Comparative metavirome analysis in polytransfused patients.

Due to the high transfusion volume, polytransfused patients with sickle cell disease (SCD) and beta-thalassemia are constantly exposed to parenterally transmitted infections. Currently, we have little information about the virome of such patients and how the virological composition might be influenced by the hemotherapy procedures that these patients receive. The objective of this study was to compare the viral diversity between these two groups with respect to the viral abundance and how it might be affected by the specific conditions of these groups. We sequenced by next-generation sequencing (NGS) and compared the virome of 30 patients with beta-thalassemia major, 45 with SCD, and 16 blood donors from the Blood Center of Ribeirão Preto, Brazil. Predominantly, commensal viruses including Torque teno virus (TTV) genotypes and human pegiviris-1 (HPgV-1) were identified in each group. Strikingly, while HPgV-1 reads were dominant in the SCD group, thalassemic patients showed high TTV abundance, expressed both in viral reads and genotypes. We speculated that the commensal virome of polytransfused patients might be influenced by the transfusion frequency and disease characteristics and that commensal viruses might be used as important genetic biomarkers for these hematological disturbances. Nevertheless, more specific studies are necessary to confirm a relationship between blood virome and transfusion treatment.

Comparative metavirome analysis in polytransfused patients

Due to the high transfusion volume, polytransfused patients with sickle cell disease (SCD) and beta-thalassemia are constantly exposed to parenterally transmitted infections. Currently, we have little information about the virome of such patients and how the virological composition might be influenced by the hemotherapy procedures that these patients receive. The objective of this study was to compare the viral diversity between these two groups with respect to the viral abundance and how it might be affected by the specific conditions of these groups. We sequenced by next-generation sequencing (NGS) and compared the virome of 30 patients with beta-thalassemia major, 45 with SCD, and 16 blood donors from the Blood Center of Ribeirão Preto, Brazil. Predominantly, commensal viruses including Torque teno virus (TTV) genotypes and human pegiviris-1 (HPgV-1) were identified in each group. Strikingly, while HPgV-1 reads were dominant in the SCD group, thalassemic patients showed high TTV abundance, expressed both in viral reads and genotypes. We speculated that the commensal virome of polytransfused patients might be influenced by the transfusion frequency and disease characteristics and that commensal viruses might be used as important genetic biomarkers for these hematological disturbances. Nevertheless, more specific studies are necessary to confirm a relationship between blood virome and transfusion treatment.

Viral metagenomics in Brazilian multiply transfused patients with sickle cell disease as an indicator for blood transfusion safety.

BACKGROUND AND AIM: Patients with sickle cell disease (SCD) are submitted to multiple transfusions in order to increase the oxygen capacity of the blood, decrease blood viscosity, and suppress the sickling of the cells. Multiply transfused patients with SCD represent significant risk of acquiring parenterally transmitted infections. The analysis of the virome profile of high-risk multiply transfused patients with SCD can reveal the presence of parenterally transmitted viruses and therefore be used an indirect approach for evaluation of blood transfusion safety. MATERIALS AND METHODS: Blood samples were collected from 45 patients with SCD receiving multiple transfusions and analyzed by metagenomic analyses. The samples were assembled in pools f which were submitted to nucleic acids extraction and sequencing by Illumina NextSeq 550 equipment. For bioinformatic analysis, we used a specific in-house developed pipeline specialized in identification of emerging viruses. RESULTS: The virome composition of SCD patients revealed the presence of commensal viruses represented by anelloviruses and Human Pegivirus-1 (HPgV-1, GB virus C). Contaminant viral sequences belonging to human lentiviruses (rev, env genes), cytomegalovirus and murine leukemia virus were also identified and are attributed to vectors used in the laboratory practice. No novel or unsuspected pathogenic viruses were identified. CONCLUSION: This study evaluates for the first time the virome of multiply transfused patients with SCD. Exclusively genetic material of commensal viruses was annotated. Therefore, we believe that viral metagenomics applied in patients with high risk for acquiring parenterally transmitted infections can serve as a direct indicator for evaluation of transfusion safety.

Serological evidence of Borrelia circulation among blood donors in the São Paulo state, Brazil.

OBJECTIVE: The objective of this study was to examine the Borrelia seroprevalence among blood donors in Southeast Brazil. BACKGROUND: There is evidence that Borrelia spirochetes are circulating in Brazil; however, there are no studies that characterise these bacteria and investigate their seroprevalence in the Brazilian population. Such a situation, combined with a recent outbreak of tick-borne Rocky Mountain spotted fever in the São Paulo state demonstrates the increasing role of ticks as arthropod vectors in Brazil. METHODS: For the purpose of the study, 452 blood donors from Ribeirão Preto city, São Paulo state were tested using anti-Borrelia immunoglobulin G (IgG) assay. The positive results were also confirmed by Western blot for anti-borrelia IgM/IgG. RESULTS: The anti-Borrelia IgG enzyme-linked immunosorbent assay (ELISA) showed nine positive and nine borderline reactive samples, giving a total seroprevalence of 2·0% of anti-Borrelia IgG among Brazilian blood donors. The confirmation of the seropositive and borderline samples by Borrelia Western blot was demonstrated by IgG-positive results in 16 samples (a seroprevalence of 3.5%). Anti-Borrelia IgM antibodies were also detected in one sample. CONCLUSIONS: Our results demonstrate that Borrelia-like spirochetes may be circulating among blood donors from the São Paulo State and that the ticks have an important epidemiological role as vectors of bacterial infections in this Brazilian region. These results not only alert us to possible actions that might be undertaken in order to completely characterise the aetiological agents of Lyme-like syndromes in Brazil but also the possible impact that these bacterial agents might have on haemotherapy practices.

Human pegivirus-1 (HPgV-1) RNA prevalence and genotypes in volunteer blood donors from the Brazilian Amazon.

OBJECTIVES: The objectives of this study were to evaluate the prevalence of Human Pegivirus-1 (HPgV-1) viremia and genotype diversity among healthy blood donors from the Eastern Brazilian Amazon (city of Macapá, State of Amapá). There is little information for prevalence and circulation of HPgV-1 in this remote Brazilian region. MATERIALS AND METHODS: We conducted a study evaluating the HPgV-1 RNA prevalence and circulating genotypes in 431 volunteer blood donors originating from the Eastern Brazilian Amazon. The obtained HPgV-1 positive samples were submitted to sequencing and genotyping analysis in order to examine the genotype diversity of this virus in the Brazilian Amazon. RESULTS: Our results demonstrated a prevalence of HPgV-1 RNA in 9.5% of the tested blood donors. The phylogenetic analyses of the detected positive samples showed the presence of HPgV-1 genotypes 1, 2 and 3. The most frequently detected genotype was 2 (78.0% of the cases) represented by sub-genotypes 2A (39.0%) and 2B (39.0%). At lower rates, genotypes 1 (14.6%) and 3 (7.4%) were also detected. CONCLUSION: Our results revealed the presence of genotypes with European, Asiatic and African endemicity in Amazonian blood donors, probably due to the complex miscegenation processes that took place in this Brazilian region. More investigations, including information for the prevalence of HPgV-1 RNA in blood donors from other Latin American countries are needed to estimate the viremic rates and genotype distribution of this virus in a highly diverse continent like South America.

Overview of Zika virus (ZIKV) infection in regards to the Brazilian epidemic.

Zika virus (ZIKV), a mosquito-borne flavivirus, belongs to the Flaviviridae family, genus Flavivirus. ZIKV was initially isolated in 1947 from a sentinel monkey in the Zika forest, Uganda. Little clinical importance was attributed to ZIKV, once only few symptomatic cases were reported in some African and Southeast Asiatic countries. This situation changed in 2007, when a large outbreak was registered on the Yap Island, Micronesia, caused by the Asian ZIKV lineage. Between 2013 and 2014, ZIKV spread explosively and caused many outbreaks in different islands of the Southern Pacific Ocean and in 2015 autochthonous transmission was reported in Brazil. Currently, Brazil is the country with the highest number of ZIKV-positive cases in Latin America. Moreover, for the first time after the discovery of ZIKV, the Brazilian scientists are studying the possibility for the virus to cause severe congenital infection related to microcephaly and serious birth defects due to the time-spatial coincidence of the alarming increase of newborns with microcephaly and the Brazilian ZIKV epidemic. The present review summarizes recent information for ZIKV epidemiology, clinical picture, transmission, diagnosis and the consequences of this emerging virus in Brazil.

Overview of Zika virus (ZIKV) infection in regards to the Brazilian epidemic

Zika virus (ZIKV), a mosquito-borne flavivirus, belongs to the Flaviviridae family, genus Flavivirus. ZIKV was initially isolated in 1947 from a sentinel monkey in the Zika forest, Uganda. Little clinical importance was attributed to ZIKV, once only few symptomatic cases were reported in some African and Southeast Asiatic countries. This situation changed in 2007, when a large outbreak was registered on the Yap Island, Micronesia, caused by the Asian ZIKV lineage. Between 2013 and 2014, ZIKV spread explosively and caused many outbreaks in different islands of the Southern Pacific Ocean and in 2015 autochthonous transmission was reported in Brazil. Currently, Brazil is the country with the highest number of ZIKV-positive cases in Latin America. Moreover, for the first time after the discovery of ZIKV, the Brazilian scientists are studying the possibility for the virus to cause severe congenital infection related to microcephaly and serious birth defects due to the time-spatial coincidence of the alarming increase of newborns with microcephaly and the Brazilian ZIKV epidemic. The present review summarizes recent information for ZIKV epidemiology, clinical picture, transmission, diagnosis and the consequences of this emerging virus in Brazil.

Late emergence of A594V and L595W mutations related to ganciclovir resistance in a patient with HCMV retinitis and long-term HIV progression.

The emergence of ganciclovir (GCV) resistance during the treatment of human cytomegalovirus (HCMV) infection is a serious clinical challenge, and is associated with high morbidity and mortality. In this case report, we describe the emergence of two consecutive mutations (A594V and L595W) related to GCV resistance in a patient with HCMV retinitis and long-term HIV progression after approximately 240 days of GCV use. Following the diagnosis of retinitis, the introduction of GCV did not result in viral load reduction. The detected mutations appeared late in the treatment, and we propose that other factors (high initial HCMV load, previous GCV exposure, low CD4+ cell count), in addition to the presence of resistance mutations, may have contributed to the treatment failure of HCMV infection in this patient.

[Problems and neonatal outcome of very low birth weight newborn infants after in vitro fertilization].

UNLABELLED: The use of assisted reproduction technologies is undoubtedly successful in the treatment of sterility. However it sets up numerous of issues for the obstetricians and neonatologists. AIMS: To evaluate the incidence, the specific problems and the neonatal outcome of newborns with very low birth weight (VLBW) <1500 g born from pregnancies after in vitro fertilization (IVF). METHODS: The study enrolled all 563 VLBW infants admitted in the NICU of the "Maichin Dom" hospital from 01.01.2008 to 30.06.2010. 119 (21.1%) of them were conceived with assisted reproduction technology (IVF- group), and 444 (78.9%) were conceived naturally (control group). All infants were followed up till their discharge home or death. Poor outcome measures were in hospital neonatal death or morbidities with long term sequels: severe congenital malformations, bronchopulmonary dysplasia, severe brain injuries (intraventricular haemorrhages gr. Ill-IV periventricular leucomalacia), retinopathy of prematurity gr. Ill-V. RESULTS: There were no significant differences in terms of mean birth weight (BW) and gestational age (GA) between the groups (1170 g and 1173 g, 29,8 and 30,0 weeks of gestation respectively). Intrauterine growth retardation (BW of <10 percentile for GA) was observed in 42% in the IVF-group, versus 38.5% (NS) The frequency of the babies from multiple pregnancies was significantly higher in the IVF-group: 88.2% versus 27.5%, and the triplets were 48% versus only 0.9% in the control group. In the IVF-babies more active obstetric approach was carried out: caesarean section in 85% versus 57%, and completed antenatal corticosteroid course in 80% versus 41% in the control group. There were no significant differences of in hospital neonatal mortality rate - 14.3% in the IVF-group versus 14.9%; congenital malformations or severe morbidities at discharge - 22.7% versus 27.5%, discharged in good health - 63% versus 57.6%. CONCLUSIONS: The major problems of VLBW-newborns after IVF result from the higher incidence of multiple pregnancies with their corresponding risks. Nevertheless, strict pregnancy follow-up, more frequently use of antenatal steroids, cesarean delivery such as intensive neonatal resuscitation ensure a clinical outcome and prognosis which do not differ from the naturally conceived VLBW-newborns.

[A case of cervical myoma and pregnancy].

We are presenting a case of women in 31 gestational week of her pregnancy with prematurely ruptured membranes and 10.5 cm myoma in the vagina. The cervix was elevated with well-defined external orifice, dilated to 10.5 cm. C-section with myomectomy was performed. The myoma had penetrated in the vagina with a lot of adhesions to the rectum and cavum Douglais. The vaginal vault and the posterior cervical wall were restored with single sutures. Smooth postoperative period.

[Pregnancy outcome after excision therapy of cervical precancerous lesions].

Since inception, screening programs that use Papanicolaou smear tests have decreased the incidence of cervical cancer up to 50% in the developed countries. The unprecedented success of this program hinges not only on the sensitivity of the Papanicolaou smear test, but also on the ability to eliminate successfully the precancerous lesions that are detected by the screening test. Cervical precancerous lesions are encountered most commonly among women of reproductive age; a peak incidence occurs among women in their twenties. Because of the 5-12% chance of progression to squamous cell cancer, management guidelines recommend aggressive treatment for women with moderate-to-severe lesions. Because many women in this age group have not yet completed childbearing at the time of diagnosis, treatment for these cervical abnormalities has potentially significant reproductive consequences. This review will summarize a part of the published literature regarding the effects of excision therapies of the uterine cervix and the risk of spontaneous abortion and preterm delivery (PTD) in future pregnancies and provide some reasonable treatment recommendations for women with cervical dysplasia.

[Laterally extended parametrectomy in cases with cervical carcinoma].

INTRODUCTION: Clinical staging of cervical carcinoma is not always an easy task, especially in cases with parametrial involvement, stage IIB. Therapeutic approach may differ in such patients- conservative (radio-, chemoradiation) therapy versus operative treatment. Research shows that the limits of the typical class III-IV radical hysterectomy may prove to be inadequate in relation to the dissection of the lateral parametria. A new surgical technique called laterally extended parametrectomy was published in 1993 and provides the means of total removal of the connective and lymphatic tissue up to the level of the true pelvic sidewall. MATERIAL AND METHODS: A retro- and prospective study was performed during the period between Jan 2008 and Feb 2011. A total of 17 radical hysterectomies with unilateral LEP were enrolled. Median age of patients was 51 years. Mean duration of surgery is 3 hours 57 min. Laterally extended parametrectomy was initiated in cases with clinically predetermined stage IB to II/IA cervical cancer. 3 patients were diagnosed with adenosquamous type of carcinoma, while the other 14 were with squamous cell cervical cancer. RESULTS: Positive iliac lymph nodes were found bilaterally in 7 patients, unilaterally also in 7 patients, while 3 cases proved to be negative. Paraaortic lymph node dissection was performed in 6 surgeries, with positive histology in 4 cases. Parametrial infiltration was found in 6 patients, 3 of them bilateral. 8 of the patients had unilateral hydroureter. CONCLUSION: Laterally extended parametrectomy is a surgical technique which provides the means for full removal of the lymphatics and connective tissue up to the pelvic sidewall, during radical hysterectomy in patients with cervical cancer, stage IB and IIA, with positive iliac lymph nodes, as well as in stage IIB cases. It offers an efficient alternative for surgical treatment of locally advanced cervical cancer, opposed to the other therapeutic approaches.

[Solifenacin in treatment of the overactive bladder syndrome--diagnosis, clinical management and results].

UNLABELLED: The overactive bladder syndrome (OAB) is a pathological condition that affects millions of people round the world. Its incidence increases with ageing. The main therapeutic option for OAB nowadays is anti-muscarinic drug therapy. AIM: To define and implement the diagnostic criteria, clinical guidelines in our country and to evaluate the results of Solifenacin therapy in patients with OAB. MATERIAL AND METHODS: Along a three years period (2006-2009) 163 OAB patients were analyzed followed and treated. In our group of patients 106 (65.2%) are females and 57 (34.8%)--males. The diagnosis is based mainly on the complaints of the patient, evaluated thoroughly with a detail anamnesis and questionnaires. Urodynamic studies were performed In some of the patients. The efficacy of Solifenacin therapy has been validated with the same methods used in SUNRISE and VENUS studies. RESULTS AND DISCUSSION: The mean age of female patients in our group is 63.8 years, and 65.6 years for males. The most commonly observed symptoms of OAB are urgency--88.3% and frequency--92%. Urge-incontinence is observed in 48.8% of the cases. Therapy with Solifenacin 5 and 10 mg has alleviated urgency in 82% and urge-incontinence in 88.9%. Mean number of pads used for 24 hours has decreased from 2.6 to 0.4 after therapy. CONCLUSION: Urgency is the main mandatory symptom for the diagnosis OAB. Therapy with Solifenacin 5 and 10 mg guarantees very good clinical results, and high degree of compliance of the patients.

The role of micro-ribonucleic acids in normal hematopoiesis and leukemic T-lymphogenesis

Micro-ribonucleic acids (microRNAs) are small molecules containing 20-23 nucleotides. Despite their small size, it is likely that almost every cellular process is regulated by them. Moreover, aberrant microRNA expression has been involved in the development of various diseases, including cancer. Although many data are available about the role of microRNAs in various lymphoproliferative disorders, their impact on the development of acute lymphoblastic leukemia of T-cell progenitors is largely unknown. In this review, we present recent information about how specific microRNAs are expressed and regulated during malignant T-lymphopoiesis and about their role during normal hematopoiesis.

The role of micro-ribonucleic acids in normal hematopoiesis and leukemic T-lymphogenesis.

Micro-ribonucleic acids (microRNAs) are small molecules containing 20-23 nucleotides. Despite their small size, it is likely that almost every cellular process is regulated by them. Moreover, aberrant microRNA expression has been involved in the development of various diseases, including cancer. Although many data are available about the role of microRNAs in various lymphoproliferative disorders, their impact on the development of acute lymphoblastic leukemia of T-cell progenitors is largely unknown. In this review, we present recent information about how specific microRNAs are expressed and regulated during malignant T-lymphopoiesis and about their role during normal hematopoiesis.

Chlorsulfuron resistant transgenic tobacco as a tool for broomrape control.

Broomrape (Orobanche ramosa L.) is the most important parasitic plant that infests tobacco (Nicotiana tabacum L.). Chemical treatment of the soil is not effective and crop rotation is not acceptable to solve this problem because of the long viability period of Orobanche seeds in the soil. Application of systemic herbicides in the field with herbicide resistant tobacco could be a successful tool for broomrape control. Several tobacco cultivars were transformed with a mutant ahas3R gene for resistance to the herbicide chlorsulfuron (Glean, DuPont). Transformed plants were selfed and the segregation of resistance was followed in the next generation. The efficiency of the herbicide was demonstrated in greenhouse and field trials. An Orobanche/tobacco growth system was used in order to prove the lethal effect of the herbicide to the attached broomrape plants.

[A case of advanced genital tuberculosis--differential diagnostics and surgical aspects].

The authors report a case of advanced genital tuberculosis. The diagnostic difficulties and treatment are discussed.