Characteristic Cytological Findings of Lobular Endocervical Glandular Hyperplasia Associated with Adenocarcinoma of the Uterine Cervix.

OBJECTIVE: To investigate the cytological findings of lobular endocervical glandular hyperplasia (LEGH) associated with adenocarcinoma and to clarify its characteristics and the coexisting adenocarcinoma using histochemistry and immunohistochemistry. METHODS: Eighteen surgical cases of LEGH of the uterine cervix were retrospectively reviewed and classified into 3 groups: pure (pure type), atypical (atypical type), and LEGH with adenocarcinoma (mixed type). The mixed type is defined as LEGH or atypical LEGH with in situ or invasive adenocarcinoma. Cytological findings of conventional endocervical smear specimens (Papanicolaou stain) were analyzed. Histochemistry (periodic acid-Schiff reaction) and immunohistochemistry (M-GGMC-1, Muc-6 glycoprotein, and Ki-67) were performed using tissue specimens. RESULTS: Cytologically, the pure type (7 cases) is characterized by glandular cell clusters that tended to form monolayered sheets with uniformly small nuclei and contain golden-yellowish mucin, whereas atypical (5 cases) and mixed (6 cases) types are characterized by glandular cell clusters similar to those of the pure type, but with complex glandular structures and mucin localization on the surface of glandular cell clusters. Ki-67 labeling index was significantly higher in atypical and mixed types than that in the pure type. Gastric-type mucinous carcinoma (MC-G) was observed in 2 out of 6 cases with mixed type. CONCLUSIONS: LEGH is found to be associated with adenocarcinoma types other than MC-G. Complex glandular structures or mucin localization on the surface of glandular cell clusters may be useful cytological findings to detect atypical and mixed types of LEGH.

Clinical management of the status of atypical endometrial cells using the descriptive reporting format for endometrial cytology.

OBJECTIVE: We aimed to develop and reinforce a clinical management regimen for atypical endometrial cell (ATEC) categories within the descriptive reporting format for endometrial cytology. METHODS: Between January 2013 and December 2014, 215 samples, for which histological examination was performed immediately or within 3 months after cytology, were cytologically diagnosed as ATEC. For these samples, the medical records were retrospectively reviewed to identify risk factors for malignancy. RESULTS: Among 152 samples diagnosed as ATEC, of undetermined significance, 19 (12.5%) were malignant. In the younger group (age <55 years), the χ2 values of body mass index (BMI) ≥25 kg/m2 (5.85), gravidity (5.64) and parity (5.15) were relatively high, suggesting that these were risk factors for malignancy. Of the nulligravida patients, those with BMI ≥25 kg/m2 , 28% were diagnosed with malignant disease. In the older group (≥55 years), endometrial thickening (6.84), atypical genital bleeding (6.43) and BMI ≥25 kg/m2 (3.79) were found to be risk factors for malignancy. Of the patients with endometrial thickening and atypical genital bleeding, 67% were diagnosed with malignant disease. Among 63 samples diagnosed as ATEC, cannot exclude atypical endometrial hyperplasia or more, 35 (55.6%) samples were positive for malignancy. CONCLUSIONS: High-risk patients diagnosed with ATEC, of undetermined significance were identified. Endometrial biopsy should be considered for nulligravida patients aged <55 years with a BMI ≥25 kg/m2 .

A Multi-Institutional Feasibility Study on the Use of Automated Screening Systems for Quality Control Rescreening of Cervical Cytology.

OBJECTIVE: To evaluate the efficacy of the automated screening system FocalPoint for cervical cytology quality control (QC) rescreening. False-negative rates (FNRs) were evaluated by a multi-institutional retrospective study. STUDY DESIGN: Cervical cytology slides that had already been reported as negative for intraepithelial lesion or malignancy (NILM) were chosen arbitrarily for FocalPoint rescreening. Slides stratified into the highest 15% probability of being abnormal were rescreened by a cytotechnologist. The slides that were abnormal were reevaluated by a cytopathologist to be false negatives. RESULTS: Rescreening of 12,000 slides, i.e. 9,000 conventional slides and 3,000 liquid-based cytology (LBC) slides, was performed; 9,826 (7,393 conventional and 2,433 LBC) were satisfactory for FocalPoint (2,174 were determined unsatisfactory) and those within the highest 15% of probability (1,496, i.e. 1,123 conventional and 373 LBC) were rescreened. As a result, 117 (96 conventional and 21 LBC) were determined as abnormal (other than NILM) and the FNR was 1.19%. Among these 117 slides, 40 (35 conventional and 5 LBC) were determined as high-grade squamous intraepithelial lesion and greater (HSIL+). CONCLUSION: Of 117 (1.19%) abnormal slides detected, 40 (0.41%) were determined to be HSIL+. This result suggests that FocalPoint is effective for QC rescreening of cervical cytology.

Touch cytology smear of an inflammatory hepatocellular adenoma displaying an unusual pattern: A case report.

The cytological diagnosis of hepatocellular adenoma (HCA) is difficult since it is a very rare tumor and lacks characteristic cytological features. We have just reported a case of inflammatory HCA that displayed an unusual histological pattern (Clin J Gastroenterol 8:426-434, 2015). A touch cytology smear sample was obtained from the surgical specimen, and it also exhibited very unique features. A 56-year-old male underwent partial hepatectomy for an inflammatory HCA (diameter: 1.4 cm) in the right posterior lobe of the liver. The cytological sample displayed a characteristic two-cell pattern. One type of cells contained thick cytoplasm, a high nucleus/cytoplasmic (N/C) ratio, and well-defined cytoplasmic borders. The other type demonstrated small pyknotic nuclei and a lower N/C ratio. The immunohistochemical staining pattern of the histological specimen suggested that the latter cells might have been undergoing apoptosis. We report a case of inflammatory HCA with characteristic features. To diagnose this type of variant, it is important to recognize the unique pattern described in this study. Diagn. Cytopathol. 2016;44:1074-1077. © 2016 Wiley Periodicals, Inc.

[Current Situation and Tasks for International Activities of BLS -Presentation for International Congress or Study Abroad-].

Nowadays, global consciousness is important for the human race. Many Japanese businesspersons and even students try to improve their foreign language skills, and try to get a chance to join international activities. International Activities of BLSs are as follows: 1. Apply for the Japan International Cooperation Agency (JICA) business for supporting developing countries as a technical expert. Since JICA activity is supported by the Japanese Government, it must be worthwhile and will complete large-scale business. After the completion, it might be difficult to find a good work place to utilize JICA ex- perience. 2. Study abroad such as joining an exchange program, or working as a researcher or trainee. Some Japanese institutes provide systems for studying abroad for BLSs. We should study language and improve our professional skills. An employer might give permission for a short stay easily. Support from a supervisor is important. 3. Overseas occupation. We will need certification or a license when we work abroad. Study or work abroad is a precious experience. We should be clear on the purpose and select a suitable place with a long-term plan. Among international activities, it is easy to be a presenter at the World Congress. There are many inter- national congresses for BLSs such as the IFBLS World Congress, AAMLS Congress, or Scientific Congress for each special filed. We should learn not only language, but also different cultures and international man- ners when we participate in international activities. Supervisors can help to improve the global perspective of young BLSs. [Review].

[Qualification System for BLS -Domestically and Internationally-].

The scope of the biomedical laboratory scientist (BLS) differs in the world. The qualification systems for BLS are also different. Even the name of our profession is not standardized, like "BLS", "Medical Technologist (MT) ", "Medical Laboratory Scientist (MLS) ", "Medical Technician", and so on. We have only the "BLS" license, which is recognized by the Ministry of Health, Labour and Welfare in Japan. Many Asian countries have a qualification system similar to Japan. In the USA, BLSs are qualified by certain scientific organizations, and they are called "MT", "BLS", or "Specialist" in consideration of their education level. The International Association of Medical Laboratory Technologists (IAMLT) was founded by Ms. Elizabeth Pletscher and her Swiss colleagues in order to cooperate with international BLSs in 1954. Since the BLS education system has been improved, chief delegates from the world decided that we should call ourselves "BLS", and IAMLT was revised to the "International Federation of Biomedical Laboratory Science (IFBLS) " at the general assembly in 2002. IFBLS has tried to spread our name and occupation to the public though our international cooperation using the same name "BLS". IFBLS shares the core competence, but does not provide certification. The International Academy of Clinical Cytology (IAC) has provided international certification for cytolo- gists and cytotechnologists. The American Society of Clinical Pathology (ASCP) has certified overseas BLSs (ASCPi), and they are able to work in the USA as BLSs after passing the examination. These certifi- cations will be valid in limited areas. As the current situation, if someone would like to work in another country as a BLS, he/she will have to pass the domestic BLS examination. [Review].

Engineered Streptomyces avermitilis host for heterologous expression of biosynthetic gene cluster for secondary metabolites.

An industrial microorganism, Streptomyces avermitilis, which is a producer of anthelmintic macrocyclic lactones, avermectins, has been constructed as a versatile model host for heterologous expression of genes encoding secondary metabolite biosynthesis. Twenty of the entire biosynthetic gene clusters for secondary metabolites were successively cloned and introduced into a versatile model host S. avermitilis SUKA17 or 22. Almost all S. avermitilis transformants carrying the entire gene cluster produced metabolites as a result of the expression of biosynthetic gene clusters introduced. A few transformants were unable to produce metabolites, but their production was restored by the expression of biosynthetic genes using an alternative promoter or the expression of a regulatory gene in the gene cluster that controls the expression of biosynthetic genes in the cluster using an alternative promoter. Production of metabolites in some transformants of the versatile host was higher than that of the original producers, and cryptic biosynthetic gene clusters in the original producer were also expressed in a versatile host.

Structural basis for matrix metalloproteinase-2 (MMP-2)-selective inhibitory action of ß-amyloid precursor protein-derived inhibitor.

Unlike other synthetic or physiological inhibitors for matrix metalloproteinases (MMPs), the ß-amyloid precursor protein-derived inhibitory peptide (APP-IP) having an ISYGNDALMP sequence has a high selectivity toward MMP-2. Our previous study identified amino acid residues of MMP-2 essential for its selective inhibition by APP-IP and demonstrated that the N to C direction of the decapeptide inhibitor relative to the substrate-binding cleft of MMP-2 is opposite that of substrate. However, detailed interactions between the two molecules remained to be clarified. Here, we determined the crystal structure of the catalytic domain of MMP-2 in complex with APP-IP. We found that APP-IP in the complex is indeed embedded into the substrate-binding cleft of the catalytic domain in the N to C direction opposite that of substrate. With the crystal structure, it was first clarified that the aromatic side chain of Tyr(3) of the inhibitor is accommodated into the S1' pocket of the protease, and the carboxylate group of Asp(6) of APP-IP coordinates bidentately to the catalytic zinc of the enzyme. The Ala(7) to Pro(10) and Tyr(3) to Ile(1) strands of the inhibitor extend into the nonprime and the prime sides of the cleft, respectively. Therefore, the decapeptide inhibitor has long range contact with the substrate-binding cleft of the protease. This mode of interaction is probably essential for the high MMP-2 selectivity of the inhibitor because MMPs share a common architecture in the vicinity of the catalytic center, but whole structures of their substrate-binding clefts have sufficient variety for the inhibitor to distinguish MMP-2 from other MMPs.

Cerebral state index versus bispectral index during propofol-fentanyl-nitrous oxide anesthesia.

PURPOSE: The aim of this study was to compare the cerebral state index (CSI) and bispectral index (BIS) during propofol-fentanyl-nitrous oxide anesthesia. METHODS: Thirty patients scheduled for abdominal surgery, with a mean age of 30-70 years, were enrolled. Anesthesia was induced with propofol and fentanyl and was maintained with propofol, fentanyl, epidural mepivacaine, and nitrous oxide in oxygen. During surgery, the propofol infusion rate was adjusted to try to keep BIS at 40 + or - 3 for 10 min and then decreased to keep the BIS at 60 + or - 3 for 10 min. RESULTS: The BIS had a larger value for the time between switching on the apparatus and starting to measure at a signal quality index >75%. The recovery time from disturbance by an electric cautery event was 41 + or - 14 s for the BIS and 3 + or - 1 s for the CSI (P < 0.05). The absolute values of the BIS and CSI were not significantly different and they showed a good correlation. The bias (mean of the differences, BIS - CSI) was negative at all measurement points, but the limits of agreement and percentage error were small. CONCLUSIONS: The absolute values of the BIS and CSI were not significantly different during propofol-fentanyl-nitrous oxide anesthesia. The start of the measurement was faster with the CSI than with the BIS after switch-on, and measurement was less disturbed by electric cautery with the CSI.

Coexistent atypical polypoid adenomyoma and complex atypical endometrial hyperplasia in the uterus.

We report a case of atypical polypoid adenomyoma (APA) concomitantly identified with complex atypical endometrial hyperplasia (CAH) in the uterus. Since an initial endometrial smear revealed atypical endometrial cells, a diagnosis of CAH was made. Even though a concomitantly performed uterine cervical smear contained both atypical epithelial and stromal cells, the diagnosis of APA was not initially made because the cytological criteria for APA had not been established. Histologically, we recognized both CAH in the uterine corpus and APA in the lower uterine segment in the hysterectomy material. Retrospectively, the cells in the first cervical smear were interpreted as part of APA because the same types of cells were observed in the intraoperative cytology sample. Although the APA and CAH lesions were interposed by normal endometrium, estrogen was suspected to be the common etiological factor. Reports regarding the cytology of APA are currently scarce. To our knowledge, this is the first report showing cytological presentation of association of APA with CAH in addition to the first cervical smear of APA containing both epithelial and stromal components. Identification of abnormal proliferation of epithelium and stromal cells of smooth muscle origin is useful in the cytological diagnosis of APA.

Application of liquid-based preparation to fine needle aspiration cytology in breast cancer.

OBJECTIVE: To examine the performance of liquid-based cytology (LBC) in breast cytology to confirm the diagnosis of carcinoma. STUDY DESIGN: Using cell clusters directly scratched from surgically removed tumor masses, we examined the immunocytochemistry, molecular biology and cytomorphology of the specimens. RESULTS: LBC was very useful for gene analysis and evaluating the immunocytochemistry. The cytologic features of LBC were slightly different from those ofa conventional aspiration cytology smear. CONCLUSION: LBC is a promising method for improving the standardization ofpreparations in breast cytology, although care should be taken to account for its characteristic cytologic features. The quantitative analysis of HER-2 mRNA correlated with the results of immunohistochemistry.

Expression and quantitative analysis of matrix metalloproteinase-2 and -9 in human gliomas.

The matrix metalloproteinase (MMP) family members catalyze extracellular proteolysis. Recent reports have suggested that expression of MMP-2 and -9 might play a critical role in neoplastic tissue invasion or metastasis. In this study, the relationship between the expression of MMP-2 and -9 and the histological features of tissues from 21 cases of human glioma were investigated. MMP-2 and -9 proteins were detected by immnohistochemical studies. Amplification of MMP-2 and -9 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) assay. MMP-2 and -9 mRNA was measured quantitatively by the real-time RT-PCR method. Immunohistochemically, 38% of the cases were positive for MMP-2. Amplification of MMP-2 mRNA by RT-PCR was detected in 62% of the cases. There was no significant relationship between the expression of MMP-2 protein or mRNA and the biological nature of the tumors, including aggressiveness and histologic classification. The quantity of MMP-2 mRNA was 0.035 +/- 0.113 (MMP-2/GAPDH %), which was significantly elevated in cases of neoplastic dissemination or recurrence (P < 0.05). Tumor cells were immunohistochemically positive for MMP-9 in 81% of the samples. A positive reaction was found not only in neoplastic cells but also in endothelial cells, suggesting that the expression of MMP-9 protein might be associated with tumoral angiogenesis. The expression of mRNA in MMP-9 was detected in 91% of the cases, suggesting a close relationship between expression of MMP-9 and malignancy. The quantity of MMP-9 was 0.097 +/- 0.113 (MMP-9/GAPDH %) in all samples, which was significantly elevated in cases of glioblastoma (P < 0.05). The average Ki-67 labeling index was 8.14 +/- 5.26 in samples from G2 glioma, 19.92 +/- 11.29 in samples from G3 glioma, and 23.52 +/- 10.14 in samples from glioblastoma. All of the cases with elevated indices had recurrence or dissemination. The results of our study suggest that quantity analyses of MMP-2 and -9 mRNA and Ki-67 labeling index should be useful for discerning tumoral behaviors such as invasion, dissemination, and recurrence.

Comparison of hemodynamic and anesthetic effects of hyperbaric bupivacaine and tetracaine in spinal anesthesia.

PURPOSE: To compare the anesthetic and hemodynamic effects and the predictive factor of anesthesia level of commonly used preparations of hyperbaric bupivacaine and tetracaine in spinal anesthesia. METHODS: Two hundred patients aged 40 to 75 years with ASA physical status I or II were anesthetized spinally via the L4-5 interspace using 0.5% hyperbaric bupivacaine in 7.27% glucose (Bupivacaine group, n = 100) or 0.5% hyperbaric tetracaine dissolved in a 10% glucose solution (Tetracaine group, n = 100) in a lateral position. The volume of anesthetic used was decided by the resident according to the surgical procedure. Patients were returned to the supine position immediately after drug injection. Blood pressure, heart rate, and anesthesia level tested by cold sensation were measured for 30 min. RESULTS: Blood pressure and heart rate decreased significantly but without any differences between the groups. The volume of drug used was significantly larger in the Bupivacaine group (2.6 +/- 0.5 ml) than in the Tetracaine group (2.1 +/- 0.4 ml) to obtain the same maximum anesthesia level. The time to reach the maximum anesthesia level was significantly longer in the Bupivacaine group (18 +/- 7 min) than in the Tetracaine group (15 +/- 6 min). The volume of the drug was the only predictive factor of the maximum anesthesia level in both groups: Level (as expressed by the number of anesthetized segments from S5 to cephalad) = 1.55 x (volume in ml) + 13.06 in the Bupivacaine group, and 2.59 x (volume) + 11.46 in the Tetracaine group. CONCLUSION: In spinal anesthesia, hyperbaric tetracaine in 10% glucose induced a faster and higher spread of anesthesia than hyperbaric bupivacaine in 7.27% glucose without any differences in hemodynamics.