Screening of Spinal Muscular Atrophy Carriers and Prenatal Diagnosis in Pregnant Women in Yancheng, China.

Spinal muscular atrophy (SMA) is a neuromuscular disorder with an autosomal recessive inheritance pattern. Patients with severe symptoms may suffer respiratory failure, leading to death. The homozygous deletion of exon 7 in the SMN1 gene accounts for nearly 95% of all cases. Population carrier screening for SMA and prenatal diagnosis by amniocentesis for high-risk couples can assist in identifying the risk of fetal disease. We provided the SMA carrier screening process to 55,447 pregnant women in Yancheng from October 2020 to December 2022. Among them, 8185 participated in this process, with a participation rate of around 14.76% (95% CI 14.47-15.06%). Quantitative real-time polymerase chain reaction (qPCR) was used to detect deletions of SMN1 exons 7 and 8 (E7, E8) in screened pregnant women. 127 were identified as carriers (111 cases of E7 and E8 heterozygous deletions, 15 cases of E7 heterozygous deletions, and 1 case of E7 heterozygous deletions and E8 homozygous deletions), resulting in a carrying rate of around 1.55% (95% CI 1.30-1.84%). After genetic counseling, 114 spouses of pregnant women who tested positive underwent SMA carrier screening; three of them were screened as SMA carriers. Multiplexed ligation-dependent probe amplification (MLPA) was used for the prenatal diagnosis of the fetuses of high-risk couples. Two of them exhibited two copies of SMN1 exon 7 (normal), and the pregnancy was continued; one exhibited no copies of SMN1 exon 7 and exon 8 (SMA patient), and the pregnancy was terminated. Analyzing SMN1 mutations in Yancheng and provide clinical evidence for SMA genetic counseling and birth defect prevention. Interventional prenatal diagnosis for high-risk families can promote informed reproductive selection and prepare for the fetus's early treatment.

Sequential respiratory support in septic patients undergoing continuous renal replacement therapy: A study based on MIMIC-III database.

Objective: Oxygen and hemodynamic management are important for providing a sufficient adequate oxygen-containing blood to the organs for septic patients. In present study, we aimed to explore the application of sequential respiratory support (SRS) and the association of SRS with the outcome of septic patients who needed continuous renal replacement therapy (CRRT). Methods: We extracted the medical information of septic patients who received CRRT within 24 h of intensive care unit (ICU) admission from the MIMIC-III v1.4. SRS was defined as receiving firstly oxygen therapy followed by mechanical ventilation (MV) within 24 h of admission to ICU. The propensity score matching (PSM) was performed to compare the differences in clinical characteristics and outcomes of patients with or without SRS. Finally, we developed logistic regression models to analyze the effects of SRS on hospital mortality. Results: A total of 181 patients entered in this study, and there were 80 patients undergoing MV including SRS group (n = 61) and non-SRS group (n = 19). In the multivariate logistic regression, the value of SRS was associated with the lower risk of hospital mortality adjusted by minimum systolic BP (SBP), maximum lactate, vasopressor use, and sequential organ failure assessment (SOFA) score or Logistic Organ Dysfunction System (LODS) scores within the first 24 h of ICU stay. After PSM adjusted by SBP, maximum lactate, vasopressor use, SOFA, and LODS, there were 31 patients in SRS group with a and 18 cases in non-SRS group, displaying a significantly lower hospital mortality in SRS group than that in patients without SRS (19.4 % vs. 83.3 %, P < 0.001). In addition, age, qSOFA, necessitating the administration of vasopressor, and duration of vasopressor were significantly correlated with the hospital mortality in septic patients undergoing CRRT and SRS. Conclusions: Receiving SRS within the first 24 h upon admission to the ICU was independently associated with the hospital mortality in patient with sepsis undergoing CRRT, and patients who were directly received MV had a high risk of death.

A novel homozygous missense variant in LRP4 causing Cenani-Lenz syndactyly syndrome and literature review.

BACKGROUND: Cenani-Lenzsyndactyly syndrome (CLSS; OMIM 212780) is a rare autosomal recessive acral deformity, which is mainly manifested in the fusion of fingers or toes, disordered phalangeal structure, shortening or fusion of the radius and ulna, and renal hypoplasia. CASE PRESENTATION: Our report described an individual with mild phenotypes from China. His parents were not consanguineous. The affected individual was non-dysmorphic. Standard X-ray showed that the both hands have only four metacarpal bones. The distal end of the first metacarpal bone on the right was relatively slender, and the distal phalanx was absent. Multiple phalanges and some soft tissues of both hands were fused. Exome sequencing revealed a novel biallelic c.282C⟩Avariant in low-density lipoprotein receptor-related protein 4 (LRP4; OMIM604270; NM_002334.4) causing p. (Asn94Lys) change in the encoded protein. This variant is predicted to be potentially pathogenic, affecting protein structure and function. CONCLUSION: We report a novel missense variant present in homozygosity in LRP4 to broaden the pathogenic spectrum of LRP4 in syndactyly, and exome sequencing technology is a powerful tool for genetic analysis in prenatal diagnosis and medical research, as a preferred method for the diagnosis of syndactyly and related phenotypes.

Direct 1,6-Difluoromethylation of 3-Methyl-4-nitro-5-styryl Isoxazoles by Photoredox-Promoted Radical Addition.

The photoredox-catalyzed 1,6-difluoromethylation of 3-methyl-4-nitro-5-styrylisoxazole with HCF2SO2Na has been developed. Structurally diverse difluoromethylated products were obtained in good yields, and their further transformations were also investigated. The di-, tri-, and monofluoromethylation of the substrates were compared, and the yield of the difluoromethylation was the highest. DFT calculations revealed that in the difluoromethylation reaction the CF2H radical was nucleophilic, and the transition state activation energy was the lowest.

A small fishing vessel recognition method using transfer learning based on laser sensors.

The management of small vessels has always been key to maritime administration. This paper presents a novel method for recognizing small fishing vessels based on laser sensors. Using four types of small fishing vessels as targets, a recognition method for small fishing vessels based on Markov transition field (MTF) time-series images and VGG-16 transfer learning is proposed. In contrast to conventional methods, this study uses polynomial fitting to obtain the contours of a fishing vessel and transforms one-dimensional vessel contours into two-dimensional time-series images using the MTF coding method. The VGG-16 model is used for the recognition process, and migration learning is applied to improve the results. The UCR time-series public dataset is used as a transfer learning dataset for the MTF time-series image encoding. The experiment demonstrates that the proposed method exhibits higher accuracy and performance than 1D-CNN and other general neural network models, and the highest accuracy rate is 98.92%.

[The value of chromosomal microarray analysis and fluorescence in situ hybridization for the prenatal diagnosis of chromosomal mosaicisms].

OBJECTIVE: To assess the value of chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) for the prenatal diagnosis of chromosomal mosaicisms. METHODS: A total of 775 pregnant women who had visited the Prenatal Diagnosis Center of Yancheng Maternal and Child Health Care Hospital from January 2018 to December 2020 were selected as study subjects. Chromosome karyotyping analysis and CMA were carried out for all women, and FISH was used to validate the suspected mosaicism cases. RESULTS: Among the 775 amniotic fluid samples, karyotyping has identified 13 mosaicism cases, which yielded a detection rate of 1.55%. Respectively, there were 4, 3, 4 and 2 cases for sex chromosome number mosaicisms, abnormal sex chromosome structure mosaicisms, abnormal autosomal number mosaicisms and abnormal autosomal structure mosaicisms. CMA has only detected only 6 of the 13 cases. Among 3 cases verified by FISH, 2 cases were consistent with the karyotyping and CMA results, and clearly showed low proportion mosaicism, and 1 case was consistent with the result of karyotyping but with a normal result by CMA. Eight pregnant women had chosen to terminate the pregnancy (5 with sex chromosome mosaicisms and 3 with autosomal mosaicisms). CONCLUSION: For fetuses suspected for chromosomal mosaicisms, CMA, FISH and G-banding karyotyping should be combined to determine the type and proportion of mosaicisms more precisely in order to provide more information for genetic counseling.

Genome-wide identification of agronomically important genes in outcrossing crops using OutcrossSeq.

Many important crops (e.g., tuber, root, and tree crops) are cross-pollinating. For these crops, no inbred lines are available for genetic study and breeding because they are self-incompatible, clonally propagated, or have a long generation time, making the identification of agronomically important genes difficult, particularly in crops with a complex autopolyploid genome. In this study, we developed a method, OutcrossSeq, for mapping agronomically important loci in outcrossing crops based on whole-genome low-coverage resequencing of a large genetic population, and designed three computation algorithms in OutcrossSeq for different types of outcrossing populations. We applied OutcrossSeq to a tuberous root crop (sweet potato, autopolyploid), a tree crop (walnut tree, highly heterozygous diploid), and hybrid crops (double-cross populations) to generate high-density genotype maps for the outcrossing populations, which enable precise identification of genomic loci underlying important agronomic traits. Candidate causative genes at these loci were detected based on functional clues. Taken together, our results indicate that OutcrossSeq is a robust and powerful method for identifying agronomically important genes in heterozygous species, including polyploids, in a cost-efficient way. The OutcrossSeq software and its instruction manual are available for downloading at www.xhhuanglab.cn/tool/OutcrossSeq.html.

Cox-LASSO Analysis for Hospital Mortality in Patients With Sepsis Received Continuous Renal Replacement Therapy: A MIMIC-III Database Study.

BACKGROUND: Sepsis remains the leading cause of mortality in-hospital in the intensive care unit (ICU). Continuous renal replacement therapy (CRRT) is recommended as an adjuvant therapy for hemodynamics management in patients with sepsis. The aim of this study was to develop an adaptive least absolute shrinkage and selection operator (LASSO) for the Cox regression model to predict the hospital mortality in patients with Sepsis-3.0 undergoing CRRT using Medical Information Martin Intensive Care (MIMIC)-III v1.4. METHODS: Patients who met the Sepsis-3.0 definition were identified using the MIMIC-III v1.4. Among them, patients who received CRRT during ICU hospitalization were included in this study. According to the survival status, patients were split into death or survival group. Adaptive LASSO for the Cox regression model was constructed by STATA software. At last, nomogram and Kaplan-Meier curves were drawn to validate the model. RESULTS: A total of 181 patients who met Sepsis 3.0 criteria received CRRT were included in the study, in which, there were 31 deaths and 150 survivals during hospitalization, respectively. The overall in-hospital mortality was 17.1%. According to the results of multivariate Cox-LASSO regression analysis, use of vasopressor, international normalized ratio (INR) ≥1.5, and quick sequential organ failure assessment (qSOFA) score were associated with hospital mortality in patients with sepsis who underwent CRRT, but lactate level, mechanical ventilation (MV) support, PaO2/FiO2, platelet count, and indicators of acute kidney injury (AKI), such as blood urea nitrogen (BUN) and creatinine, were not independently associated with hospital mortality after adjusted by qSOFA. The risk nomogram and Kaplan-Meier curves verified that the use of vasopressor and INR ≥1.5 possess significant predictive value. CONCLUSIONS: Using the Cox-LASSO regression model, use of vasopressor, INR ≥1.5, and qSOFA score are found to be associated with hospital mortality in patients with Sepsis-3.0 who received CRRT. This finding may assist clinicians in tailoring precise management and therapy for these patients who underwent CRRT.

The Clinical Utility of Non-invasive Prenatal Testing for Pregnant Women With Different Diagnostic Indications.

BACKGROUND: Our aim was to evaluate the clinical utility of non-invasive prenatal testing for pregnant women with different diagnostic indications. METHODS: In eight counties and districts of Yancheng, we studied 13,149 pregnant women with different indications who were offered NIPT for fetal screening, including for sex chromosomal aneuploidies (SCAs), rare autosomal trisomies (RATs), and subchromosomal copy number variations (CNVs). The purpose was to compare the detection of positive predictive values (PPVs) of different indications with the use of NIPT. The results were validated by karyotyping, chromosomal microarray analysis (CMA), or follow-up of pregnancy outcomes. RESULTS: 13,149 maternal plasma samples were sequenced, among which 28 samples (0.2%) failed the sequencing quality control. The remaining 13,121 samples were analyzed, and birth follow-up missed 2,192 samples (16.7%). The PPVs of NIPT results for trisomy 21 (T21) and trisomy 18 (T18) and SCAs were 96.67, 63.64, and 31.34%, respectively. Among the advanced maternal age (AMA), serum screening high risk (SSHR), serum screening intermediate risk (SSIR), and voluntary screening (VS) groups, the PPVs for the common trisomies were 81.25, 85.71, 100, and 70%, respectively; the PPVs for total chromosomal abnormalities were 55.82, 65.22, 23.08, and 36.59%, respectively. CONCLUSION: NIPT for T21 and T18 and SCAs screening were ideal, and the PPVs for trisomy 13 (T13), RATs, and CNVs were low. For the AMA and VS groups, NIPT could be used as a first-line screening program; for SSHR and SSIR groups, NIPT could be used as a second-line supplementary screening program.

Prenatal diagnosis of sex chromosome mosaicism with two marker chromosomes in three cell lines and a review of the literature.

The present study described the diagnosis of a fetus with sex chromosome mosaicism in three cell lines and two marker chromosomes. A 24­year­old woman underwent amniocentesis at 21 weeks and 4 days of gestation due to noninvasive prenatal testing identifying that the fetus had sex chromosome abnormalities. Amniotic cell culture revealed a karyotype of 45,X[13]/46,X,+mar1[6]/46,X,+mar2[9], and prenatal ultrasound was unremarkable. The woman underwent repeat amniocentesis at 23 weeks and 4 days of gestation for molecular detection. Single nucleotide polymorphism (SNP) microarray analysis on uncultured amniocytes revealed that the fetus had two Y chromosomes and 7.8­Mb deletions in Yq11.222q12. The deletion regions included DAZ, RBMY and PRY genes, which could cause spermatogenesis obstacle and sterility. Interphase fluorescence in situ hybridization (FISH) using centromeric probes DXZ1/DYZ3/D18Z1 was performed on uncultured amniocytes to verify the two marker chromosomes to be Y chromosome derivatives. According to these examinations, the mar1 was identified as a derivative of the Y chromosome with a deletion in Yq11.222q12, and the mar2 was identified as a dicentric derivative of the Y chromosome. The molecular karyotype was therefore 45,X,ish(DXZ1+, DYZ3­,D18Z1++)[5]/46,X,del(Y)(q11.222),ish(DXZ1+,DYZ3+,D18Z1++)[11]/46, X,idic(Y)(q11.222),ish(DXZ1+,DYZ3++,D18Z1++)[14]. The comprehensive use of cytogenetic, SNP array and FISH detections was advantageous for accurately identifying the karyotype, identifying the origin of the marker chromosome and preparing effective genetic counseling.

Haplotype-resolved sweet potato genome traces back its hexaploidization history.

Here we present the 15 pseudochromosomes of sweet potato, Ipomoea batatas, the seventh most important crop in the world and the fourth most significant in China. By using a novel haplotyping method based on genome assembly, we have produced a half haplotype-resolved genome from ~296 Gb of paired-end sequence reads amounting to roughly 67-fold coverage. By phylogenetic tree analysis of homologous chromosomes, it was possible to estimate the time of two recent whole-genome duplication events as occurring about 0.8 and 0.5 million years ago. This half haplotype-resolved hexaploid genome represents the first successful attempt to investigate the complexity of chromosome sequence composition directly in a polyploid genome, using sequencing of the polyploid organism itself rather than any of its simplified proxy relatives. Adaptation and application of our approach should provide higher resolution in future genomic structure investigations, especially for similarly complex genomes.

Temperature compensation of the SAW yarn tension sensor.

The objective of this research was to investigate the possibility of the temperature compensation for the surface acoustic wave (SAW) yarn tension sensor. The motivation for this work was prompted by the oscillation frequency of the SAW yarn tension sensor varying with the temperature. In this paper, we deduce the functional relationship between the temperature variation and the oscillation frequency shift caused by the temperature. This functional relationship and the temperature sensor are used to get the oscillation frequency shift caused by the temperature, so that we can use the oscillation frequency shift caused by the temperature to implement the temperature compensation of the SAW yarn tension sensor. In this paper, we also get the relative error of the temperature compensation. The theoretical and experimental results confirm that this temperature compensation method can implement the temperature compensation of the SAW yarn tension sensor.

Resistance to Ditylenchus destructor Infection in Sweet Potato by the Expression of Small Interfering RNAs Targeting unc-15, a Movement-Related Gene.

Stem nematode (Ditylenchus destructor) is one of most serious diseases that limit the productivity and quality of sweet potato (Ipomoea batatas), a root crop with worldwide importance for food security and nutrition improvement. Hence, there is a global demand for developing sweet potato varieties that are resistant to the disease. In this study, we have investigated the interference of stem nematode infectivity by the expression of small interfering RNAs (siRNAs) in transgenic sweet potato that are homologous to the unc-15 gene, which affects the muscle protein paramyosin of the pathogen. The production of double-stranded RNAs and siRNAs in transgenic lines with a single transgene integration event was verified by Northern blot analysis. The expression of unc-15 was reduced dramatically in stem nematodes collected from the inoculated storage roots of transgenic plants, and the infection areas of their storage roots were dramatically smaller than that of wild-type (WT). Compared with the WT, the transgenic plants showed increased yield in the stem nematode-infested field. Our results demonstrate that the expression of siRNAs targeting the unc-15 gene of D. destructor is an effective approach in improving stem nematode resistance in sweet potato, in adjunct with the global integrated pest management programs.

Impact of amylose content on starch physicochemical properties in transgenic sweet potato.

The intrinsic relationship between amylose content and starch physicochemical properties was studied using six representative starch samples (amylose content 0-65%) produced from transgenic sweet potato (cultivar Xushu22). The transgenic lines (waxy and high-amylose) and wild-type (WT) sweet potatoes were analyzed for amylose content, particle size and chain length distribution, X-ray diffraction analysis, thermal characteristics, pasting and rheological property. Compared to the WT starch, the waxy and high-amylose starches showed larger average granule sizes and had fewer short chains and more medium and long chains. X-ray diffractogram analysis revealed that high-amylose starches show a type-B crystal form with a markedly decreased degree of crystallinity in contrast to the type-A crystal form of the WT and waxy starches. In the high-amylose sweet potato starches, the rise of setback value and the reduction of breakdown value led to the high shear resistance as indicated by the higher G', G", and tanδ from the oscillation test. ΔH was not found to be decreased with the reduction of crystallinity. The shear stress resistance of starch gel after gelatinization was also enhanced as amylose content increased. Principal component analysis also confirmed that the amylose content greatly influenced the starch structure and properties, e.g., storage modulus, setback value, and average chain length. Thus, our study not only shed light on how amylose content affects starch properties but also identified novel starches that are available for various applications.

[Research on medical instrument information integration technology based on IHE PCD].

Integrating medical instruments with medical information systems becomes more and more important in healthcare industry. To make medical instruments without standard communication interface possess the capability of interoperating and sharing information with medical information systems, we developed a medical instrument integration gateway based on Integrating the Healthcare Enterprise Patient Care Device (IHE PCD) integration profiles in this research. The core component is an integration engine which is implemented according to integration profiles and Health Level Seven (HL7) messages defined in IHE PCD. Working with instrument specific Javascripts, the engine transforms medical instrument data into HL7 ORU message. This research enables medical instruments to interoperate and exchange medical data with information systems in a standardized way, and is valuable for medical instrument integration, especially for traditional instruments.

[Research on measuring the velocity and displacement of the coxa and knee based on video image processing].

Based on repeated experiments as well as continuous researching and improving, an efficient scheme to measure velocity and displacement of the coxa and knee movements based on video image processing technique is presented in this paper. The scheme performed precise and real-time quantitative measurements of 2D velocity or displacement of the coxa and knee using a video camera mounted on one side of the healing and training beds. The beds were based on simplified pinhole projection model. In addition, we used a special-designed auxiliary calibration target, composed by 24 circle points uniformly located on two concentric circles and two straight rods which can rotate freely along the concentric center within the vertical plane, to do the measurements. Experiments carried out in our laboratory showed that the proposed scheme could basically satisfy the requirements about precision and processing speed of such kind of system, and would be very suitable to be applied to smart evaluation/training and healing system for muscles/balance function disability as an advanced and intuitional helping method.

Angiotensin II-mediated vascular changes in aged offspring rats exposed to perinatal nicotine.

This study determined the long-term influence of prenatal nicotine exposure (PN) on blood pressure and vascular functions in the aged offspring rats. PN did not affect body weight and plasma adrenocorticotropic hormone level; however, it significantly reduced plasma angiotensin I and angiotensin II in both sexes. Systolic pressure in the male aged PN offspring was significantly higher. Angiotensin II-increased mean arterial pressure was higher in the aged PN offspring than that in the control regardless of sex. AT1 receptor blocker losartan, not AT2 receptor antagonist PD123319, reduced blood pressure in the aged PN rats more than that in the control. In the aged PN offspring, angiotensin II-increased vessel contraction and intracellular calcium level were higher in small mesenteric arteries. Acetylcholine-mediated vascular relaxation was weaker, and nitric oxide-related endothelial functions were damaged in aortic rings of PN offspring. Thickness of the wall of mesenteric arteries was increased in the male aged PN offspring. Ratio of AT1/AT2 receptors was significantly increased in the vessel of the PN group regardless of sex. These data provide new information on the very long term influence of PN on vascular structures and functions in the aged offspring, demonstrate that the aged PN female rats were not free of vascular risks after menopause, and suggest that multiple pathways may be involved in the detrimental alterations of the cardiovascular system of the PN rats.

[Achievements on tuberculosis control in Shandong province -- a population-based survey in 2010].

OBJECTIVE: To study the prevalence and trend of tuberculosis for related policy development in Shandong. METHODS: The population under the current study was randomly sampled, using both stratified clustering sampling and proportional population sampling methods, following the national survey protocol. A total of 35 clusters including about 1500 subjects per cluster were established, representing a population of 9.31 million. Questionnaire interview and chest X-ray exam were applied to all inhabitants above 15 years of age. Sputum microscopy and culture were given to all suspected cases with cough longer than 2 weeks or having abnormal X-ray results. RESULTS: In total, 54 279 subjects were interviewed and examined, accounting for 95.78% of eligible population. Of them, 183 active pulmonary tuberculosis cases were identified, with 60.11% asymptomatic. Two of the 35 (5.71%) clusters had no active tuberculosis cases found, and 24 (68.57%) did not show smear positive results. The standardized prevalence rates of active, smear positive and bacteriologic positive pulmonary tuberculosis cases were 270.87/10(5), 17.45/10(5) and 29.57/10(5), with the estimated case numbers as 211 900 (170 100 - 253 600), 13 600 (5800 - 21 500) and 23 100 (13 200 - 33 000) respectively. Compared to the survey in 2000, the rates on smear positive and bacteriologic positive tuberculosis had decreased significantly, at a rate of 81.63%, and 75.56% respectively. The rates in urban areas and in women decreased quickly than those in rural areas and in men. People living in the rural areas, being elderly or males, had significantly higher prevalence rates of tuberculosis. CONCLUSION: Remarkable reduction of tuberculosis prevalence had been achieved despite the fact that tuberculosis remained a major public health problem in Shandong province. Symptomatic patients should be under more serious concern in order to improve the detection of early cases. More efforts should be given to rural population, especially elderly, male population.

Composition and stability of anthocyanins from purple Solanum tuberosum and their protective influence on Cr(VI) targeted to bovine serum albumin.

Anthocyanins from the purple Solanum tuberosum newly cultivated by the Taian Academy of Agricultural Sciences were extracted and analyzed using high-performance liquid chromatography (HPLC) and UV-vis spectroscopy. Four individual anthocyanins were detected as the major components, and the total anthocyanin content was 273.5 ± 14.3 mg of cyanidin-3-glucoside equiv/100 g of dry seeds. Results of color stability showed that the purple S. tuberosum anthocyanins (PSTAs) are more stable under low pH and temperatures. Heat and general food additives have fine compatibility with PSTAs; however, they are very sensitive with oxidant and reduction. The influence of PSTAs on Cr(VI) targeted to bovine serum albumin (BSA) was also studied. The quenching of BSA fluorescence caused by Cr(VI) could be delayed by PSTAs. UV-vis and circular dichroism (CD) data suggested that PSTAs can protect the secondary and tertiary structures of BSA by probably interacting with Cr(VI) in advance.

Isolation from Ochrobactrum anthropi of a novel class II 5-enopyruvylshikimate-3-phosphate synthase with high tolerance to glyphosate.

Applying the genomic library construction process and colony screening, a novel aroA gene encoding 5-enopyruvylshikimate-3-phosphate synthase from Ochrobactrum anthropi was identified, cloned, and overexpressed, and the enzyme was purified to homogeneity. Furthermore, site-directed mutagenesis was employed to assess the role of single amino acid residues in glyphosate resistance.