Linked-Read Whole Genome Sequencing Solves a Double DMD Gene Rearrangement

Next generation sequencing (NGS) has changed our approach to diagnosis of genetic disorders. Nowadays, the most comprehensive application of NGS is whole genome sequencing (WGS) that is able to detect virtually all DNA variations. However, even after accurate WGS, many genetic conditions remain unsolved. This may be due to the current NGS protocols, based on DNA fragmentation and short reads. To overcome these limitations, we applied a linked-read sequencing technology that combines single-molecule barcoding with short-read WGS. We were able to assemble haplotypes and distinguish between alleles along the genome. As an exemplary case, we studied the case of a female carrier of X-linked muscular dystrophy with an unsolved genetic status. A deletion of exons 16–29 in DMD gene was responsible for the disease in her family, but she showed a normal dosage of these exons by Multiplex Ligation-dependent Probe Amplification (MLPA) and array CGH. This situation is usually considered compatible with a “non-carrier” status. Unexpectedly, the girl also showed an increased dosage of flanking exons 1–15 and 30–34. Using linked-read WGS, we were able to distinguish between the two X chromosomes. In the first allele, we found the 16–29 deletion, while the second allele showed a 1–34 duplication: in both cases, linked-read WGS correctly mapped the borders at single-nucleotide resolution. This duplication in trans apparently restored the normal dosage of exons 16–29 seen by quantitative assays. This had a dramatic impact in genetic counselling, by converting a non-carrier into a double carrier status prediction. We conclude that linked-read WGS should be considered as a valuable option to improve our understanding of unsolved genetic conditions.

Clinical Features and Management of Snakebite Envenoming in French Guiana

The management of snakebite (SB) envenoming in French Guiana (FG) is based on symptomatic measures and antivenom (AV) administration (Antivipmyn Tri®; Instituto Bioclon—Mexico). Our study aimed to assess clinical manifestations, the efficacy, and safety of Antivipmyn Tri® in the management of SB. Our study is a prospective observational work. It was conducted in the Intensive Care Unit (ICU) of Cayenne General Hospital between 1 January 2016 and 31 December 2019. We included all patients hospitalized for SB envenoming. Our study contained three groups (without AV, three vials, and six vials Antivipmyn Tri®). During the study period, 133 patients were included. The main clinical symptoms were edema (98.5%), pain (97.7%), systemic hemorrhage (18%), blister (14.3%), and local hemorrhage (14.3%). AV was prescribed for 83 patients (62.3%), and 17 of them (20%) developed early adverse reactions. Biological parameters at admission showed defibrinogenation in 124 cases (93.2%), International Normalized Ratio (INR) > 2 in 104 cases (78.2%), and partial thromboplastin time (PTT) > 1.5 in 74 cases (55.6%). The time from SB to AV was 9:00 (5:22–20:40). The median time from SB to achieve a normal dosage of fibrinogen was 47:00 vs. 25:30, that of Factor II was 24:55 vs. 15:10, that of Factor V was 31:42 vs. 19:42, and that of Factor VIII was 21:30 vs. 10:20 in patients without and with AV, respectively, (p < 0.001 for all factors). Patients receiving Antivipmyn Tri® showed a reduction in the time to return to normal clotting tests, as compared to those who did not. We suggest assessing other antivenoms available in the region to compare their efficacy and safety with Antivipmyn Tri® in FG.

Mechanical Behavior of Fine Recycled Concrete Aggregate Concrete with the Mineral Admixtures

The paper describes the mechanical behavior of fine recycled concrete aggregate (FRCA) concrete according to the mineral admixtures. Three types of the mineral admixtures, i.e., fly ash (FA), ground-granulated blast-furnace slag (GGBS), and silica fume (SF), are used and the replacement ratios of FRCA are 50% and 100%. The dosages of the admixtures of FA, GGBS, and SF are determined with the normal dosage (30%, 40%, and 5.0%, respectively) based on the ACI committee reports (No. 232, 233, and 234) and half-normal dosage. The mechanical performance is investigated with the compressive and splitting tensile strength, and elastic modulus. Additionally, the total porosity is measured in natural fine aggregate (NFA) and FRCA 100% replaced specimens by mercury intrusion porosimetry (MIP) for investigating the relationship with the compressive strength. Based on the experimental test results, the mineral admixtures improve the mechanical performance of FRCA concrete. The effective dosages of FA, GGBS, and SF for FRCA concrete are investigated according to the replacement ratio of the FRCA. In particular, FRCA 100% replaced concrete may be possible to be used for the structural concrete members with the specific dosage of the mineral admixtures. The prediction of the splitting tensile strength and the elastic modulus by the codes or previous formulas exhibits underestimated and overestimated results, respectively. The relationship between the total porosity and the compressive strength of the FRCA concrete should be modified with more experimental tests.

Liver damage in severe COVID-19 patients from Sichuan area

Background: COVID-19 has spread worldwide, which becomes a huge threat to human beings.Materials: Severe COVID-19 patients from Sichuan area admitted to department of critical care medicine in Chengdu Public Health Clinical Medical Center were retrospectively enrolled. The liver function during the ICU hospitalization were record and analyzed.Results: The severe COVID-19 patients mainly presented with respiratory symptoms such as fever, cough and dyspnea, and the incidence was higher in the elderly patients and males. ALT, AST, TB, and PT increased to varying degrees during the course of the disease, and ALB decreased. The incidence of liver dysfunction in patients taking Lopinavir/Ritonavi was significantly higher than patient who did not have it, but there was no statistical difference (p<0.05). Patients taking low-dose Lopinavir/Ritonavi had a smaller effect on liver function than patients receiving normal dosage.Conclusion: Severe COVID-19 patients have obvious liver damage early in the course of the disease and have a slower recovery. Pay attention to avoid using drugs that can aggravate liver damage while treating the primary disease. If there is no alternative drug, we can give some liver protection treatment appropriately.

Enhanced Treatment Effects of Tilmicosin Against Staphylococcus aureus Cow Mastitis by Self-Assembly Sodium Alginate-Chitosan Nanogel

The Staphylococcus aureus (S. aureus) cow mastitis causes great losses to the cow industry. In order to improve the treatment effect of tilmicosin against cow mastitis, the combination of solid lipid nanoparticle (SLN) technology with in situ hydrogel technology was used to prepare the self-assembly tilmicosin nanogel (TIL-nanogel). The physicochemical characteristics, in vitro release, antibacterial activity and in vivo treatment efficacy of TIL-SLNs and TIL-nanogel were studied, respectively. The results showed the loading capacity (LC), encapsulation efficiency (EE), size, zeta potential and poly dispersion index (PDI) of TIL-nanogel were 23.33 ± 0.77%, 67.89 ± 3.01%, 431.57 ± 12.87 nm, 8.3 ± 0.06 mv and, 0.424 ± 0.032, respectively. The TIL-nanogel showed stronger sustained release in vitro than TIL-SLNs and commercial injection. The cure rate of half dosage and normal dosage of TIL-nanogel was 58.3% and 75.0%, which was higher than that of commercial injection (50.0%) at normal dosage. The results suggest that the treatment dosage of tilmicosin for cow mastitis could be reduced by TIL-nanogel. The novel TIL-nanogel will be beneficial by decreasing the usage of tilmicosin and the treatment costs of cow mastitis.

Assessing Risk and Adverse Events

This chapter provides an overview of safety assessment in clinical trials, including challenges for designing and reporting studies measuring safety. Adverse events are undesirable, unfavorable, harmful, and unexpected effects resulting from a normal dosage of drug, medication, therapy, or any other intervention such as surgery. Monitoring of adverse events is critical to the patient’s safety (i.e., human subject’s protection) and data integrity in clinical trials. Moreover, this chapter reviews the regulatory aspects related to adverse events in clinical trials. Finally it presents a case discussion in which an unexpected adverse effect is detected and investigators need to determine whether this event is or is not related to the intervention and define a plan of action.

Herbicide tolerance of maize genotypes in the wet 2016 year

The herbicide tolerance levels of 49 Martonvásár inbred parents were examined in Martonvásár in a herbicide susceptibility trial in 2016. The normal dosage recommended in the permit documentations and double dosage were used for the 12 small-plot herbicide treatments performed in two repetitions. Spraying of early post-emergent herbicides was carried out in the 1–2-leaf stage, while post-emergent treatments were applied in the 7–8-leaf stage of maize. The extent of phytotoxicity was scored for the early post-emergent herbicides two and four weeks after treatments and for the post-emergent herbicides two weeks after treatments, respectively. Some of the herbicides examined are not approved in seed production; however it is important to know the reaction of maize parent genotypes for every type of herbicides. The active agent topramezone was withdrawn from the market in 2015, but it was included in the trials as its usage was allowed until stocks run out in 2016. The herbicide agents were examined as follows: mesotrione + S-metolachlor + terbutylazine; isoxaflutol + tiencarbazon methyl + cyprosulfamide; isoxaflutol + cyprosulfamide; mesotrione + terbuthylazine; tembotrione + isoxidifen-ethyl; mesotrione + nicosulfuron; prosulfu ron; nicosulfuron +prosulfuron + dicamba; bentazone + dicamba; nicosulfuron; topramezone; foramsulfuron + isoxadifen-ethyl.Among early post-emergent herbicides, isoxaflutol + cyprosulfamide caused the less phytotoxic damage in the genotypes. The large amount of precipitation during the spring facilitated the infiltration of the active ingredient S-metolachlor, used regularly and successfully also in seed production, into the root zone, resulting in phytotoxic symptoms on susceptible inbred lines at the time of the first inspection. These genotypes recovered by the end of the vegetation period. The spring weather was cooler than usual, retarding the development of maize and thus led to the slower fermentation of herbicide active ingredients, accordingly, all of the post-emergent herbicides caused visible phytotoxic symptoms on some of genotypes. The most severe damages were generally caused by the double dosage of nicosulfuron + prosulfuron + dicamba, nicosulfuron, and foramsulfuron + isoxadifen-ethyl.

Environment Affects Cotton and Velvetleaf Response to Pyrithiobac

Growth chamber experiments evaluated the influence of ambient temperature and soil moisture on cotton and velvetleaf response to pyrithiobac. Additional studies determined the basis for observed plant responses to14C pyrithiobac. Cotton injury from six times the normal dosage was < 20% at 2 wk for all temperatures and soil moistures. Pyrithiobac injured velvetleaf less at lower soil moistures. Both species absorbed more14C-pyrithiobac at 30/28 or 35/33 C than at 25/23 C. Cotton absorbed more herbicide than velvetleaf at all temperatures and soil moistures. Velvetleaf translocated < 16% of absorbed14C out of the treated leaf while cotton translocated < 3% of absorbed material. At warmer temperatures, velvetleaf translocated less14C when soil was dry (–1.0 MPa) than when plants were watered to field capacity (–0.03 MPa). This decreased absorption and translocation may affect pyrithiobac activity on velvetleaf growing in dry soil. Translocation differences did not fully explain whole plant effects. The metabolism difference may account for cotton tolerance.

Taste sensitivity to trehalose and its alteration by gene dosage in Drosophila melanogaster.

The taste sensitivity to the disaccharide trehalose of Drosophila melanogaster is under the genetic control by the Tre gene on the X chromosome. The gene is genetically dimorphic for high and low sensitivity and is likely to be functioning in the primary step of chemoreception. We have determined the cytological localization of the Tre gene to be between 5A10 and 5B1-3 by analyzing the sensitivity to trehalose in flies which are segmentally aneuploid bearing either deficiencies or duplicated fragments of T(X;Y) translocations. We also constructed flies which are aneuploidy and thus carry different dosage of Tre and/or Tre(+) alleles in order to examine the gene dosage effect on trehalose sensitivity and to deduce the nature of the gene's action. Trehalose sensitivity decreased in females carrying half the normal dosage of a given Tre allele, but a proportional increase in sensitivity was not observed in flies bearing a duplication of the Tre alleles. The changes in sensitivity in various aneuploid flies suggest that there is an upper limit to the number of molecules that can be incorporated into the receptor membrane. Genetic evidence strongly suggests that Tre is the structural gene for the trehalose receptor. We present a model to account for the mechanism of genetical control on the sensitivity to trehalose.

Clinical Experience with Intravenous Augmentin in the Treatment of Paediatric Infections

The clinical efficacy of intravenous Augmentin (a formulation containing amoxycillin plus clavulanic acid) was investigated in an open study in fifty-eight children with a mean age of 6 years (range 1–15 years). The normal dosage was in the range 100–200 mg/kg/day Augmentin, administered parenterally by short i.v. infusion in 3 or 4 divided doses. Most patients were hospitalised for lower respiratory tract infections. Complete clinical cure or distinct clinical improvement was achieved in all assessable cases. Bacteriological success was obtained in 92% of the assessable cases. In two patients, mild, transient exanthema was noted after i.v. Augmentin was replaced by oral Augmentin. No additional therapeutic measures were required.