Esterase activity and isoenzymes in relation to morphogenesis in Mammillaria gracillis Pfeiff. tissue culture

Cactus Mammillaria gracillis Pfeiff. (Cactaceae), cultivated in vitro, spontaneously switches from an organised to unorganised way of growth, producing a habituated organogenic callus which regenerates normal and hyperhydric shoots without the addition of any growth regulators. Tumour tissues, induced by A. tumefaciens wild strain B6S3 (tumour TW) and rooty mutant GV3101 (tumour TR), do not express any organogenic potential. The esterase (arylesterase EC 3.1.1.2 and carboxylesterase EC 3.1.1.1) activity and isoenzyme pattern of morphologically different cactus tissues: shoot, callus, hyperhydric regenerant, tumours TW and TR, were compared. Tissue samples were frozen at –80 °C and lyophilized before protein extraction. Two esterase substrates, 1- and 2-naphthylacetate, were used. Esterase activity of all tissues varied during the period of one subculture. In shoots and tumours, the highest esterase activity for both substrates was measured on the 21st day, while in the callus and hyperhydric regenerants the highest activity was on the 7th day. Esterases were separated electrophoretically in polyacrylamide gradient gels under non-denaturating conditions. In total, 13 isoesterases, reacting with both substrates, were resolved. No differences in isoenzyme profile were noticed in correlation with the age of tissues, but the esterase activity varied among tissues. The significance of these results is discussed

Esterase activity and isoenzymes in relation to morphogenesis in Mammillaria gracillis Pfeiff. tissue culture

Cactus Mammillaria gracillis Pfeiff. (Cactaceae), cultivated in vitro, spontaneously switches from an organised to unorganised way of growth, producing a habituated organogenic callus which regenerates normal and hyperhydric shoots without the addition of any growth regulators. Tumour tissues, induced by A. tumefaciens wild strain B6S3 (tumour TW) and rooty mutant GV3101 (tumour TR), do not express any organogenic potential. The esterase (arylesterase EC 3.1.1.2 and carboxylesterase EC 3.1.1.1) activity and isoenzyme pattern of morphologically different cactus tissues: shoot, callus, hyperhydric regenerant, tumours TW and TR, were compared. Tissue samples were frozen at –80 °C and lyophilized before protein extraction. Two esterase substrates, 1- and 2-naphthylacetate, were used. Esterase activity of all tissues varied during the period of one subculture. In shoots and tumours, the highest esterase activity for both substrates was measured on the 21st day, while in the callus and hyperhydric regenerants the highest activity was on the 7th day. Esterases were separated electrophoretically in polyacrylamide gradient gels under non-denaturating conditions. In total, 13 isoesterases, reacting with both substrates, were resolved. No differences in isoenzyme profile were noticed in correlation with the age of tissues, but the esterase activity varied among tissues. The significance of these results is discussed

Coulomb-nuclear interference in the breakup of 11^{11}Be

Within a theory of breakup reactions formulated in the framework of the post form distorted wave Born approximation, we calculate contributions of the pure Coulomb and the pure nuclear breakup as well as those of their interference terms to a variety of cross sections in breakup reactions of the one-neutron halo nucleus $^{11}$Be on a number of target nuclei. In contrast to the assumption often made, the Coulomb-nuclear interference terms are found to be non-negligible in case of exclusive cross sections of the fragments emitted in this reaction on medium mass and heavy target nuclei. The consideration of the nuclear breakup leads to a better description of such data.Comment: 9 pages, latex, 2 figures, to be published in Phys. Rev. C (Rapid Communication

Development of a patient decision aid for children and adolescents following anterior cruciate ligament rupture: an international mixed-methods study

AIM: To develop and user test an evidence-based patient decision aid for children and adolescents who are considering anterior cruciate ligament (ACL) reconstruction.DESIGN: Mixed-methods study describing the development of a patient decision aid.SETTING: A draft decision aid was developed by a multidisciplinary steering group (including various types of health professionals and researchers, and consumers) informed by the best available evidence and existing patient decision aids.PARTICIPANTS: People who ruptured their ACL when they were under 18 years old (ie, adolescents), their parents, and health professionals who manage these patients. Participants were recruited through social media and the network outreach of the steering group.PRIMARY AND SECONDARY OUTCOMES: Semistructured interviews and questionnaires were used to gather feedback on the decision aid. The feedback was used to refine the decision aid and assess acceptability. An iterative cycle of interviews, refining the aid according to feedback and further interviews, was used. Interviews were analysed using reflexive thematic analysis.RESULTS: We conducted 32 interviews; 16 health professionals (12 physiotherapists, 4 orthopaedic surgeons) and 16 people who ruptured their ACL when they were under 18 years old (7 were adolescents and 9 were adults at the time of the interview). Parents participated in 8 interviews. Most health professionals, patients and parents rated the aid's acceptability as good-to-excellent. Health professionals and patients agreed on most aspects of the decision aid, but some health professionals had differing views on non-surgical management, risk of harms, treatment protocols and evidence on benefits and harms.CONCLUSION: Our patient decision aid is an acceptable tool to help children and adolescents choose an appropriate management option following ACL rupture with their parents and health professionals. A clinical trial evaluating the potential benefit of this tool for children and adolescents considering ACL reconstruction is warranted.</p

Mouse nuclear myosin I knock-out shows interchangeability and redundancy of myosin isoforms in the cell nucleus.

Nuclear myosin I (NM1) is a nuclear isoform of the well-known "cytoplasmic" Myosin 1c protein (Myo1c). Located on the 11(th) chromosome in mice, NM1 results from an alternative start of transcription of the Myo1c gene adding an extra 16 amino acids at the N-terminus. Previous studies revealed its roles in RNA Polymerase I and RNA Polymerase II transcription, chromatin remodeling, and chromosomal movements. Its nuclear localization signal is localized in the middle of the molecule and therefore directs both Myosin 1c isoforms to the nucleus. In order to trace specific functions of the NM1 isoform, we generated mice lacking the NM1 start codon without affecting the cytoplasmic Myo1c protein. Mutant mice were analyzed in a comprehensive phenotypic screen in cooperation with the German Mouse Clinic. Strikingly, no obvious phenotype related to previously described functions has been observed. However, we found minor changes in bone mineral density and the number and size of red blood cells in knock-out mice, which are most probably not related to previously described functions of NM1 in the nucleus. In Myo1c/NM1 depleted U2OS cells, the level of Pol I transcription was restored by overexpression of shRNA-resistant mouse Myo1c. Moreover, we found Myo1c interacting with Pol II. The ratio between Myo1c and NM1 proteins were similar in the nucleus and deletion of NM1 did not cause any compensatory overexpression of Myo1c protein. We observed that Myo1c can replace NM1 in its nuclear functions. Amount of both proteins is nearly equal and NM1 knock-out does not cause any compensatory overexpression of Myo1c. We therefore suggest that both isoforms can substitute each other in nuclear processes

Efficacy, safety, and tolerability of antidepressants for pain in adults : overview of systematic reviews

Objective To provide a comprehensive overview of the efficacy, safety, and tolerability of antidepressants for pain according to condition. Design Overview of systematic reviews. Data sources PubMed, Embase, PsycINFO, and the Cochrane Central Register of Controlled Trials from inception to 20 June 2022. Eligibility criteria for selecting studies Systematic reviews comparing any antidepressant with placebo for any pain condition in adults. Data extraction and synthesis Two reviewers independently extracted data. The main outcome measure was pain; for headache disorders it was frequency of headaches. Continuous pain outcomes were converted into a scale of 0 (no pain) to 100 (worst pain) and were presented as mean differences (95% confidence intervals). Dichotomous outcomes were presented as risk ratios (95% confidence intervals). Data were extracted from the time point closest to the end of treatment. When end of treatment was too variable across trials in a review, data were extracted from the outcome or time point with the largest number of trials and participants. Secondary outcomes were safety and tolerability (withdrawals because of adverse events). Findings were classified from each comparison as efficacious, not efficacious, or inconclusive. Certainty of evidence was assessed with the grading of recommendations assessment, development, and evaluation framework. Results 26 reviews (156 unique trials and >25 000 participants) were included. These reviews reported on the efficacy of eight antidepressant classes covering 22 pain conditions (42 distinct comparisons). No review provided high certainty evidence on the efficacy of antidepressants for pain for any condition. 11 comparisons (nine conditions) were found where antidepressants were efficacious, four with moderate certainty evidence: serotonin-norepinephrine reuptake inhibitors (SNRIs) for back pain (mean difference −5.3, 95% confidence interval −7.3 to −3.3), postoperative pain (−7.3, −12.9 to −1.7), neuropathic pain (−6.8, −8.7 to −4.8), and fibromyalgia (risk ratio 1.4, 95% confidence interval 1.3 to 1.6). For the other 31 comparisons, antidepressants were either not efficacious (five comparisons) or the evidence was inconclusive (26 comparisons). Conclusions Evidence of efficacy of antidepressants was found in 11 of the 42 comparisons included in this overview of systematic reviews—seven of the 11 comparisons investigated the efficacy of SNRIs. For the other 31 comparisons, antidepressants were either inefficacious or evidence on efficacy was inconclusive. The findings suggest that a more nuanced approach is needed when prescribing antidepressants for pain conditions. Systematic review registration PROSPERO CRD42022311073

Caspase Inhibition with XIAP as an Adjunct to AAV Vector Gene-Replacement Therapy: Improving Efficacy and Prolonging the Treatment Window

AAV-mediated gene therapy in the rd10 mouse, with retinal degeneration caused by mutation in the rod cyclic guanosine monophosphate phosphodiesterase β-subunit (PDEβ) gene, produces significant, but transient, rescue of photoreceptor structure and function. This study evaluates the ability of AAV-mediated delivery of X-linked inhibitor of apoptosis (XIAP) to enhance and prolong the efficacy of PDEβ gene-replacement therapy.Rd10 mice were bred and housed in darkness. Two groups of animals were generated: Group 1 received sub-retinal AAV5-XIAP or AAV5-GFP at postnatal age (P) 4 or 21 days; Group 2 received sub-retinal AAV5-XIAP plus AAV5- PDEβ, AAV5-GFP plus AAV5- PDEβ, or AAV- PDEβ alone at age P4 or P21. Animals were maintained for an additional 4 weeks in darkness before being moved to a cyclic-light environment. A subset of animals from Group 1 received a second sub-retinal injection of AAV8-733-PDEβ two weeks after being moved to the light. Histology, immunohistochemistry, Western blots, and electroretinograms were performed at different times after moving to the light.Injection of AAV5-XIAP alone at P4 and 21 resulted in significant slowing of light-induced retinal degeneration, as measured by outer nuclear thickness and cell counts, but did not result in improved outer segment structure and rhodopsin localization. In contrast, co-injection of AAV5-XIAP and AAV5-PDEβ resulted in increased levels of rescue and decreased rates of retinal degeneration compared to treatment with AAV5-PDEβ alone. Mice treated with AAV5-XIAP at P4, but not P21, remained responsive to subsequent rescue by AAV8-733-PDEβ when injected two weeks after moving to a light-cycling environment.Adjunctive treatment with the anti-apoptotic gene XIAP confers additive protective effect to gene-replacement therapy with AAV5-PDEβ in the rd10 mouse. In addition, AAV5-XIAP, when given early, can increase the age at which gene-replacement therapy remains effective, thus effectively prolonging the window of opportunity for therapeutic intervention

Standardization of molecular monitoring of CML: results and recommendations from the European treatment and outcome study

Standardized monitoring of BCR::ABL1 mRNA levels is essential for the management of chronic myeloid leukemia (CML) patients. From 2016 to 2021 the European Treatment and Outcome Study for CML (EUTOS) explored the use of secondary, lyophilized cell-based BCR::ABL1 reference panels traceable to the World Health Organization primary reference material to standardize and validate local laboratory tests. Panels were used to assign and validate conversion factors (CFs) to the International Scale and assess the ability of laboratories to assess deep molecular response (DMR). The study also explored aspects of internal quality control. The percentage of EUTOS reference laboratories (n = 50) with CFs validated as optimal or satisfactory increased from 67.5% to 97.6% and 36.4% to 91.7% for ABL1 and GUSB, respectively, during the study period and 98% of laboratories were able to detect MR4.5 in most samples. Laboratories with unvalidated CFs had a higher coefficient of variation for BCR::ABL1(IS) and some laboratories had a limit of blank greater than zero which could affect the accurate reporting of DMR. Our study indicates that secondary reference panels can be used effectively to obtain and validate CFs in a manner equivalent to sample exchange and can also be used to monitor additional aspects of quality assurance.</p

Autistic children's language imitation shows reduced sensitivity to ostracism

In dialogue, speakers tend to imitate, or align with, a partner’s language choices. Higher levels of alignment facilitate communication and can be elicited by affiliation goals. Since autistic children have interaction and communication impairments, we investigated whether a failure to display affiliative language imitation contributes to their conversational difficulties. We measured autistic children’s lexical alignment with a partner, following an ostracism manipulation which induces affiliative motivation in typical adults and children. While autistic children demonstrated lexical alignment, we observed no affiliative influence on ostracised children’s tendency to align, relative to controls. Our results suggest that increased language imitation—a potentially valuable form of social adaptation—is unavailable to autistic children, which may reflect their impaired affective understanding