Evaluation of Vineyard Cropping Systems Using On-Board RGB-Depth Perception.

A non-destructive measuring technique was applied to test major vine geometric traits on measurements collected by a contactless sensor. Three-dimensional optical sensors have evolved over the past decade, and these advancements may be useful in improving phenomics technologies for other crops, such as woody perennials. Red, green and blue-depth (RGB-D) cameras, namely Microsoft Kinect, have a significant influence on recent computer vision and robotics research. In this experiment an adaptable mobile platform was used for the acquisition of depth images for the non-destructive assessment of branch volume (pruning weight) and related to grape yield in vineyard crops. Vineyard yield prediction provides useful insights about the anticipated yield to the winegrower, guiding strategic decisions to accomplish optimal quantity and efficiency, and supporting the winegrower with decision-making. A Kinect v2 system on-board to an on-ground electric vehicle was capable of producing precise 3D point clouds of vine rows under six different management cropping systems. The generated models demonstrated strong consistency between 3D images and vine structures from the actual physical parameters when average values were calculated. Correlations of Kinect branch volume with pruning weight (dry biomass) resulted in high coefficients of determination (R2 = 0.80). In the study of vineyard yield correlations, the measured volume was found to have a good power law relationship (R2 = 0.87). However due to low capability of most depth cameras to properly build 3-D shapes of small details the results for each treatment when calculated separately were not consistent. Nonetheless, Kinect v2 has a tremendous potential as a 3D sensor in agricultural applications for proximal sensing operations, benefiting from its high frame rate, low price in comparison with other depth cameras, and high robustness.

Comparing UAV-Based Technologies and RGB-D Reconstruction Methods for Plant Height and Biomass Monitoring on Grass Ley.

Pastures are botanically diverse and difficult to characterize. Digital modeling of pasture biomass and quality by non-destructive methods can provide highly valuable support for decision-making. This study aimed to evaluate aerial and on-ground methods to characterize grass ley fields, estimating plant height, biomass and volume, using digital grass models. Two fields were sampled, one timothy-dominant and the other ryegrass-dominant. Both sensing systems allowed estimation of biomass, volume and plant height, which were compared with ground truth, also taking into consideration basic economical aspects. To obtain ground-truth data for validation, 10 plots of 1 m² were manually and destructively sampled on each field. The studied systems differed in data resolution, thus in estimation capability. There was a reasonably good agreement between the UAV-based, the RGB-D-based estimates and the manual height measurements on both fields. RGB-D-based estimation correlated well with ground truth of plant height ( R 2 > 0.80 ) for both fields, and with dry biomass ( R 2 = 0.88 ), only for the timothy field. RGB-D-based estimation of plant volume for ryegrass showed a high agreement ( R 2 = 0.87 ). The UAV-based system showed a weaker estimation capability for plant height and dry biomass ( R 2 < 0.6 ). UAV-systems are more affordable, easier to operate and can cover a larger surface. On-ground techniques with RGB-D cameras can produce highly detailed models, but with more variable results than UAV-based models. On-ground RGB-D data can be effectively analysed with open source software, which is a cost reduction advantage, compared with aerial image analysis. Since the resolution for agricultural operations does not need fine identification the end-details of the grass plants, the use of aerial platforms could result a better option in grasslands.

CXCR5-Dependent Entry of CD8 T Cells into Rhesus Macaque B-Cell Follicles Achieved through T-Cell Engineering.

Follicular helper CD4 T cells, TFH, residing in B-cell follicles within secondary lymphoid tissues, are readily infected by AIDS viruses and are a major source of persistent virus despite relative control of viral replication. This persistence is due at least in part to a relative exclusion of effective antiviral CD8 T cells from B-cell follicles. To determine whether CD8 T cells could be engineered to enter B-cell follicles, we genetically modified unselected CD8 T cells to express CXC chemokine receptor 5 (CXCR5), the chemokine receptor implicated in cellular entry into B-cell follicles. Engineered CD8 T cells expressing human CXCR5 (CD8hCXCR5) exhibited ligand-specific signaling and chemotaxis in vitro Six infected rhesus macaques were infused with differentially fluorescent dye-labeled autologous CD8hCXCR5 and untransduced CD8 T cells and necropsied 48 h later. Flow cytometry of both spleen and lymph node samples revealed higher frequencies of CD8hCXCR5 than untransduced cells, consistent with preferential trafficking to B-cell follicle-containing tissues. Confocal fluorescence microscopy of thin-sectioned lymphoid tissues demonstrated strong preferential localization of CD8hCXCR5 T cells within B-cell follicles with only rare cells in extrafollicular locations. CD8hCXCR5 T cells were present throughout the follicles with some observed near infected TFH In contrast, untransduced CD8 T cells were found in the extrafollicular T-cell zone. Our ability to direct localization of unselected CD8 T cells into B-cell follicles using CXCR5 expression provides a strategy to place highly effective virus-specific CD8 T cells into these AIDS virus sanctuaries and potentially suppress residual viral replication.IMPORTANCE AIDS virus persistence in individuals under effective drug therapy or those who spontaneously control viremia remains an obstacle to definitive treatment. Infected follicular helper CD4 T cells, TFH, present inside B-cell follicles represent a major source of this residual virus. While effective CD8 T-cell responses can control viral replication in conjunction with drug therapy or in rare cases spontaneously, most antiviral CD8 T cells do not enter B-cell follicles, and those that do fail to robustly control viral replication in the TFH population. Thus, these sites are a sanctuary and a reservoir for replicating AIDS viruses. Here, we demonstrate that engineering unselected CD8 T cells to express CXCR5, a chemokine receptor on TFH associated with B-cell follicle localization, redirects them into B-cell follicles. These proof of principle results open a pathway for directing engineered antiviral T cells into these viral sanctuaries to help eliminate this source of persistent virus.

DNA prime/protein boost vaccination elicits robust humoral response in rhesus macaques using oligomeric simian immunodeficiency virus envelope and Advax delta inulin adjuvant.

The partial success of the RV144 trial underscores the importance of envelope-specific antibody responses for an effective HIV-1 vaccine. Oligomeric HIV-1 envelope proteins delivered with a potent adjuvant are expected to elicit strong antibody responses with broad neutralization specificity. To test this hypothesis, two SIV envelope proteins were formulated with delta inulin-based adjuvant (Advax) and used to immunize nonhuman primates. Oligomeric gp140-gp145 from SIVmac251 and SIVsmE660 was purified to homogeneity. Oligomers showed high-affinity interaction with CD4 and were highly immunogenic in rabbits, inducing Tier 2 SIV-neutralizing antibodies. The immunogenicity of an oligomeric Env DNA prime and protein boost together with Advax was evaluated in Chinese rhesus macaques. DNA administration elicited antibodies to both envelopes, and titres were markedly enhanced following homologous protein boosts via intranasal and intramuscular routes. Strong antibody responses were detected against the V1 and V2 domains of gp120. During peak immune responses, a low to moderate level of neutralizing activity was detected against Tier 1A/1B SIV isolates, with a moderate level noted against a Tier 2 isolate. Increased serum antibody affinity to SIVmac251 gp140 and generation of Env-specific memory B cells were observed in the immunized macaques. Animals were subjected to low-dose intravaginal challenge with SIVmac251 one week after the last protein boost. One out of three immunized animals was protected from infection. Although performed with a small number of macaques, this study demonstrates the utility of oligomeric envelopes formulated with Advax in eliciting broad antibody responses with the potential to provide protection against SIV transmission.

Adoptive Transfer of Engineered Rhesus Simian Immunodeficiency Virus-Specific CD8+ T Cells Reduces the Number of Transmitted/Founder Viruses Established in Rhesus Macaques.

AIDS virus infections are rarely controlled by cell-mediated immunity, in part due to viral immune evasion and immunodeficiency resulting from CD4+ T-cell infection. One likely aspect of this failure is that antiviral cellular immune responses are either absent or present at low levels during the initial establishment of infection. To test whether an extensive, timely, and effective response could reduce the establishment of infection from a high-dose inoculum, we adoptively transferred large numbers of T cells that were molecularly engineered with anti-simian immunodeficiency virus (anti-SIV) activity into rhesus macaques 3 days following an intrarectal SIV inoculation. To measure in vivo antiviral activity, we assessed the number of viruses transmitted using SIVmac239X, a molecularly tagged viral stock containing 10 genotypic variants, at a dose calculated to transmit 12 founder viruses. Single-genome sequencing of plasma virus revealed that the two animals receiving T cells expressing SIV-specific T-cell receptors (TCRs) had significantly fewer viral genotypes than the two control animals receiving non-SIV-specific T cells (means of 4.0 versus 7.5 transmitted viral genotypes; P = 0.044). Accounting for the likelihood of transmission of multiple viruses of a particular genotype, the calculated means of the total number of founder viruses transmitted were 4.5 and 14.5 in the experimental and control groups, respectively (P = 0.021). Thus, a large antiviral T-cell response timed with virus exposure can limit viral transmission. The presence of strong, preexisting T-cell responses, including those induced by vaccines, might help prevent the establishment of infection at the lower-exposure doses in humans that typically transmit only a single virus. IMPORTANCE: The establishment of AIDS virus infection in an individual is essentially a race between the spreading virus and host immune defenses. Cell-mediated immune responses induced by infection or vaccination are important contributors in limiting viral replication. However, in human immunodeficiency virus (HIV)/SIV infection, the virus usually wins the race, irreversibly crippling the immune system before an effective cellular immune response is developed and active. We found that providing an accelerated response by adoptively transferring large numbers of antiviral T cells shortly after a high-dose mucosal inoculation, while not preventing infection altogether, limited the number of individual viruses transmitted. Thus, the presence of strong, preexisting T-cell responses, including those induced by vaccines, might prevent infection in humans, where the virus exposure is considerably lower.

African green monkey TRIM5α restriction in simian immunodeficiency virus-specific rhesus macaque effector CD4 T cells enhances their survival and antiviral function.

UNLABELLED: The expression of xenogeneic TRIM5α proteins can restrict infection in various retrovirus/host cell pairings. Previously, we have shown that African green monkey TRIM5α (AgmTRIM5α) potently restricts both human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus mac239 (SIV(mac239)) replication in a transformed human T-cell line (L. V. Coren, et al., Retrovirology 12:11, 2015, http://dx.doi.org/10.1186/s12977-015-0137-9). To assess AgmTRIM5α restriction in primary cells, we transduced AgmTRIM5α into primary rhesus macaque CD4 T cells and infected them with SIV(mac239). Experiments with T-cell clones revealed that AgmTRIM5α could reproducibly restrict SIV(mac239) replication, and that this restriction synergizes with an intrinsic resistance to infection present in some CD4 T-cell clones. AgmTRIM5α transduction of virus-specific CD4 T-cell clones increased and prolonged their ability to suppress SIV spread in CD4 target cells. This increased antiviral function was strongly linked to decreased viral replication in the AgmTRIM5α-expressing effectors, consistent with restriction preventing the virus-induced cytopathogenicity that disables effector function. Taken together, our data show that AgmTRIM5α restriction, although not absolute, reduces SIV replication in primary rhesus CD4 T cells which, in turn, increases their antiviral function. These results support prior in vivo data indicating that the contribution of virus-specific CD4 T-cell effectors to viral control is limited due to infection. IMPORTANCE: The potential of effector CD4 T cells to immunologically modulate SIV/HIV infection likely is limited by their susceptibility to infection and subsequent inactivation or elimination. Here, we show that AgmTRIM5α expression inhibits SIV spread in primary effector CD4 T cells in vitro. Importantly, protection of effector CD4 T cells by AgmTRIM5α markedly enhanced their antiviral function by delaying SIV infection, thereby extending their viability despite the presence of virus. Our in vitro data support prior in vivo HIV-1 studies suggesting that the antiviral CD4 effector response is impaired due to infection and subsequent cytopathogenicity. The ability of AgmTRIM5α expression to restrict SIV infection in primary rhesus effector CD4 T cells now opens an opportunity to use the SIV/rhesus macaque model to further elucidate the potential and scope of anti-AIDS virus effector CD4 T-cell function.

Potent restriction of HIV-1 and SIVmac239 replication by African green monkey TRIM5α.

BACKGROUND: The TRIM5α protein is a principal restriction factor that contributes to an HIV-1 replication block in rhesus macaque CD4+ T cells by preventing reverse transcription. HIV-1 restriction is induced in human CD4+ T cells by expression of rhesus TRIM5α as well as those of other old world monkeys. While TRIM5α restriction has been extensively studied in single-round infection assays, fewer studies have examined restriction after extended viral replication. RESULTS: To examine TRIM5α restriction of replication, we studied the ability of TRIM5α proteins from African green monkey (AgmTRIM5α) and gorilla (gorTRIM5α) to restrict HIV-1 and SIVmac239 replication. These xenogeneic TRIM5α genes were transduced into human Jurkat-CCR5 cells (JR5), which were then exposed to HIV-1 or SIVmac239. In our single-round infection assays, AgmTRIM5α showed a relatively modest 4- to 10-fold restriction of HIV-1 and SIVmac239, while gorTRIM5α produced a 2- and 3-fold restriction of HIV-1 and SIVmac239, respectively, consistent with the majority of previously published single-round studies. To assess the impact of these modest effects on infection, we tested restriction in replication systems initiated with either cell-free or cell-to-cell challenges. AgmTRIM5α powerfully restricted both HIV-1 and SIVmac239 replication 14 days after cell-free infection, with a ≥ 3-log effect. Moreover, expression of AgmTRIM5α restricted HIV-1 and SIVmac239 replication by 2-logs when co-cultured with infected JR5 cells for 12 days. In contrast, neither expression of gorTRIM5α nor rhesus TRIM5α induced significant resistance when co-cultured with infected cells. Follow up experiments showed that the observed differences between replication and infection were not due to assembly defects as xenogeneic TRIM5α expression had no effect on either virion production or specific infectivity. CONCLUSIONS: Our results indicate that AgmTRIM5α has a much greater effect on extended replication than on any single infection event, suggesting that AgmTRIM5α restriction acts cumulatively, building up over many rounds of replication. Furthermore, AgmTRIM5α was able to potently restrict both HIV-1 and SIV replication in a cell-to-cell infection challenge. Thus, AgmTRIM5α is unique among the TRIM5α species tested to date, being able to restrict even at the high multiplicities of infection presented by mixed culture with nonrestrictive infected cells.

Src-kinase inhibitors sensitize human cells of myeloid origin to Toll-like-receptor-induced interleukin 12 synthesis.

Src-kinase inhibitors hold great potential as targeted therapy against malignant cells. However, such inhibitors may also affect nonmalignant cells and cause pronounced off-target effects. We investigated the role of the dual kinase inhibitor dasatinib on human myeloid cells. Dasatinib is clinically used for the treatment of bcr/abl⁺ leukemias because it blocks the mutated tyrosine kinase abl. To understand its effect on the development of antigen-specific T-cell responses, we assessed antigen-specific priming of human, naïve T cells. In surprising contrast to the direct inhibition of T-cell activation by dasatinib, pretreatment of maturing dendritic cells (DCs) with dasatinib strongly enhanced their stimulatory activity. This effect strictly depended on the activating DC stimulus and led to enhanced interleukin 12 (IL-12) production and T-cell responses of higher functional avidity. Src-kinase inhibitors, and not conventional tyrosine kinase inhibitors, increased IL-12 production in several cell types of myeloid origin, such as monocytes and classical or nonclassical DCs. Interestingly, only human cells, but not mouse or macaques DCs, were affected. These data highlight the potential immunostimulatory capacity of a group of novel drugs, src-kinase inhibitors, thereby opening new opportunities for chemoimmunotherapy. These data also provide evidence for a regulatory role of src kinases in the activation of myeloid cells.

Triosephosphate isomerase of Taenia solium (TTPI): phage display and antibodies as tools for finding target regions to inhibit catalytic activity.

Previous studies demonstrated that antibodies against triosephosphate isomerase of Taenia solium (TTPI) can alter its enzymatic catalysis. In the present study, we used antibodies produced against the NH2-terminal region of TTPI (1/3NH2TTPI) and the phage display technology to find target regions to inhibit TTPI activity. As a first step, we obtained polyclonal antibodies against non-conserved regions from the 1/3NH2TTPI, which had an inhibitory effect of about 74 % on catalytic activity. Afterward, they were used to screen a library of phage-displayed dodecapeptides; as a result, 41 phage mimotope clones were isolated and grouped according to their amino acid sequence, finding the consensus A1 (VPTXPI), A2 (VPTXXI), B (LTPGQ), and D (DPLPR). Antibodies against selected phage mimotope clones were obtained by rabbit's immunization; these ones clearly recognized TTPI by both Western blot and ELISA. However, only the mimotope PDTS16 (DSVTPTSVMAVA) clone, which belongs to the VPTXXI consensus, raised antibodies capable of inhibiting the TTPI catalytic activity in 45 %. Anti-PDTS16 antibodies were confronted to several synthetic peptides that encompass the 1/3NH2TTPI, and they only recognized three, which share the motif FDTLQK belonging to the helix-α1 in TTPI. This suggests that this motif is the main part of the epitope recognized by anti-PDTS16 antibodies and revealed its importance for TTPI catalysis.

A non-chemical system for online weed control.

Non-chemical weed control methods need to be directed towards a site-specific weeding approach, in order to be able to compete the conventional herbicide equivalents. A system for online weed control was developed. It automatically adjusts the tine angle of a harrow and creates different levels of intensity: from gentle to aggressive. Two experimental plots in a maize field were harrowed with two consecutive passes. The plots presented from low to high weed infestation levels. Discriminant capabilities of an ultrasonic sensor were used to determine the crop and weed variability of the field. A controlling unit used ultrasonic readings to adjust the tine angle, producing an appropriate harrowing intensity. Thus, areas with high crop and weed densities were more aggressively harrowed, while areas with lower densities were cultivated with a gentler treatment; areas with very low densities or without weeds were not treated. Although the weed development was relatively advanced and the soil surface was hard, the weed control achieved by the system reached an average of 51% (20%-91%), without causing significant crop damage as a result of harrowing. This system is proposed as a relatively low cost, online, and real-time automatic harrow that improves the weed control efficacy, reduces energy consumption, and avoids the usage of herbicide.

Development and testing of a decision making based method to adjust automatically the harrowing intensity.

Harrowing is often used to reduce weed competition, generally using a constant intensity across a whole field. The efficacy of weed harrowing in wheat and barley can be optimized, if site-specific conditions of soil, weed infestation and crop growth stage are taken into account. This study aimed to develop and test an algorithm to automatically adjust the harrowing intensity by varying the tine angle and number of passes. The field variability of crop leaf cover, weed density and soil density was acquired with geo-referenced sensors to investigate the harrowing selectivity and crop recovery. Crop leaf cover and weed density were assessed using bispectral cameras through differential images analysis. The draught force of the soil opposite to the direction of travel was measured with electronic load cell sensor connected to a rigid tine mounted in front of the harrow. Optimal harrowing intensity levels were derived in previously implemented experiments, based on the weed control efficacy and yield gain. The assessments of crop leaf cover, weed density and soil density were combined via rules with the aforementioned optimal intensities, in a linguistic fuzzy inference system (LFIS). The system was evaluated in two field experiments that compared constant intensities with variable intensities inferred by the system. A higher weed density reduction could be achieved when the harrowing intensity was not kept constant along the cultivated plot. Varying the intensity tended to reduce the crop leaf cover, though slightly improving crop yield. A real-time intensity adjustment with this system is achievable, if the cameras are attached in the front and at the rear or sides of the harrow.

Discriminating crop, weeds and soil surface with a terrestrial LIDAR sensor.

In this study, the evaluation of the accuracy and performance of a light detection and ranging (LIDAR) sensor for vegetation using distance and reflection measurements aiming to detect and discriminate maize plants and weeds from soil surface was done. The study continues a previous work carried out in a maize field in Spain with a LIDAR sensor using exclusively one index, the height profile. The current system uses a combination of the two mentioned indexes. The experiment was carried out in a maize field at growth stage 12-14, at 16 different locations selected to represent the widest possible density of three weeds: Echinochloa crus-galli (L.) P.Beauv., Lamium purpureum L., Galium aparine L.and Veronica persica Poir.. A terrestrial LIDAR sensor was mounted on a tripod pointing to the inter-row area, with its horizontal axis and the field of view pointing vertically downwards to the ground, scanning a vertical plane with the potential presence of vegetation. Immediately after the LIDAR data acquisition (distances and reflection measurements), actual heights of plants were estimated using an appropriate methodology. For that purpose, digital images were taken of each sampled area. Data showed a high correlation between LIDAR measured height and actual plant heights (R2 = 0.75). Binary logistic regression between weed presence/absence and the sensor readings (LIDAR height and reflection values) was used to validate the accuracy of the sensor. This permitted the discrimination of vegetation from the ground with an accuracy of up to 95%. In addition, a Canonical Discrimination Analysis (CDA) was able to discriminate mostly between soil and vegetation and, to a far lesser extent, between crop and weeds. The studied methodology arises as a good system for weed detection, which in combination with other principles, such as vision-based technologies, could improve the efficiency and accuracy of herbicide spraying.

Immune correlates analysis of a phase 3 trial of the AZD1222 (ChAdOx1 nCoV-19) vaccine.

In the phase 3 trial of the AZD1222 (ChAdOx1 nCoV-19) vaccine conducted in the U.S., Chile, and Peru, anti-spike binding IgG concentration (spike IgG) and pseudovirus 50% neutralizing antibody titer (nAb ID50) measured four weeks after two doses were assessed as correlates of risk and protection against PCR-confirmed symptomatic SARS-CoV-2 infection (COVID-19). These analyses of SARS-CoV-2 negative participants were based on case-cohort sampling of vaccine recipients (33 COVID-19 cases by 4 months post dose two, 463 non-cases). The adjusted hazard ratio of COVID-19 was 0.32 (95% CI: 0.14, 0.76) per 10-fold increase in spike IgG concentration and 0.28 (0.10, 0.77) per 10-fold increase in nAb ID50 titer. At nAb ID50 below the limit of detection (< 2.612 IU50/ml), 10, 100, and 270 IU50/ml, vaccine efficacy was -5.8% (-651%, 75.6%), 64.9% (56.4%, 86.9%), 90.0% (55.8%, 97.6%) and 94.2% (69.4%, 99.1%). These findings provide further evidence towards defining an immune marker correlate of protection to help guide regulatory/approval decisions for COVID-19 vaccines.

[Social and cultural barriers in pregnancy adolescents]. / El embarazo en adolescentes. Factores socioculturales.

OBJECTIVE: to compare pregnant adolescents of 15 and 19 years old to non-pregnant adolescents of the same age group with active sexual life in order to explore the existing relationship of the social and cultural barriers and the accessing of methods of birth control. METHODS: a case-control study was performed in a six month period at Ciudad Juárez, Chihuahua, Mexico. The cases included pregnant adolescents of 15-19 years of age; the controls were non-pregnant adolescents with an active sexual life of the same age group. The size of the sample was of 96 adolescents, 48 cases and 48 controls. It was calculated RM to obtain associations of a 95 % IC. RESULTS: the non-utilization of birth control methods was associated with pregnancy (OR = 2.78, CI 95 % = 1.21-6.36), low education (OR = 2.66, 95 % CI = 0.97-7.29), and no medical service access (OR = 2.23, CI 95 % = 0.96-5.18). CONCLUSIONS: the social and cultural barriers were associated with pregnancy during the adolescence.

Paraplegia due to spinal meningioma during the third trimester of pregnancy: case report and literature review.

INTRODUCTION: Spinal meningiomas represent 25-45% of intradural spinal tumors and ~2% of meningiomas of the central nervous system (CNS), and their occurrence during pregnancy is unusual. We present an updated literature review. CASE REPORT: A 36-year-old woman, at 32.6 weeks of gestation, was hospitalized for urinary tract infection and urinary retention. One month earlier, she had decreased strength in lower limbs, and this weakness rapidly progressed to flaccid paraplegia without sphincters control. Magnetic resonance imaging (MRI) revealed a well-defined intradural extramedullary lesion in T3-T4. Using a posterolateral approach, the tumor was completely removed; however, there was no clinical improvement, and the patient was discharged with an impairment scale (AIS) grade A. Histopathology examination indicated a psammomatous meningioma. DISCUSSION: Meningiomas are benign tumors that are slowly progressive; however, the hemodynamic and hormonal changes of pregnancy are related to their accelerated growth. Reports show that the onset of the symptoms during the third trimester of pregnancy, including early neurological symptoms or signs of spinal cord compression, can be easily attributed to those of pregnancy by both the patient and the doctor. The time to diagnosis and medulla compression time are thus prolonged, which can be further compounded in middle-high income countries due to limitations in obtaining images for evaluation. Although rare, spinal meningiomas should be considered in the differential diagnosis of patients with neurological symptoms during pregnancy. Their early recognition is important to avoid irreversible neurological damage.

Clinical and epidemiological characteristics of probable cases of congenital Zika syndrome and dengue antibody levels, Tuxtla Gutierrez, Chiapas, Mexico

Introduction: We previously found that the occurrence of congenital Zika syndrome was under-reported in Mexico. It was postulated that high dengue virus antibody levels found at the population-level in endemic countries might have contributed to the occurrence of the regional pandemic of Zika disease. A case series of suspected cases of congenital Zika syndrome in a maternity hospital in Tuxtla Gutierrez, Chiapas, Mexico was assembled to assess why they were not reported and to explore the hypothesis of dengue virus antibody-dependent enhancement of Zika disease. Methods: We used a quantitative approach to describe clinical and imaging records and used data from interviews of a total of 13 suspected cases of congenital Zika syndrome. We also quantitated dengue 1 and 2 antibodies using an 80% plaque reduction neutralization test of sera specimens obtained from the mothers of these 13 cases and compared them to those of a group of mothers who delivered normal newborns in the same hospital. Results: Only one of the suspected cases was laboratory-confirmed because appropriate specimens were not collected from the newborns as required by the case definition. We found 1) microcephaly, 2) hypoplasia/hypogeneses, thinning or absence of brain structures, 3) multiple birth defects, 4) calcifications, and cysts, 5) meningocele/encephalocele, and 6) hydrocephalus in 100 %, 76.9 %, 38.5 %, 38.5 %, 30.8 %, and 23.1 %, respectively of the case series. The cases clustered geographically, and 77 % occurred between May 2016 to March 2017 and recalled or were told by a doctor they had Zika fever. There was a four-fold increased risk of congenital Zika syndrome among those with dengue 1 antibody as compared to those with dengue 2 antibodies (odds ratio = 3.6; 95 % confidence interval: 0.7, 20.5), reaching only borderline statistical significance. Conclusions: We found in the largest maternal facility of the capital of the State of Chiapas, in Mexico, that only 7.7 % of suspected cases were confirmed, and that the rather complex requirement of cerebrospinal fluid specimens or serological specimens of newborns for suspected cases of congenital Zika syndrome used during the pandemic resulted in low sensitivity of the surveillance system. The finding of higher levels of dengue 1 than dengue 2 antibodies in cases than the referent population, requires further evaluation and may suggest a role for dengue antibody-dependent response in Zika disease.

Introducción: Previamente los autores habían encontrado evidencia de sub-notificación de la ocurrencia del síndrome congénito por Zika en México. Se ha postulado que niveles elevados de anticuerpos contra los virus del dengue a nivel poblacional en los países endémicos hubiese contribuido a la ocurrencia de la pandemia regional de enfermedad por Zika. Ensamblamos una serie de casos sospechosos de síndrome congénito por Zika en un hospital de maternidad en Tuxtla Gutiérrez, Chiapas, México, para evaluar por qué no fueron notificados y explorar la hipótesis de enfermedad por Zika incrementada por anticuerpos anti-dengue. Métodos: Utilizamos un enfoque cuantitativo para describir 13 casos sospechosos de síndrome congénito por revisamos registros clínicos e imágenes, entrevistas. También cuantificamos los niveles de anticuerpos para los virus dengue 1 y 2 en suero de las madres de los casos comparados con los de mujeres que tuvieron recién nacidos normales en el mismo hospital. Resultados: Solamente uno de los 12 casos sospechosos fue confirmado por laboratorio, porque en los demás no se recolectaron especímenes adecuados de los neonatos como lo requería la definición de casos. Encontramos 1) microcefalia, 2) hipoplasia y adelgazamiento de las estructuras cerebrales, 3) malformaciones múltiples, 4) calcificaciones o quistes, 5) meningocele/encefalocele, y 6) hidrocefalia en: 100 %, 76.9 %, 38.5 %, 38.5 %, 30.8 %, y 23.1 %, en ese orden entre los casos sospechosos. Los casos se aglutinaron geográficamente y 77 % ocurrieron entre Mayo del 2016 y Marzo del 2017, y sus madres recordaban que tuvieron o que un profesional de la salud les dijo que tuvieron fiebre por Zika. Encontramos un incremento de casi 4 veces en el riesgo de síndrome congénito por Zika para aquellos con altos niveles de anticuerpos anti-dengue 1 comparado con anticuerpos anti-dengue 2 (cociente de suertes = 3.6; intervalo de confianza del 95 %: 0.7, 20.5), alcanzando solamente una significancia estadística limítrofe. Conclusiones: Encontramos en el establecimiento de atención a la maternidad más grande en la capital de Chiapas, México, que solamente 7.7 % de los casos sospechosos de síndrome congénito por Zika fueron confirmados y que los relativamente complejos requerimientos de la definición de casos de muestras serológicas o de líquido cefalorraquídeo resultó en una baja sensibilidad del sistema de vigilancia. El hallazgo de niveles más altos de anticuerpos a dengue 1 que dengue 2 requiere más evaluación y pudiera sugerir un papel de la respuesta dependiente de anticuerpos al dengue en Zika.

T-cell responses to KSHV infection: a systematic approach.

Prior studies of T-cell responses to KSHV have included relatively few participants and focused on relatively few KSHV antigens. To provide a more comprehensive analysis, we investigated T-cell responses to the whole KSHV proteome using IFN-γ ELISpot. Using ∼7,500 overlapping 15mer peptides we generated one to three peptide pools for each of the 82 KSHV ORFs. IFN-γ ELISpot analysis of PBMCs from 19 patients with a history of KSHV-associated disease and 24 healthy donors (11 KSHV seropositive) detected widely varied responses. Fifty six of the 82 ORFs were recognized by at least one individual but there was little overlap between participants. Responses to at least one ORF pool were observed in all 19 patients and in 7 seropositive donors. Four seropositive donors and 10 seronegative donors had no detectable responses while 3 seronegative donors had weak responses to one ORF. Patients recognised more ORFs than the donors (p=0.04) but the response intensity (spot forming units: SFU per million cells) was similar in the two groups. In four of the responding donors, individual peptides eliciting the predominant responses were identified: three donors responded to only one peptide per ORF, while one recognized five. Using intracellular cytokine staining in four participant samples, we detected peptide-induced IFN-γ, MIP1-ß, and TNF-α as well as CD107a degranulation, consistent with multifunctional effector responses in CD8+ and CD4+ T cells. Sequence analysis of TCRs present in peptide specific T-cell clones generated from two participants showed both mono- and multi-clonotypic responses. Finally, we molecularly cloned the KSHV specific TCRs and incorporated the sequences into retroviral vectors to transfer the specificities to fresh donor cells for additional studies. This study suggests that KSHV infected individuals respond to diverse KSHV antigens, consistent with a lack of shared immunodominance and establishes useful tools to facilitate KSHV immunology studies.

A novel SIV gag-specific CD4(+)T-cell clone suppresses SIVmac239 replication in CD4(+)T cells revealing the interplay between antiviral effector cells and their infected targets.

To study CD4(+)T-cell suppression of AIDS virus replication, we isolated nine rhesus macaque SIVGag-specific CD4(+)T-cell clones. One responding clone, Gag68, produced a typical cytotoxic CD8(+)T-cell response: induction of intracellular IFN-γ, MIP-1α, MIP-1ß, and CD107a degranulation. Gag68 effectively suppressed the spread of SIVmac239 in CD4(+)T cells with a corresponding reduction of infected Gag68 effector cells, suggesting that CD4(+)effectors need to suppress their own infection in addition to their targets to be effective. Gag68 TCR cloning and gene transfer into CD4(+)T cells enabled additional experiments with this unique specificity after the original clone senesced. Our data supports the idea that CD4(+)T cells can directly limit AIDS virus spread in T cells. Furthermore, Gag68 TCR transfer into CD4(+)T-cell clones with differing properties holds promise to better understand the suppressive effector mechanisms used by this important component of the antiviral response using the rhesus macaque model.

Cambios clínicos de la esclerosis múltiple según modificación de los criterios de McDonald. Hospital Almenara, 2001-2015 / Clinical changes of multiple sclerosis according to modification of the McDonald criteria. Hospital Almenara, 2001-2015

Objetivos. Determinar si las modificaciones en los criterios de McDonald en los últimos 15 años produjeron cambios en las características clínicas y el tiempo de diagnóstico de esclerosis múltiple. Métodos. Se revisaron 91 historias clínicas de pacientes con esclerosis múltiple de los últimos 15 años en el Servicio de Neurología del Hospital Nacional Guillermo Almenara Irigoyen, Lima. Se registraron características clínicas y demográficas de la enfermedad. Los datos se agruparon según la fecha del diagnóstico y los criterios de McDonald vigentes en ese momento: grupo 1 (2001 a 2005), grupo 2 (2005 a 2009) y grupo 3 (2010 a 2015). Resultados. La edad promedio de los pacientes fue 36,2 años, 51,7% fueron mujeres. Las manifestaciones clínicas más frecuentes fueron motoras. El 69,23% fue tipo remitente recurrente (EMRR), 26,37% secundaria progresiva (EMSP) y 4,40% primaria progresiva (EMPP). La discapacidad en EMRR fue leve y en EMSP fue moderada. Los pacientes con EMPP tuvieron discapacidad severa en el grupo 2 y moderada en el grupo 3. El tiempo de enfermedad fue mayor para el grupo 1 y similar en los grupos 2 y 3. Conclusiones. Las manifestaciones clínicas de la enfermedad no han cambiado durante los últimos 15 años. La EMRR fue el tipo más común. En los últimos 5 años hubo un aumento en el diagnóstico de la EMPP y disminución de EMSP por el diagnóstico precoz de la enfermedad. El tiempo de diagnóstico de la enfermedad disminuyó en los últimos 10 años.

Objective. Define if modifications in the McDonald criteria in the last 15 years produced changes in the clinical characteristics and the time of diagnosis of multiple sclerosis. Methods. We revised 91 clinical histories from patients with multiple sclerosis in the last 15 years from the Neurology Service of Hospital Nacional Guillermo Almenara Irigoyen, Lima. We registered clinical and demographical characteristics of the disease. The clinical data were grouped according the date of the diagnosis and McDonald criteria in force at that time: group 1 (2001 to 2005), group 2 (2005 to 2009) and group 3 (2010 to 2015). Results. The average age of the patients was 36,2 years. 51,7% were female. The most common clinical manifestations were motor. 69,23% was a recurrent sender type, 26,37% progressive secondary and 4,40% progressive primary. Disability in recurrent sender type was mild and in the progressive secondary group was moderate. The progressive primary patients had severe disability in group 2 and moderate in group 3. The average disease duration was higher for group 1 and similar in the 2 and 3 groups. Conclusions. The initial manifestations of the disease of patients with multiple sclerosis have not changed during this past 15 years. The predominance of recurrent remitent forms remains the most common but with an increase in progressive primary forms and decrease of progressive secondary group in the last 5 years due to earlier diagnosis of the disease. The time of diagnosis of the disease has decreased in the last 10 years.

Potential use of ground-based sensor technologies for weed detection.

Site-specific weed management is the part of precision agriculture (PA) that tries to effectively control weed infestations with the least economical and environmental burdens. This can be achieved with the aid of ground-based or near-range sensors in combination with decision rules and precise application technologies. Near-range sensor technologies, developed for mounting on a vehicle, have been emerging for PA applications during the last three decades. These technologies focus on identifying plants and measuring their physiological status with the aid of their spectral and morphological characteristics. Cameras, spectrometers, fluorometers and distance sensors are the most prominent sensors for PA applications. The objective of this article is to describe-ground based sensors that have the potential to be used for weed detection and measurement of weed infestation level. An overview of current sensor systems is presented, describing their concepts, results that have been achieved, already utilized commercial systems and problems that persist. A perspective for the development of these sensors is given.