Morphological and molecular characterization of a new autochthonous Trichoderma sp. isolate and its biocontrol efficacy against Alternaria sp.

The development of agriculture requires the use of microorganisms in the management of phytopathogens as a way to compensate for the use of chemical pesticides, in order to produce healthy crops. The objective of this study was to characterize a new isolate of Trichoderma sp. based on morphological and molecular features, and its potential ability to control the pathogen Alternaria sp. The antagonistic isolate was isolated from soil samples of potato fields in Guasave Sinaloa, Mexico, whereas the pathogen was collected from infected apple leaves in the orchard "La Escondida" in Guerrero County, Chihuahua, Mexico. For morphological characterization both fungi were grown on solid PDA medium. DNA of Trichoderma sp. was isolated using the CTAB method and PCR analyses were done using ITS1, ITS4 primers resulting in amplified products of 600 bp. These were sequenced, submitted to Genbank (acc. no. MN950427) and used for further phylogenetic analysis through Bayesian inference approach. Five clades were identified and the polytome topography recovered from clade 4 indicates a high genetic similarity with T. asperellum. A BLAST examination of the resulting sequence in GenBank showed 98.11% similarity with T. asperellum. This result together with the morphological and the phylogenetic analyses indicates that the isolate belongs to Trichoderma asperellum Samuels, Lieckfeldt & Nirenberg. Biocontrol tests of this isolate showed inhibition of Alternaria sp. between 50% and 93%. These results are essential for biodiversity research and give some new possibilities for pest management.

Tumor histology is an independent prognostic factor in locally advanced cervical carcinoma: A retrospective study.

BACKGROUND: Even with different histologic origins, squamous cell carcinoma (SCC) and adenocarcinoma (AC) are considered a single entity, and the first-line treatment is the same. Locally advanced disease at the diagnosis of cervical cancer is the most important prognostic factor, the recurrence rate is high, making it necessary to evaluate prognostic factors other than clinical or radiological staging; histology could be one of them but continues to be controversial. The aim of this study was to evaluate tumor histology as a prognostic factor in terms of treatment outcomes, disease-free survival (DFS) and overall survival (OS) in a retrospective cohort of patients with Locally Advanced Cervical Carcinoma (LACC). METHODS: The records of 1291patients with LACC were reviewed, all of them were treated with 45-50 Gy of external beam radiotherapy with concurrent chemotherapy and brachytherapy. A descriptive and comparative analysis was conducted. Treatment response was analyzed by the chi-square test; DFS and OS were calculated for each histology with the Kaplan-Meier method and compared with the log-rank test; and the Cox model was applied for the multivariate analysis. RESULTS: We included 1291 patients with LACC treated from 2005 to 2014, of which 1154 (89·4%) had SCC and 137 (10·6%) had AC. Complete response to treatment was achieved in 933 (80·8%) patients with SCC and 113 (82·5%) patients with AC. Recurrence of the disease was reported in 29·9% of SCC patients and 31·9% of AC patients. Five-year DFS was 70% for SCC and 62·2% for AC. The five-year OS rates were 74·3% and 60% for SCC and AC, respectively. The mean DFS was 48·8 months for SCC vs 46·10 for AC (p = 0·043), the mean OS was 50·8 for SCC and 47·0 for AC (p = 0·002). CONCLUSION: Our findings support the hypothesis that SCC and AC are different clinical entities. TRIAL REGISTRATION: NCT04537273 .

Radiation oncology in Mexico: Current status according to Mexico's Radiation Oncology Certification Board.

AIM: Describe the results of the first national census of radiotherapy in Mexico in order to make a situational diagnosis of radiotherapy availability, offer more accurate information to radiation oncologists, and promote an adequate scientific based investment for the country. BACKGROUND: According to the Organisation for Economic Co-operation and Development (OECD), the density of radiotherapy (RT) machines per million habitants in Mexico is approximately 1.7-1.8. Other international organizations such as DIRAC-IAEA report 1.15 per million habitants. National organizations collect data indirectly and previous surveys had a low accrual rate (32.5%). Therefore, a precise census is required. MATERIAL AND METHODS: The Mexican Radiation Oncology Certification Board (CMRO for its acronym in Spanish) conducted a nationwide census from January through November 2019. Gathered information was combined with CMRO database for sociodemographic information and human resources. RESULTS: The study included 103 RT centers [95.1% answered the survey], with a median of 2 centers by state (ranging from 0 in Tlaxcala to 20 in Mexico City) and with a report of only 1 center in 11 states (34.4%). Fifty-six (54.3%) of the centers are public. Fourteen centers (13.6%) have residency-training programs. The total number of RT machines is 162 [141 clinical and linear accelerators (87%) and 21 radionuclide units (13%)] with a median of 3 machines by state (0 in Tlaxcala to 46 in Mexico City) and with ≤3 machines in 18 states (56.25%). The overall calculated density of RT machines per million habitants is 1.32, varying from 0 in Tlaxcala to 5.16 in Mexico City. The density of linear and clinical accelerators per million population is 1.19. The total number of brachytherapy units is 66, with a median of 1 center with brachytherapy unit per state and 29 states with ≤3 centers with a brachytherapy unit (90.6%). Thirty-seven brachytherapy units (56.1%) have automated afterload high-dose rate. The overall rate of brachytherapy units per million inhabitants is 0.55, varying from 0 in 5 states (15.6%), 0.1-0.49 in 8 states (25%), 0.5-0.99 in 13 states (40.6%), 1-1.49 in 5 states (15.6%) and 1.5-1.99 in Mexico City (3.1%). The Mexican CMRO has 368 radiation oncologists certified (99 women and 269 men), of whom only 346 remain as an active part of Mexico's workforce. CONCLUSIONS: This is the first time the CMRO conducts a national census for a radiotherapy diagnostic situation in Mexico. The country currently holds a density of clinical and linear accelerators of 1.19 per million habitants. Brachytherapy density is 0.55 devices per million habitants, and 57% of radiotherapy centers have brachytherapy units.

Radiotherapy for the treatment of pituitary adenomas: A dosimetric comparison of three planning techniques.

AIM: Our goal was to compare conformal 3D (C3D) radiotherapy (RT), modulated intensity RT (IMRT), and volumetric modulated arc therapy (VMAT) planning techniques in treating pituitary adenomas. BACKGROUND: RT is important for managing pituitary adenomas. Treatment planning advances allow for higher radiation dosing with less risk of affecting organs at risk (OAR). MATERIALS AND METHODS: We conducted a 5-year retrospective review of patients with pituitary adenoma treated with external beam radiation therapy (C3D with flattening filter, flattening filter-free [FFF], IMRT, and VMAT). We compared dose-volume histogram data. For OARs, we recorded D2%, maximum, and mean doses. For planning target volume (PTV), we registered V95%, V107%, D95%, D98%, D50%, D2%, minimum dose, conformity index (CI), and homogeneity index (HI). RESULTS: Fifty-eight patients with pituitary adenoma were included. Target-volume coverage was acceptable for all techniques. The HI values were 0.06, IMRT; 0.07, VMAT; 0.08, C3D; and 0.09, C3D FFF (p < 0.0001). VMAT and IMRT provided the best target volume conformity (CI, 0.64 and 0.74, respectively; p < 0.0001). VMAT yielded the lowest doses to the optic pathway, lens, and cochlea. The position of the neck in extreme flexion showed that it helps in planning mainly with VMAT by allowing only one arc to be used and achieving the desired conformity, decreasing the treatment time, while allowing greater protection to the organs of risk using C3D, C3DFFF. CONCLUSIONS: Our results confirmed that EBRT in pituitary adenomas using IMRT, VMAT, C3D, C3FFF provide adequate coverage to the target. VMAT with a single arc or incomplete arc had a better compliance with desired dosimetric goals, such as target coverage and normal structures dose constraints, as well as shorter treatment time. Neck extreme flexion may have benefits in treatment planning for better preservation of organs at risk. C3D with extreme neck flexion is an appropriate treatment option when other treatment techniques are not available.

Pelvic radiation therapy with volumetric modulated arc therapy and intensity-modulated radiotherapy after renal transplant: A report of 3 cases.

AIM: Describe characteristics and outcomes of three patients treated with pelvic radiation therapy after kidney transplant. BACKGROUND: The incidence of pelvic cancers in kidney transplant (KT) recipients is rising. Currently it is the leading cause of death. Moreover, treatment is challenging because anatomical variants, comorbidities, and associated treatments, which raises the concern of using radiotherapy (RT). RT has been discouraged due to the increased risk of urethral/ureteral stricture and KT dysfunction. MATERIALS AND METHODS: We reviewed the electronic health records and digital planning system of patients treated with pelvic RT between December 2013 and December 2018 to identify patients with previous KT. CASES DESCRIPTION: We describe three successful cases of KT patients in which modern techniques allowed full standard RT for pelvic malignances (2 prostate and 1 vaginal cancer) with or without elective pelvic nodal RT, without allograft toxicity at short and long follow-up (up to 60 months). CONCLUSION: When needed, RT modern techniques remain a valid option with excellent oncologic results and acceptable toxicity. Physicians should give special considerations to accomplish all OAR dose constraints in the patient's specific setting. Recent publications recommend KT mean dose <4 Gy, but graft proximity to CTV makes this unfeasible. We present 2 cases where dose constraint was not achieved, and to a short follow-up of 20 months renal toxicity has not been documented. We recommend the lowest possible mean dose to the KT, but never compromising the CTV coverage, since morbimortality from recurrent or progressive cancer disease outweighs the risk of graft injury.

The importance of image guided radiotherapy in small cell lung cancer: Case report and review of literature.

AIM: Describe the anatomical changes and tumor displacement due to a rapid response of a patient's small cell lung cancer (SCLC) during definitive chemoradiotherapy (CRT). BACKGROUND: The treatment for SCLC is based on CRT. If interfractional changes during RT are incorrectly assessed they might compromise adequate coverage of the tumor or increase dose to organs at risk. Image guided RT with cone-beam computed tomography (CBCT) allows to identify daily treatment variations. MATERIAL AND METHODS: Describe a SCLC case with rapid changes in size, shape and location of the primary tumor during RT. CASE REPORT: A 62-year-old woman was diagnosed with SCLC with complete obstruction of the anterior and lingular bronchi and incomplete left thorax expansion due to a 12 × 15 cm mass. During CRT (45 Gy in 1.5 Gy per fraction, twice daily) the patient presented rapid tumor response, leading to resolution of bronchi obstruction and hemithorax expansion. Tumor shifted up to 4 cm from its original position. The identification of variations led to two new simulations and planning in a 3-week treatment course. CONCLUSIONS: The complete radiological response was possible due to systematic monitoring of the tumor during CRT. We recommend frequent on-site image verification. Daily CBCT should be considered with pretreatment tumor obstruction, pleural effusion, atelectasis, large volumes or radiosensitive histology that might resolve early and rapidly and could lead to a miss of the tumor or increased toxicity. Further research should be made in replanning effect in coverage of microscopic disease since it increases uncertainty in this scenario.

Reduced BUCY 2 and G-CSF-primed bone marrow associates with low graft-versus-host-disease and transplant-related mortality in allogeneic HSCT.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the ideal treatment for several diseases. However, the morbidity and mortality associated with the procedure might limit its widespread use; therefore, we implemented reduced BUCY2 as conditioning method along with the use of G-CSF-primed bone marrow (G-BM) in order to reduce complications, including graft-versus-host-disease (GVHD), and to improve survival in these patients. An analysis of transplant characteristics, complications, and survival of patients undergoing an allo-HSCT using this conditioning regimen (busulfan 12 mg/kg and cyclophosphamide 80 mg/kg) plus G-BM was performed. Forty patients were included from 1999 to 2015. All of them had a HLA-matched donor, with a median age of 32 years (range 16-59), and 55% were male. The most frequent diagnosis was myelodysplastic syndrome (MDS) in 14 patients (35%), followed by acute lymphoid leukemia (ALL) in 12 (30%). The mean of CD34+ was 2.09 × 106/kg. The mean time to neutrophil and platelet recovery was 20 and 18 days, respectively. The most common toxicity was mucositis (75%) with grade III-IV in 53% of cases. Acute GVHD appeared in 12.5 and 35% of patients developed chronic GVHD. Transplant-related mortality (TRM) was 10%. Five-year relapse-free survival was 69%, and the 5-year overall survival was 69.5%. Our conditioning method along with G-BM preserves an immunosuppressive and myeloablative effect allowing eradication of the malignant clone and achieving adequate bone marrow engraftment with acceptable toxicity, low incidence of GVHD, and low TRM, representing a favorable alternative for allo-HSCT.

Caracterización molecular de los genes pr1 y tpt1 en especies y variedades cubanas de tabaco (Nicotiana tabacum L. ) / Molecular characterization of pr1 and tpt1 proteins in cuban tobacco species and varieties (Nicoatiana tabacum L. )

La identificación de genes nuevos relacionados con la respuesta de Nicotiana tabacum L. al estrés biótico y abiótico contribuye al mejoramiento genético del cultivo del tabaco en todo el mundo. El objetivo de este trabajo fue detectar la presencia de los genes tpt1 y pr1 en el genoma de algunas especies y variedades cubanas de tabaco. Se identificaron 265 etiquetas de secuencias expresadas (ESTs-expressed sequences tags) que nunca se habían informado con anterioridad en especies vegetales, y el gen que codifica para la proteí­na tumoral controlada durante la transcripción (Transcriptional Controlled Tumor Protein) (tpt1) nunca se había informado en N. tabacum, pues los estudios del mismo se han centrado en su mayoría en animales y humanos. Los resultados del microarreglo se confirmaron utilizando la RT-PCR cuantitativa en tiempo real. Los genes tpt1 y proteí­na relacionada con la patogénesis (pr1) se utilizaron para diseñar cebadores para la caracterización molecular de algunas especies y variedades de tabaco cubano. El gen tpt1 solamente se expresó en dos especies (N. glutinosa y N. tomentosiformis) y el pr1, después de la digestión, mostró diferentes bandas entre las variedades susceptibles y resistentes. La presencia de estos genes en una parte del germoplasma analizado constituye un paso inicial para la utilización de este conocimiento en el mejoramiento genético del tabaco.

The identification of new genes related with Nicotiana tabacum L. response to biotic and abiotic stress contribute to the genetic improvement of tobacco plants around the world. The aim of this research was to identify tpt1 and pr1 genes in the genomic of some cuban tobacco varieties and species. The microarray technology has been used with ESTs for the construction of a cDNA library. 265 expressed sequences tags (ESTs) that have never been reported before in vegetables species were identified and the gene that codified for the Transcriptional Controlled Tumor Protein (tpt1), has never been reported before in N. tabacum, with the studies about this gene being focused on human and animals mostly. The microarray results were confirmed using quantitative RT- PCR. TCTP and pathogenesis-related protein 1 (PR-1) ESTs were used to design primers for the molecular characterization of some species and cuban tobacco varieties. The tpt1 gene was only expressed in two species (N. glutinosa y N. tomentosiformis) and pr1 gen, after a digestion, exhibited different bands for susceptible and resistant varieties. The detection of these genes in some species and cuban tobacco varieties represents one step to go deeply in the molecular characterization of cuban germoplasm.

Unusual Metastases in Renal Cell Carcinoma: A Single Institution Experience and Review of Literature.

BACKGROUND: To report location and management of atypical metastases from renal cell carcinoma (RCC) in the Instituto Nacional de Ciencias Medicas e Investigacion Salvador Zubiran (INCMNSZ) in Mexico City. METHODS: Between 1987 to 2009, 545 patients with RCC were retrospectively identified at the INCMNSZ. Patients with unusual metastases confirmed by histopathology were analyzed. Epidemiological, clinical, diagnosis, treatment and outcome data were reviewed. RESULTS: Sixty patients developed 98 unusual metastases secondary to RCC. The group was comprised of 35 men (58.3%), with a median age of 60 years at diagnosis. Metachronous unusual metastases with primary renal cancer were observed in 37 individuals (61.7%). Median time from primary RCC diagnosis to the first unusual metastasis was 16.5 months. Median survival from diagnosis of the first unusual metastasis to death was 5.0 months (CI 95%: 2.8-7.2 months). Patients with an initial solitary metastatic lesion in an unusual site (28.3%) had a better survival compared to patients who primarily presented with multiple metastases, 17.0 (CI 95%: 6.1-27.9) Vs 3.0 months (CI 95%: 0.9-5.1), p = 0.001. Unusual metastasis resection (21 patients) improved survival, 25.0 (CI 95%: 5.1-44.9) Vs 3.0 months (CI 95%: 0.8-5.2), p < 0.0001. No survival difference was observed between localization of unsual metastases (p = 0.72). CONCLUSIONS: In patients with advanced RCC we suggest an individual diagnostic and surgical approach to achieve complete resection with disease-free margins, even in the presence of unusual metastatic sites, multifocality, or history of metastasectomy. These strategy might provide not only palliation for symptoms, but an opportunity for meaningful disease free and overall survival.

Emergence of a new satellite RNA from cucumber mosaic virus isolate P1.

The cucumber mosaic virus (CMV) isolate P1 caused very mild symptoms on many plant species. After serial passages by mechanical inoculation over five years, CMV P1 caused severe symptoms on several tobacco cultivars and tomato. A specific band of approximately 0.3 kb in length was amplified by RT-PCR with primers synthesized based on reported CMV satellite RNA (satRNA) sequences. Sequence analysis showed there were two satRNAs (Sat-P1-1 and Sat-P1-2). Sat-P1-1 contained 335 nucleotides, and Sat-P1-2 contained 394 nucleotides. These two satRNAs shared 64% overall nucleotide sequence homology, and differences between the two satRNAs included mutations as well as deletions. Sat-P1-1 was identical to a satRNA (Z96099) reported in 1995 in CMV P1. Based on differences in the sequence and secondary structure between these two satRNAs, we conclude that Sat-P1-2 represents the emergence of a new satellite (necrotic satellite) from attenuated satRNA populations. The possible effect of the emergence of this new satRNA is discussed.