Microinjection of the CRISPR/Cas9 editing system through the germ pore of a wheat microspore induces mutations in the target Ms2 gene.

BACKGROUND: Microinjection is a direct procedure for delivering various compounds via micropipette into individual cells. Combined with the CRISPR/Cas9 editing technology, it has been used to produce genetically engineered animal cells. However, genetic micromanipulation of intact plant cells has been a relatively unexplored area of research, partly due to the cytological characteristics of these cells. This study aimed to gain insight into the genetic micromanipulation of wheat microspores using microinjection procedures combined with the CRISPR/Cas9 editing system targeting the Ms2 gene. METHODS AND RESULTS: Microspores were first reprogrammed by starvation and heat shock treatment to make them structurally suitable for microinjection. The large central vacuole was fragmented and the nucleus with cytoplasm was positioned in the center of the cell. This step and an additional maltose gradient provided an adequate source of intact single cells in the three wheat genotypes. The microcapillary was inserted into the cell through the germ pore to deliver a working solution with a fluorescent marker. This procedure was much more efficient and less harmful to the microspore than inserting the microcapillary through the cell wall. The CRISPR/Cas9 binary vectors injected into reprogrammed microspores induced mutations in the target Ms2 gene with deletions ranging from 1 to 16 bp. CONCLUSIONS: This is the first report of successful genome editing in an intact microspore/wheat cell using the microinjection technique and the CRISPR/Cas9 editing system. The study presented offers a range of molecular and cellular biology tools that can aid in genetic micromanipulation and single-cell analysis.

Establishing femoral component rotation using a dynamic tensioner does not improve patellar position after total knee replacement with use of anatomic implants: A case-control study.

BACKGROUND: Anterior knee pain (AKP) is one of the reasons for dissatisfaction after total knee replacement (TKR). It may result from patellofemoral joint dysfunction, caused by improper rotation of implant components. The aim of this study was to analyze patella positioning in patients after standard measured resection TKR and TKR with a use of a dynamic tensioner, and to assess the frequency of AKP, range of motion (ROM), and patient-reported outcome measures 6 weeks and 3 months postoperatively. METHODS: The study consisted of 127 patients who underwent TKR. Eighty-nine of them treated with use of the dynamic tensioner FUZION formed the study group; the remainder formed the control group. All participants received cemented PERSONA MC without patella resurfacing. All patients had a standard anteroposterior, lateral weight-bearing, long-leg view X-ray and computed tomography examination in 30° of knee flexion following the procedure. RESULTS: There were no significant differences between the study and the control groups regarding: posterior condylar axis (PCA)-patella angle, surgical transepicondylar axis (sTEA)-patella angle, PCA-sTEA angle, deviation from 90° in PCA-patella, sTEA-patella and PCA-sTEA angles. No significant difference was found in a ratio of obtaining PCA-patella angle deviation of more than 3°. Regarding clinical parameters, statistically and questionablly clinically significant difference in favor of the Study Group was found in Forgotten Joint Score 6 weeks and 3 months post-operativley and ROM 6 weeks post-operatively. However, such difference was not found by analyzing ROM 3 months post-operatively, AKP and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores. CONCLUSION: Compared with the standard 3° of femoral component external rotation, use of a dynamic tensioner does not allow for more accurate restoration of the patellar facet position with reference to the PCA.

Evaluation of CSF 8-iso-prostaglandin F2α and erythrocyte anisocytosis as prognostic biomarkers for delayed cerebral ischemia after aneurysmal subarachnoid hemorrhage.

Delayed cerebral ischemia (DCI) is a serious, life-threatening, complication affecting patients who have survived the initial bleeding from a ruptured intracranial aneurysm. Due to the challenging diagnosis, potential DCI prognostic markers should be of value in clinical practice. According to recent reports isoprostanes and red blood cell distribution (RDW) showed to be promising in this respect. We conducted a prospective study of 27 aSAH patients and control group (n = 8). All patients from the study group were treated within the first day of the initial bleeding. We collected data regarding clinical status and results of biochemical, and radiological examinations. We measured cerebrospinal fluid (CSF) concentration of 8-iso-prostaglandin F2α (F2-IsoP) and RDW on days 1, 3, and 5. Both CSF F2-IsoP level and RDW-SD measured on day 1 were significant predictors of DCI. The receiver operating characteristics curve for DCI prediction based on the multivariate model yielded an area under the curve of 0.924 (95% CI 0.824-1.000, p < 0.001). In our study, the model based on the combination of RDW and the level of isoprostanes in CSF on the first day after the initial bleeding showed a prognostic value for DCI prediction. Further studies are required to validate this observation.

Predictors of 30-Day Mortality for Surgically Treated Patients with Spontaneous Supratentorial Intracerebral Hemorrhage and Validation of the Surgical Swedish Intracerebral Hemorrhage Score: A Retrospective Single-Center Analysis of 136 Cases.

OBJECTIVE: We aimed to identify independent risk factors of 30-day mortality in patients with surgically treated spontaneous supratentorial intracerebral hemorrhage (ICH), validate the Surgical Swedish ICH (SwICH) score within Polish healthcare system, and compare the SwICH score to the ICH score. METHODS: We carried out a single-center retrospective analysis of the medical data juxtaposed with computed tomography scans of 136 ICH patients treated surgically between 2008 and 2022. Statistical analysis was performed using the same characteristics as in the SwICH score and the ICH score. Backward stepwise logistic regression with both 5-fold crossvalidation and 1000× bootstrap procedure was used to create new scoring system. Finally predictive potential of these scales were compared. RESULTS: The most important predictors of 30-day mortality were: ICH volume (P < 0.01), Glasgow Coma Scale at admission (P < 0.01), anticoagulant status (P = 0.03), and age (P < 0.01). The SwICH score appears to have a better predictive potential than the ICH score, although this did not reach statistical significance [area under the curve {AUC}: 0.789 (95% confidence interval {CI}: 0.715-0.863) vs. AUC: 0.757 (95% CI: 0.677-0.837)]. Moreover, based on the analyzed characteristics, we developed our score (encompassing: age, ICH volume, anticoagulants status, Glasgow Coma Scale at admission), [AUC of 0.872 (95% CI: 0.815-0.929)]. This score was significantly better than previous ones. CONCLUSIONS: Differences in health care systems seem to affect the accuracy of prognostic scales for patients with ICH, including possible differences in indications for surgery and postoperative care. Thus, it is important to validate assessment tools before they can be applied in a new setting and develop population-specific scores. This may improve the effectiveness of risk stratification in patients with ICH.

Global contribution of invertebrates to forest litter decomposition.

Forest litter decomposition is an essential component of global carbon and nutrient turnover. Invertebrates play important roles in litter decomposition, but the regional pattern of their effects is poorly understood. We examined 476 case studies across 93 sites and performed a meta-analysis to estimate regional effects of invertebrates on forest litter decomposition. We then assessed how invertebrate diversity, climate and soil pH drive regional variations in invertebrate-mediated decomposition. We found that (1) invertebrate contributions to litter decomposition are 1.4 times higher in tropical and subtropical forests than in forests elsewhere, with an overall contribution of 31% to global forest litter decomposition; and (2) termite diversity, together with warm, humid and acidic environments in the tropics and subtropics are positively associated with forest litter decomposition by invertebrates. Our results demonstrate the significant difference in invertebrate effects on mediating forest litter decomposition among regions. We demonstrate, also, the significance of termites in driving litter mass loss in the tropics and subtropics. These results are particularly pertinent in the tropics and subtropics where climate change and human disturbance threaten invertebrate biodiversity and the ecosystem services it provides.

The role of family and individual factors in going through adolescence having a disabled sibling - analysis of a healthy child's functioning. A case-control study protocol.

BACKGROUND: Having a disabled sibling can be a source of extremely important and enriching experiences that foster the development of social and emotional competences, as well as broadening the child's perspective and knowledge. On the other hand, it also poses many challenges for the siblings, especially in adolescence. Thus, the aim of the proposed project is to investigate the specificity of the growing up process in young people with disabled siblings. PARTICIPANTS AND PROCEDURE: A total of 160 dyads (320 participants) - an adolescent and one of his/her parents - will take part in this cross-sectional case-control study. Participants will be assigned to four groups, having a sibling with: 1) intellectual disability, 2) motor disability, 3) chronic somatic disease, and 4) the control group - having a sibling without any disability. Both the healthy adolescent and one of the parents will fill out a set of questionnaires regarding the study variables: Questionnaire of Relationships with Siblings, Parental Attitude Scale-2, KidScreen-27, Parentification Questionnaire for Youth, Teenage Rebellion Questionnaire, Child Behavior Checklist 6-18. RESULTS: The primary outcomes include investigating the healthy adolescent functioning in three environments: family, peers and school. The proposed research model focuses both on the difficulties associated with having a disabled sibling (e.g. possible occurrence of disorders) and resources (e.g. higher quality of relationships in the family, especially in the sibling subsystem). CONCLUSIONS: The proposed comprehensive approach to the issue of disability in the family from the perspective of a healthy child will allow for a better understanding of the mechanisms underlying the process of growing up with disabled siblings.

Meckel's Diverticulum Injuries after Blunt Trauma.

Background: The complications associated with Meckel's diverticulum (MD) are well-known. However, blunt injuries regarding MD have not been widely described in the literature. This was the reason for undertaking research on MD lesions. Materials and Methods: The materials consisted of 28 cases of damage to MD after blunt trauma published during the years 1921-2022. The collected data were subjected to statistical analysis. Results: The following MD injuries have been identified, starting with the most common isolated perforation of MD, bleeding from the area of MD, perforation of MD with concomitant tearing of the mesentery intestines and bleeding, intussusception of MD, and intramural hematoma of MD with adjacent mesenteric hematoma. Most injuries were caused by a traffic accident, followed by sports, accidents at work, and then violence. Almost all the cases (27) involved men. Several possible mechanisms may contribute to post-traumatic damage to MD. First of all, they are associated with abdominal compression and a secondary increase in intra-abdominal pressure as well as with the action of shearing forces during deceleration. Conclusions: Traumatic MD injuries are differentiated and very rare. They can coexist with other serious injuries to the abdominal organs.

The Anterior Inferior Cerebral Artery Variability in the Context of Neurovascular Compression Syndromes: A Narrative Review.

The anterior inferior cerebellar artery (AICA) is situated within the posterior cranial fossa and typically arises from the basilar artery, usually at the pontomedullary junction. AICA is implicated in various clinical conditions, encompassing the development of aneurysms, thrombus formation, and the manifestation of lateral pontine syndrome. Furthermore, owing to its close proximity to cranial nerves within the middle cerebellopontine angle, AICA's pulsatile compression at the root entry/exit zone of cranial nerves may give rise to specific neurovascular compression syndromes (NVCs), including hemifacial spasm (HFS) and geniculate neuralgia concurrent with HFS. In this narrative review, we undertake an examination of the influence of anatomical variations in AICA on the occurrence of NVCs. Significant methodological disparities between cadaveric and radiological studies (CTA, MRA, and DSA) were found, particularly in diagnosing AICA's absence, which was more common in radiological studies (up to 36.1%) compared to cadaver studies (less than 5%). Other observed variations included atypical origins from the vertebral artery and basilar-vertebral junction, as well as the AICA-and-PICA common trunk. Single cases of arterial triplication or fenestration have also been documented. Specifically, in relation to HFS, AICA variants that compress the facial nerve at its root entry/exit zone include parabola-shaped loops, dominant segments proximal to the REZ, and anchor-shaped bifurcations impacting the nerve's cisternal portion.

The role of multidisciplinary diagnostic and therapeutic model of care in Lamb-Shaffer syndrome - case report.

This case study illustrates a multidisciplinary diagnostic and therapeutic model of care for a 7-year-old male with Lamb-Shaffer syndrome (LAMSHF). LAMSHF is an ultra-rare genetic neurodevelopmental disorder, caused by heterozygous alterations in the SOX5 gene. An integrative model of therapy of cognitive functions and speech is described. The presented approach allows the development of language competences through stimulation of basic cognitive functions, which allows the learning of the abstract rules of an inflected language. A surprising, unexpected improvement in the cognitive functioning of the child was observed (both in terms of reasoning and speech), as well as an increase in his independence. The clinically important problem of the need for continued stimulation of cognitive development, in spite of the unfavourable prognosis associated with LAMSHF, is highlighted.

Reaction Kinetics using a Chemputable Framework for Data Collection and Analysis.

Automated chemistry platforms have been widely explored, but many focus on fixed tasks for chemical synthesis or analysis. However, a typical synthetic chemistry workflow utilizes both, such as kinetic measurements for reaction development and optimization. Due to their repetitive and time-consuming nature, kinetic measurements are often omitted, which limits the mechanistic investigation of reactions. Herein, we present a "Chemputer" platform with on-line analytics (UV/Vis, NMR) which automates routine kinetic measurements. The system's capabilities are showcased by exploring an inverse electron-demand Diels-Alder using initial rate measurements, a metal complexation using variable time normalization analysis (VTNA), and formation of a series of tosylamide derivatives using Hammett analysis. Over 60 individual experiments are presented which required minimal intervention, highlighting the significant time savings of automation. Owing to the modular design of the platform, which facilitates rapid integration of commercial analytical tools, our approach is widely accessible and adjustable to the reaction under investigation. The platform is operated using the chemical programming language, XDL, hence experimental procedures and results are stored in a precise, computer-readable format. We propose that widespread adoption of this reporting protocol in the chemical community could build a database of validated kinetic data beneficial for Machine Learning.

Developmental learning disorders in children with prenatal/perinatal exposure to hypoxia: A systematic review protocol.

BACKGROUND: Developmental learning disorder (DLD) belongs to neurodevelopmental disorders because it results from the developmental neurodiversity of the brain. The main causes of DLD are genetics, but environmental factors, like inadequate supply of oxygen during pregnancy or labor, are considered. METHODS: Our search strategy will consist of electronic databases (PubMed, PsycINFO, Web of Science, EMBASE, and Cochrane Library) and hand searching. The observational studies including cohort and case-control studies will be included. The primary outcome will be (DLD). Screening and eligibility will be done independently by two reviewers based on pre-specified eligibility criteria. Data extraction will be based on a pre-pilot data extraction form, and conducted by two authors independently. Study quality will be assessed by two authors independently. Any discrepancies identified at any stage of the review will be resolved by discussion or/and consultation with another reviewer. We plan a narrative and tabular summary of the findings. DISCUSSION: This systematic review of aetiology follows the traditional approach to evidence-based healthcare. This secondary research will assess the association between hypoxia and DLD by assessing the relationship of health-related event and outcome and examining the association between them. This review can provide information for healthcare professionals and policymakers indicating whether taking into account information about hypoxia should be permanently included in the diagnostic ontogenetic interview in the process of diagnosing neurodevelopmental disorders. SYSTEMATIC REVIEW REGISTRATION: PROSPERO: CRD42022371387.

The Superior Cerebellar Artery: Variability and Clinical Significance.

The superior cerebellar artery (SCA) arises from the distal part of the basilar artery and passes by the oculomotor, trochlear, and trigeminal nerves. SCA is known to play a crucial role in the development of trigeminal neuralgia. However, due to its anatomical variability, it may also trigger other neurovascular compression (NVC), including hemifacial spasm, oculomotor nerve palsy, and ocular neuromyotonia. Additionally, it may be associated with ischemic syndromes and aneurysm development, highlighting its clinical significance. The most common anatomical variations of the SCA include duplication, a single vessel origin from the posterior cerebral artery (PCA), and a common trunk with PCA. Rarely observed variants include bifurcation and origin from the internal carotid artery. Certain anatomical variants such as early bifurcation and caudal course of duplicated SCA trunk may increase the risk of NVC. In this narrative review, we aimed to examine the impact of the anatomical variations of SCA on the NVCs based on papers published in Pubmed, Scopus, and Web of Science databases with a snowballing approach. Our review emphasizes the importance of a thorough understanding of the anatomical variability of SCA to optimize the management of patients with NVCs associated with this artery.

Differences in joint line level and posterior condylar offset during total knee replacement with use of gap-balancing and measured resection techniques-matched cohort study.

BACKGROUND: Total knee replacement (TKR) is considered one of the most common elective orthopaedic procedures. The main focus of TKR is to offer patient's symptomatic relief from persistent knee pain. To achieve this it is crucial to restore joint biomechanics by performing proper bone cuts. Some surgeons favor the measured resection technique, others prefer gap balancing technique. The researchers of the presented study performed TKR using these two techniques. The aim of this study was to compare the postoperative change in joint line and posterior condylar offset after TKR with use of anatomic knee design implants between gap balancing and measured resection techniques. METHODS: Two hundred twenty-five X-rays of patients who underwent TKR performed by a single surgeon between 2020 and 2021 were analyzed. The first group of patients (101) was operated with the use of gap balancing technique and the second group (124) was operated with the use of measured resection technique. Patients included in the study were > 50 years of age, had confirmed primary knee osteoarthritis, underwent primary TKR with a PS (posterior stabilized) knee implants without patella resurfacing and had at least 15 degree flexion contracture. T-student test and U Mann-Whitney test were used in statistical analysis of results, according to the normality of distribution examined with the Shapiro-Wilk test. Post-hoc analysis was performed using the Dwass-Steel-Crichtlow-Fligner test (DSCF). RESULTS: The postoperative analysis showed a significantly elevated joint line level in the gap balancing group (-2.6 ± 4.1 vs -0.7 ± 4.8, p < 0.0005). In the gap balancing group significantly more patients had joint lines elevated > 2 mm comparing to measured resection technique. The difference between pre- and postoperative PCO (posterior condylar offset) and PCOR (posterior condylar offset ratio) results had no significant differences (100.8 ± 11.8 vs 101 ± 12.5, p > 0.05) between the groups. CONCLUSIONS: The results of the study suggest that when it comes to restoring joint line level measured resection technique seems to be superior in comparison to the gap balancing technique. What is more, results indicate measured resection is equal in terms of restoring posterior condylar offset to the gap balancing technique. TRIAL REGISTRATION: NCT04164147, date of registration: November 14, 2019.

Sociocultural predictors of obligatory exercise in young men: A Polish-Chinese comparison.

Background: Obligatory exercise has been shown to have negative physical and mental effects on exercisers and is more prevalent among young people. However, there is limited research on obligatory exercise among young men. Social comparison theory offers a novel perspective to explore the relationship between sociocultural factors and obligatory exercise among young men, which offers an opportunity to understand potential factors contributing to obligatory exercise among young men across different cultures. Method: We recruited a purposive sample of young people aged 18-30 from Poland (n=79) and China (n=194). Participants completed self-report measures including the Sociocultural Attitudes Toward Appearance Questionnaire3, Inventory of Physical Activity Objectives, and Obligatory Exercise Questionnaire. In the data analysis stage, we examined the strength of the relationships between the independent variables and the dependent variable through multiple regression analysis, and tested the role of the mediating variables. Results: The main analyses revealed that Internalization-Athlete was a common direct predictor of obligatory exercise for both Polish and Chinese young men; that there were direct sociocultural predictors of obligatory exercise that were only used in relation to Polish or Chinese young men; and that social adaptation goals for motivation for physical activity mediated the development of obligatory exercise for Polish and Chinese young men, and that there were cross-cultural differences. Conclusion: Attention should be paid to their attitudes towards the idea of a muscular and athletic body and socially adapted physical activity motivations when understanding young men's obligatory exercise, while also considering cross-cultural differences.

Comparison of Femoral Component Rotation between Robotic-Assisted vs. Soft-Tissue Tensor Total Knee Arthroplasty with Anatomic Implants.

Background and Objectives: Total knee arthroplasty (TKA) is the most effective treatment method for end-stage osteoarthritis. One of the most important aspects of this surgery is adequate implant positioning, as it guarantees the desired outcome of restoring limb biomechanics. Surgical technique is being continuously improved along with hardware development. There are two novel devices designed to help establish proper femoral component rotation: soft-tissue tensor and robotic-assisted TKA (RATKA). This study compared the femoral component rotation achieved with the use of three methods: RATKA, soft tissue tensioner and the conventional measured-resection technique, all of them utilizing anatomical design prosthesis components. Materials and Methods: A total of 139 patients diagnosed with end-stage osteoarthritis underwent total knee arthroplasty between December 2020 and June 2021. After the surgery, they were divided into three groups depending on procedure technique and implant type: Persona (Zimmer Biomet) + Fuzion Balancer, RATKA + Journey II BCS or conventional TKA + Persona/Journey. Postoperatively, a computed tomography examination was performed in order to measure femoral component rotation. All three groups were compared independently during statistical analysis. Fisher's exact, Kruskal-Wallis and Dwass-Steel-Crichtlow-Fligner tests were used for particular calculations. Results: Statistically significant differences in femoral component rotation between groups were noticed. However, in terms of values other than 0° in external rotation, no significant variance was revealed. Conclusions: Additional total knee arthroplasty instruments seem to improve the outcomes of the surgery, providing better component positioning than in the conventional measured-resection technique based only on bone landmarks.

Internalization of athletic body ideal as a mediating variable between family influence and body image of young women. A cross-cultural study of polish, Italian, and Ukrainian women.

Introduction: Our aim was to analyze the strength of the family's influence on the internalization of the ideal of an athletic figure and, consequently, on the multifactorial image of the body, from the perspective of intercultural differences. Methods: A total of 488 healthy women aged 19-26; of Polish (154), Ukrainian (228), and Italian (106) took part in the study. The Sociocultural Attitudes Towards Appearance Questionnaire (SATAQ-4) and the Multidimensional Body-Self Relations Questionnaire (MBSRQ 69) were used to measure athletic ideal internalization and family pressure. The body image of Ukrainian, Polish, and Italian women depends both on the degree of internalization the ideal of an athletic figure and influence of the family. Results and Discussion: The research also suggests significant differences between the three populations, which may suggest cultural differences between young women living in Eastern, Central, and South European countries.

Socio-cultural attitudes toward the body as a predictor of motivation for physical activity in young people brought up in Asian and European culture-Chinese-Polish comparison.

The influence of sociocultural attitudes toward the body on young people's physical activity has received increasing attention. However, there is a lack of cross-cultural research in this area. The main aim of this research was to identify the similarities and differences in the sociocultural attitudes toward the body of Polish and Chinese young people who grew up in European and Asian cultures and to analyze their effect on the motivation for physical activity. A cross-sectional research study was conducted among 18- to 30-year-old Polish (n = 259) and Chinese (n = 208) young people. The variables were measured using the Sociocultural Attitudes towards Appearance Questionnaire 3 (SATAQ 3) and the Inventory of Physical Activity Objectives (IPAO). Descriptive and comparative statistics, Spearman's rho, and the stepwise multiple regression analysis were used. The main analysis showed There are both similarities and significant differences in the performance of young Polish and Chinese men and women on the variables studied; Internalization-Athlete, Pressures, and Internalization-General are universal sociocultural predictors of motivation for physical activity among young people in Poland and China; Information is a specific sociocultural predictor of motivation for physical activity in Polish young people. The cultural nuances need to be considered in understanding young people's Motivation for undertaking physical activity.

Psychological Functioning of Women Diagnosed with Lichen Planus and Other Diseases of the Oral Cavity-Explorative Study.

The role of psychosocial factors in the development of changes in lichen planus and other diseases of the oral cavity has been implicated, but is still understudied. Therefore, the aim of our study was to describe the specific profile of psychological functioning of patients with these diseases, including the role of temperamental traits, action-oriented personality components, and self-esteem. In total, 94 adult women participated in the study: (1) with lichen planus (LP; n = 46; Mage = 54.80, SD = 12.53), (2) with other oral conditions (n = 25; Mage = 34.76, SD = 16.03), (3) without chronic disease (n = 24; Mage = 40.96, SD = 13.33). The following questionnaires were used: ZKA-PQ/SF, Polish Adaptive and Maladaptive Perfectionism Questionnaire, ACS-90, PROCOS, and MSEI. Results indicated no significant differences in temperament dimensions between studied groups. However, women diagnosed with LP presented lower levels of maladaptive perfectionism and social support than healthy women. Furthermore, women with LP also obtained lower scores for social resourcefulness and higher scores for moral self-approval than healthy women. Summarizing, patients with LP often use compensatory mechanisms that negatively affect their social functioning; thus diagnostic/therapeutic programs directed towards those group should be holistic, including psychologists and psychiatrists who support patients' psychological well-being.

A bifunctional selectable marker for wheat transformation contributes to the characterization of male-sterile phenotype induced by a synthetic Ms2 gene.

KEY MESSAGE: An engineered selectable marker combining herbicide resistance and yellow fluorescence contributes to the characterization of male-sterile phenotype in wheat, the severity of which correlates with expression levels of a synthetic Ms2 gene. Genetic transformation of wheat is conducted using selectable markers, such as herbicide and antibiotic resistance genes. Despite their proven effectiveness, they do not provide visual control of the transformation process and transgene status in progeny, which creates uncertainty and prolongs screening procedures. To overcome this limitation, this study developed a fusion protein by combining gene sequences encoding phosphinothricin acetyltransferase and mCitrine fluorescent protein. The fusion gene, introduced into wheat cells by particle bombardment, enabled herbicide selection, and visual identification of primary transformants along with their progeny. This marker was then used to select transgenic plants containing a synthetic Ms2 gene. Ms2 is a dominant gene whose activation in wheat anthers leads to male sterility, but the relationship between the expression levels and the male-sterile phenotype is unknown. The Ms2 gene was driven either by a truncated Ms2 promoter containing a TRIM element or a rice promoter OsLTP6. The expression of these synthetic genes resulted in complete male sterility or partial fertility, respectively. The low-fertility phenotype was characterized by smaller anthers than the wild type, many defective pollen grains, and low seed sets. The reduction in the size of anthers was observed at earlier and later stages of their development. Consistently, Ms2 transcripts were detected in these organs, but their levels were significantly lower than those in completely sterile Ms2TRIM::Ms2 plants. These results suggested that the severity of the male-sterile phenotype was modulated by Ms2 expression levels and that higher levels may be key to activating total male sterility.

Curve-on-curve technique does not improve tibial coverage in total knee arthroplasty in comparison to tibial tuberosity technique with use of anatomical implants: randomized controlled trial.

INTRODUCTION: During the last years, main attention while performing total knee replacement was paid to femoral component alignment; however, there is still lack of studies concerning tibial baseplate rotational alignment, especially in terms of anatomical designs of knee prosthesis. Some recent studies proved that tibial baseplate malrotation might be a cause of knee pain and patients' dissatisfaction. The aim of this study was to compare tibial component rotation and its coverage on the tibial plateau achieved with curve-on-curve and tibial tuberosity techniques (t-t technique) with use of anatomic knee designs with asymmetric tibial baseplate. MATERIALS AND METHODS: A total of 88 patients were randomly assigned in a 1:1 ratio to undergo total knee arthroplasty with use of the PERSONA PS (Zimmer Biomet) knee design with an asymmetric baseplate. The rotation of the tibial component was assessed and performed with two different techniques: curve-on-curve technique and tibial tuberosity technique. Tibial component rotation was measured on computed tomography (CT) scans using the method suggested by Benazzo et al. and designed for asymmetrical implants. For the measurement of the tibial bone coverage, the component surface area was outlined and measured on a proper CT section, then the tibial cut surface area was outlined and measured on a section just below the cement level. Pre- and post-operative range of motion was measured by another independent researcher 12 months post-operatively during follow-up visit. RESULTS: There was a statistically significant difference between both groups in median value of tibial rotation angle: 7° (interquartile range (IQR) = 0-12) in curve-on-curve technique group vs 2° (IQR-1-7) in tibial tuberosity technique group, probability value (p) = 0.0041, with values above 0 meaning external rotation of the component. There was no statistically significant difference between both groups in terms of range of motion (ROM) with average values of 124.3° ± 13.0° for curve-on-curve technique and 125.6° ± 12.8° for t-t technique with p = 0.45. There was a statistically insignificant difference between both groups in terms of coverage percentage in slight favor for curve-on-curve technique (85.9 ± 4.2 vs 84.5 ± 4.8, p = 0.17). CONCLUSION: In this study, no difference between the groups in terms of tibial bone coverage and range of motion was proved, even though both techniques differed significantly with values of tibial rotation. Future studies should be focused on influence of specific values of tibial rotation on patient-reported outcomes and survivorship of anatomic knee implants.