Apmis

doi: 10.1111/apm.12868pmid: N/A

Travaglino, Antonio; Raffone, Antonio; Saccone, Gabriele; Mascolo, Massimo; Pignatiello, Sara; Mollo, Antonio; De Placido, Giuseppe; Insabato, Luigi; Zullo, Fulvio

doi: 10.1111/apm.12938pmid: 30803040

Guidelines recommend protein phosphatase and tensin homolog (PTEN) immunohistochemistry for differentiating between benign endometrial hyperplasia (BEH) and atypical endometrial hyperplasia/endometrioid intraepithelial neoplasia (AEH/EIN). However, it is unclear when PTEN expression should be defined as ‘lost’ and thus suggestive of AEH/EIN. We aimed to determine the optimal immunohistochemical criteria to define PTEN loss in endometrial hyperplasia, through a systematic review and meta‐analysis of diagnostic accuracy. Electronic databases were searched for studies assessing immunohistochemical expression of PTEN in both BEH and AEH/EIN specimens. PTEN status (‘loss’ or ‘presence’) was the index test; histological diagnosis (‘AEH/EIN’ or ‘BEH’) was the reference standard. Accuracy was quantified based on the area under the curve (AUC) on summary receiver operating characteristic (SROC) curves, for several different thresholds of PTEN expression. Eighteen studies with 1362 hyperplasias were included. Six different criteria to define PTEN loss were assessed. Low diagnostic accuracy was found for complete loss of expression (AUC = 0.71), presence of any null gland (AUC = 0.63), positive cells <10% (AUC = 0.64), positive cells <50% (AUC = 0.71) and moderate‐to‐null intensity (AUC = 0.64). Barely moderate diagnostic accuracy was only found for the subjective criterion ‘weak‐to‐null intensity’ (AUC = 0.78). Therefore, the clinical usefulness of PTEN immunohistochemistry in this field should be further investigated.

Lin, Jen‐Tai; Chan, Ti‐Chun; Li, Chien‐Feng; Huan, Steven K.H.; Tian, Yu‐Feng; Liang, Peir‐In; Pan, Cheng‐Tang; Shiue, Yow‐Ling

doi: 10.1111/apm.12939pmid: 30803053

The objective of this study was to examine the expression level of cytochrome P450 4B1 (CYP4B1) protein and its clinical significance in specimens from patients with urothelial carcinomas (UC) including upper tract urothelial carcinoma (UTUC, n = 340) and urinary bladder urothelial carcinoma (UBUC, n = 295). Data mining on public domains identified five potential candidate transcripts which were downregulated in advanced UBUCs, indicating that it might implicate in UC progression. Immunohistochemistry was performed to analyze the CYP4B1 protein levels on 635 tissues from UC patients retrospectively. Immunoexpression of CYP4B1 was further estimated using the H‐score method. Correlations between CYP4B1 H‐score and important clinicopathological factors, as well as the significance of CYP4B1 expression level for disease‐specific and metastasis‐free survivals were evaluated. In UTUCs and UBUCs, 118 (34.7%) and 92 (31.2%) patients, respectively, were identified to be of CYP4B1 downregulation. The CYP4B1 expression level was found to be associated with several clinicopathological factors and patient survivals. Downregulation of CYP4B1 protein was correlated to advanced primary tumor (p < 0.001), nodal metastasis (p < 0.001), high histological grade (p = 0.001), vascular invasion (p < 0.001), perineural invasion (p = 0.017) and mitotic rate (p = 0.036) in UTUCs and/or UBUCs. Low CYP4B1 protein level independently predicted inferior disease‐specific (p = 0.009; p < 0.001) and metastasis‐free (p = 0.035; p < 0.001) survivals in UTUC and UBUC patients. Our findings showed that downregulation of CYP4B1 protein level is an independent unfavorable prognosticator. Loss of the CYP4B1 gene expression may play an important role in UC progression.

Gideskog, Maria; Melhus, Åsa

doi: 10.1111/apm.12929pmid: 30908773

The objective of this study was to investigate a sudden increase in methicillin‐resistant Staphylococcus aureus (MRSA) cases primarily in one maternity ward at the Center for Children's and Women's Health at Linköping University Hospital, Sweden. Approximately 300 individuals including patients, their family members, and healthcare workers were screened for MRSA. The antibiotic susceptibility was tested and isolates polymerase chain reaction (PCR)‐positive for the mecA gene were spa typed. Isolates with the same antibiogram and spa type were further whole genome sequenced. Compliance to current cleaning and hygiene routines was also controlled, and environmental samples collected. The results showed that a total of 13 individuals were involved in the outbreak. It was caused by a t386 MRSA strain (ST‐1, NCBI‐accession AB505628) with additional resistance to erythromycin and clindamycin. All cases were epidemiologically connected to the index patient, who had recently emigrated from a high‐endemic area for MRSA. With improved cleaning and better compliance to basic hygiene routines, no further cases were reported. This study demonstrates how rapid an MRSA strain can disseminate in a ward with susceptible patients and insufficient cleaning and hygiene. For a better control of MRSA, clinical cultures and screening samples need to be obtained early and more extensively than according to the current recommendations.

Kugaji, Manohar S.; Kumbar, Vijay M.; Peram, Malleswara Rao; Patil, Sanjivini; Bhat, Kishore G.; Diwan, Prakash V.

doi: 10.1111/apm.12930pmid: 30861212

Periodontal disease is an oral inflammatory disease that destroys the tooth supporting periodontal tissues resulting in tooth loss. Porphyromonas gingivalis is a keystone pathogen that plays a significant role in periodontitis. In previous studies, resveratrol has shown significant results by targeting inflammatory and adhesive markers. Virulence factors of P. gingivalis play an important role in the bacterial adhesion and colonization. In this study, we aimed to demonstrate the anti‐biofilm and anti‐bacterial activity of resveratrol and also study the effect of resveratrol on the expression of virulence factor genes of P. gingivalis using reverse transcriptase polymerase chain reaction (RT‐PCR). The anti‐microbial and anti‐biofilm activity of resveratrol on P. gingivalis was carried out by broth microdilution assay and biofilm adhesion reduction–crystal violet assay, respectively. We carried out the gene expression analysis by RT‐PCR with the P. gingivalis treated compound to analyze the change in the expression of virulence factors: fimbriae and gingipain. Minimal inhibitory concentrations (MIC) of resveratrol against P. gingivalis and other clinical strains are in the range of 78.12–156.25 μg/mL. Resveratrol dose‐dependently prevented the biofilm formation and also attenuated the virulence of P. gingivalis by reducing the expression of virulence factor genes such as fimbriae (type II and IV) and proteinases (kgp and rgpA). Resveratrol demonstrated superior anti‐bacterial and anti‐biofilm activity against P. gingivalis. There was significant reduction in the expression of fimbriae and gingipain with the resveratrol‐treated compound. The results suggest that resveratrol, due to its multiple actions, may become a simple and inexpensive therapeutic strategy for treating periodontal disease.

Tisi, Giancarlo; Gargiulo, Franco; Gozzini, Elisa; Baronchelli, Carla; Odicino, Franco; Salinaro, Federica; Sartori, Enrico; Caruso, Arnaldo; Facchetti, Fabio; De Francesco, Maria Antonia

doi: 10.1111/apm.12931pmid: 30815926

Xiao, Guohui; Yi, Yusi; Che, Rongbo; Zhang, Qinchao; Imran, Muhammad; Khan, Abidullah; Yan, Jie; Lin, Xu'ai

doi: 10.1111/apm.12935pmid: 30908774

Leptospirosis is a zoonotic disease caused by pathogenic Leptospira. However, understanding of the pathogenic mechanism of Leptospira is still elusive due to the limited number of genetic tools available for this microorganism. Currently, the reason for the genetic inaccessibility of Leptospira is still unknown. It is well known that as an acquired immunity of bacteria, Clustered Regularly Interspaced Short Palindromic Repeat‐CRISPR‐associated gene (CRISPR‐Cas) systems can help bacteria against invading mobile genetic elements. In this study, the occurrence and diversity of CRISPR‐Cas systems in 41 genomes of Leptospira strains were investigated. Three subtypes (subtype I‐B, subtype I‐C and subtype I‐E) of CRISPR‐Cas systems were identified in both pathogenic and intermediate Leptospira species but not in saprophytic species. Noteworthy, the majority of pathogenic species harbor two different types of CRISPR‐Cas systems (subtype I‐B and subtype I‐E). Furthermore, Cas2 protein of subtype I‐C in L. interrogans exhibited a metal‐dependent DNase activity in a nonspecific manner. CRISPR spacers in subtype I‐B are highly conserved within the same serovars and hypervariable across different serovars of L. interrogans. Based on the subtype I‐B CRISPR arrays, the serotypes of different L. interrogans strains were easily identified. Investigation of the origin of CRISPR spacers showed that 192 spacers (23.5%) matched to mobile genetic elements, indicating CRISPR‐Cas systems may play an important role in the defense of foreign invading DNA.

Bellanger, Anne‐Pauline; Gbaguidi‐Haore, Houssein; Liapis, Eleni; Scherer, Emeline; Millon, Laurence

doi: 10.1111/apm.12936pmid: 30803048

Rapid identification of Candida species is important for appropriate antifungal therapy of fungemia. The matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) system is a useful tool to identify bacteria and yeasts. In this study, we evaluated the feasibility of identifying yeasts after a short‐term incubation on a solid medium. We tested 24 strains of eight Candida species. Blood culture bottles were spiked with a calibrated suspension of each Candida strain. Three different culture media, two types of blood culture bottles and three different incubation time points were tested. A multivariable random‐effects logistic regression analysis was performed for determining factors independently associated with a successful MALDI‐TOF MS identification. One‐hundred and thirty‐one out of 432 MALDI‐TOF MS analyses (30%) exhibited a score ≥ 1.7. The performance of the technique varied across Candida species. Factors associated with a successful identification were the use of a chromogenic Candida medium and the time points 4 and 5 h. Using the factors ‘chromogenic Candida medium’ and time point 5 h the global performance of identification reached 60% and a mean MALDI‐TOF score of 1.78. Identifying yeasts after a short‐term incubation on a solid medium seems possible, especially when using a chromogenic Candida medium and respecting at least 5 h of incubation. This assay was a first step and needs to be completed using more strains, various chromogenic Candida medium and maybe also testing a longer culture time such as 6 h.

Kouhsari, Ebrahim; Douraghi, Masoumeh; Fakhre Yaseri, Hashem; Talebi, Malihe; Ahmadi, Alireza; Sholeh, Mohammad; Amirmozafari, Nour

doi: 10.1111/apm.12937pmid: 30803047

Clostridioides difficile is a major cause of nosocomial infectious diarrhea in hospitalized patients throughout the world. We aimed to characterize C. difficile isolates among hospitalized patients, hospital staffs, and hospital environment samples obtained in three tertiary care hospitals of Iran with regard to their molecular types between June 2016 and November 2017. The toxigenicity of C. difficile isolates was determined by toxigenic culture and multiplex‐PCR. Toxigenic C. difficile isolates collected were ribotyped using capillary gel electrophoresis‐based PCR and the database of WEBRIBO (http://webribo.ages.at). Of 500 clinical and non‐clinical samples, toxigenic C. difficile were identified in 35 of 250 stool samples (14%) and in 3 of 250 swabs (1.2%). The most frequently found ribotypes (RTs) were 039, AI‐12, and AI‐21 (15.8, 10.52, and 10.52% of all isolates, respectively). Further RTs were: 017, 001, AI‐3, AI‐15, AI‐18, AI‐10, AI‐4, and PR21195 (as new ribotype). The epidemic RTs (027 and 078) seen in the Europe, North America, and Asia were completely absent in this study.

Dias, Ana Luisa Abrahão; da Silva, Raquel Gomes; Cunha, Fernanda Gonçalves Pereira; Morcillo, André Moreno; Lorand‐Metze, Irene; Vilela, Maria Marluce dos Santos; Riccetto, Adriana Gut Lopes

doi: 10.1111/apm.12932pmid: 30908772

Our aim was to evaluate the cost‐effectiveness of a minimal lymphocyte subset quantification (LSQ) by flow cytometry as the first screening in children with clinically suspected primary immunodeficiency (PID). Two hundred sixty‐eight Brazilian patients (0–21 years old) were studied. They were divided by clinical and phenotypical features into those fulfilling criteria for PID (PID phenotype) according to the 2017 International Union of Immunological Societies (IUIS) classification and those not fulfilling these criteria (non‐PID phenotype). We evaluated how many patients had values below the 10th percentile for five lymphocyte subsets in peripheral blood, (suggestive of PID) according to reference values for Brazil, Italy and USA. Three lymphocyte subsets (T CD3/CD4, B CD19 and NK CD16/CD56) had p‐value < 0.05 and Odds Ratio (OR) indicating a risk at least two times higher for the diagnosis of a PID phenotype. The application of Kappa coefficient (k) on Brazilian vs Italian and Brazilian vs US data sets resulted in k compatible with strong or excellent level of agreement between the three classification systems. The authors conclude that a number of CD3+/CD4+, CD19+ and CD16+/CD56+ (NK) cells in peripheral blood <10th percentile represented a significant risk for the diagnosis of PID in this cohort. Natural killer (NK) deficiency is quite rare and has a very specific clinical profile. So, the analysis of these cells could be requested only in some cases, saving even more costs. The minimal immunophenotyping, with quantification of T CD4+, CD19+ and in some cases CD16+/CD56+ cells, may be a useful tool for the first screening of PID, saving costs, especially in developing countries.