Table of Contents | Genet. Mol. Res. 2021 (1)
Strategies for cervical cancer screening in the scope of the Brazilian Unified National Health System
Cervical cancer (CC) is the third most diagnosed malignancy among women in Latin America. CC control focuses on prevention methods: primary prevention is carried out to prevent Human Papillomavirus (HPV) infections, such as vaccination, while secondary prevention methods aim to detect and treat cervical premalignant lesions. In addition, cytology is used for triage, and many countries, including Brazil, still use it for this purpose. However, this method has significant limitations, such as low sensitivity and poor reproducibility. Some molecular assays are relevant, such as HPV testing. In this pilot study, a micro-costing analysis was done to compare cytology and HPV testing for cervical cancer screening in the Brazilian Unified Health System. For this assessment, a cross-sectional study was performed involving 500 women aged 18–55 years old without cervical precancer or cancer. The participants underwent cervical cytology and HPV testing. The overall prevalence of HPV infection was 25.2%. Among these HPV-infected women, 13.6% presented normal or inflammatory, cytology and were HPV positive by molecular assay. Most of these patients were infected with high-risk-HPV 16 and 18 and were asymptomatic. Oncotic cytology was the most cost-effective in the comparison between these two screening strategies. However, the molecular assay had a higher sensitivity and specificity when compared with cytology. HPV testing, despite the higher costs, may still be economically advantageous for CC control programs as it could reduce the cost of CC treatment in the public health system. In conclusion, although HPV testing is currently more expensive than cytology assay, its use would still be justified as it can reduce the economic and social impact due to cases not detected by cytology.
Safflower genetic diversity based on agronomic characteristics in Mato Grosso state, Brazil, for a crop improvement program
Safflower, Carthamus tinctorius (Asteraceae), is an oilseed plant with good adaptability to warm and dry climatic conditions. It is used for biodiesel production, human food, animal feed, and in the pharmaceutical industry. Recently, the crop has been highlighted, mainly for its oil quality and for biofuel production. We compared 124 safflower genotypes, which are a part of the State University of Mato Grosso germplasm collection, based on their agronomic characteristics, to provide the initial guidelines for a breeding program. Evaluations were carried out during the crop cycle and parameters were defined according to the descriptions recommended by International Board for Plant Genetic Resources and by the Ministry of Agriculture, Livestock and Supply. Multivariate analysis was used to assess the divergence among genotypes, by using Average Euclidean Distance, which ranged from 0.07 to 0.57, showing considerable genetic diversity among safflower genotypes for the agronomic characteristics that were evaluated (flowering, plant cycle, number of branches per plant, plant height, number of chapters per plant, number of seeds per chapter, chapter diameter, stem diameter, weight of 100 seeds, seed size and plant yield. Tocher and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) clustering methods were partially consistent in ordering similar genotypes. The agronomic characteristics: plant yield, number of chapters per plant and plant height provided the greatest contribution to genetic divergence among the genotypes. According to the groupings established with each methodology and depending on the variation structures within each group, several genotypes stood out in terms of agronomic performance and may be indicated for future crosses aiming to obtain improved safflower cultivars for Brazil.
Various theories have been developed regarding the pathology and etiology of hypertensive disorders of pregnancy (HDP). The impaired placenta secretes antiangiogenic factors into maternal blood, resulting in the development of pregnancy-induced hypertension (PIH). Both µ-calpain and m-calpain are proteins known to be involved in placentation, with the catalytic subunits encoded by the CAPN1 and CAPN2 genes, respectively. The purpose of this study was to find disease susceptibility genes for PIH by conducting an association analysis for HDP using specific gene markers comprising individual single nucleotide variants (SNVs) and haplotypes within the CAPN2 gene. We selected five SNVs in the human CAPN2 gene and performed an association study with 95 HDP patients and 177 age-matched non-HDP subjects. In the analyses conducted for the CAPN2 SNVs, the recessive model of rs1153968 differed significantly between the gestational hypertension (GH, a category of HDP) and control groups. However, rs1153968 deviated from Hardy-Weinberg equilibrium and was deemed unlikely to be a significant factor. No significant differences were seen in any of the other SNVs analyzed. However, association analyses involving the rs1892077-rs98041140-rs17599-rs1153954 haplotype in the CAPN2 gene haplotypes revealed that G-A-A-T was found at a significantly lower frequency, and G-A-C-T and G-G-A-A at significantly greater frequencies, in patients with PIH. These findings may be useful in preventing PIH onset, as well as for the early discovery and treatment of patients who are carriers of the haplotype likely to be associated with this condition.
Resolving a human identification case for the Rio de Janeiro Police with massively parallel sequencing of mtDNA using a proposed pipeline
The forensic community is concerned about the quality control of mitotypes reported, as probability estimates may be biased to the disadvantage of a suspect when there is incomplete data. This is particularly important when using new technology such as Massively Parallel Sequencing (MPS), which performs simultaneous analysis of millions of DNA fragments, generating vast amounts of data from small amounts of sample, providing more extensive genetic information. The use of MPS for complex forensic evidence analysis could save time and resources, and provide reliable data for extremely degraded DNA samples. However, data analysis methods can be challenging to implement in a forensic labs routine. Here, we propose a reliable and straightforward pipeline for mtDNA data analysis in forensic samples using MPS, and we used this pipeline to solve a human identification case by Rio de Janeiro Police who had recovered a severely putrefied body that could not be identified by conventional autosomal STR analysis. Mitochondrial DNA (mtDNA) from samples were sequenced on Ion Torrent PGM. Raw data from the sequencing was analyzed in two main pipeline protocols: quality control and forensic analysis. The first includes the analyses of raw data and contamination control. The second generates haplotypes, allowing haplogroup classification, and biogeographic inference, as well as verification of a matrilineal relationship. This straightforward freeware pipeline for mtDNA data analysis should facilitate the implementation of this type of system in forensic lab routine. Using our pipeline, the definition of the samples being of African origin was more precise when considering the whole mtGenome and not only the control region, which is usually used for routine forensic analysis. The final analysis was consistent with the existence of a matrilineal relationship between the alleged son (bone sample recovered from the sea) and the mother.
Evaluation of resistance to Cowpea aphid-borne mosaic virus in passion fruit backcrosses for recurrent selection and development of resistant cultivars
The cultivation of passion fruit occupies an important place in Brazilian fruit culture; however, there have been successive declines in production. The main cause of this retraction in production is an increase in the incidence of Cowpea aphid-borne mosaic virus. This virus can cause severe deformation of the fruit and make passion fruit production inviable. Due to a lack of cultivars resistant to this virus, the Universidade Estadual do Norte Fluminense Darcy Ribeiro passion fruit breeding program has sought to develop resistant cultivars. The objective of this work was to evaluate the resistance to CABMV of a passion fruit second generation backcross (BC2) segregating population via the REML/BLUP procedure aiming at selecting resistant genotypes with agronomic characteristics to start a recurrent selection program and possibly produce a new passion fruit cultivar. Virus resistance was measured by the average area under the disease progress curve using a score scale, evaluating the percentage of symptoms in the plant and young leaves. The total fruit mass produced per plant was used to assess fruit production. Based on this information, the components of variance and genetic value were estimated via REML/BLUP. The highest values estimated for genetic parameters, for the two characteristics evaluated, were based on the average of each family. The BC2 - 17 family showed the best additive genetic gain concerning resistance to CABMV and the worst for fruit production. The BC2 - 293 family, had the highest estimated value of genetic gain for fruit production. The genetic variability found in the BC2 segregating population allows us to select superior genotypes. Twenty-nine genotypes were selected to start the recurrent selection program aimed at resistance to CABMV. The - Two genotypes (BC2 - 89 and BC2 - 323) showed potential to be launched as CABMV resistant passion fruit cultivars.
The pigeonpea is an important grain legume in various regions of the world. It is a pulse that is cultivated mainly by small farmers in semi-arid tropical regions for various purposes, including food security, livestock feed, and agroforestry systems. We examined the genetic diversity and structure of 73 pigeonpea accessions from 10 origins, including regions of Brazil, Central America, and India. Allelic diversity and polymorphism, molecular analysis of variance (AMOVA), Bayesian analysis (Structure) and a dendrogram were estimated based on data concerning 11 microsatellite loci. AMOVA revealed wide genetic differentiation among the pigeonpea populations, with FST ranging from 0.22 to 0.25. Expected heterozygosity in the accessions was greater than the observed heterozygosity in eight of the 11 loci, with the observed heterozygosity ranging from 0.06 to 0.55. Polymorphic information content ranged from 0.13 to 0.67 among the 11 loci. Genetic similarity among pigeonpea accessions ranged from 0.26 to 0.93, indicating considerable variability among them, with the formation of three groups, one of which included all Brazilian accessions. Two groups were formed in the Structure analysis, indicating reduced allele sharing between the populations of Northeast Brazil and other populations. This information highlights the importance of incorporating germplasm from the Brazilian Northeast into pigeonpea breeding programs in Brazil, to increase the variability in germplasm collections.
Prognosis of sinonasal squamous cell carcinoma (SNSCC) depends on TNM (Tumor, Node, Metastasis) staging, histological manifestations, and activity of cell-surface receptors (p53, Ki67, EGFR − Epidermal Growth Factor Receptor (ErbB-1; Her1). EGFR is the best- studied transmembrane tyrosine kinase receptor. The intensity of tumor cells division and the rate of their metastasis depend on EGFR. It also regulates the processes of angiogenesis. All this determines the severity of the course of the disease and the ability to predict its outcome. Depending on the speed of EGFR expression, targeted therapy may or may not be prescribed. The clinical picture and prognosis of SNSCC depends on the rate of EGFR expression. We conducted a prospective case study on 32 SNSCC patients treated at the National Ear Nose and Throat Hospital during two years (2011-2012). Clinical presentations and computed tomography scan imaging were analyzed and biopsy samples were examined to investigate EGFR activity (Her1). A semi quantification technique was used. The level of EGFR expression was correlated with disease staging. The rate of positive EGFR expression in SNSCC was 17/32; 8/32 scored 3+. EGFR was positive in 16/32 stage III patients. The EGFR-positive rate was higher in exophytic, advanced stage cancer, with neurological deficits and lymphadenopathy of the neck. EGFR expression was increased in 17/32 of sinonasal squamous cell carcinoma samples. Patients with more severe manifestations, including neurological symptoms, facial deformities and exophytic tumor growth had a significantly higher EGFR expression rate.
We evaluated the results of laparoscopic nephrectomy for the treatment of renal tumors in a provincial hospital. Subjects and methods: 30 patients were diagnosed with renal tumors and underwent laparoscopic nephrectomy at a General Hospital, in Vietnam. The mean age was 53.23 years, the rate of incidental detection of tumors was 8/30. Average tumor size was 5.33cm. Angiomyolipoma was found int 7/30 and renal cell carcinoma (RCC) in 23/30. The transperitoneal approach was used in 13/30. Most (21/30) patients had one renal artery and one renal vein. The mean operative time was 116 minutes, the average blood loss 161 ml, the drainage time five days and the hospital stay six days. Operative complications: minor hemorrhage 2/30; major haemorrhage 1/30. Postoperative complications: 3/30. Conversion to open surgery 1/30. Most (18/30) patients were examined after six months; one case had liver metastases. Laparoscopic nephrectomy (LRN) in the treatment of renal tumors is a safe, effective method and can be done in provincial hospitals. However, in order to make a more accurate conclusion, there is a need for large-scale studies with a longer follow-up time.
Laparoscopic partial nephrectomy in the treatment of renal tumors in Thanh Hoa General Hospital, Vietnam
We evaluated the initial results of laparoscopic partial nephrectomy (LPN) in the treatment of renal tumors in Thanh Hoa General Hospital. During examination, 10 patients (6 females, 4 males) were diagnosed with renal tumors on CT scans. Later they underwent LPN at Thanh Hoa General Hospital from November 2016 to March 2018. The mean age was 54.5 years. Average tumor size was 3.5 cm. There were seven patients in the pT1a stage and 3 in the pT1b stage. The R.E.N.A.L. nephrometry scoring: two patients in 4a; two patients in 4p; three patients in 5a; two patients in 6p; and one patient in 8x. Average operative time was 125.6 minutes. Average warm-ischemic time was 29.4 minutes. Blood loss average was 85.2 mL. Hemorrhagic complication in one patient. Convert to laparoscopic radical nephrectomy in one patient. Patients were followed for one to three months without complications, metastases, or local recurrence.
Results of 300 сases of mini - percutaneous nephrolithotomy at the Thanh Hoa General Hospital in Vietnam
We evaluated the results of mini–percutaneous nephrolithotomy (mini-PCNL) under ultrasonic guidance for treatment of kidney stones at Thanh Hoa General Hospital. This descriptive study examined, from July 2016 to September 2018, 300 cases of kidney stones performed mini-PCNL under ultrasonic guidance. Nephrolithotripsy was done by 30 and 100w Homium lasers. The 300 patients included 63.3% men and 36.7% women. The mean age was 42.13 ± 33.4 years. Recurrent stones were found in 15%, renal pelvis and one calyx stone in 51.7%. Hydronephrosis before surgery at level 1 was found in 46.7%. Stone average length was 3.11 ± 0.63cm and average area was 5.16 ± 2.41cm2. Tunnel position in the middle calyx was found in 68.3%. Average surgery time averaged 58.42 ± 27.8 minutes. Bleeding during surgery was reported for 15 patients, hematuria after surgery in two patients, and fever after surgery in 46 patients. The average hospitalization duration was 4.6 ± 2.3 days. Early stones clearance rate was 78.3%. Mini–PCNL under ultrasound guidance with a 30 – 100w Holmium Laser energy source is a useful method of treating kidney stones safely, achieving an early stone clearance efficiency of 82.4%,and a low rate of postoperative complications of 16.7%. This method can effectively replace open surgery for the treatment of kidney stones.
Evaluation of the effectiveness of arthroscopic rotator cuff repair at the Thong Nhat Hospital in Vietnam
We evaluated the results of arthroscopic rotator cuff repair. Thirty cases of rotator cuff tear were treated by arthroscopic rotator cuff repair at the Thong Nhat hospital from 1/3/2012 to 30/6/2016. All patients were over 18 years old, had positive rotator cuff tear tests, and underwent X-ray and magnetic resonance imaging. The arthroscopic images showed rotator cuff tears, with indication for repair. The functional status of each patient was evaluated using the Constant-Murley Scale and the criteria of the University of California at Los Angeles (UCLA). During postoperative rehabilitation, we applied Cohen's protocol. Seventeen patients were female and 13 were male. The median age was 54 years (21–83). Sixteen patients had partial thickness tears and 14 patients had full-thickness tears. Repair technique: single-row in 13 patients, suture-bridge in 15 patients, transosseous-equivalent in two patients. Four patients with SLAP lesions were treated by debridement labrum for type I, and labral repair for more serious lesions. Five patients with long head biceps tendon rupture were treated with tenodesis. After surgery, 100% of patients were subjected to fluid extravasation through the shoulder joint, but did not have any compartment syndrome; all incisions healed good without infection or neurapraxia. The mean UCLA score was 32.4 (14 patients had excellent results, 14 good, and two fair). The mean Constant score was 38.5; the mean postoperative Constant score was 88.77 (p = 0.001). Among the patients, 90% achieved 80% function compared to a normal shoulder. All patients gave their consent to the study and were prepared for the full course of treatment and arthroscopic surgery. Repair of the rotator cuff was for some participants the only effective treatment and a way to restore the shoulder. Postoperative reconstruction of the rotator cuff of the shoulder resulted in a significant improvement in the indicators of the functional state of the shoulder joint.
We assessed the effectiveness of Aclasta for osteoporosis in patients with femoral head fractures at Thong Nhat Hospital in Ho Chi Minh City. A total of 156 patients with femoral head fractures and osteoporosis in the orthopedic department at Thong Nhat in Ho Chi Minh City were assigned to a randomized controlled trial from 01 Jan 2013 to 01 Jan 2014, divided into two groups: 75 patients were treated with Aclasta; 81 patients received a placebo. Most of them were followed for at least one year for new bone fractures and T-score index before and one year after treatment. New fractures were found at a rate of about 8% in the Aclasta group and 13.6% in the placebo group, which was significantly different (P < 0.001). Vertebral fractures were different in Aclasta (1.3%) and Placebo groups (3.7%), and the incidence of femoral head fracture was 2.6 and 3.7%. T-score improved significantly after one year in the Aclasta group; the proportion of patients with a T-score in this group changed due to bone deficiency 33.3 and 8%, while in the placebo group, it was 7.4 and 0%; this difference was significant (p <0.001). Using Aclasta for treatment of osteoporosis after femoral head fracture surgery reduces the incidence of new fractures and gives T-score improvement in patients with femoral head fractures, especially in elderly patients.
We evaluated the results of treatment of meniscus tear injury by arthroscopy to determine the influence of age, gender and cause of injury in patients undergoing endoscopic suturing. This was a prospective study conducted from 7/2006 to 7/2013. We followed 90 patients, from 17 to 50 years old attended at Thong Nhat hospital (Ho Chi Minh City). All of them were diagnosed with a torn meniscus and were treated with arthroscopic meniscus suturing by outside-in, inside-out and all inside techniques. Early results, including evaluating the side effects during surgery and after surgery and results after 6 months were collected. Among the 90 patients in our study, men accounted for 62%, almost two times higher than percentage of women. The main reason was due to sports injuries, accounting for 65%. The most common age was between 21 and 30 (46.7%). Results after surgery: 32/90 were very good, 52/90 (good, 4/90 medium, and 2/90 bad. The indication of suturing meniscus tear due to trauma gives superior results for knee function for patients, reducing long-term complications such as osteoarthritis.
Stroke is one of the leading causes of death both globally and in Southeast Asia, which is confirmed by high values: 5 million and 500 thousand deaths annually, respectively. The situation in Vietnam is also critical. We have systematized and studied the risk factors in patients with ischemic stroke in Phu Tho Province General Hospital. This was a prospective cohort study of 192 patients. Transient ischemic attack was described in 13% of all cases, and the risk of progression to severe disease or death had an odds ratio (OR) = 5.2; 63.5% of all patients with hypertension had a risk of severe complications or death with OR 3.2. Previous cardiovascular events were found during the history of the disease in 28.1% of patients. Hyperlipidemia had a significant prevalence among the patients; the risk of exacerbation was OR = 2.6. Tobacco smoking and alcohol abuse also had a high prevalence among patients (72.6%). The risk of death among smokers and alcohol abusers was 2.4 times higher compared with patients without bad habits. Ischemic cerebrovascular accident is one of the most common health problems in Vietnam and needs more attention.
We investigated independent predictors of acute ischemic stroke (AIS) in older elderly (>75 years) patients in comparison with those of the elderly (from 60-75 years) in Vietnam. Data are for 308 consecutive AIS patients aged ≥ 60 years (138 aged >75 years) admitted to the stroke unit of Phu Tho Hospital from (2014-2017). An assessment of symptoms was also carried out using the NIH Stroke Scale (NIHSS), symptoms, instrumental and laboratory parameters during hospitalization and complications of the underlying disease. The existing predictors of hospital death, trauma, and length of hospital stay were also taken into account. When evaluating the parameters, logistic regression was used. Risk profiles before and after age 75 were compared. Age is a proven, unmodifiable, universal predisposing risk factor for stroke. Poor outcomes were more frequent in the oldest (>75 years) compared to the younger patients (≤75 years). NIHSS score and clinical parameters of AIS severity, including need for oxygen, indwelling catheter, or nasogastric tube), disability incidents, and medical complications predicted most of the outcomes in both age groups. After age 75, AIS etiology and primary clinical signs and symptoms were additional independent determinants for most outcomes, along with age, sex, and pre-stroke functional and health statusThe key factors were mechanical ventilation and pneumonia, and pre-stroke signs during the first three days in the hospital for AIS in the oldest- old (>75 years).; in patients ≤75 years, atrial fibrillation was associated with poor outcomes and death at discharge.