Role of synaptic input remodeling of corticospinal motor neurons after spinal cord injury / 中国组织工程研究

BACKGROUND:The recovery of function after spinal cord injury depends on the functional remodeling of the motor cortex.However,the anatomical evidence underlying the functional remodeling of the motor cortex is still illusive.Analyzing the anatomical changes in the motor cortex after spinal cord injury can provide new ideas and research directions for regulating functional recovery and rehabilitation after spinal cord injury. OBJECTIVE:To analyze the neural circuit structural basis of functional remodeling of the primary motor cortex after spinal cord injury. METHODS:C57BL/6J mice were randomly divided into a sham operation group and a spinal cord injury group.The adeno-associated virus vectors expressing the fusion protein of Cre recombinase were injected into C4 of mice of both groups.The adeno-associated virus vectors with Cre recombinase-inducible expression of avian sarcoma/leukosis envelope glycoprotein receptor TVA and rabies glycoprotein were injected into the primary motor cortex.Fourteen days later,a C6 dorsal hemisection mice model was established in the spinal cord injury group.The pseudotyped rabies virus was injected into the primary motor cortex of mice of both groups.After 7 days,brain samples were collected and frozen sections were made.The distribution of input neurons innervating corticospinal motor neurons in the brain was observed and analyzed quantitatively. RESULTS AND CONCLUSION:Fluorescence microscopy observation and quantitative analysis found that input neurons innervating corticospinal motor neurons of the primary motor cortex in mice of both groups were distributed in the cerebral cortex,thalamus and midbrain.Among them,in the sham operation group,the number of input neurons in the mouse cerebral cortex accounted for(84.0±3.6)%of total brain input neurons;that in the thalamus accounted for(10.6±2.3)%,and that in the midbrain accounted for(0.7±0.4)%.Direct synaptic input neurons in the spinal cord injury group accounted for(81.7±1.0)%,(13.1±0.5)%,and(1.6±0.8)%in the cerebral cortex,thalamus and midbrain,respectively.The proportion and number of primary motor cortex input neurons in the three regions of the spinal cord injury group did not differ significantly from that of the sham operation group.After spinal cord injury,the number of input neurons innervating corticospinal pyramidal motor neurons in various brain regions did not change significantly,suggesting that functional remodeling of the motor cortex after spinal cord injury may not only depend on changes in synaptic input related to injured corticospinal motor neurons,but also on transcriptional regulation changes within the injured neurons themselves.

Effect of bioreactor scale-up culture of micro-carrier Vero cells after extra-tank trypsinization on virus-producing ability of rabies virus CTN-1Ⅴ strain / 中国生物制品学杂志

@#Objective To investigate the effect of amplification culture of micro-carrier Vero cells from 30 L bioreactor to 300 L bioreactor after extra-tank trypsinization on the virus-producing ability of rabies virus(RABV)CTN-1Ⅴ strain.Methods The 140-passage of Vero cells were cultured at 37 ℃ for 72-120 h,then amplified by passaging at a cell density ratio of 1∶4 into the 10 × cell factory. After incubation at 37 ℃ for 72-120 h,the monolayer cells were detached and inoculated into the 30 L bioreactor with micro-carriers 7-10 g/L,culture temperature 37 ℃,pH 7. 0-7. 4,dissolved oxygen 30%-80%,stirring speed 10-50 r/min,and continuous perfusion culture 72-120 h. Total three batches of microcarrier Vero cells were cultured,which were amplified to the 300 L bioreactor after extra-tank trypsinization,with microcarrier5-8 g/L,culture temperature 37 ℃,pH 7. 0-7. 4,dissolved oxygen 30%-80%,stirring speed 30-80 r/min,and perfusion culture 72-120 h. RABV CTN-1Ⅴ was inoculated at the MOI of 0. 05,and the virus solution was harvested every 24 h and detected for the virus titer and antigen content. Results The cell density was about 1 × 10~7 cells/mL after culture for 96 h in the 30 L bioreactor,and was about 7. 4 × 10~6 cells/mL after culture for 96 h in the 300 L bioreactor. At 96 h after virus inoculation,the virus harvest solution reached the peak potency,with the average virus titer of 6. 8 lgLD_(50)/mL and the average antigen content of 2. 58 IU/mL. Conclusion The scale-up culture process of micro-carrier Vero cells after extra-tank trypsinization from 30 L bioreactor to 300 L bioreactor is stable and feasible,with no significant effect on the virus-producing ability of RABV CTN-1Ⅴ strain,which provides a reference for the large-scale production of inactivated RABV vaccine

Development of large-scale amplification process of rabies virus in 150 L bioreactor / 中国生物制品学杂志

@#Objective To develop a large-scale culture process for rabies virus(RABV)in 150 L bioreactor,and lay a foundation for the further development of a larger-scale and high-density microcarrier reactor process.Methods Vero cells and RABV strain CTN-1V were cultured in 30 L(model:C30-2)and 150 L(model:VESSEL FERMENTER 300L)bioreactors by perfused culture with 20 g/L Cytodex-1 microcarrier and DO 20%-60%,at culture temperature 36-38 ℃ and pH 7.0-7.4.During the culture process,the cell density and virus titer were measured.The virus culture media was harvested for consecutive 13 d and detected for the sterility,mycoplasma,and the residues of antigen,host cell protein(HCP),bovine serum albumin(BSA)and DNA.Results The density of cultured cells in 30 L and 150 L bioreactors all reached above 1.2 ×10~7cell/mL.There was no significant difference in cell density at different time points during the culture(t = 0.225-2.173,P = 0.096-0.833).The highest virus titer(8.5 lgLD_(50)/mL)was found in the both bioreactors 6 d after infection with no significant difference(t = 1.000,P = 0.374).The residues of antigen,HCP,BSA and DNA in the virus suspension from the two bioreactors were basically the same.Conclusion 150 L bioreactor can be used for the large-scale culture of RABV,and the harvested virus conformed to the relevant standards in Chinese Pharmacopoeia(Volume Ⅲ,2020 edition).

Preparation,identification and comparison of murine and rabbit polyclonal antibodies against matrix protein of rabies virus strain CVS-11 / 中国生物制品学杂志

@#Objective To prepare murine and rabbit polyclonal antibodies against rabies virus(RV) matrix(M) protein and compare their reactivity.Methods The prokaryotic expression vector pET-28a-M was constructed by using the cDNA of cells infected with RV CVS-11 strain as template,then transformed into E.coli BL21(DE3),and the induced by IPTG to express M protein.After nickel column affinity chromatography and dialysis renaturation,female BALB/c mice and New Zealand white rabbits were immunized with the M protein,and the whole blood was taken to separate the serum.The titers of the murine and rabbit polyclonal antibodies were detected by ELISA,and the reactivity was measured by Western blot,indirect immunofluorescence assay(IFA) and immunoprecipitation(IP).Results The plasmid pET-28a-M was constructed correctly as identified by sequencing.The titers of murine and rabbit polyclonal antibodies were 1:100 and 1:256 000respectively,and the polyclonal antibodies had reactivity with different RV strains.Conclusion The murine and rabbit polyclonal antibodies against M protein were successfully prepared,which provides important biological tools for exploring the interaction between M protein and host protein as well as studying the pathogenesis of RV.

Human Rabies during the COVID-19 Pandemic: Insights into Rabies Worldwide and Brazil

ABSTRACT Human Rabies (HR) is a fatal zoonotic disease caused by lyssaviruses, with the rabies virus (RABV) identified as the causative agent. While the incidence of HR transmitted by dogs has decreased in Latin America, there has been a corresponding rise in transmission via wild animals. Given the lack of effective treatments and specific therapies, the management of HR relies on the availability of post-exposure prophylaxis and animal control measures. This review examines the dynamics and spread of HR during the global pandemic.

Detection of rabies virus in Callithrix penicillata (Geoffroy, 1812) in Montes Claros, Minas Gerais State, Brazil

ABSTRACT This report describes the occurrence of the rabies virus in two species of wild animals in the urban area of Montes Claros (MOC), Minas Gerais State, Brazil, in May 2023. The virus has been detected in frugivorous chiropterans (Artibeus sp) and marmosets (Callithrix penicillata). This is the first notified case of the rabies virus in the species C. penicillata in the urban area of MOC. Our findings show that the rabies virus is circulating in the urban area of MOC; therefore, permanent preventive measures must be adopted to avoid infection of other animals and humans.

Virología molecular de la rabia: un enfoque clínico / Rabies molecular virology: a clinical approach

Introducción: La rabia es una enfermedad zoonótica asociada al virus RABV, el cual tiene características neurotrópicas. El virus se transmite por el contacto con saliva de animales infectados; la mordedura de un perro es la causa más común. Es un virus que causa la muerte de miles de personas cada año. Objetivo: Describir a profundidad los principios moleculares de la infección por rabia, así como su patogenia, diagnóstico y tratamiento. Métodos: Se realizó una búsqueda de bibliografía en PubMed, SciELO, Scopus, Researchgate; se consultaron 163 referencias y se seleccionaron 51 fuentes que contenían la información más relevante para cumplir con el objetivo del trabajo. Conclusión: Actualmente es posible entender de mejor manera los mecanismos de transmisión y propagación del virus en el organismo; existe nuevo conocimiento sobre los receptores involucrados, así como la función de estos en la replicación viral. Sin embargo, el objetivo de la erradicación de la rabia a corto plazo es complejo. La invasión de territorios selváticos vuelve a la rabia un posible patógeno reemergente; la vacunación de especies transmisoras es el medio ideal para conseguir el control de la enfermedad.

Introduction: Rabies is a zoonotic disease associated with the RABV virus, which has neurotropic characteristics. The virus is transmitted by contact with saliva from infected animals; a dog's bite is the most common cause. This virus causes the death of thousands of people every year. Objective: To describe in depth the molecular principles of rabies infection, as well as its pathogenesis, diagnosis and treatment. Methods: A literature search was conducted in PubMed, SciELO, Scopus, and Researchgate. A total of 163 references were consulted, and 51 sources containing the most relevant information were selected to fulfill the objective of the work. Conclusions: It is currently possible to better understand the mechanisms of transmission and spread of the virus in the organism; there is new knowledge about the receptors involved, as well as their function in viral replication. However, the goal of eradicating rabies in the short term is complex. The invasion of wild territories makes rabies a possible re-emerging pathogen; vaccination of transmitting species is the ideal means to achieve disease control.

Construction and verification of recombinant chimeric vesicular stomatitis virus expressing rabies virus G protein / 中国生物制品学杂志

@#Objective To construct recombinant chimeric vesicular stomatitis virus(VSV)expressing G protein of rabies virus(RV)using VSV as vector.MethodsTo rescue the recombinant virus,G gene of VSV antigenome was replaced with G gene of RV vaccine CTN-1 strain,and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and proteins N,P and L of VSV. The expression of RV G gene and G protein was detected by RT-PCR,immunofluorescence assay and Western blot. The recombinant virus was subcultured in Vero cells,the virus titer of different generations was detected and the virus growth curve was drawn.ResultsThe recombinant virus VSV-RVG was successfully rescued. RTPCR results demonstrated that the RV G gene was successfully inserted into the genome of the recombinant virus,and the expression of RVG protein was detected by immunofluorescence assay and Western blot. The recombinant virus was continuously passaged for 5 generations,and the virus titer was stable within 7. 5 ～ 8. 5 lgTCID50/mL.ConclusionThe recombinant chimeric VSV expressing RV G protein was successfully constructed with good genetic stability,which lays a foundation of the construction of reverse genetics technology platform based on VSV vector.

Comparison of immunization strategies for human rabies pre-exposure prophylaxis in laboratory animals / 中华微生物学和免疫学杂志

Objective:To compare the differences in the safety, efficacy and protective effects of rabies vaccine using the current pre-exposure prophylaxis schedule in China (0-7-21 or 28) and the newly recommended immunization program of WHO (0-7), aiming to provide data support for modifying the related content of Technical Guideline for Human Rabies Prevention and Control. Methods:The mice were randomly divided into five groups, namely 0-7-21 group (3-injection regimen), 0-7 group (2-injection regimen), 0-14 group, 0-21 group and control group, according to the current 3-injection regimen (0-7-21) in China and the 2-injection regimen (0-7) recommended by WHO. The survival status of the mice was observed. The mice were weighed every five days to compare the safety of different immunization procedures. Rabies virus neutralizing antibodies (RVNA) were detected 7, 14, 21, 28 and 35 d after the initial immunization. On day 35, the mice in each group were challenged with lethal dose of CVS-11 rabies virus to evaluate the protective effects of different pre-exposure immunization procedures.Results:There was no significant difference in weight gain of mice after vaccination. The positive rate of RVNA was 100% in all immunized groups from day 14, which could provide complete protection to mice. There was a significant difference in RVNA levels between 0-7-21 and 0-7 groups at 35 d( P<0.05), but there was no statistical difference at other time points ( P>0.05). RVNA level had a significant difference between 0-7 and 0-21 groups at 21 d and 35 d ( P<0.05). There was no statistical difference in RVNA level between other groups at each time point ( P>0.05). In the protective test, the survival rates of mice in all immune groups were 100%. Conclusions:The current 3-injection pre-exposure immunization procedure for rabies vaccine (0-7-21) and the newly recommended 2-injection immunization procedure (0-7) had similar efficacy and protective effects in animal tests. In view of the cost saving and better compliance of the 2-injection immunization procedure, it was recommended that relevant departments should conduct clinical trials as soon as possible to promote the implementation of this program.

Rabies virus viability in vitro / 中华微生物学和免疫学杂志

Objective:To investigate the in vitro viability of rabies virus in tissues and body fluid samples. Methods:The viability of rabies virus in tissues and suspensions was analyzed by virus titer determination method, direct immunofluorescence, RT-PCR and laboratory techniques for virus isolation.Results:With the increase of temperature, the viability of rabies virus in brain tissues and suspensions decreased gradually. Rabies virus lost infectivity after 30 min at 56℃, but remained viable in tissues for 7 d at 37℃. The virus showed no viability after 1 h at pH9.6. The rabies virus in suspensions could be completely inactivated after the stimulation with ethanol at a final concentration above 30%, sodium hypochlorite above 500 mg/L or benzalkonium bromide above 100 mg/L for 3 min. It was found that 80% acetone had the strongest inactivation effect on rabies virus in tissues, and no virus could be isolated after soaking for 4 h.Conclusions:Rabies virus was not tolerant to high temperature and relatively stable in the environment with pH6.8-7.4. Common disinfectants could kill the virus. This study provided detailed data about the viability of rabies virus in vitro, which would be conducive to the prevention and control of rabies.

Effects of histidine 74 in rabies virus matrix protein on mitochondrial localization / 中国人兽共患病学报

This study investigated the effect of histidine(H)at position 74 on the localization of full-length viral matrix protein(M)in mitochondria.A eukaryotic expression vector expressing the N-terminus or C-terminus of wild-type M protein or its histidine 74 mutant protein(H→P,H74P)fused with V5(tag protein)-APEX2(modified ascorbate peroxidase)or A-PEX2-V5 was constructed and transfected into 293T cells.Fluorescence staining and laser confocal microscopy were used to an-alyze the effect of histidine 74 mutation on the localization of full-length M protein in mitochondria.Immunoblotting showed that the eukaryotic expression vector expressed the fusion protein normally.Fluorescence staining indicated that the H→P mu-tation at position 74 did not affect the ability of the full-length viral M protein to localize to mitochondria.Our findings regard-ing the effect of histidine 74 on the mitochondrial localization of full-length virus M protein provide a foundation for future study of the biological function of M protein and the replication mechanism of rabies virus.

Inhibitory effects of Galidesivir(BCX4430)on rabies virus in vitro / 中国人兽共患病学报

This study analyzed the inhibitory effects of BCX4430 on rabies virus with different virulence(CVS-11 and SC16)in vitro.Direct immunofluorescence(DFA),quantitative real-time PCR and half tissue culture infective dose(TCID50)assays were used.BCX4430(250 pmol/L),compared with T-705,significantly decreased the viral titer and the relative expres-sion level of viral mRNA,similarly to the inhibitory effects of ribavirin.Thus,our study demonstrated that BCX4430 signifi-cantly inhibits the replication of rabies virus in vitro.

Establishment and preliminary application of dual fluorescent quantitative PCR for detection of RV and CAV2 / 公共卫生与预防医学

Objective To investigate the shedding of CAV2-ΔE3-CGS after immunization and the background of canine adenovirus (CAV) infection, and to establish a dual fluorescent quantitative PCR detection method for rabies virus (RV) and canine adenovirus type 2 (CAV2). Methods A dual fluorescent quantitative PCR detection method was established by designing specific primers and probes for E1 gene of CAV and G gene of RV for the detection of CAV2-ΔE3-CGS. Oral swabs, anal swabs and environmental samples of stray dogs from experimental animal farm and dog detention center were tested. Results The standard curves generated by this method were Y=-3.351 × logX + 44.895, R2 = .999 and Y=-3.413 × logX + 45.192, R2=0.996, respectively. The linear relationships were good, and the minimum detection limits were both 102 copies/μL. CAV2-ΔE3-CGS was not detected in experimental animal farm. CAV was detected in dog detention center, and the positive rates were 5.88% (5/85) in oral swabs, 8.24% (7/85) in anal swabs, and 4% (1/25) in environmental samples. Conclusion The dual fluorescent quantitative PCR method can be used for the detection of CAV2-ΔE3-CGS after immunization and the investigation of CAV infection. The present study has shown that no CAV2-ΔE3-CGS has been detected after immunization and CAV infection rate of stay dogs is low in Shanghai. CAV2-ΔE3-CGS oral immunization meets requirement and is applicable.

Factors related to vaccines for both patients exposed with rabies and dogs

ABSTRACT Introduction: Rabies is a viral disease that can be transmitted from animals to humans. All warm-blooded animals, including humans, can host rabies's virus. Vaccination of dogs is an effective method of preventing rabies in humans. Objectives: To evaluate knowledge, attitudes and practices towards rabies prevention and factors related to vaccines for both patients exposed with rabies and dogs. Methods: A cross-sectional descriptive study, using the questionnaire on rabies prevention by interviewing 1 484 households. Multivariable logistic regression analysis was used to identify factors related to vaccines for both patients exposed to rabies and dogs. Results: Of all 67.32 % of the people surveyed with high knowledge, only 43.8 % and 31.47 % were assessed as having a good attitude and practice. Interviewees with poor knowledge and practice are independently related to low-rate vaccines provision for pets. Factors associated with low vaccination rates after pet bites were poor attitudes and practices. Conclusion: People with a high level of education, knowledge and practice, the rabies vaccination rate on pets is high. People with good attitudes and practices had a high rate of rabies vaccination after being bitten by an animal.

RESUMEN Introducción: La rabia es una enfermedad viral que puede transmitirse de animales a humanos. Todos los animales de sangre caliente, incluidos los humanos, pueden albergar el virus de la rabia. La vacunación de perros es un método eficaz para prevenir la rabia en humanos. Objetivos: Evaluar conocimientos, actitudes y prácticas de la comunidad hacia la prevención de la rabia y los factores relacionados con las vacunas, tanto para pacientes expuestos como para perros. Métodos: Estudio descriptivo transversal, se utilizó un cuestionario sobre prevención de la rabia, mediante entrevistas en 1 484 hogares. Se realizó un análisis de regresión logística multivariable para identificar los factores relacionados con las vacunas, tanto para pacientes expuestos como para los perros. Resultados: Del 67,32 % de encuestados, con alto conocimiento, solo el 43,8 % y el 31,47 % fueron evaluados como de buena actitud y práctica. Los entrevistados con insuficiente conocimiento y prácticas se relacionan de forma independiente con la baja provisión de vacunas para las mascotas. Las razones de bajas tasas de vacunación después de las mordeduras causadas por mascotas fueron las malas actitudes y prácticas. Conclusión: Las personas con nivel alto de práctica, educación y conocimientos, tienen elevados niveles de vacunación antirrábica de sus mascotas. Después de la mordedura de un animal, las personas con buenas prácticas y actitudes, tienen más altos niveles de vacunación antirrábica.

Encefalitis por rabia o encefalitis post-vacunal en mujer gestante con neonato prematuro sobreviviente

La rabia humana es una infección grave que requiere aplicación de suero antirrábico y vacuna, la cual puede producir encefalomielitis desmielinizante aguda (EDMA) post-vacunal. Se reporta una gestante de 27 años mordida por un perro en mano derecha. No recibió suero antirrábico pero sí 5 vacunas. Con última vacuna presentó adormecimiento de mano, malestar, fiebre, hidrofobia, alteración de conciencia. Se realizó cesárea e indujo al coma terapéutico. La biopsia de piel de nuca e inoculación en ratón fueron negativos mientras que la seroneutralización en ratón y detección de anticuerpos antirrábicos fueron positivos. En cordón umbilical, la seroneutralización en ratón fue positiva. La resonancia magnética evidenció lesiones compatibles con EDMA. Según algunos criterios, podría tratarse de rabia; sin embargo, el cuadro sería más compatible con EDMA.

Human rabies is a serious infection that requires application of rabies serum and vaccine, which can cause post-vaccination acute demyelinating encephalomyelitis (ADEM). We report a 27-year-old pregnant woman who has been bitten by a dog on her right hand. She didn't receive rabies serum but she received 5 vaccinations. With last vaccination she presents hand numbness, malaise, fever, hydrophobia, altered consciousness. A cesarean section was performed and the patient was induced a therapeutic coma. Nape skin biopsy and inoculation in mice were negative, while seroneutralization in mice and detection of anti-rabies antibodies were positive. In umbilical cord, mouse seroneutralization was positive. Magnetic resonance imaging revealed lesions compatible with ADEM. By some criteria, it could be rabies; however, the chart would be more compatible with ADEM.

Educação em saúde para prevenção da raiva humana / Health education for prevention of human rabies / Educación en salud para prevención de la rabia humana

A raiva é uma antropozoonose viral que se desenvolve de forma progressiva e aguda podendo apresentar até 100% de letalidade. O seu agente etiológico é o vírus rábico do gênero Lyssavirus pertencente à família Rhabdoviridae. O presente trabalho teve como objetivo divulgar informações acerca da ocorrência da raiva em humanos em virtude da sua expressiva importância para saúde pública e analisar a percepção da população sobre a raiva humana, de forma a obter dados relacionados a conhecimentos básicos sobre a doença. Para isso, foi criada uma página informativa denominada "@contraraiva_" na rede social Instagram para a realização de postagens interativas sobre o tema abordado, e uso de um questionário criado a partir da plataforma Google Forms em diferentes mídias sociais para a obtenção e coleta de dados. Foram obtidas 1.075 respostas, provenientes de diferentes localidades. O questionário alcançou todas as regiões brasileiras, todos 26 estados e o Distrito Federal. As informações publicadas pela página criada ajudaram a sanar dúvidas relacionadas aos principais aspectos da doença. Os dados obtidos a partir do questionário contribuem para o planejamento de ações voltadas para a educação em saúde de forma mais estratégica, visando contribuir para os pontos em que a população tem menos conhecimentos.(AU)

Rabies is a viral anthropozoonosis that is developed in a progressive and acute way and can present up to 100% lethality. Its etiologic agent is the rabies virus of the Lyssavirus gene belonging to the Rhabdoviridae family. This study aimed at disseminating information about the occurrence of rabies in humans due to its expressive importance for public health, and at analyzing the population perception on human rabies in order to obtain data related to basic knowledge about the disease. For that purpose, an information page called "@ contraraiva _" was created on the social network Instagram for providing interactive posts on the topic, and a questionnaire was created from the Google Forms platform on different social media to obtain and collect data. A total of 1,075 responses were obtained from different locations. The questionnaire included all Brazilian regions, all 26 states and the Federal District. The information published on the created page helped to clarify doubts related to the main aspects of the disease. The data obtained from the questionnaire contribute towards the planning of actions aimed at health education in a more strategic way, aiming at contributing to the points where the population is less knowledgeable.(AU)

La rabia es una antropozoonosis viral que se desarrolla de forma progresiva y aguda y puede presentar hasta un 100% de letalidad. Su agente etiológico es el virus de la rabia del género Lyssavirus perteneciente a la familia Rhabdoviridae. Este estudio tuvo como objetivo difundir informaciones sobre la ocurrencia de la rabia en humanos en virtud de su importancia expresiva para la salud pública, y analizar la percepción de la población sobre la rabia humana, con el fin de obtener datos relacionados a los conocimientos básicos sobre la enfermedad. Para ello, se creó una página de información denominada "@ contraraiva _" en la red social Instagram para realizar publicaciones interactivas sobre el tema abordado, y utilizar un cuestionario creado a partir de la plataforma Google Forms en diferentes redes sociales para la obtención y recolección de datos. Se obtuvieron 1.075 respuestas de diferentes lugares. El cuestionario llegó a todas las regiones brasileñas, a los 26 estados y al Distrito Federal. Las informaciones publicadas en el sitio web creado, ayudaron a sanar dudas relacionadas a los principales aspectos de la enfermedad. Los datos obtenidos del cuestionario contribuyen a la planificación de acciones orientadas a la educación para la salud de forma más estratégica, con el objetivo de contribuir a los puntos donde la población tiene menos conocimientos.(AU)

Phylogenetic analysis of nucleoprotein gene of Rabies virus in Malaysia from 2015 to 2018

@#Rabies is a fatal zoonotic disease caused by rabies virus (RABV) and remains a public health problem in Malaysia. Malaysia was declared rabies-free in 2012, however rabies outbreaks occurred at few states in Peninsular Malaysia three years later; and for the first time, in Sarawak (East Malaysia) in 2017 which has caused more than 20 human deaths. This study describes the phylogenetic analysis of the complete nucleoprotein (N) gene of RABV from animal samples in Malaysia from year 2015 to 2018. The N gene of 17 RABVs from Perlis, Kedah and Sarawak were amplified and sequenced. The nucleotide and deduced amino acid similarities of N gene analysis indicated that there is high similarity among the local RABVs. Phylogenetic analysis of the N gene revealed that all Malaysia RABVs belonged to the Asian clade. Among these, RABVs from Peninsular Malaysia were clustered together with RABVs from Thailand, Vietnam and other Southeast Asia countries except Indonesia. However, RABVs from Sarawak were grouped together with Indonesian strains from Kalimantan. Our study provides baseline genetic information of the potential origins of the circulating RABVs in Malaysia. This crucial information helped the authority in policies making and strategies to be taken in outbreak control. Continuous surveillance program to monitor the disease trend, strict border control, vaccination of dog and cat population and public awareness are important steps to control the spread of the RABV.

Algunos aspectos clínicos y epidemiológicos relacionados con mordeduras de perro en niños / Some clinical and epidemiological aspects related to dog bites in children

Introducción: Las mordeduras de perro afectan fundamentalmente a pacientes en la edad pediátrica, estas engendran peligros como secuelas estéticas y la posibilidad implícita de transmisibilidad del virus de la rabia a seres humanos. Objetivo: caracterizar el comportamiento clínico-epidemiológico de las mordeduras de perro en niños. Métodos: Se realizó un estudio observacional, longitudinal y prospectivo en pacientes con diagnóstico de mordedura de perro con necesidad de inmunización pasiva y activa, ingresados en el Servicio de Miscelánea del Hospital Infantil Sur Antonio María Béguez César de Santiago de Cuba, del 1ro de enero de 2017 al 31 de diciembre de 2018. Resultados: En los pacientes incluidos en el estudio predominaron los del sexo masculino (57,7 por ciento), los grupos de edades de 1 a 5 años 11 meses y 29 días, y de 6 a 10 años 11 meses y 29 días fueron los de mayor incidencia. Aunque no existió asociación estadística entre la localización anatómica de las lesiones y la edad, fueron las de miembros inferiores las más frecuentes en todas las edades con 63 pacientes (40,4 por ciento); destacándose como complicaciones la celulitis, las lesiones óseas y las secuelas estéticas; ningún niño presentó infección rábica. En el 50 por ciento de los casos la mordedura ocurrió en el horario nocturno y los perros de otro domicilio causaron la mayor parte de las agresiones. Conclusiones: El diagnóstico de mordeduras de perro se realizó con mayor frecuencia en niños del sexo masculino de uno y más años, en el horario nocturno y la principal complicación fue la celulitis(AU)

Introduction: Dog bites mainly affect patients in pediatric ages. They pose dangers such as esthetic sequels and the implicit possibility of transmission of the rabies virus to human beings. Objective: Characterize the clinical-epidemiological behavior of dog bites in children. Methods: An observational longitudinal prospective study was conducted of patients diagnosed with dog bite requiring passive and active immunization admitted to the Miscellany Service of Antonio María Béguez César South Children's Hospital in Santiago de Cuba from 1 January 2017 to 31 December 2018. Results: Among the patients included in the study there was a predominance of the male sex (57.7 percent). The age groups 1 to 5 years 11 months and 29 days and 6 to 10 years 11 months and 29 days showed the greatest incidence. Though no statistical association was found between anatomical location of the injuries and age, bites on the lower limbs were the most common at all ages with 63 patients (40.4 percent). Outstanding complications were cellulitis, bone lesions and esthetic sequels. No child had rabies infection. In 50 percent of the cases the bite occurred in the night hours. Most of the attacks were performed by dogs from other households. Conclusions: Dog bite diagnosis was more common among male children aged one year and over. Most bites occurred in the night hours. The main complication was cellulitis(AU)

Whole-Brain Monosynaptic Inputs to Hypoglossal Motor Neurons in Mice / 神经科学通报·英文版

Hypoglossal motor neurons (HMNs) innervate tongue muscles and play key roles in a variety of physiological functions, including swallowing, mastication, suckling, vocalization, and respiration. Dysfunction of HMNs is associated with several diseases, such as obstructive sleep apnea (OSA) and sudden infant death syndrome. OSA is a serious breathing disorder associated with the activity of HMNs during different sleep-wake states. Identifying the neural mechanisms by which the state-dependent activities of HMNs are controlled may be helpful in providing a theoretical basis for effective therapy for OSA. However, the presynaptic partners governing the activity of HMNs remain to be elucidated. In the present study, we used a cell-type-specific retrograde tracing system based on a modified rabies virus along with a Cre/loxP gene-expression strategy to map the whole-brain monosynaptic inputs to HMNs in mice. We identified 53 nuclei targeting HMNs from six brain regions: the amygdala, hypothalamus, midbrain, pons, medulla, and cerebellum. We discovered that GABAergic neurons in the central amygdaloid nucleus, as well as calretinin neurons in the parasubthalamic nucleus, sent monosynaptic projections to HMNs. In addition, HMNs received direct inputs from several regions associated with respiration, such as the pre-Botzinger complex, parabrachial nucleus, nucleus of the solitary tract, and hypothalamus. Some regions engaged in sleep-wake regulation (the parafacial zone, parabrachial nucleus, ventral medulla, sublaterodorsal tegmental nucleus, dorsal raphe nucleus, periaqueductal gray, and hypothalamus) also provided primary inputs to HMNs. These results contribute to further elucidating the neural circuits underlying disorders caused by the dysfunction of HMNs.

In Vivo Two-photon Calcium Imaging in Dendrites of Rabies Virus-labeled V1 Corticothalamic Neurons / 神经科学通报·英文版

Monitoring neuronal activity in vivo is critical to understanding the physiological or pathological functions of the brain. Two-photon Ca imaging in vivo using a cranial window and specific neuronal labeling enables real-time, in situ, and long-term imaging of the living brain. Here, we constructed a recombinant rabies virus containing the Ca indicator GCaMP6s along with the fluorescent protein DsRed2 as a baseline reference to ensure GCaMP6s signal reliability. This functional tracer was applied to retrogradely label specific V1-thalamus circuits and detect spontaneous Ca activity in the dendrites of V1 corticothalamic neurons by in vivo two-photon Ca imaging. Notably, we were able to record single-spine spontaneous Ca activity in specific circuits. Distinct spontaneous Ca dynamics in dendrites of V1 corticothalamic neurons were found for different V1-thalamus circuits. Our method can be applied to monitor Ca dynamics in specific input circuits in vivo, and contribute to functional studies of defined neural circuits and the dissection of functional circuit connections.