Induction of Male Sterility by Targeted Mutation of a Restorer-of-Fertility Gene with CRISPR/Cas9-Mediated Genome Editing in Brassica napus L.

Brassica napus L. (canola, oil seed rape) is one of the world's most important oil seed crops. In the last four decades, the discovery of cytoplasmic male-sterility (CMS) systems and the restoration of fertility (Rf) genes in B. napus has improved the crop traits by heterosis. The homologs of Rf genes, known as the restoration of fertility-like (RFL) genes, have also gained importance because of their similarities with Rf genes. Such as a high non-synonymous/synonymous codon replacement ratio (dN/dS), autonomous gene duplications, and a possible engrossment in fertility restoration. B. napus contains 53 RFL genes on chromosomes A9 and C8. Our research aims to study the function of BnaRFL11 in fertility restoration using the CRISPR/Cas9 genome editing technique. A total of 88/108 (81.48%) T0 lines, and for T1, 110/145 (75%) lines carried T-DNA insertions. Stable mutations were detected in the T0 and T1 generations, with an average allelic mutation transmission rate of 81%. We used CRISPR-P software to detect off-target 50 plants sequenced from the T0 generation that showed no off-target mutation, signifying that if the designed sgRNA is specific for the target, the off-target effects are negligible. We also concluded that the mutagenic competence of the designed sgRNAs mediated by U6-26 and U6-29 ranged widely from 31% to 96%. The phenotypic analysis of bnarfl11 revealed defects in the floral structure, leaf size, branch number, and seed production. We discovered a significant difference between the sterile line and fertile line flower development after using a stereomicroscope and scanning electron microscope. The pollen visibility test showed that the pollen grain had utterly degenerated. The cytological observations of homozygous mutant plants showed an anther abortion stage similar to nap-CMS, with a Orf222, Orf139, Ap3, and nad5c gene upregulation. The bnarfl11 shows vegetative defects, including fewer branches and a reduced leaf size, suggesting that PPR-encoding genes are essential for the plants' vegetative and reproductive growth. Our results demonstrated that BnaRFL11 has a possible role in fertility restoration. The current study's findings suggest that CRISPR/Cas9 mutations may divulge the functions of genes in polyploid species and provide agronomically desirable traits through a targeted mutation.

Committee II: Guidelines for cytologic sampling techniques of lung and mediastinal lymph nodes.

The Papanicolaou Society of Cytopathology has developed a set of guidelines for pulmonary cytology including indications for bronchial brushings, washings, and endobronchial ultrasound guided transbronchial fine-needle aspiration (EBUS-TBNA), technical recommendations for cytological sampling, recommended terminology and classification schemes, recommendations for ancillary testing and recommendations for post-cytological management and follow-up. All recommendations are based on the expertise of the authors, an extensive literature review and feedback from presentations at national and international conferences. This document selectively presents the results of these discussions. The present document summarizes recommendations regarding techniques used to obtain cytological and small histologic specimens from the lung and mediastinal lymph nodes including rapid on-site evaluation (ROSE), and the triage of specimens for immunocytochemical and molecular studies.

Committee I: Indications for pulmonary cytology sampling methods.

The Papanicolaou Society of Cytopathology has developed a set of guidelines for pulmonary cytology including indications for bronchial brushings, washings and endobronchial ultrasound-guided fine needle aspiration, technical recommendations for cytologic sampling, recommended terminology and classification scheme, recommendations for ancillary testing and recommendations for postcytologic diagnosis management and follow-up. All recommendation documents are based on the expertise of the authors, extensive literature review and feedback from presentations at national and international conferences. This document selectively presents the results of these discussions. The present document summarizes the recommendations for clinical and imaging evaluation of pulmonary lesions along with the indications for cytologic studies regarding these abnormalities. Preprocedural requirements regarding brushing, washing and needle aspiration procedures are discussed also. Diagn. Cytopathol. 2016;44:1010-1023. © 2016 Wiley Periodicals, Inc.

Características citohistológicas de lesiones premalignas y malignas del cuello uterino / Cytohistological characteristics of premalignant and malignant lesions of cervix

Se realizó un estudio descriptivo y retrospectivo de 738 pacientes con lesiones premalignas o malignas del cuello uterino, diagnosticados mediante biopsias y estudios citológicos archivados en fichas en el Departamento de Anatomía Patológica del Hospital General Docente "Dr. Juan Bruno Zayas Alfonso" de Santiago de Cuba, durante el bienio 2011-2012, con vistas a determinar algunas características de dichas formaciones neoplásicas. En la serie se obtuvo que más de la mitad de las muestras hísticas cervicouterinas mostraran alguna lesión premaligna o maligna, predominantemente en pacientes con edades de 25 a 54 años, y el diagnóstico más frecuente fue la displasia moderada; de igual modo, el virus de papiloma humano estuvo presente en 64,8 % de los estudios citológicos y en 75,3 % de las biopsias, y la efectividad del estudio citológico en el laboratorio fue de 92,0 %.

A descriptive and retrospective study of 738 patients with premalignant and malignant lesions of cervix, diagnosed by means of biopsies and cytological studies preserved from records in the Pathology Department of "Dr. Juan Bruno Zayas Alfonso" Teaching General Hospital in Santiago de Cuba was carried out during the biennium 2011-2012, with the aim of determining some characteristics of these neoplastic disorders. In the series it was obtained that more than half of the tissue cervical samples showed some premalignant and malignant lesion, predominantly in patients with ages from 25 to 54 years, and the most frequent diagnosis was moderate dysplasia; likely, the human papillomavirus was present in 64.8% of the cytological studies and in 75.3% of the biopsies, and the effectiveness of the cytological study in the laboratory was 92.0%.