, in situ hybridization (ISH). Multiple ISH tests are sometimes required for multiple primary or metastatic tumors. A method for multiplex ISH test on tissues from multiple blocks is helpful in these situations. OBJECT: To evaluate the clinical application of transferred-tissue microarray (TTM) followed by a dual-probe HER2 fluorescence in situ hybridization (FISH). DESIGN: A 3×3 TTM technique was successfully established using 152 invasive mammary carcinoma tissue fragments. To evaluate detection of HER2 positive tumors, this cohort was enriched with tumors with IHC scores of 2 and 3. RESULTS: The HER2 FISH analyses revealed that all transferred-tissue fragments were adequate for determining HER2 amplification. Tissue loss was minimal and had no major adverse effects on interpretation of the test results. Of the 81 tumors with IHC scores of 3, 72 (88.8%) were positive for HER2 FISH. The remaining tumors were negative for HER2 FISH in both TTM and reflex whole tissue section. Finally, FISH results for tumors with IHC scores of 2 were compared between TTM and whole tissue section. Concordance was high in overall positivity/negativity (100%), HER2 copy number (97.5%), and HER2/CEP17 ratio (100%). CONCLUSIONS: This novel technique is a reliable option for performing multiple HER2 FISH tests simultaneously in clinical and research-oriented settings with less tissue damage compared with conventional tissue microarray techniques.

Molecular Structures and Mechanisms of Waterborne Biodegradable Polyurethane Nanoparticles.

Biodegradable hydrogels have become promising materials for many biological applications in the past years. Recently, novel waterborne biodegradable polyurethane (WDPU) nanoparticles have been synthesized by a green water-based process, and serve as fundamental building blocks to form materials with great biocompatibility, biodegradability, and mechanical properties. However, the molecular structures and mechanisms of the WDPU nanoparticles and the relationship between the chemical compositions of the polymer segments and the material properties of the biodegradable hydrogels at macro-scale are still not well understood. In this study, we explore the fundamental mechanisms of WDPU nanoparticles through a full atomistic simulation approach to understand how the chemical compositions at the molecular level affect the molecular structures and material properties of WDPU nanoparticles. Specifically, we compare two WDPUs, i.e. PCL75LL25 and PCL75DL25, of the same hard segment composition and very similar soft segment composition [75% poly(e-caprolatone) and 25% polylactide], except the lactide in the former is L-form and in the latter is D,L-form. Our results show that the material properties of the biodegradable hydrogel can be designed by tuning the chemical compositions of the polymer segments. We find that the PCL75DL25 and PCL75LL25 have distinct molecular structures and physical crosslinks within the nanoparticles. The molecular structure of WDPU with PDLLA as soft segments is more extended, leading to more physical crosslinks between PCL segments. This study provide fundamental insights into the molecular structures and mechanisms of WDPU nanoparticles and help enabling the design of material properties of biocompatible hydrogel.

Differential CD133 expression distinguishes malignant from benign papillary lesions of the breast.

Malignant papillary lesions are, in contrast to their benign counterpart, rare malignant tumors in the breast. To differentiate between benign or atypical and malignant papillary lesions can be difficult, especially in core biopsy specimens. In the present study, excisional or more extensive surgical specimens of 33 papillary carcinomas, 2 micropapillary carcinomas, and 17 atypical papillomas of the breast were reviewed and classified according to the latest WHO classification. Thirty-three intraductal papillomas and 49 invasive carcinomas, no special type (NST), were included in the study for comparison. CD133 expression in papillary carcinomas was significantly lower than that in benign and atypical papillomas (p < 0.001). CD133 expression in invasive carcinoma NST was also significantly higher than that in papillary carcinomas. Our data suggests that absence of expression of CD133 can be a useful marker in the differential diagnosis between malignant papillary lesions and their benign or atypical mimics. The characteristic loss of CD133 expression in papillary carcinomas of the breast also indicates that these lesions are distinct from other types of breast cancer.

[Resilience in cancer patients and related nursing interventions].

Cancer is a critical health problem in Taiwan. The range of physical, psychological, social, and existential stressors associated with cancer diagnosis and treatment can cause significant distress in cancer patients and survivors. The focus of cancer research has broadened in the past decade from the disease itself to factors that can have a positive influence on the health and life quality of cancer patients. However, few studies have explored how patients adapt and become resilient to the life challenges of their disease. This article introduces the concept of resilience and its influence factors. We analyze study findings and introduce four nursing interventions that have been used to nurture resilience in cancer patients. The authors hope findings help strengthen nurse competencies in order to enhance cancer patient quality of care.

Improved technique for manually constructing tissue microarrays for large-core arrays.

Tissue microarrays were originally developed to enable alignment of multiple tissue cores in a single paraffin block and to enable high-throughput laboratory analysis. However, a major drawback is the loss of tissue cores during slide preparation, especially when sectioning the tissue block. Tissue cylinders directly aligned in the metal box without preheating tend to detach from the surrounding paraffin, which results in incomplete or folded tissue sections. The proposed solution is preheating all tissue cylinders on a hot plate to facilitate fusion between the paraffin within the core and the paraffin surrounding the core. In this study, 6 tissue microarray blocks were constructed from 528 tissue cores extracted from various formalin-fixed, paraffin-embedded human tissue samples. The tissue cores in the arrays revealed good homogenization with the surrounding paraffin wax, and the tissue sections were obtained intact. Both hematoxylin-eosin and immunohistochemical staining confirmed satisfactory results. This simple and economical method is easily performed in the laboratory without expensive instrumentation.

Utility of the capsule-based technique for cell block preparation - in body fluids and Liqui-PREP™ specimens.

OBJECTIVE: The aim of this study was to find a better technique for the cell block preparation. STUDY DESIGN: We developed a novel capsule-based technique and applied it to different cell block preparation materials including plasma thromboplastin (PT) clot and agarose gel-embedding medium, in order to concentrate the cells in a limited field, then compared the result with the widely used, modified alcohol fixative preparation technique based on specimen adequacy and diagnostic accuracy. Twenty-eight nongynecologic body fluids and 41 gynecologic Liqui-PREP™ (LGM International Inc., Fort Lauderdale, Fla., USA) cytology specimens were collected for cell block preparation. We performed routine hematoxylin and eosin staining on the cell block sections, which were scored according to four specimen adequacy parameters: background, cellularity, nuclear preservation and cytoplasmic preservation. The differences between the diagnostic results of cell block slide and conventional cytology smear were also presented. RESULTS: The capsule-based PT clot technique in nongynecologic body fluids provided more cellularity in the cell block sections and reduced atypical diagnosis when compared to conventional smears. CONCLUSION: We suggest the capsule-based PT clot technique is a better technique for nongynecologic body fluids in the preparation of a satisfactory cell block.

High-Glucose Environment Inhibits p38MAPK Signaling and Reduces Human ß-Defensin-3 Expression [corrected] in Keratinocytes.

Diabetes mellitus is characterized by elevated plasma glucose and increased rates of skin infections. Altered immune responses have been suggested to contribute to this prevalent complication, which involves microbial invasion. In this study we explored the effects of a high-glucose environment on the innate immunity of keratinocytes by focusing on ß defensin-3 (BD3) using in vivo and in vitro models. Our results demonstrated that the perilesional skins of diabetic rats failed to show enhanced BD3 expression after wounding. In addition, high-glucose treatment reduced human BD3 (hBD3) expression of cultured human keratinocytes. This pathogenic process involved inhibition of p38MAPK signaling, an event that resulted from increased formation of advanced glycation end products. On the other hand, toll-like receptor-2 expression and function of cultured keratinocytes were not significantly affected by high-glucose treatment. In summary, high-glucose conditions inhibited the BD3 expression of epidermal keratinocytes, which in turn contributed to the frequent occurrences of infection associated with diabetic wounding.

A simple and economical method for the manual construction of frozen tissue arrays.

Tissue microarray has been developed to enable multiple cores of tissue in one or more new paraffin blocks. Currently, almost all tissue microarrays are made by coring cylindrical tissues from formalin-fixed and paraffin-embedded tissues. The disadvantages of formalin-fixed and paraffin-embedded tissues include the poor preservation of antigenicity of certain proteins and mRNA degradation induced by the fixation and embedding process. However, frozen tissue array construction presents technical difficulties, and tissue array devices are expensive, particularly for small- and medium-sized laboratories. We describe a simple manual method for producing well-aligned tissue arrays by a capsule freeze method that allows us to successfully perform hematoxylin-eosin and immunohistochemical stain. All 120 tissue samples were collected and constructed into blocks by this capsule freeze method. The capsules were not affected during the sectioning process, and the capsule material always disappeared during the aqueous steps of the stain processing. The frozen tissue arrays were smoothly sectioned without the use of a tape transfer system and immunohistochemical study was performed with satisfactory results. This alternative method can be applied in any laboratory, and is both simple and economical.

Diagnostic value of computerized nuclear morphometry for the prediction of malignancy in liver fine needle aspiration cytology.

OBJECTIVE: To evaluate the diagnostic value of nuclear parameters using computerized morphometry in the differential diagnosis of well-differentiated hepatocellular carcinoma (WD-HCC) and regenerative atypia in liver fine needle aspiration cytology (FNAC). STUDY DESIGN: All 78 cases, including 30 negative, 30 positive and 18 suspicious liver FNA cytology smears were examined with regard to nuclear area, nuclear perimeter, largest to smallest diameter ratio of the nuclei (L/S ratio) and coefficient of variation of the nuclear area (NACV). The diagnostic value was assessed using receiver operating characteristics (ROC) curves. Cutoff points were determined by the area under the ROC curve (AUC), which provides the highest combined sensitivity and specificity. RESULTS: The nuclear area and perimeter in liver FNA were more helpful than L/S ratio and NACV in discriminating between hepatocytes with regenerative atypia and WD-HCC. Cutoff points of 56.67 for nuclear area and 27.16 for perimeter were suggested to help shift the suspicious diagnoses to WD-HCC. CONCLUSION: When faced with difficult diagnoses on liver FNA, we suggest application of computerized morphometry may be a helpful ancillary diagnostic tool in the further classification of such cases.

[Children's accidents: emergency care intervention and preventive strategies].

The injuries sustained by children in accidents have been a global public health topic because their influence is not confined to the child and his or her family, but extends to the wider society. This paper explains the emergency care intervention involved in treating child accident victims, the physical and mental changes that follow both trauma and care, and the adult's role when children have injuries. In addition, a systemic analysis is provided of the emergency medical system and emergency room triage, and ethical issues are discussed, in order that preventive strategies may be adopted as quickly as possible and to ensure good medical care for children suffering injury in accidents.

Application of the microarray technique to cell blocks.

OBJECTIVE: To evaluate whether the technique ofmicroarray construction can be applied to paraffin-embedded cell block specinmens. STUDY DESIGN: We used a manual microarray method for construction of a well-aligned cell microarray. First we evaluated the cellularity of the cell block and assigned the cases to the null, low, moderate or high cellularity category. Second, based on the different cellularity, we constructed 25 specimen cylinders into 1 block. We used routine hematoxylin-eosin (H-E) stain and immunocytochemistry (ICC) on the slide. RESULTS: All cell microarray paraffin block shaping and specimen cylinder arraying were finished in 1 step, so the specimen cylinders and the paraffin blocks of the cell microarray could very easily be incorporated. No sample was lost during the H-E staining process. However, a few samples fell off partially during the ICC process. Additionally, we observed that the higher cellularity groups yielded a better outcome as compared to the lower cellularity groups. CONCLUSION: This study introduced a very simple and economical technique for the creation of cell microarrays from cell blocks. This procedure should acquaint cytopathologists with microarray technology and encourage its use.