Streptococcus pneumoniae serotype 19A in hospitalized children with invasive pneumococcal disease after the introduction of conjugated vaccines in Lima, Peru.

BACKGROUND: The Pneumococcal conjugate vaccine (PCV) has decreased cases of invasive pneumococcal disease (IPD) worldwide. However, the impact of PCVs introduction may be affected by the serotype distribution in a specific context. METHODS: Cross-sectional multicenter passive surveillance study of IPD cases in pediatric patients hospitalized in Lima, Peru between 2016 and 2019 (after PCV13 introduction) to determine the serotype distribution and antimicrobial resistance of Streptococcus pneumoniae. Serotyping was performed by a sequential multiplex PCR and confirmed by whole genome sequencing. RESULTS: Eighty-five S. pneumoniae isolates were recovered (4.07/100,000 among children <60 months of age). Serotype 19A was the most common (49.4%). Children infected with serotype 19A in comparison with children infected with other serotypes were younger, had a lower rate of meningitis and higher rates of pneumonia, complicated pneumonia and antimicrobial resistance; 28.6% of patients with serotype 19A have received at least one dose of PCV13 vs. 62.8% of patients with other serotypes. Using MIC-breakpoints, 81.2% (56/69) of non-meningitis strains and 31.2% (5/16) of meningitis strains were susceptible to penicillin; 18.8% (3/16) of meningitis strains had intermediate resistance to ceftriaxone. Resistance to azithromycin was 78.8% (67/85). Serotype 19A frequency increased over time in the same study population, from 4.2% (4/96) in 2006-2008, to 8.6% (5/58) in 2009-2011, to 49.4% (42/85) in the current study (2016-2019) (p < 0.001). CONCLUSIONS: After PCV13 introduction in Peru, serotype 19A remains the most prevalent; however, the vaccination coverage is still not optimal. Therefore, additonal surveillance studies are needed to determine the remaining IPD burden.

Risks of low vaccination coverage and strategies to prevent the resurgence of vaccine-preventable diseases in infants in the COVID-19 pandemic scenario: recommendations for Latin America and the Caribbean by the group of experts on infant immunization for Latin America.

INTRODUCTION: The WHO 2030 Immunization Agenda (IA-2030) harmonizes immunization activity plans at community, national, regional and global levels. Additionally, medical societies play an important role. The Latin American Group of Experts on Infant Immunization, established in 2018, advises on the harmonization, update, and optimization of infant vaccination programs in Latin America and the Caribbean (LAC). In September 2021, 41 such experts from 13 LAC countries met to develop recommendations for increasing regional vaccination coverage to avoid the reemergence of vaccine-preventable diseases and/or the occurrence of outbreaks. AREAS COVERED: The following items were evaluated: (i) immunization challenges before and during the COVID-19 pandemic; (ii) the status of current immunization programs, particularly infant pertussis and polio vaccination; (iii) possible solutions for overcoming vaccination challenges and achieving regional vaccination coverage targets. EXPERT OPINION/COMMENTARY: Medical societies provide valuable recommendations to guide and update vaccination schedules. In the LAC region, possible strategies to achieve target vaccination rates include the use of combination vaccines, strengthening surveillance systems, improving school attendance, advancing vaccine education and confidence, striving for vaccination equity, widening operational capacity, creating strategic alliances, and strengthening the role of medical groups. It is hoped that these recommendations will be implemented in the LAC region.

Pneumococcal serotypes and antibiotic resistance in healthy carriage children after introduction of PCV13 in Lima, Peru.

OBJECTIVE: To determinate the frequency of Streptococcus pneumoniae nasopharyngeal carriers, serotypes and antimicrobial resistance in healthy children in Lima, Peru, post-PCV13 introduction and to compare the results with a similar study conducted between 2006 and 2008 before PCV7 introduction (pre-PCV7). METHODS: A cross-sectional multicenter study was conducted between January 2018 and August 2019 in 1000 healthy children under two years of age. We use standard microbiological methods to determinate S. pneumoniae from nasopharyngeal swab, Kirby Bauer and minimum inhibitory concentration methods to determinate antimicrobial susceptibility and whole genomic sequencing to determinate pneumococcal serotypes. RESULTS: The pneumococcal carriage rate was 20.8 % vs. 31.1 % in pre-PCV7 (p < 0.001). The most frequent serotypes were 15C, 19A and 6C (12.4 %, 10.9 % and 10.9 % respectively). The carriage of PCV13 serotypes after PCV13 introduction decreased from 59.1 % (before PCV7 introduction) to 18.7 % (p < 0.001). Penicillin resistance was 75.5 %, TMP/SMX 75.5 % and azithromycin 50.0 %, using disk diffusion. Penicillin resistance rates using MIC breakpoint for meningitis (MIC ≥ 0.12) increased from 60.4 % to 74.5 % (p = 0.001). CONCLUSION: The introduction of PCV13 in the immunization program in Peru has decreased the pneumococcal nasopharyngeal carriage and the frequency of PCV13 serotypes; however, there has been an increase in non-PCV13 serotypes and antimicrobial resistance.

Recommendations for modernizing infant vaccination schedules with combination vaccines in Colombia and Peru.

The objective of this article was to consider the vaccination challenges in Colombia and Peru and the role of pediatric combination vaccines in overcoming these challenges. Barriers to including new vaccines with more antigens remain apparent in parts of these countries, where vaccine-preventable diseases in infants continue to be a major problem. The challenges include the heterogeneity of vaccine coverage within each country and in neighboring countries, which can contribute to poor rates of vaccination coverage; the adverse impact of the inward migration of unvaccinated individuals, which has favored the re-emergence of vaccine-preventable diseases; vaccine shortages; and the impact of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic and the associated shifts in health care resources. To improve the coverage of pediatric vaccines in Colombia and Peru, it will be necessary to ensure the widespread integration into vaccine schedules of combination vaccines containing diphtheria, tetanus, acellular pertussis, inactivated poliovirus, Haemophilus influenzae type b and hepatitis B antigens with a three-dose primary series delivered at 2, 4 and 6 months of age followed by a booster at 18 months of age. Such vaccines play important roles in preventing diphtheria, tetanus and pertussis; eradicating polio; and providing boosting against H. influenzae type b.

El objetivo de este artículo es considerar los desafíos que se enfrentan en Colombia y Perú con respecto a la vacunación y el papel de las vacunas combinadas pediátricas para superar estos desafíos. Los obstáculos para incluir vacunas nuevas con más antígenos siguen siendo evidentes en algunos lugares de estos países, donde las enfermedades prevenibles por vacunación en menores de 1 año continúan siendo un grave problema. Entre los desafíos se incluye la heterogeneidad de la cobertura de vacunación en cada país y en los países vecinos, lo que puede contribuir con que se registren tasas bajas de cobertura de vacunación; el impacto adverso de la migración interna de personas no vacunadas, lo que ha favorecido la reaparición de enfermedades prevenibles por vacunación; la escasez de vacunas, y el impacto de la pandemia del coronavirus de tipo 2 causante del síndrome respiratorio agudo grave (SARS-CoV-2) y los consiguientes cambios en los recursos de atención médica. Para mejorar la cobertura de las vacunas pediátricas en Colombia y Perú será necesario integrar de manera generalizada en los calendarios de vacunación vacunas combinadas con antígenos de difteria, tétanos, tos ferina acelular, poliovirus inactivados, Haemophilus influenzae tipo b y hepatitis B con una serie primaria de tres dosis administradas a los 2, 4 y 6 meses de edad, seguida de un refuerzo a los 18 meses de edad. Esas vacunas desempeñan un papel esencial en la prevención de la difteria, el tétanos y la tos ferina; la erradicación de la polio; y el refuerzo contra H. influenzae tipo b.

O objetivo deste artigo foi avaliar os desafios da vacinação na Colômbia e no Peru e o papel das vacinas pediátricas combinadas na superação de tais desafios. Os obstáculos para incluir novas vacinas com mais antígenos permanecem visíveis em partes desses países, onde doenças imunopreveníveis em lactentes continuam a ser um grande problema. Os desafios incluem a heterogeneidade da cobertura vacinal dentro de cada país e nos países vizinhos, o que pode contribuir para baixas taxas de cobertura vacinal; o impacto adverso da migração interna de pessoas não vacinadas, o que favoreceu o ressurgimento de doenças imunopreveníveis; a escassez de vacinas; e o impacto da pandemia de síndrome respiratória aguda grave do coronavírus 2 (SARS-CoV-2) e mudanças relacionadas nos recursos de atenção à saúde. Para melhorar a cobertura das vacinas pediátricas na Colômbia e no Peru, será necessário assegurar sua integração generalizada em esquemas de vacinas combinadas contendo antígenos de difteria, tétano, pertussis acelular, poliovírus inativado, Haemophilus influenzae tipo B e hepatite B, com uma série primária de três doses aplicadas aos 2, 4 e 6 meses de idade seguidas de um reforço aos 18 meses de idade. Tais vacinas desempenham papéis importantes na prevenção da difteria, tétano e coqueluche; na erradicação da poliomielite; e no reforço contra H. influenzae tipo b.

Recommendations for modernizing infant vaccination schedules with combination vaccines in Colombia and Peru

[ABSTRACT]. The objective of this article was to consider the vaccination challenges in Colombia and Peru and the role of pediatric combination vaccines in overcoming these challenges. Barriers to including new vaccines with more antigens remain apparent in parts of these countries, where vaccine-preventable diseases in infants continue to be a major problem. The challenges include the heterogeneity of vaccine coverage within each country and in neighboring countries, which can contribute to poor rates of vaccination coverage; the adverse impact of the inward migration of unvaccinated individuals, which has favored the re-emergence of vaccine-preventable diseases; vaccine shortages; and the impact of the severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2) pandemic and the associated shifts in health care resources. To improve the coverage of pediatric vaccines in Colombia and Peru, it will be necessary to ensure the widespread integration into vaccine sched- ules of combination vaccines containing diphtheria, tetanus, acellular pertussis, inactivated poliovirus, Haemophilus influenzae type b and hepatitis B antigens with a three-dose primary series delivered at 2, 4 and 6 months of age followed by a booster at 18 months of age. Such vaccines play important roles in preventing diphtheria, tetanus and pertussis; eradicating polio; and providing boosting against H. influenzae type b.

[RESUMEN]. El objetivo de este artículo es considerar los desafíos que se enfrentan en Colombia y Perú con respecto a la vacunación y el papel de las vacunas combinadas pediátricas para superar estos desafíos. Los obstáculos para incluir vacunas nuevas con más antígenos siguen siendo evidentes en algunos lugares de estos países, donde las enfermedades prevenibles por vacunación en menores de 1 año continúan siendo un grave pro- blema. Entre los desafíos se incluye la heterogeneidad de la cobertura de vacunación en cada país y en los países vecinos, lo que puede contribuir con que se registren tasas bajas de cobertura de vacunación; el impacto adverso de la migración interna de personas no vacunadas, lo que ha favorecido la reaparición de enfermedades prevenibles por vacunación; la escasez de vacunas, y el impacto de la pandemia del corona- virus de tipo 2 causante del síndrome respiratorio agudo grave (SARS-CoV-2) y los consiguientes cambios en los recursos de atención médica. Para mejorar la cobertura de las vacunas pediátricas en Colombia y Perú será necesario integrar de manera generalizada en los calendarios de vacunación vacunas combinadas con antígenos de difteria, tétanos, tos ferina acelular, poliovirus inactivados, Haemophilus influenzae tipo b y hepatitis B con una serie primaria de tres dosis administradas a los 2, 4 y 6 meses de edad, seguida de un refuerzo a los 18 meses de edad. Esas vacunas desempeñan un papel esencial en la prevención de la difteria, el tétanos y la tos ferina; la erradicación de la polio; y el refuerzo contra H. influenzae tipo b.

[RESUMO]. O objetivo deste artigo foi avaliar os desafios da vacinação na Colômbia e no Peru e o papel das vacinas pediátricas combinadas na superação de tais desafios. Os obstáculos para incluir novas vacinas com mais antígenos permanecem visíveis em partes desses países, onde doenças imunopreveníveis em lactentes con- tinuam a ser um grande problema. Os desafios incluem a heterogeneidade da cobertura vacinal dentro de cada país e nos países vizinhos, o que pode contribuir para baixas taxas de cobertura vacinal; o impacto adverso da migração interna de pessoas não vacinadas, o que favoreceu o ressurgimento de doenças imu- nopreveníveis; a escassez de vacinas; e o impacto da pandemia de síndrome respiratória aguda grave do coronavírus 2 (SARS-CoV-2) e mudanças relacionadas nos recursos de atenção à saúde. Para melhorar a cobertura das vacinas pediátricas na Colômbia e no Peru, será necessário assegurar sua integração gen- eralizada em esquemas de vacinas combinadas contendo antígenos de difteria, tétano, pertussis acelular, poliovírus inativado, Haemophilus influenzae tipo B e hepatite B, com uma série primária de três doses aplica- das aos 2, 4 e 6 meses de idade seguidas de um reforço aos 18 meses de idade. Tais vacinas desempenham papéis importantes na prevenção da difteria, tétano e coqueluche; na erradicação da poliomielite; e no reforço contra H. influenzae tipo b.

Recommendations for modernizing infant vaccination schedules with combination vaccines in Colombia and Peru / Recomendaciones para modernizar los calendarios de vacunación infantil con vacunas combinadas en Colombia y Perú / Recomendações para modernizar os esquemas de vacinação infantil com vacinas combinadas na Colômbia e no Peru

ABSTRACT The objective of this article was to consider the vaccination challenges in Colombia and Peru and the role of pediatric combination vaccines in overcoming these challenges. Barriers to including new vaccines with more antigens remain apparent in parts of these countries, where vaccine-preventable diseases in infants continue to be a major problem. The challenges include the heterogeneity of vaccine coverage within each country and in neighboring countries, which can contribute to poor rates of vaccination coverage; the adverse impact of the inward migration of unvaccinated individuals, which has favored the re-emergence of vaccine-preventable diseases; vaccine shortages; and the impact of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic and the associated shifts in health care resources. To improve the coverage of pediatric vaccines in Colombia and Peru, it will be necessary to ensure the widespread integration into vaccine schedules of combination vaccines containing diphtheria, tetanus, acellular pertussis, inactivated poliovirus, Haemophilus influenzae type b and hepatitis B antigens with a three-dose primary series delivered at 2, 4 and 6 months of age followed by a booster at 18 months of age. Such vaccines play important roles in preventing diphtheria, tetanus and pertussis; eradicating polio; and providing boosting against H. influenzae type b.

RESUMEN El objetivo de este artículo es considerar los desafíos que se enfrentan en Colombia y Perú con respecto a la vacunación y el papel de las vacunas combinadas pediátricas para superar estos desafíos. Los obstáculos para incluir vacunas nuevas con más antígenos siguen siendo evidentes en algunos lugares de estos países, donde las enfermedades prevenibles por vacunación en menores de 1 año continúan siendo un grave problema. Entre los desafíos se incluye la heterogeneidad de la cobertura de vacunación en cada país y en los países vecinos, lo que puede contribuir con que se registren tasas bajas de cobertura de vacunación; el impacto adverso de la migración interna de personas no vacunadas, lo que ha favorecido la reaparición de enfermedades prevenibles por vacunación; la escasez de vacunas, y el impacto de la pandemia del coronavirus de tipo 2 causante del síndrome respiratorio agudo grave (SARS-CoV-2) y los consiguientes cambios en los recursos de atención médica. Para mejorar la cobertura de las vacunas pediátricas en Colombia y Perú será necesario integrar de manera generalizada en los calendarios de vacunación vacunas combinadas con antígenos de difteria, tétanos, tos ferina acelular, poliovirus inactivados, Haemophilus influenzae tipo b y hepatitis B con una serie primaria de tres dosis administradas a los 2, 4 y 6 meses de edad, seguida de un refuerzo a los 18 meses de edad. Esas vacunas desempeñan un papel esencial en la prevención de la difteria, el tétanos y la tos ferina; la erradicación de la polio; y el refuerzo contra H. influenzae tipo b.

RESUMO O objetivo deste artigo foi avaliar os desafios da vacinação na Colômbia e no Peru e o papel das vacinas pediátricas combinadas na superação de tais desafios. Os obstáculos para incluir novas vacinas com mais antígenos permanecem visíveis em partes desses países, onde doenças imunopreveníveis em lactentes continuam a ser um grande problema. Os desafios incluem a heterogeneidade da cobertura vacinal dentro de cada país e nos países vizinhos, o que pode contribuir para baixas taxas de cobertura vacinal; o impacto adverso da migração interna de pessoas não vacinadas, o que favoreceu o ressurgimento de doenças imunopreveníveis; a escassez de vacinas; e o impacto da pandemia de síndrome respiratória aguda grave do coronavírus 2 (SARS-CoV-2) e mudanças relacionadas nos recursos de atenção à saúde. Para melhorar a cobertura das vacinas pediátricas na Colômbia e no Peru, será necessário assegurar sua integração generalizada em esquemas de vacinas combinadas contendo antígenos de difteria, tétano, pertussis acelular, poliovírus inativado, Haemophilus influenzae tipo B e hepatite B, com uma série primária de três doses aplicadas aos 2, 4 e 6 meses de idade seguidas de um reforço aos 18 meses de idade. Tais vacinas desempenham papéis importantes na prevenção da difteria, tétano e coqueluche; na erradicação da poliomielite; e no reforço contra H. influenzae tipo b.

Increase of Macrolide-Resistance in Streptococcus pneumoniae Strains After the Introduction of the 13-Valent Pneumococcal Conjugate Vaccine in Lima, Peru.

Streptococcus pneumoniae upper respiratory infections and pneumonia are often treated with macrolides, but recently macrolide resistance is becoming an increasingly important problem. The 13-valent pneumococcal conjugate vaccine (PCV13) was introduced in the National Immunization Program of Peru in 2015. This study aimed to evaluate the temporal evolution of macrolide resistance in S. pneumoniae isolates collected in five cross-sectional studies conducted before and after this vaccine introduction, from 2006 to 2019 in Lima, Peru. A total of 521 and 242 S. pneumoniae isolates recovered from nasopharyngeal swabs from healthy carrier children < 2 years old (2 carriage studies) and samples from normally sterile body areas from pediatric patients with invasive pneumococcal disease (IPD) (3 IPD studies), respectively, were included in this study. Phenotypic macrolide resistance was detected using the Kirby-Bauer method and/or MIC test. We found a significant increase in macrolide resistance over time, from 33.5% to 50.0% in carriage studies, and from 24.8% to 37.5% and 70.8% in IPD studies. Macrolide resistance genes [erm(B) and mef(A/E)] were screened using PCR. In carriage studies, we detected a significant decrease in the frequency of mef(A/E) genes among macrolide-resistant S. pneumoniae strains (from 66.7% to 50.0%) after introduction of PCV13. The most common mechanism of macrolide-resistant among IPD strains was the presence of erm(B) (96.0%, 95.2% and 85.1% in the 3 IPD studies respectively). Macrolide resistance was more common in serotype 19A strains (80% and 90% among carriage and IPD strains, respectively) vs. non-serotype 19A (35.5% and 34.4% among carriage and IPD strains, respectively). In conclusion, S. pneumoniae macrolide resistance rates are very high among Peruvian children. Future studies are needed in order to evaluate macrolide resistance trends among pneumococcal strains, especially now after the COVID-19 pandemic, since azithromycin was vastly used as empiric treatment of COVID-19 in Peru.

Cerebral dopamine neurotrophic factor transfection in dopamine neurons using neurotensin-polyplex nanoparticles reverses 6-hydroxydopamine-induced nigrostriatal neurodegeneration.

Overexpression of neurotrophic factors in nigral dopamine neurons is a promising approach to reverse neurodegeneration of the nigrostriatal dopamine system, a hallmark in Parkinson's disease. The human cerebral dopamine neurotrophic factor (hCDNF) has recently emerged as a strong candidate for Parkinson's disease therapy. This study shows that hCDNF expression in dopamine neurons using the neurotensin-polyplex nanoparticle system reverses 6-hydroxydopamine-induced morphological, biochemical, and behavioral alterations. Three independent electron microscopy techniques showed that the neurotensin-polyplex nanoparticles containing the hCDNF gene, ranging in size from 20 to 150 nm, enabled the expression of a secretable hCDNF in vitro. Their injection in the substantia nigra compacta on day 21 after the 6-hydroxydopamine lesion resulted in detectable hCDNF in dopamine neurons, whose levels remained constant throughout the study in the substantia nigra compacta and striatum. Compared with the lesioned group, tyrosine hydroxylase-positive (TH+) nigral cell population and TH+ fiber density rose in the substantia nigra compacta and striatum after hCDNF transfection. An increase in ßIII-tubulin and growth-associated protein 43 phospho-S41 (GAP43p) followed TH+ cell recovery, as well as dopamine and its catabolite levels. Partial reversal (80%) of drug-activated circling behavior and full recovery of spontaneous motor and non-motor behavior were achieved. Brain-derived neurotrophic factor recovery in dopamine neurons that also occurred suggests its participation in the neurotrophic effects. These findings support the potential of nanoparticle-mediated hCDNF gene delivery to develop a disease-modifying treatment against Parkinson's disease. The Institutional Animal Care and Use Committee of Centro de Investigación y de Estudios Avanzados approved our experimental procedures for animal use (authorization No. 162-15) on June 9, 2019.

Radiogenomics: Hunting Down Liver Metastasis in Colorectal Cancer Patients.

Radiomics is a developing new discipline that analyzes conventional medical images to extract quantifiable data that can be mined for new biomarkers that show the biology of pathological processes at microscopic levels. These data can be converted into image-based signatures to improve diagnostic, prognostic and predictive accuracy in cancer patients. The combination of radiomics and molecular data, called radiogenomics, has clear implications for cancer patients' management. Though some studies have focused on radiogenomics signatures in hepatocellular carcinoma patients, only a few have examined colorectal cancer metastatic lesions in the liver. Moreover, the need to differentiate between liver lesions is fundamental for accurate diagnosis and treatment. In this review, we summarize the knowledge gained from radiomics and radiogenomics studies in hepatic metastatic colorectal cancer patients and their use in early diagnosis, response assessment and treatment decisions. We also investigate their value as possible prognostic biomarkers. In addition, the great potential of image mining to provide a comprehensive view of liver niche formation is examined thoroughly. Finally, new challenges and current limitations for the early detection of the liver premetastatic niche, based on radiomics and radiogenomics, are also discussed.

Correction: Earl et al. Somatic Mutation Profiling in the Liquid Biopsy and Clinical Analysis of Hereditary and Familial Pancreatic Cancer Cases Reveals KRAS Negativity and a Longer Overall Survival. Cancers 2021, 13, 1612.

The authors wish to make the following corrections to this paper [...].

Somatic Mutation Profiling in the Liquid Biopsy and Clinical Analysis of Hereditary and Familial Pancreatic Cancer Cases Reveals KRAS Negativity and a Longer Overall Survival.

Pancreatic ductal adenocarcinoma (PDAC) presents many challenges in the clinic and there are many areas for improvement in diagnostics and patient management. The five-year survival rate is around 7.2% as the majority of patients present with advanced disease at diagnosis that is treatment resistant. Approximately 10-15% of PDAC cases have a hereditary basis or Familial Pancreatic Cancer (FPC). Here we demonstrate the use of circulating free DNA (cfDNA) in plasma as a prognostic biomarker in PDAC. The levels of cfDNA correlated with disease status, disease stage, and overall survival. Furthermore, we show for the first time via BEAMing that the majority of hereditary or familial PDAC cases (around 84%) are negative for a KRAS somatic mutation. In addition, KRAS mutation negative cases harbor somatic mutations in potentially druggable genes such as KIT, PDGFR, MET, BRAF, and PIK3CA that could be exploited in the clinic. Finally, familial or hereditary cases have a longer overall survival compared to sporadic cases (10.2 vs. 21.7 months, respectively). Currently, all patients are treated the same in the clinic with cytotoxic agents, although here we demonstrate that there are different subtypes of tumors at the genetic level that could pave the way to personalized treatment.

Intranigral Administration of ß-Sitosterol-ß-D-Glucoside Elicits Neurotoxic A1 Astrocyte Reactivity and Chronic Neuroinflammation in the Rat Substantia Nigra.

Chronic consumption of ß-sitosterol-ß-D-glucoside (BSSG), a neurotoxin contained in cycad seeds, leads to Parkinson's disease in humans and rodents. Here, we explored whether a single intranigral administration of BSSG triggers neuroinflammation and neurotoxic A1 reactive astrocytes besides dopaminergic neurodegeneration. We injected 6 µg BSSG/1 µL DMSO or vehicle into the left substantia nigra and immunostained with antibodies against tyrosine hydroxylase (TH) together with markers of microglia (OX42), astrocytes (GFAP, S100ß, C3), and leukocytes (CD45). We also measured nitric oxide (NO), lipid peroxidation (LPX), and proinflammatory cytokines (TNF-α, IL-1ß, IL-6). The Evans blue assay was used to explore the blood-brain barrier (BBB) permeability. We found that BSSG activates NO production on days 15 and 30 and LPX on day 120. Throughout the study, high levels of TNF-α were present in BSSG-treated animals, whereas IL-1ß was induced until day 60 and IL-6 until day 30. Immunoreactivity of activated microglia (899.0 ± 80.20%) and reactive astrocytes (651.50 ± 11.28%) progressively increased until day 30 and then decreased to remain 251.2 ± 48.8% (microglia) and 91.02 ± 39.8 (astrocytes) higher over controls on day 120. C3(+) cells were also GFAP and S100ß immunoreactive, showing they were neurotoxic A1 reactive astrocytes. BBB remained permeable until day 15 when immune cell infiltration was maximum. TH immunoreactivity progressively declined, reaching 83.6 ± 1.8% reduction on day 120. Our data show that BSSG acute administration causes chronic neuroinflammation mediated by activated microglia, neurotoxic A1 reactive astrocytes, and infiltrated immune cells. The severe neuroinflammation might trigger Parkinson's disease in BSSG intoxication.

Synthetic Monopartite Peptide That Enables the Nuclear Import of Genes Delivered by the Neurotensin-Polyplex Vector.

Neurotensin (NTS)-polyplex is a multicomponent nonviral vector that enables gene delivery via internalization of the neurotensin type 1 receptor (NTSR1) to dopaminergic neurons and cancer cells. An approach to improving its therapeutic safety is replacing the viral karyophilic component (peptide KPSV40; MAPTKRKGSCPGAAPNKPK), which performs the nuclear import activity, by a shorter synthetic peptide (KPRa; KMAPKKRK). We explored this issue and the mechanism of plasmid DNA translocation through the expression of the green fluorescent protein or red fluorescent protein fused with KPRa and internalization assays and whole-cell patch-clamp configuration experiments in a single cell together with importin α/ß pathway blockers. We showed that KPRa electrostatically bound to plasmid DNA increased the transgene expression compared with KPSV40 and enabled nuclear translocation of KPRa-fused red fluorescent proteins and plasmid DNA. Such translocation was blocked with ivermectin or mifepristone, suggesting importin α/ß pathway mediation. KPRa also enabled NTS-polyplex-mediated expression of reporter or physiological genes such as human mesencephalic-derived neurotrophic factor (hMANF) in dopaminergic neurons in vivo. KPRa is a synthetic monopartite peptide that showed nuclear import activity in NTS-polyplex vector-mediated gene delivery. KPRa could also improve the transfection of other nonviral vectors used in gene therapy.

Unilateral intranigral administration of ß-sitosterol ß-D-glucoside triggers pathological α-synuclein spreading and bilateral nigrostriatal dopaminergic neurodegeneration in the rat.

The spreading and accumulation of α-synuclein and dopaminergic neurodegeneration, two hallmarks of Parkinson's disease (PD), have been faithfully reproduced in rodent brains by chronic, oral administration of ß-sitosterol ß-D-glucoside (BSSG). We investigated whether a single injection of BSSG (6 µg BSSG/µL DMSO) in the left substantia nigra of Wistar rats causes the same effects. Mock DMSO injections and untreated rats formed control groups. We performed immunostainings against the pathological α-synuclein, the dopaminergic marker tyrosine hydroxylase (TH), the neuroskeleton marker ß-III tubulin, the neurotensin receptor type 1 (NTSR1) as non-dopaminergic phenotype marker and Fluro-Jade C (F-J C) label for neurodegeneration. Using ß-galactosidase (ß-Gal) assay and active caspase-3 immunostaining, we assessed cell death mechanisms. Golgi-Cox staining was used to measure the density and types of dendritic spines of striatal medium spiny neurons. Motor and non-motor alterations were also evaluated. The study period comprised 15 to 120 days after the lesion. In the injured substantia nigra, BSSG caused a progressive α-synuclein aggregation and dopaminergic neurodegeneration caused by senescence and apoptosis. The α-synuclein immunoreactivity was also present within microglia cells. Decreased density of dopaminergic fibers and dendritic spines also occurred in the striatum. Remarkably, all the histopathological changes also appeared on the contralateral nigrostriatal system, and α-synuclein aggregates were present in other brain regions. Motor and non-motor behavioral alterations were progressive. Our data show that the stereotaxic BSSG administration reproduces PD α-synucleinopathy phenotype in the rat. This approach will aid in identifying the spread mechanism of α-synuclein pathology and validate anti-synucleinopathy therapies.

A sequential methodology for integral evaluation of motor and non-motor behaviors in parkinsonian rodents.

An animal model, suitable for resembling Parkinson's disease (PD) progress, should show both, motor and non-motor alterations. However, these features have been scarcely evaluated or developed in parkinsonian models induced by neurotoxins. This protocol provides modifications to original methods, allowing six different motor and non-motor behavior tests, which adequately and timely emulate the main parkinsonian sensorimotor alterations in the rat or mouse: (1) bilateral sensorimotor alterations, examined by the vibrissae test; (2) balance and motor coordination, evaluated by the uncoordinated gait test; (3) locomotor asymmetry, analyzed by the cylinder test; (4) bradykinesia, as a locomotor alteration evidenced by the open field test; (5) depressive-like behavior, judged by the forced swimming test; and (6) hyposmia, assessed by the olfactory asymmetry test. Some advantages of using these behavioral tests over others include:•No sophisticated materials or equipment are required for their application and evaluation.•They are used in rodent models for parkinsonian research, but they can also be helpful for studying other movement disorders.•These tests can accurately discriminate the affected side from the healthy one, after unilateral injury of one hemisphere, resulting in sensorimotor, olfactory or locomotor asymmetry.

A comprehensive analysis of candidate genes in familial pancreatic cancer families reveals a high frequency of potentially pathogenic germline variants.

BACKGROUND: The 5-year survival rate of patients with pancreatic ductal adenocarcinoma (PDAC) is around 5% due to the fact that the majority of patients present with advanced disease that is treatment resistant. Familial pancreatic cancer (FPC) is a rare disorder that is defined as a family with at least two affected first degree relatives, with an estimated incidence of 4%-10%. The genetic basis is unknown in the majority of families although around 10%-13% of families carry germline mutations in known genes associated with hereditary cancer and pancreatitis syndromes. METHODS: Panel sequencing was performed of 35 genes associated with hereditary cancer in 43 PDAC cases from families with an apparent hereditary pancreatic cancer syndrome. FINDINGS: Pathogenic variants were identified in 19% (5/26) of PDAC cases from pure FPC families in the genes MLH1, CDKN2A, POLQ and FANCM. Low frequency potentially pathogenic VUS were also identified in 35% (9/26) of PDAC cases from FPC families in the genes FANCC, MLH1, PMS2, CFTR, APC and MUTYH. Furthermore, an important proportion of PDAC cases harboured more than one pathogenic, likely pathogenic or potentially pathogenic VUS, highlighting the multigene phenotype of FPC. INTERPRETATION: The genetic basis of familial or hereditary pancreatic cancer can be explained in 21% of families by previously described hereditary cancer genes. Low frequency variants in other DNA repair genes are also present in 35% of families which may contribute to the risk of pancreatic cancer development. FUNDING: This study was funded by the Instituto de Salud Carlos III (Plan Estatal de I + D + i 2013-2016): ISCIII (PI09/02221, PI12/01635, PI15/02101 and PI18/1034) and co-financed by the European Development Regional Fund ''A way to achieve Europe'' (ERDF), the Biomedical Research Network in Cancer: CIBERONC (CB16/12/00446), Red Temática de investigación cooperativa en cáncer: RTICC (RD12/0036/0073) and La Asociación Española contra el Cáncer: AECC (Grupos Coordinados Estables 2016).

A single intranigral administration of ß-sitosterol ß-d-glucoside elicits bilateral sensorimotor and non-motor alterations in the rat.

Parkinson's disease (PD) is a progressive neuropathology characterized by motor and non-motor alterations. ß-sitosterol ß-d-glucoside (BSSG) is a neurotoxin whose prolonged oral administration in rats has been proposed as a new PD model. Herein, we demonstrate that a single, unilateral, and intranigral administration of BSSG also elicits bilateral sensorimotor alterations in the rat. Six behavioral tests evaluated the effect of different concentrations of BSSG (3, 6, 9, and 12 µg/µL DMSO) from 15 to 120 days after administration. The first behavioral alterations, which appeared on day 15, were unbalanced and uncoordinated gaits and a decrease in the sensorimotor cortex activity, as evidenced by the beam-walking and the vibrissae tests, respectively. After 30 days, the corridor test revealed hyposmia and a decreased locomotor activity in the open field. The last alteration was a depressive-like behavior, as shown by the forced swim test on days 60 and 120. According to the cylinder test, no locomotor asymmetry was observed over time with any BSSG concentrations tested. Also, a mesencephalic TH(+) cell loss (p < 0.05) was shown on day 30 when compared with the mock condition, and such a loss was even higher on day 120. At this time, the presence of pathological α-synuclein aggregates in the mesencephalon was documented. Our results show that the stereotaxic intranigral administration of BSSG reproduces some characteristics of oral administration, such as the progression of behavioral alterations, dopaminergic neurons loss, and the presence of Lewy body-like synuclein aggregations, in less time and resources.

Acute Neuroinflammatory Response in the Substantia Nigra Pars Compacta of Rats after a Local Injection of Lipopolysaccharide.

Models of Parkinson's disease with neurotoxins have shown that microglial activation does not evoke a typical inflammatory response in the substantia nigra, questioning whether neuroinflammation leads to neurodegeneration. To address this issue, the archetypal inflammatory stimulus, lipopolysaccharide (LPS), was injected into the rat substantia nigra. LPS induced fever, sickness behavior, and microglial activation (OX42 immunoreactivity), followed by astrocyte activation and leukocyte infiltration (GFAP and CD45 immunoreactivities). During the acute phase of neuroinflammation, pro- and anti-inflammatory cytokines (TNF-α, IL-1ß, IL-6, IL-4, and IL-10) responded differentially at mRNA and protein level. Increased NO production and lipid peroxidation occurred at 168 h after LPS injection. At this time, evidence of neurodegeneration could be seen, entailing decreased tyrosine hydroxylase (TH) immunoreactivity, irregular body contour, and prolongation discontinuity of TH+ cells, as well as apparent phagocytosis of TH+ cells by OX42+ cells. Altogether, these results show that LPS evokes a typical inflammatory response in the substantia nigra that is followed by dopaminergic neurodegeneration.

Resistencia antibiótica y distribución de serotipos en cepas neumocócicas invasivas en adultos hospitalizados en Lima, Perú / Antibiotic resistance and distribution of serotypes of invasive pneumococcal strains isolated from hospitalized adults in Lima, Peru

RESUMEN Objetivos. Describir las características clínicas, resistencia antibiótica y distribución de serotipos de cepas causantes de enfermedad neumocócica invasiva (ENI) en adultos. Materiales y métodos. Estudio tipo serie de casos. Se recolectaron cepas de neumococo de pacientes adultos hospitalizados con ENI en cinco hospitales nacionales y dos laboratorios de Lima durante los años 2009-2011. Resultados. Se estudiaron datos de 43 pacientes con ENI, el 58,2% fueron mayores de 60 años. Los diagnósticos fueron neumonía 39,5%, meningitis 30,2%, bacteriemia 13,9%, peritonitis 11,6%, artritis séptica 4,8%. El porcentaje de fallecidos fue 28,9%, de los cuales el 72,7% fueron mayores de 60 años. Las cepas de neumococo presentaron la siguiente resistencia: penicilina 0% en cepas no meningitis y 30,8% en cepas meningitis; ceftriaxona 4,5% y 16,7% de resistencia intermedia en cepas no meningitis y cepas meningitis respectivamente; 69% a trimetoprim/sulfametoxazol y 35,7% a eritromicina. Los serotipos más comunes fueron 19F, 23F, 6B, 14 y 6C. El porcentaje de cepas vacunales fue 44,2% para la vacuna conjugada siete-valente (PCV7) y para la PCV10, 51,2% para PCV13 y 60,4% para la vacuna polisacárida veintitrés-valente (PPV23). Conclusiones. El neumococo es un patógeno relevante en adultos, en especial en los adultos mayores, debido a su elevada mortalidad.

ABSTRACT Objectives. To describe the clinical characteristics, antibiotic resistance, and distribution of serotypes of bacterial strains that cause invasive pneumococcal disease (IPD) in adults. Materials and methods. Case series. Pneumococcal strains were isolated from 2009 to 2011 from hospitalized adult patients with IPD in five hospitals and two laboratories located in Lima. Results. The analysis of data from 43 patients with IPD indicated that 58.2% were older than 60 years. The most common complications were pneumonia (39.5%), meningitis (30.2%), bacteremia (13.9%), peritonitis (11.6%), and septic arthritis (4.8%). The mortality rate was 28.9%, and 72.7% of cases involved patients older than 60 years. The pneumococcal strains were resistant to the following antibiotics: penicillin, 0% and 30.8% in non-meningitis and meningitis strains, respectively; ceftriaxone, 4.5% and 16.7% in non-meningitis and meningitis strains, respectively; trimethoprim/sulfamethoxazole, 69.0%; and erythromycin, 35.7%. The most common serotypes were 19F, 23F, 6B, 14, and 6C. The percentage of vaccine strains was 44.2% for the 7-valent conjugate pneumococcal vaccine (PCV7) and PCV10, 51.2% for PCV13, and 60.4% for the 23-valent polysaccharide vaccine (PPV23). Conclusions. Pneumococcus is an important pathogen in adults, particularly in older adults, owing to its high mortality rate.

Key features of invasive pneumococcal isolates recovered in Lima, Peru determined through whole genome sequencing.

Before PCV7 introduction, invasive pneumococcal disease (IPD) was responsible for approximately 12,000-18,000 deaths annually among children <5years in Latin America. In Peru, PCV7 was introduced in 2009. We used whole genome sequencing to deduce key features of invasive strains collected in Lima, Peru from 2006 to 2011. We sequenced 212 IPD isolates from 16 hospitals in Lima pre (2006-2009; n=133) and post (2010-2011; n=79) PCV7 introduction; 130 (61.3%) isolates were from children≤5years old. CDC's Streptococcus lab bioinformatics pipeline revealed serotypes, sequence types (STs), pilus genes, PBP types and other resistance determinants. During the pre-PCV7 period, serotype 14 was the most common serotype (24.8%), followed by 6B (20.3%), 19F (10.5%), and 23F (6.8%). Post-PCV7, the proportion of PCV7 serotype 6B decreased significantly (to 6.3%), while 19F (16.3%), 14 (15.0%), 23F (7.5%), and 19A (7.5%) were the most common serotypes; only serotypes 3 and 10A increased significantly. Overall, 82% (n=173) of all isolates carried at least one resistance determinant, including 72 (34%) isolates that carried resistance determinants against 3 or more antimicrobial classes; of these 72 isolates, 56 (78%) belonged to a PCV7 serotype. Eighty-two STs were identified, with 53 of them organized in 14 clonal complexes. ST frequencies were distributed differently pre and post-PCV7 introduction, with only 18 of the 57 STs identified in years 2006-2009 isolates also observed in years 2010-2011 isolates. The apparent expansion of a 19F/ST1421 lineage with predicted ß-lactam resistance (PBP type 13:16:20) and carrying resistance determinants against four additional antimicrobial classes was observed.