Autism spectrum disorder (ASD) currently affects nearly 1 in 160 children worldwide. In over two-thirds of evaluations, no validated diagnostics are used and gold standard diagnostic tools are used in less than 5% of evaluations. Currently, the diagnosis of ASD requires lengthy and expensive tests, in addition to clinical confirmation. Therefore, fast, cheap, portable, and easy-to-administer screening instruments for ASD are required. Several studies have shown that children with ASD have a lower preference for social scenes compared with children without ASD. Based on this, eye-tracking and measurement of gaze preference for social scenes has been used as a screening tool for ASD. Currently available eye-tracking software requires intensive calibration, training, or holding of the head to prevent interference with gaze recognition limiting its use in children with ASD. In this study, we designed a simple eye-tracking algorithm that does not require calibration or head holding, as a platform for future validation of a cost-effective ASD potential screening instrument. This system operates on a portable and inexpensive tablet to measure gaze preference of children for social compared to abstract scenes. A child watches a one-minute stimulus video composed of a social scene projected on the left side and an abstract scene projected on the right side of the tablet’s screen. We designed five stimulus videos by changing the social/abstract scenes. Every child observed all the five videos in random order. We developed an eye-tracking algorithm that calculates the child’s gaze preference for the social and abstract scenes, estimated as the percentage of the accumulated time that the child observes the left or right side of the screen, respectively. Twenty-three children without a prior history of ASD and 8 children with a clinical diagnosis of ASD were evaluated. The recorded video of the child´s eye movement was analyzed both manually by an observer and automatically by our algorithm. This study demonstrates that the algorithm correctly differentiates visual preference for either the left or right side of the screen (social or abstract scenes), identifies distractions, and maintains high accuracy compared to the manual classification. The error of the algorithm was 1.52%, when compared to the gold standard of manual observation. This tablet-based gaze preference/eye-tracking algorithm can estimate gaze preference in both children with ASD and without ASD to a high degree of accuracy, without the need for calibration, training, or restraint of the children. This system can be utilized in low-resource settings as a portable and cost-effective potential screening tool for ASD.

Norovirus is a major cause of acute gastroenteritis worldwide. Over 30 different genotypes, mostly from genogroup I (GI) and II (GII), have been shown to infect humans. Despite three decades of genome sequencing, our understanding of the role of genomic diversification across continents and time is incomplete. To close the spatiotemporal gap of genomic information of human noroviruses, we conducted a large-scale genome-wide analyses that included the nearly full-length sequencing of 281 archival viruses circulating since the 1970s in over 10 countries from four continents, with a major emphasis on norovirus genotypes that are currently underrepresented in public genome databases. We provided new genome information for 24 distinct genotypes, including the oldest genome information from 12 norovirus genotypes. Analyses of this new genomic information, together with those publicly available, showed that (i) noroviruses evolve at similar rates across genomic regions and genotypes; (ii) emerging viruses evolved from transiently-circulating intermediate viruses; (iii) diversifying selection on the VP1 protein was recorded in genotypes with multiple variants; (iv) non-structural proteins showed a similar branching on their phylogenetic trees; and (v) contrary to the current understanding, there are restrictions on the ability to recombine different genomic regions, which results in co-circulating populations of viruses evolving independently in human communities. This study provides a comprehensive genetic analysis of diverse norovirus genotypes and the role of non-structural proteins on viral diversification, shedding new light on the mechanisms of norovirus evolution and transmission.

SARS-CoV-2 main protease is a common target for inhibition assays due to its high conservation among coronaviruses. Since flavonoids show antiviral activity, several in silico works have proposed them as potential SARS-CoV-2 main protease inhibitors. Nonetheless, there is reason to doubt certain results given the lack of consideration for flavonoid promiscuity or main protease plasticity, usage of short library sizes, absence of control molecules and/or the limitation of the methodology to a single target site. Here, we report a virtual screening study where dorsilurin E, euchrenone a11, sanggenol O and CHEMBL2171598 are proposed to inhibit main protease through different pathways. Remarkably, novel structural mechanisms were observed after sanggenol O and CHEMBL2171598 bound to experimentally proven allosteric sites. The former drastically affected the active site, while the latter triggered a hinge movement which has been previously reported for an inactive SARS-CoV main protease mutant. The use of a curated database of 4.8 k flavonoids, combining two well-known docking software (AutoDock Vina and AutoDock4.2), molecular dynamics and MMPBSA, guaranteed an adequate analysis and robust interpretation. These criteria can be considered for future screening campaigns against SARS-CoV-2 main protease.

Tuberculosis is an infectious disease that causes ill health and death in millions of people each year worldwide. Timely diagnosis and treatment is key to full patient recovery. The Microscopic Observed Drug Susceptibility (MODS) is a test to diagnose TB infection and drug susceptibility directly from a sputum sample in 7–10 days with a low cost and high sensitivity and specificity, based on the visual recognition of specific growth cording patterns of M. Tuberculosis in a broth culture. Despite its advantages, MODS is still limited in remote, low resource settings, because it requires permanent and trained technical staff for the image-based diagnostics. Hence, it is important to develop alternative solutions, based on reliable automated analysis and interpretation of MODS cultures. In this study, we trained and evaluated a convolutional neural network (CNN) for automatic interpretation of MODS cultures digital images. The CNN was trained on a dataset of 12,510 MODS positive and negative images obtained from three different laboratories, where it achieved 96.63 +/- 0.35% accuracy, and a sensitivity and specificity ranging from 91% to 99%, when validated across held-out laboratory datasets. The model's learned features resemble visual cues used by expert diagnosticians to interpret MODS cultures, suggesting that our model may have the ability to generalize and scale. It performed robustly when validated across held-out laboratory datasets and can be improved upon with data from new laboratories. This CNN can assist laboratory personnel, in low resource settings, and is a step towards facilitating automated diagnostics access to critical areas in developing countries.

Tuberculosis control efforts are hampered by a mismatch in diagnostic technology: modern optimal diagnostic tests are least available in poor areas where they are needed most. Lack of adequate early diagnostics and MDR detection is a critical problem in control efforts. The Microscopic Observation Drug Susceptibility (MODS) assay uses visual recognition of cording patterns from Mycobacterium tuberculosis (MTB) to diagnose tuberculosis infection and drug susceptibility directly from a sputum sample in 7–10 days with a low cost. An important limitation that laboratories in the developing world face in MODS implementation is the presence of permanent technical staff with expertise in reading MODS. We developed a pattern recognition algorithm to automatically interpret MODS results from digital images. The algorithm using image processing, feature extraction and pattern recognition determined geometrical and illumination features used in an object-model and a photo-model to classify TB-positive images. 765 MODS digital photos were processed. The single-object model identified MTB (96.9% sensitivity and 96.3% specificity) and was able to discriminate non-tuberculous mycobacteria with a high specificity (97.1% M. avium, 99.1% M. chelonae, and 93.8% M. kansasii). The photo model identified TB-positive samples with 99.1% sensitivity and 99.7% specificity. This algorithm is a valuable tool that will enable automatic remote diagnosis using Internet or cellphone telephony. The use of this algorithm and its further implementation in a telediagnostics platform will contribute to both faster TB detection and MDR TB determination leading to an earlier initiation of appropriate treatment.

Tuberculosis (TB) is a major global health problem and drug resistance compromises the efforts to control this disease. Pyrazinamide (PZA) is an important drug used in both first and second line treatment regimes. However, its complete mechanism of action and resistance remains unclear. We genotyped and sequenced the complete genomes of 68 M. tuberculosis strains isolated from unrelated TB patients in Peru. No clustering pattern of the strains was verified based on spoligotyping. We analyzed the association between PZA resistance with non-synonymous mutations and specific genes. We found mutations in pncA and novel genes significantly associated with PZA resistance in strains without pncA mutations. These included genes related to transportation of metal ions, pH regulation and immune system evasion. These results suggest potential alternate mechanisms of PZA resistance that have not been found in other populations, supporting that the antibacterial activity of PZA may hit multiple targets.