Generation and persistence of S1 IgG and neutralizing antibodies in post-COVID-19 patients.

BACKGROUND: Antibody-mediated immune response plays an important role in protection against reinfection. In the case of SARS-CoV-2 infection, the maximum duration of antibody response is still unknown. In this work, the generation of neutralizing  antibodies (NAbs) and IgG antibodies against the S1 subunit (S1 IgG ) of SARS-CoV-2 and their possible duration were determined through decay models. METHODS: 132 participants with SARS-CoV-2 infection were classified according to the severity of the disease. Seroconversion and persistence of S1 IgG antibodies and NAbs were determined by ELISA, samples were taken at two different times post-infection and duration of those antibodies was estimated using Linear Mixed Models (LMMs). RESULTS: The highest amount of S1 IgGs antibodies was associated with age (41 years or older), greater severity of COVID-19 and male gender. NAbs production was associated with the same variables, except for age. The percentage of NAbs decay is higher in the asymptomatic group (P = 0.033), while in S1 IgG antibodies decay, no statistical difference was found between the 4 severity groups. An exponential decay model was built by using a LMM and similarly, two dispersion regions where constructed. The duration of S1 IgG antibodies was 744 days (668-781) for first region and 744 days (453-1231) for the second. Regarding NAbs, an adaptative LMM was used to model a logistic function, determining a duration of 267 days (215-347). CONCLUSION: Humoral immunity to SARS-CoV-2 infection depends on the severity of the disease, gender and age. This immune response could be long-lasting as for other coronaviruses.

Genetic diversity and drug susceptibility of Mycobacterium tuberculosis in a city with a high prevalence of drug resistant tuberculosis from Southeast of Mexico.

BACKGROUND: Mexico is on the top five countries with the highest number of TB cases in America continent, nevertheless, information about genotypes circulating is practically unknown. Considering the above this study aims to characterize the genetic diversity of TB in the city of Veracruz, México. METHODS: A cross-sectional study was conducted among positive smear samples from patients living in Veracruz City, samples were cultured, and first-line drug profiles determined. Genotyping was made by spoligotyping and MIRU-VNTR 24 loci. Associations of lineages, clusters, and variables were also analyzed. RESULTS: Among the 202 isolates analyzed resistance to at least one drug was observed in 60 (30%) isolates and 41(20%) were multidrug-resistant. Three major lineages were identified: L4/Euro-American (88%), L1/Indo-Oceanic (9%), and L2/East Asian (3%). The Euro-American lineage included more than six sublineages, the most abundant were: H (32%), T (23%), LAM (18%), and X (12%). 140 isolates (70%) were placed in 42 SITs patterns. CONCLUSIONS: These results provide the first baseline data on the genetic structure of TB in the city of Veracruz. Sublineages H, X and LAM were predominant; however, it was founded an important diversity of genotypes that could contribute to the dispersion of TB and explain the high prevalence. This information might be useful for the development of further interventions to reduce impact of TB.

The knockdown of eIF4AI interferes with the respiratory syncytial virus replication cycle.

The respiratory syncytial virus (RSV) is one of the main etiological agents in acute respiratory infections. To date, the replicative cycle of this virus is not completely known, and the events as well as the role of cellular and viral proteins that participate in the infectious cycle of RSV are still a matter of intense research. An important protein that is a control point for many viruses is the helicase eIF4AI, which participates at the beginning of the cap-dependent translation of eukaryotes and cap-independent translation of certain viral mRNAs. Recently, eIF4AI has been considered as a potential viral therapeutic target. In order to understand the role of eIF4AI during the infectious cycle of RSV, we evaluated the effect of eIF4AI knockdown on the amount of positive-strand viral RNA and viral progeny of this virus. Our results showed a decrease for both parameters, suggesting a possible involvement of eIF4AI during replicative cycle of RSV. In addition, using confocal microscopy, it was observed that eIF4AI colocalized with RSV viral protein, supporting the possible participation of eIF4AI during the replicative cycle of RSV. Keywords: eIF4AI; RSV; translation; antiviral.

Activity of matrix metalloproteinase 2 and 9 isoforms in sputum samples from individuals infected with M. tuberculosis.

M. tuberculosis stimulates matrix metalloproteinases secretion in the host; however, the patterns of matrix metalloproteinase 9 (MMP-9) and 2 (MMP-2) isoforms are unknown. Therefore, the aim of this study was to evaluate the activities of these isoforms in sputum samples of patients with varying degrees of mycobacterial load. Sputum samples were recovered from individuals with positive and negative diagnosis of tuberculosis. Bacillary load was determined by bacilloscopy, and presence and activity of MMP-2 and MMP-9 isoforms determined by gelatin zymography. Of the recovered samples; 39 were from individuals without tuberculosis and 48 with tuberculosis; 12 had low bacillary and 36 high bacillary load. Participation of MMP-2 isoforms were not observed, but in MMP-9, there was a decrease in the latent, associated with lipocalin and homodimeric isoforms. In contrast, there was a rise in the active isoform, all with statistical significance. In conclusion our results show that MMP-2 isoforms do not seems to be directly involved with tuberculosis evolution of infection. However, variations in MMP-9 isoforms were observed, which coincided with the progression of tuberculosis infection. These results raise questions regarding how MMP-9 isoforms might influence the formation and progression of granuloma, and their use as markers of progression of tuberculosis infection.

Eukaryotic initiation factor 4A (eIF4A) during viral infections.

The helicase eIF4A is part of the cellular eIF4F translation initiation complex. The main functions of eIF4A are to remove secondary complex structures within the 5'-untranslated region and to displace proteins attached to mRNA. As intracellular parasites, viruses regulate the processes involved in protein synthesis, and different mechanisms related to controlling translation factors, such as eIF4A, have been found. The inhibitors of this factor are currently known; these substances could be used in the near future as part of antiviral pharmacological therapies in instances of replication cycles in which eIF4A is required. In this review, the particularities of how some viruses make use of this initiation factor to synthesize their proteins are discussed.

Risk factors associated with gastric cancer in Mexico: education, breakfast and chili.

BACKGROUND AND AIM: the aim of the study was to use a validated questionnaire to identify factors associated with the development of gastric cancer (GC) in the Mexican population. METHODS: the study included cases and controls that were paired by sex and ± 10 years of age at diagnosis. In relation to cases, 46 patients with a confirmed histopathological diagnosis of adenocarcinoma-type GC, as reported in the hospital records, were selected, and 46 blood bank donors from the same hospital were included as controls. The previously validated Questionnaire to Find Factors Associated with Gastric Cancer (QUFA-GC©) was used to collect data. Odds ratio (OR) and 95% confidence interval (IC) were estimated via univariate analysis (paired OR). Multivariate analysis was performed by logistic regression. A decision tree was constructed using the J48 algorithm. RESULTS: an association was found by univariate analysis between GC risk and a lack of formal education, having smoked for ≥ 10 years, eating rapidly, consuming very hot food and drinks, a non-suitable breakfast within two hours of waking, pickled food and capsaicin. In contrast, a protective association against GC was found with taking recreational exercise and consuming fresh fruit and vegetables. No association was found between the development of GC and having an income that reflected poverty, using a refrigerator, perception of the omission of breakfast and time period of alcoholism. In the final multivariate analysis model, having no formal education (OR = 17.47, 95% CI = 5.17-76.69), consuming a non-suitable breakfast within two hours of waking (OR = 8.99, 95% CI = 2.85-35.50) and the consumption of capsaicin ˃ 29.9 mg capsaicin per day (OR = 3.77, 95% CI = 1.21-13.11) were factors associated with GC. CONCLUSIONS: an association was found by multivariate analysis between the presence of GC and education, type of breakfast and the consumption of capsaicin. These variables are susceptible to intervention and can be identified via the QUFA-GC

Prevalence of respiratory viruses in wheezing children not older than 24 months of age.

INTRODUCTION: Wheezing in children not older than 24 months of age is a frequent event, and viruses are usually the causative agents. The aim of the study was to estimate the prevalence of respiratory viruses in wheezing children who were not older than 24 months of age and who had no history of asthma. METHODS: Fifty-five Mexican children were included in an analytical cross-sectional study. Nasal secretions were obtained by using sterile rayon-tipped applicators to identify the virus by polymerase chain reaction or reverse transcription-polymerase chain reaction: adenovirus, bocavirus, human rhinovirus, influenza virus type A, human metapneumovirus, parainfluenza, rhinovirus, and respiratory syncytial virus. The prevalence of viral etiology was estimated by dividing the frequency of the identified virus by the number of participants. Ninety-five percent confidence intervals for proportions were calculated. RESULTS: Most of the patients were male (35/55, 63.6%). The average time of evolution of wheezing episode was 3 days. The third part of enrolled population were receiving antibiotics. Respiratory viruses were detected in 33 (60%; 95% CI: 46.8-71.9%) out of 55 cases, and viral coinfection was detected in five cases (9.1%; 95% CI: 3.5-19.9%). Human metapneumovirus was the most frequently identified virus (23.6%), followed by bocavirus (14.5%), respiratory syncytial virus and rhinovirus (12.7% each), and to a lesser extent influenza virus type A and parainfluenza. Rhinovirus was the predominant virus in outpatient children (28.6%). In the inpatient emergency room and inhalotherapy room, human metapneumovirus predominated (41.2 and 16.1%, respectively). CONCLUSION: bocavirus and human metapneumovirus were the most frequently identified viruses in Mexican children who were < 2 years of age, suffered from wheezing, and had no history of asthma.

4EBP1 Is Dephosphorylated by Respiratory Syncytial Virus Infection.

Respiratory syncytial virus (RSV) requires protein biosynthesis machinery to generate progeny. There is evidence that RSV might alter some translation components since stress granules are formed in their host cells. Consistent with these observations, we found that RSV induces dephosphorylation of 4EBP1 (eIF4E-binding protein), an important cellular translation factor. Our results show no correlation between the 4EBP1 dephosphorylation time and the decrease in the global rate of protein synthesis. Interestingly, treatment with rapamycin stimulates virus generation. The results suggest that RSV is a virus that still contains unknown mechanisms involved in the translation of their mRNAs through the alteration or modification of some translation factors, such as 4EBP1, possibly to favor its replicative cycle.

Annexins A2 and A5 are potential early biomarkers of hepatocarcinogenesis.

Hepatocellular carcinoma (HCC) is a highly lethal liver cancer with late diagnosis; therefore, the identification of new early biomarkers could help reduce mortality. We determine the tissue and plasma status of five annexins during hepatocarcinogenesis by diethylnitrosamine-induced cirrhosis-HCC. We found that Anxa5 was the earliest upregulated gene at week 12 after HCC initiation, while Anxa1 and Anxa2 were upregulated in advanced HCC stages (weeks 18 and 22). Furthermore, the protein level of Annexin A1, A2, A5 and A10 was increased from the early stages. Immunofluorescence and subcellular fractionation revealed Annexin A1, A2, and A5 in the cytoplasm and nuclei of tumor cells. Notably, increased plasma levels of Annexin A5 significantly (r2 = 0.8203) correlated with Annexin A5 levels in liver tissue from week 12 and gradually increased until week 22. Using the TCGA database, we found that the expression of ANXA2 (HR = 1.7, p = 0.0046) and ANXA5 (HR = 1.8, p = 0.00077) was associated with poor survival in HCC patients. In conclusion, we have identified Annexin A1 and A5 as potentially useful early biomarkers for poor prognosis in HCC patients.

[Review of new evidence about the possible vertical transmission of coronavirus disease-2019]. / Revisión de nuevas evidencias acerca de la posible transmisión vertical de la COVID-19.

OBJECTIVE: To conduct a systematic review of original peer-reviewed studies, containing data on the identification of SARS-CoV-2 in clinical samples of amniotic fluid, placenta or membranes, umbilical cord blood, and human milk, from women with a clinically or confirmed diagnosis of COVID-19. These studies should have been published after the guide for the management of patients with COVID-19 from World Health Organization guide (available in March 13, 2020). RESULTS: Seventeen studies were included, in which 143 clinical samples were identified (38 of amniotic fluid; 34 of placentas or membranes; 39 from umbilical cord blood and 32 from human milk). Among the 143 samples, nine were positive for SARS-CoV-2 RNA (one amniotic fluid sample obtained before rupturing the membranes; six samples of placenta or membranes, although authors indicate the possibility of contamination by maternal blood in three of these, and two samples of human milk). CONCLUSIONS: Following our search criteria, we found no studies that demonstrate the detection of SARS-CoV-2, in conjunction with viral isolation and the evaluation of the infective capacity of viral particles, in clinical samples of amniotic fluid, placenta or membranes, umbilical cord blood and human milk, from women with a confirmed or clinical diagnosis of COVID-19. However, vertical transmission cannot be ruled out, larger studies are required that ideally locate in situ RNA and protein of SARS-CoV-2, as well as isolation that demonstrate the infective capacity of the viral particles.

Whole genome characterization, and geographical distribution of M. tuberculosis in central region of Veracruz, Mexico.

The purpose of this work was to perform by Whole Genomic Sequencing (WGS) a characterization of tuberculosis isolates circulating in the central region of Veracruz, Mexico, and to determine its geographical distribution. The genome of 25 clinical isolates of tuberculosis patients, recovered from central zone of Veracruz, Mexico, were sequenced and the information obtained was used to characterize lineage, prediction of drug resistance, identification of clonal complexes, and finally correlated with the geolocalization data. Isolates analyzed were included into seven L4 sublineages, most frequent was X3; X1 (4.1.1.3) in 35%. rpoBSer450Leu polymorphism was the most frequently found variant. Sublineage Haarlem (4.1.2) had the widest distribution, found in five municipalities. Of the of two clonal complexes found, the most abundant included eight isolates, with X3/X1 lineage, placed in two municipalities. Combination of WGS and geographic information system was very useful for the identification of sublineages, clonal complexes, and their geographical dispersion with important implications in the epidemiological surveillance and clinical control of TB.

Influence of skin-to-skin contact on breastfeeding: results of the Mexican National Survey of Demographic Dynamics, 2018.

BACKGROUND: Skin-to-skin contact and breastfeeding initiation within the first hour after birth are key recommendations to promote breastfeeding. In Mexico, the National Survey of Demographic Dynamics 2018, known by its Spanish acronym ENADID, collected information about breastfeeding practices. The ENADID survey is probabilistic and allows results to be generalized to the entire population in Mexico. METHODS: Information from a public database featuring 26,587 mother-baby pairs was analyzed by proportions, means and associations, as well as machine learning methods, to conduct a comparison among the pairs according to immediate skin-to-skin contact after delivery status. RESULTS: Skin-to-skin contact was described by 78.7% of the mothers and was associated with receiving an explanation regarding how to give breastmilk or the breast to the baby immediately following birth [Odds ratio (OR) 6.46; 95% Confidence Interval (CI) 6.02, 6.97], initiating breastfeeding in the first hour of life (OR 2.01; 95% CI (1.84, 2.18) and a breastfeeding duration of ≥ 6 months (OR 1.16; 95% CI 1.08, 1.25). The breastfeeding duration, in days, was greater in the group with skin-to-skin contact than in the group without skin contact. CONCLUSIONS: In Mexico, immediate and uninterrupted skin-to-skin contact between newborns and their mothers should be facilitated. Support should be provided to mothers to favor skin-to-skin contact and breasting initiation during the first hour of life, ideally through an empathic explanation by trained health personnel. Future research should focus on the evaluation of strategies to modify maternity services to facilitate immediate skin-to-skin contact after delivery and develop training programs for health personnel to support the initiation of breastfeeding during the first hour of life.

SNPs in Genes Related to DNA Damage Repair in Mycobacterium Tuberculosis: Their Association with Type 2 Diabetes Mellitus and Drug Resistance.

Genes related to DNA damage repair in Mycobacterium tuberculosis are critical for survival and genomic diversification. The aim of this study is to compare the presence of SNPs in genes related to DNA damage repair in sensitive and drug-resistant M. tuberculosis genomes isolated from patients with and without type 2 diabetes mellitus (T2DM). We collected 399 M. tuberculosis L4 genomes from several public repositories; 224 genomes belonging to hosts without T2DM, of which 123 (54.9%) had drug sensitive tuberculosis (TB) and 101 (45.1%) had drug resistance (DR)-TB; and 175 genomes from individuals with T2DM, of which 100 (57.1%) had drug sensitive TB and 75 (42.9%) had DR-TB. The presence of SNPs in the coding regions of 65 genes related to DNA damage repair was analyzed and compared with the resistance profile and the presence/absence of T2DM in the host. The results show the phylogenetic relationships of some SNPS and L4 sub-lineages, as well as differences in the distribution of SNPs present in DNA damage repair-related genes related to the resistance profile of the infecting strain and the presence of T2DM in the host. Given these differences, it was possible to generate two discriminant functions to distinguish between drug sensitive and drug resistant genomes, as well as patients with or without T2DM.

Evaluation of Anomalies and Neurodevelopment in Children Exposed to ZIKV during Pregnancy.

Zika virus (ZIKV) infection in pregnancy is associated with birth and developmental alterations in infants. In this study, clinical records of 47 infants whose mothers had Zika during pregnancy or clinical manifestations compatible with Zika were reviewed. A description of the infants' anomalies was established, and a neurodevelopmental assessment was performed on 18 infants, using the Evaluation of Infant Development (EDI for its initialism in Spanish) and DDST-II (Denver Developmental Screening Test II) tests. From his sample, 74.5% of the infants evaluated had major anomalies and 51.9% had minor anomalies. The incidence of major anomalies, related to trimester of pregnancy, was 84.2% for the first trimester, 77.8% for the second trimester, and 37.5% in the third trimester. A similar trend was observed in the frequency of infants without anomalies and was less evident in the incidence of minor anomalies (p = 0.016). Through neurodevelopmental assessments, EDI identified 27.8% of infants as having normal development, while 55.5% of affected infants had developmental delay, and 16.7% were at risk for developmental delay. The DDSST-II showed that 77.7% infants had delay in the gross motor and language area, 88.8% in the fine-adaptative motor area, and 72.2% in the personal-social area. In this work, children of mothers with ZIKV infection during pregnancy may have major or minor anomalies regardless of the trimester of pregnancy in which the infection occurred. The neurodevelopmental assessment shows that ZIKV can cause a developmental delay in infants with the fine-adaptative motor area being the most affected.

IgG Antibodies Generation and Side Effects Caused by Ad5-nCoV Vaccine (CanSino Biologics) and BNT162b2 Vaccine (Pfizer/BioNTech) among Mexican Population.

SARS-CoV-2 has rapidly generated a pandemic. Vaccines are currently being rolled out to control the viral spread and prevent deaths. Emergency vaccines, using new platforms, have been approved. Their effectiveness, safety and immunogenicity in different populations are not fully known. This study aimed to discover the immunogenicity of the messenger ribonucleic acid (mRNA) BNT162b2 and adenovirus vector Ad5-nCoV vaccines through IgG antibody generation against subunit 1 of protein S (S1 IgG) and assess the side effects of the vaccines. A total of 115 vaccinated people were included, 61 of whom received the BNT162b2 vaccine, while 54 received Ad5-nCoV. Measurements of S1 IgG antibodies were carried out using the enzyme-linked immunosorbent assay (ELISA) technique. The BNT162b2 vaccine generated S1 IgG antibodies in 80.3% of the participants after the first dose. The number of seropositive participants increased to 98.36% with the administration of the second dose. The Ad5-nCoV vaccine generated S1 IgG antibodies in 88.89% of those vaccinated. Women generated more antibodies when administered either vaccine. There were no serious adverse effects from vaccination. In conclusion, not all participants had detectable S1 IgG antibodies. The Ad5-nCoV vaccine presented the most seronegative cases. The studied vaccines were shown to be safe.

Potential Protection of Pre-Existent Antibodies to Human Coronavirus 229E on COVID-19 Severity.

The causes of the broad spectrum of severity in COVID-19 are unknown. A protective effect through humoral immunity from previous infections by viruses of the SARS-CoV-2 family could explain a mild form of this disease. This study aimed to address whether the presence of antibodies against human seasonal coronaviruses (HCoVs) could prevent severe manifestations of COVID-19. A cross-sectional study was carried out in 165 participants. The presence of pre-existent antibodies against the seasonal HCoV-OC43, HCoV-HKU1, HCoV-229E, and HCoV-NL63 were detected. From all of the seasonal HCoVs studied, it was only found that being seropositive to HCoV-229E presented an association (p = 0.012) with developing mild clinical symptoms of COVID-19 or being asymptomatic. Multinomial regression analysis showed that being seropositive to HCoV-229E is associated with mild or moderate clinical symptoms for COVID-19. Statistical analysis also showed that being female is associated with being asymptomatic for SARS-CoV-2 infection or developing mild COVID-19. A subgroup analysis taking only seropositive to HCoV-229E revealed that females are more likely to develop asymptomatic SARS-CoV-2 infection (OR = 27.242, 95% CI 2.092-354.706, p = 0.012). Our results suggest that previous infections by HCoV-229E could prevent more serious clinical manifestations of COVID-19, but these are not the only variables that influence this event.

Seroprevalence of Trypanosoma cruzi Infection in Pregnant Women Suggests a High Risk for Congenital Transmission in Central Veracruz, Mexico.

PURPOSE: The state of Veracruz, Mexico, is a well-recognized endemic region for Chagas disease, but congenital transmission has not been extensively studied. We estimated here the prevalence and the risk of congenital transmission of Trypanosoma cruzi in pregnant women from 27 municipalities of central Veracruz. METHODS: 528 sera from pregnant women were analyzed by ELISA and IFA assays for the detection of IgG antibodies against T. cruzi. RESULTS: The presence of anti-T. cruzi antibodies was identified in women from 17 municipalities, obtaining an overall seroprevalence of 17.0%. A higher seropositivity was observed in the municipalities of Orizaba (25.2%), Nogales (13.6%), and Río Blanco (10.5%). The results suggest that there is a high risk of congenital transmission of T. cruzi in the region. CONCLUSION: There are currently limited actions for the surveillance and control of congenital transmission of Chagas disease in Veracruz.

Rotavirus infection induces the phosphorylation of eIF2alpha but prevents the formation of stress granules.

Early during the infection process, rotavirus causes the shutoff of cell protein synthesis, with the nonstructural viral protein NSP3 playing a vital role in the phenomenon. In this work, we have found that the translation initiation factor 2alpha (eIF2alpha) in infected cells becomes phosphorylated early after virus infection and remains in this state throughout the virus replication cycle, leading to a further inhibition of cell protein synthesis. Under these restrictive conditions, however, the viral proteins and some cellular proteins are efficiently translated. The phosphorylation of eIF2alpha was shown to depend on the synthesis of three viral proteins, VP2, NSP2, and NSP5, since in cells in which the expression of any of these three proteins was knocked down by RNA interference, the translation factor was not phosphorylated. The modification of this factor is, however, not needed for the replication of the virus, since mutant cells that produce a nonphosphorylatable eIF2alpha sustained virus replication as efficiently as wild-type cells. In uninfected cells, the phosphorylation of eIF2alpha induces the formation of stress granules, aggregates of stalled translation complexes that prevent the translation of mRNAs. In rotavirus-infected cells, even though eIF2alpha is phosphorylated these granules are not formed, suggesting that the virus prevents the assembly of these structures to allow the translation of its mRNAs. Under these conditions, some of the cellular proteins that form part of these structures were found to change their intracellular localization, with some of them having dramatic changes, like the poly(A) binding protein, which relocates from the cytoplasm to the nucleus in infected cells, a relocation that depends on the viral protein NSP3.

Breastfeeding in the time of Zika: a systematic literature review.

BACKGROUND: The disease Zika is considered as emergent. The infection can be acquired through different routes: a bite from the Aedes mosquito, sexual contact, from mother to child during pregnancy and by blood transfusion. The possibility of Zika transmission through human lactation has been considered. Zika is a disease of great concern for public health because it has been associated with neonatal and postnatal microcephaly, among other birth defects. OBJECTIVES: To review published evidence of the probable transmission of Zika through human lactation. DATA SOURCES: Electronic databases: Cochrane Central Register of Controlled Trials, EBSCO, Gale, Science Direct, Scopus, US National Library of Medicine (PubMed) and Web of Science. World Health Organization and Centers for Disease Control and Prevention web pages. STUDY ELIGIBILITY CRITERIA: To be eligible, studies of any design had to provide primary data of human breast milk as a potential fluid for the transmission of Zika, or primary or secondary follow-up data of infants with at least one previous published study that complied with the first criterion of eligibility. PARTICIPANTS: Studies about women with suspected, probable or confirmed Zika during pregnancy, or the postnatal period and beyond. Studies about infants who breastfeed directly from the breast or where fed with the expressed breast milk of the suspected, probable or confirmed women with Zika. RESULTS: This study only chose data from research papers; no patients were taken directly by the authors. A total of 1,146 were screened and nine studies were included in the qualitative synthesis, from which a total of 10 cases were identified, with documented follow-up in three of these cases. Through the timing of maternal Zika infection, five cases were classified as prenatal (time before delivery), one as immediate postnatal (period from 0 to 4 days after birth); no cases were classified as medium postnatal (period from 5 days to 8 weeks after birth); two were classified as long postnatal (period from 8 weeks to 6 months after birth) and two as beyond six months after birth. CONCLUSION: Human milk may be considered as a potentially infectious fluid, but we found no currently documented studies of the long-term complications in infants up to 32 months of age, with suspected, probable or confirmed Zika through human lactation, or evidence with respect to the human pathophysiology of the infection acquired through human lactation. In the light of the studies reviewed here, the World Health Organization recommendation of June 29th 2016, remains valid: "the benefits of breastfeeding for the infant and mother outweigh any potential risk of Zika virus transmission through breast milk."

Sequencing of the entire rpob gene and characterization of mutations in isolates of Mycobacterium tuberculosis circulating in an endemic tuberculosis setting.

OBJECTIVE: To evaluate the use of a sequencing procedure for the entire rpoB gene of Mycobacterium tuberculosis to identify mutations pre-rifampicin resistance determining region (RRDR), within RRDR, and post-RRDR in isolates circulating in a region affected by tuberculosis (TB). METHODS: Five primers were designed, with which five DNA fragments of rpoB were obtained, sequenced by Sanger, and analysed in silico in order to identify mutations over the entire rpoB gene in rifampicin-sensitive and rifampicin-resistant TB. RESULTS: It was possible to analyse the entire rpoB gene in five rifampicin-sensitive and 15 rifampicin-resistant isolates. Thirty-six mutations were identified. Two mutations were found pre-RRDR, nine within-RRDR and 25 post-RRDR. The most frequent mutations within RRDR were S531L (53%), followed by S512T (20%), all of which were found in rifampicin-resistant isolates. Of the 25 mutations found post-RRDR, 14 were only in resistant isolates, and the most frequent was D853N, which was present in 85% of isolates. Mutations E818K, D836N and T882P were observed in 80% of the rifampicin-resistant and rifampicin-sensitive isolates. CONCLUSIONS: The proposed sequencing method allowed identification of mutations in the entire rpoB gene. This procedure represents a useful tool for diagnosing rifampicin resistance. The number of mutations that were found raises new questions about the diversity of mutations in the rpoB gene and their role in rifampicin resistance in regions where TB is endemic.