Nut Geometry Inspection Using Improved Hough Line and Circle Methods.

Nuts are the cornerstone of human industrial construction, especially A-grade nuts that can only be used in power plants, precision instruments, aircraft, and rockets. However, the traditional nuts inspection method is to manually operate the measuring instrument for conducting an inspection, so the quality of the A-grade nut cannot be guaranteed. In this work, a machine vision-based inspection system was proposed, which performs a real-time geometric inspection of the nuts before and after tapping on the production line. In order to automatically screen out A-Grade nuts on the production line, there are 7 inspections within this proposed nut inspection system. The measurements of parallel, opposite side length, straightness, radius, roundness, concentricity, and eccentricity were proposed. To shorten the overall detection time regarding nut production, the program needed to be accurate and uncomplicated. By modifying the Hough line and Hough circle, the algorithm became faster and more suitable for nut detection. The optimized Hough line and Hough circle can be used for all measures in the testing process.

Activated macrophages of CD 163 gene edited pigs generated by direct cytoplasmic microinjection with CRISPR gRNA/Cas9 mRNA are resistant to PRRS virus assault.

Porcine reproductive and respiratory syndrome virus (PRRSV) infects placental and lung macrophages, causing a global epidemic with economic loss. Attempts to develop an effective vaccine to control the disease have not been effective. Currently, developing PRRSV disease-resistant pigs via a gene editing (GE) strategy to mutate the PRRSV receptor or to delete the binding domain on the macrophage appears promising. In this study, we used the strategy of Edinburg University to construct two guide RNAs (gRNAs) located on the proximal front and post sites of exon 7. Directive microinjection of two gRNAs and Cas9 mRNA into the cytoplasm of pronuclear zygotes efficiently generated four piglets confirmed as CD163 knockout (KO) and/or CD163 exon 7 deleted (CD163ΔE7). In four GE piglets, three pigs carried two chromosome CD163 KO or ΔE7. Peripheral blood mononuclear cells (PBMCs) from three GE and wild-type (WT) pigs were activated into macrophages for in vitro transfection. The results showed that the activated macrophages from all GE pigs were significantly more viable than those from WT pig. Current results suggest that we have successfully generated PRRSV-resistant pigs, although in vivo challenge is needed to validate that the pigs are PRRSV resistant.

Causes of in-hospital death in patients with type 2 diabetes with microvascular and macrovascular complications in Taiwan.

AIMS: Diabetes mellitus is a major cause of death worldwide, including Taiwan. The mortality data of the subsets of patients who suffered from microvascular or macrovascular complications is limited. The aim of this study was to investigate the causes of in-hospital death of patients with type 2 diabetes, especially the patients with microvascular, macrovascular and both micro-macrovascular complications. METHODS: A total of 12 159 patients with type 2 diabetes were identified from the Taiwan National Health Insurance Research Database (NHIRD) to analyse the causes of death. Type 2 diabetic subjects with microvascular, macrovascular and both micro-macrovascular complications were further classified and compared to patients without microvascular and macrovascular complications in the logistic regression analysis of the risk of death. RESULTS: Pneumonia increased risk of in-hospital death in patients with microvascular, macrovascular and both micro-macrovascular complications, with adjusted odds ratios (AORs) of 2.13 (95% confidence interval [CI] 1.09-4.18), 3.26 (1.71-6.24) and 3.96 (2.17-7.22), respectively. Septicaemia increased risk of in-hospital death in patients with macrovascular (AOR 2.57 [1.31-5.04]) and both micro-macrovascular complications (AOR 4.69 [2.58-8.50]). CONCLUSION: Pneumonia increased risk of in-hospital death among the type 2 diabetic patients with microvascular, macrovascular and both micro-macrovascular complications. Therefore, efforts aim at preventing pneumonia or decreasing its severity may increase survival.

Optimization-Based Approaches for Minimizing Deployment Costs for Wireless Sensor Networks with Bounded Estimation Errors.

As wireless sensor networks have become more prevalent, data from sensors in daily life are constantly being recorded. Due to cost or energy consumption considerations, optimization-based approaches are proposed to reduce deployed sensors and yield results within the error tolerance. The correlation-aware method is also designed in a mathematical model that combines theoretical and practical perspectives. The sensor deployment strategies, including XGBoost, Pearson correlation, and Lagrangian Relaxation (LR), are determined to minimize deployment costs while maintaining estimation errors below a given threshold. Moreover, the results significantly ensure the accuracy of the gathered information while minimizing the cost of deployment and maximizing the lifetime of the WSN. Furthermore, the proposed solution can be readily applied to sensor distribution problems in various fields.

A PCB Alignment System Using RST Template Matching with CUDA on Embedded GPU Board.

The fiducial-marks-based alignment process is one of the most critical steps in printed circuit board (PCB) manufacturing. In the alignment process, a machine vision technique is used to detect the fiducial marks and then adjust the position of the vision system in such a way that it is aligned with the PCB. The present study proposed an embedded PCB alignment system, in which a rotation, scale and translation (RST) template-matching algorithm was employed to locate the marks on the PCB surface. The coordinates and angles of the detected marks were then compared with the reference values which were set by users, and the difference between them was used to adjust the position of the vision system accordingly. To improve the positioning accuracy, the angle and location matching process was performed in refinement processes. To overcome the matching time, in the present study we accelerated the rotation matching by eliminating the weak features in the scanning process and converting the normalized cross correlation (NCC) formula to a sum of products. Moreover, the scanning time was reduced by implementing the entire RST process in parallel on threads of a graphics processing unit (GPU) by applying hash functions to find refined positions in the refinement matching process. The experimental results showed that the resulting matching time was around 32× faster than that achieved on a conventional central processing unit (CPU) for a test image size of 1280 × 960 pixels. Furthermore, the precision of the alignment process achieved a considerable result with a tolerance of 36.4µm.

General theory of asymmetrical polarization-dependent optics in functional material-doped 90° twisted nematic liquid crystals.

General theory, which can completely describe the asymmetrical optics in a functional material (FM)-doped 90° twisted nematic liquid crystals (TNLCs), is proposed using Cayley-Hamilton theorem and Jones calculus. The FMs, whose shape and size are similar to those of the adopted NLCs, can be aligned along the long axes of the NLCs. The FMs discussed herein are dichroic dye (DD) and polymer. The experimental results of asymmetrical transmission in DD-doped 90° TNLCs are consistent with the theoretical calculation. Such asymmetrical characterization can be further used in the current applications based on 90° TNLCs in all fields to obtain new potential functions.

Generation of GGTA1 Mutant Pigs by Direct Pronuclear Microinjection of CRISPR/Cas9 Plasmid Vectors.

This study was conducted to confirm that 1-site and 4-site ppU6-GGTA1-gRNA CRISPR vectors together with the pCX-Flag2-NLS1-Cas9-NLS2 plasmid can both generate KO pigs by direct pronuclear microinjection. In total, 41 and 53 fertilized eggs were microinjected on 1-site and 4-site strategies, respectively. The 1-site construction generated a litter of 8 piglets, and 2 were mono-allelic mutant (mMt). The injection of 4-site constructions resulted in one biallelic mutant (bMt) and one mMt piglet in a litter of 7. Those 3 mMt pigs had a 4 bp deletion, 5 bp insertion, or 7 bp insertion at site I, and the bMt pig had 5 types of mutations at cleavage sites I and III. The expression of alpha-Gal on the bMt peripheral blood mononuclear cells (PBMCs) was reduced, and survival rate of bMt PBMCs was maintained as indicated by results of cultivation with sera of humans or Formosan Macaques. We concluded that mutant pigs could be generated by direct pronuclear microinjection of ppU6-GGTA1-gRNA CRISPR vectors with the pCX-Flag2-NLS1-Cas9-NLS2 plasmid and that the 4-site strategy has a better mutant efficiency. Porcine U6 promoter was firstly used to express KO vectors and effectively generate mutant pigs, worthily to adopt for future KO studies.

A galactopoiesis accordant yield of functional recombinant human factor IX from homozygous transgenic pigs requires a large amount of vitamin K supplementation.

Transgenic pigs failed to accord milk yield curve to lactate rhFIX-a vitamin K (VK) dependent protein even fed with VK enriched to 8 times higher than nutritional requirement. A further higher VK supplementation may be required. Homozygous transgenic sows (n = 4, 200 kg) at their 3rd nursing were divided into control and treatment groups and respectively received VK enriched and further menadione (soluble VK) supplemented diet (220 mg/kg VK enriched diet) for 33 days. At next lactation, control sows than received treatment and previous treated were fed on control diet. Results revealed that menadione treatment increased milk bioactivity of rhFIX from the 7th day of 73 to the 21st day of 153 IU/mL; it gradually decreased to 96 IU/mL on 35th day of lactation. Under control feeding, bioactivity remained relatively unchanged. However, milk rhFIX concentration and ratio of activated rhFIX responded little to the treatment. The menadione-induced bioactivity curve agrees with the known lactation pattern of sow means rhFIX secretion is still galactopoietic but requires high VK intake to show. The ineffectual VK spend on lactational carboxylation might be common in other mammary VK dependent expression system but can be effectively overcome by a high supplementation of menadione with a 5-folds improvement in quality.

Construction of a CRISPR-Cas9 System for Pig Genome Targeting.

A Cas9/sgRNA RNA-guided endonuclease expression system including a codon-optimized Streptococcus pyogenes A20 Cas9 recombinant protein expression vector and a spacer-guide chimeric RNA expression vector using the porcine U6 promoter was constructed for application in pigs. Only the Flag2-NLS1-Cas9-NLS2 recombinant protein in complex with sgRNA was translocated into the nucleus; the Flag2-NLS1-Cas9-NLS2 protein alone was excluded from the nucleus. Up to 13% of porcine PK1 cells targeted in vitro were observed, regardless of transfection efficiency.

A novel method for separation of caseins from milk by phosphates precipitation.

Milk protein of farm animals is difficult to isolate because of the presence of casein micelles, which are hard to separate from whey by using centrifugation or filtration. Insoluble casein micelles also create an obstacle for purification instruments to operate efficiently. The conventional method, to precipitate caseins by lowering pH to 4.6 and then recover the whey fraction for further purification using chromatography techniques, is not applicable to proteins having an isoelectric point similar to caseins. In addition, the acid condition used for casein removal usually leads to significantly poor yields and reduced biological activities. In this study, a novel method of precipitating caseins under neutral or weak acidic conditions is presented. The method employs a phosphate salt and a freeze-thaw procedure to obtain a casein-free whey protein fraction. This fraction contains more than 90% yield with little loss of bioactivity of the target protein, and is readily available for further chromatographic purification. This method was successfully applied to purify recombinant human factor IX and recombinant hirudin from the milk of transgenic pigs in the presented study. It is an efficient pretreatment approach prior to chromatographic purification of milk protein from farm animals and particularly of great value to collect those recombinants secreted from transgenic livestock.

Production of viable cloned miniature pigs by aggregation of handmade cloned embryos at the 4-cell stage.

The aim of the present study was to improve the quality of handmade cloned porcine embryos by multiple embryo aggregations. Embryos derived from aggregation of three cloned embryos (3×) had a better blastocyst rate than cloned control (1×) embryos (73.6% vs 35.1%, respectively; P<0.05), but did not differ from those produced by aggregation of two cloned embryos (2×; 63.0%). Total cell numbers differed among treatments (P<0.05), with the greatest cell numbers (126) in the 3× group and the lowest (55) in the control group. The ratio of inner cell mass:total cell number was comparable in the 2× and 3× groups (25.1% vs 26.1%, respectively) and was significantly better than that in the control group (15.3%). The proportion of apoptotic cells in 2× and 3× groups was lower than that in the control group (2.7% and 2.2% vs 4.7%, respectively; P<0.05). Expression of Oct4 and Cdx2 was higher, whereas that of Bax was lower (P<0.05), in the 3× compared with non-aggregate group. Seven piglets were born to two surrogate mothers after embryo transfer of 3× aggregated blastocysts. In conclusion, aggregated embryos had greater total cell numbers and better pluripotency gene expression, with reduced expression of the pro-apoptosis gene Bax. Collectively, these improvement may be associated with the development of cloned embryos to term.

[Development of evidence-based clinical practice guidelines for endotracheal suction in premature infants].

BACKGROUND: Endotracheal suctioning (ETS) for mechanically ventilated premature infants is a routine practice in neonatal intensive care. However, ETS is associated with hypoxemia and bradycardia, which may cause brain damage and negatively affect neurodevelopmental outcomes. PURPOSE: This study develops a set of evidence-based clinical-practice ETS guidelines for premature infants. METHODS: A multidisciplinary task group at a medical center in southern Taiwan was established. Team members analyzed current ETS practice guidelines, constructed foreground questions, reviewed each question systemically, drafted a new set of guidelines, established expert consensus, disseminated the developed guidelines within a hospital setting, and evaluated their efficacy in practice. RESULTS: The developed ETS guidelines address 13 issues with 39 recommendations. The Delphi method found that 75% of experts agreed with all of the recommendations. Issues addressed in the ETS included pre-assessment, frequency of suctioning, pre-oxygenation, suction-tube selection, infection control, suction pressure and depth decision making, humidity with normal saline, suction duration and number, containment and comforting, assessment during and after suctioning, recovery time, and documentation. The panel of clinical and methodological experts recommended that ETS be used in practice and nurses in the neonatal intensive care unit evaluated the ETS as applicable and accessible. The cardiorespiratory responses to ETS of 5 premature infants were within normal ranges without episodes of hypoxemia (oxygen saturation < 85%) or bradycardia (heart rate < 100 beat/min). CONCLUSIONS: This set of evidence-based ETS guidelines for premature infants integrated recommendations from the best available literature and obtained a high consensus among clinical experts. Thus, these guidelines are recommended for clinical application.

An efficient and mass reproducible method for vitrifying mouse embryos on a paper in cryotubes.

To date, the most successful and popular vitrification method is based on the minimum volume cooling (MVC) concept, in which embryos are vitrified in a very small volume of vitrification solution (VS) and then stored in cryotubes in liquid nitrogen (LN2). Unfortunately, these methods need special devices and may not be suitable for vitrifying a large number of embryos. Theoretically, more embryos in VS on a paper (MVC concept) in cryotubes can be vitrified effectively. Therefore, this study directly vitrifies mouse embryos on a Kimwipes tissue in an 1.8 mL cryotube. The ICR 2-celled to blastocyst embryos were used for testing this procedure. In Treatment 1, embryos transferred with 1-2 µL of VS into a cryotube. Treatment 2 was similar to Treatment 1 except that the cryotube was filled with LN2. Treatment 3 was identical to Treatment 1 except that a small piece (5 mm²) of a sterilized Kimwipes tissue was placed on the top of VS. Treatment 4 was identical to Treatment 3 except for the cryotube being filled with LN2. After each treatment, the cryotubes were capped and transferred to a LN2 tank. After warming, the recovered embryos were cultured in KSOM+AA for 1-3 days. There were no differences in the recovery rate, overnight survival rate, blastocyst rate, and birth rate after embryo transfer among all treatment groups. Our results demonstrated an alternative simple, efficient, and mass reproducible method for vitrifying mouse embryos using papers as a vehicle and cryotubes as a container.

- Invited Review - Reproductive technologies needed for the generation of precise gene-edited pigs in the pathways from laboratory to farm.

Gene editing (GE) offers a new breeding technique (NBT) of sustainable value to animal agriculture. There are 3 GE working sites covering 5 feasible pathways to generate GE pigs along with the crucial intervals of GE/genotyping, microinjection/electroporation, induced pluripotent stem cells, somatic cell nuclear transfer, cryopreservation, and nonsurgical embryo transfer. The extension of NBT in the new era of pig breeding depends on the synergistic effect of GE and reproductive biotechnologies; the outcome relies not only on scientific due diligence and operational excellence but also on the feasibility of application on farms to improve sustainability.

Neural network combining with clinical ultrasonography: A new approach for classification of salivary gland tumors.

OBJECTIVE: Little information is available about deep learning methods used in ultrasound images of salivary gland tumors. We aimed to compare the accuracy of the ultrasound-trained model to computed tomography or magnetic resonance imaging trained model. MATERIALS AND METHODS: Six hundred and thirty-eight patients were included in this retrospective study. There were 558 benign and 80 malignant salivary gland tumors. A total of 500 images (250 benign and 250 malignant) were acquired in the training and validation set, then 62 images (31 benign and 31 malignant) in the test set. Both machine learning and deep learning were used in our model. RESULTS: The test accuracy, sensitivity, and specificity of our final model were 93.5%, 100%, and 87%, respectively. There were no over fitting in our model as the validation accuracy was similar with the test accuracy. CONCLUSIONS: The sensitivity and specificity were comparable with current MRI and CT images using artificial intelligence.

Purification and characterization of hemagglutinating proteins from Poker-chip Venus (Meretrix lusoria) and Corbicula clam (Corbicula fluminea).

Hemagglutinating proteins (HAPs) were purified from Poker-chip Venus (Meretrix lusoria) and Corbicula clam (Corbicula fluminea) using gel-filtration chromatography on a Sephacryl S-300 column. The molecular weights of the HAPs obtained from Poker-chip Venus and Corbicula clam were 358 kDa and 380 kDa, respectively. Purified HAP from Poker-chip Venus yielded two subunits with molecular weights of 26 kDa and 29 kDa. However, only one HAP subunit was purified from Corbicula clam, and its molecular weight was 32 kDa. The two Poker-chip Venus HAPs possessed hemagglutinating ability (HAA) for erythrocytes of some vertebrate animal species, especially tilapia. Moreover, HAA of the HAP purified from Poker-chip Venus was higher than that of the HAP of Corbicula clam. Furthermore, Poker-chip Venus HAPs possessed better HAA at a pH higher than 7.0. When the temperature was at 4°C-10°C or the salinity was less than 0.5‰, the two Poker-chip Venus HAPs possessed better HAA compared with that of Corbicula clam.

The application of new breeding technology based on gene editing in pig industry - A review.

Genome/gene-editing (GE) techniques, characterized by a low technological barrier, high efficiency, and broad application among organisms, are now being employed not only in medical science but also in agriculture/veterinary science. Different engineered CRISPR/Cas9s have been identified to expand the application of this technology. In pig production, GE is a precise new breeding technology (NBT), and promising outcomes in improving economic traits, such as growth, lean or healthy meat production, animal welfare, and disease resistance, have already been documented and reviewed. These promising achievements in porcine gene editing, including the Myostatin gene knockout (KO) in indigenous breeds to improve lean meat production, the uncoupling protein 1 (UCP1) gene knock-in to enhance piglet thermogenesis and survival under cold stress, the generation of GGTA1 and CMP-N-glycolylneuraminic acid hydroxylase (CMAH) gene double KO (dKO) pigs to produce healthy red meat, and the KO or deletion of exon 7 of the CD163 gene to confer resistance to porcine reproductive and respiratory syndrome virus infection, are described in the present article. Other related approaches for such purposes are also discussed. The current trend of global regulations or legislation for GE organisms is that they are exempted from classification as genetically modified organisms (GMOs) if no exogenes are integrated into the genome, according to product-based and not process-based methods. Moreover, an updated case study in the EU showed that current GMO legislation is not fit for purpose in term of NBTs, which contribute to the objectives of the EU's Green Deal and biodiversity strategies and even meet the United Nations' sustainable development goals for a more resilient and sustainable agri-food system. The GE pigs generated via NBT will be exempted from classification as GMOs, and their global valorization and commercialization can be foreseen.

Estimating intraclonal heterogeneity and subpopulation changes from bulk expression profiles in CMap.

The connectivity among signatures upon perturbations curated in the CMap library provides a valuable resource for understanding therapeutic pathways and biological processes associated with the drugs and diseases. However, because of the nature of bulk-level expression profiling by the L1000 assay, intraclonal heterogeneity and subpopulation compositional change that could contribute to the responses to perturbations are largely neglected, hampering the interpretability and reproducibility of the connections. In this work, we proposed a computational framework, Premnas, to estimate the abundance of undetermined subpopulations from L1000 profiles in CMap directly according to an ad hoc subpopulation representation learned from a well-normalized batch of single-cell RNA-seq datasets by the archetypal analysis. By recovering the information of subpopulation changes upon perturbation, the potentials of drug-resistant/susceptible subpopulations with CMap L1000 were further explored and examined. The proposed framework enables a new perspective to understand the connectivity among cellular signatures and expands the scope of the CMAP and other similar perturbation datasets limited by the bulk profiling technology.

Literature review of liver injury induced by Tinospora crispa associated with two cases of acute fulminant hepatitis.

INTRODUCTION: Species of Tinospora are used as herbal remedies for the treatment of various diseases with very few toxic effects having been reported. Tinospora cordifolia (TCF) has been reported to effectively prevent hepatotoxicity. However, there are an increasing number of cases revealing that Tinospora crispa (TCP) might have the negative effect of inducing hepatotoxicity. Because of the similar leaves, people may mistake TCP for TCF, and consume it with the purpose of protecting liver function. OBJECTIVE: Find out the misusing level of TCP and TCF and which chemical compound in TCP might induce hepatotoxicity. METHODS: We report two cases of acute fulminant hepatitis associated with chronic use of TCP. Given that the two herbs were misidentified in these two reports, we investigated the frequency of erroneous identification by using three keywords ("Guduchi", "Tinospora cordifolia", "Tinospora crispa") to search images from the Google Images database. To further clarify the influence of liver function between TCP and TCF, we searched PubMed (up to 29 July 2018) for relevant publications on clinical trials or case reports. RESULTS: Based on web review, over 35 percent of websites failed to accurately identify these two herbs. The different effects on liver function between TCP and TCF were compared through literature review. It indicated that TCF exerted liver protection, TCP had a contrary effect, suggesting its cis-Clerodane-type furano-diterpenoids might be an important factor of inducing hepatotoxicity. CONCLUSIONS: We concluded that people might cause hepatic injury or even death without correctly identifying these two Tinospora species.

Growth and Behavior of Congenitally Anophthalmic Lee-Sung Pigs.

Circadian rhythm is usually regulated by the environmental light-dark cycle. Congenitally anophthalmic miniature pigs provide a valuable model for the study of factors affecting circadian rhythms in the absence of visual exposure to the light-dark cycle. This study investigated the growth and daily behavior patterns of Lee-Sung pigs with congenital anophthalmia. Growth in 5 Lee-Sung pigs (LSP) with congenital anophthalmia (LSP-A) and 10 normally developed pigs (LSP-N) was assessed when they were 1 through 6 mo old. Behavioral studies using digital video recording were completed in 6 sexually mature LSP (3 LSP-A and 3 LSP-N). MRI showed that LSP-A lose their vision because of a lack of retinal input and optic chiasm development. LSP-N and LSP-A did not differ in body weight or size at 2, 4, and 6 mo of age. Behavior and activity pattern studies showed that both LSP-A and LSP-N were active mainly during daylight, but LSP-A spent significantly more time exploring their environment during the day (28%) and night (10%) than did LSP-N. This study revealed that growth performance was similar between LSP-A and normal pigs, but their behavior and activity patterns differed. LSP-A showed circadian rhythm abnormalities similar to those in blind humans. This study provides basic data on LSP-A as a model for studying compensatory cross-modal brain plasticity and hormone regulation in the absence of retinal input is deficient and for understanding the role of circadian rhythm regulation.