Endogenous pH 6.0 β-Galactosidase Activity Is Linked to Neuronal Differentiation in the Olfactory Epithelium

The histochemical detection of β-galactosidase enzymatic activity at pH 6.0 (β-gal-pH6) is a widely used biomarker of cellular senescence in aging tissues. This histochemical assay also detects the presence of programmed cell senescence during specific time windows in degenerating structures of vertebrate embryos. However, it has recently been shown that this enzymatic activity is also enhanced in subpopulations of differentiating neurons in the developing central nervous system in vertebrates. The present study addressed the histochemical detection of β-gal-pH6 enzymatic activity in the developing postnatal olfactory epithelium in the mouse. This activity was detected in the intermediate layer of the olfactory epithelium. As development progressed, the band of β-gal-pH6 labeling in this layer increased in width. Immunohistochemistry and lectin histochemistry showed the β-gal-pH6 staining to be strongly correlated with the immunolabeling of the olfactory marker protein (OMP) that identifies mature olfactory sensory neurons. The cell somata of a subpopulation of differentiated olfactory neurons that were recognized with the Dolichos biflorus agglutinin (DBA) were always located inside this band of β-gal-pH6 staining. However, the β-gal-pH6 histochemical signal was always absent from the apical region where the cytokeratin-8 positive supporting cells were located. Furthermore, no β-gal-pH6 staining was found in the basal region of the olfactory epithelium where PCNA/pHisH3 immunoreactive proliferating progenitor cells, GAP43 positive immature neurons, and cytokeratin-5 positive horizontal basal cells were located. Therefore, β-gal-pH6 seems to be linked to neuronal differentiation and cannot be regarded as a biomarker of cellular senescence during olfactory epithelium development in mice.

WAPO-A1 is the causal gene of the 7AL QTL for spikelet number per spike in wheat

Improving our understanding of the genes regulating grain yield can contribute to the development of more productive wheat varieties. Previously, a highly significant QTL affecting spikelet number per spike (SNS), grain number per spike (GNS) and grain yield was detected on chromosome arm 7AL in multiple genome-wide association studies. Using a high-resolution genetic map, we established that the A-genome homeolog of WHEAT ORTHOLOG OF APO1 (WAPO-A1) was a leading candidate gene for this QTL. Using mutants and transgenic plants, we demonstrate in this study that WAPO-A1 is the causal gene underpinning this QTL. Loss-of-function mutants wapo-A1 and wapo-B1 showed reduced SNS in tetraploid wheat, and the effect was exacerbated in wapo1 combining both mutations. By contrast, spikes of transgenic wheat plants carrying extra copies of WAPO-A1 driven by its native promoter had higher SNS, a more compact spike apical region and a smaller terminal spikelet than the wild type. Taken together, these results indicate that WAPO1 affects SNS by regulating the timing of terminal spikelet formation. Both transgenic and wapo1 mutant plants showed a wide range of floral abnormalities, indicating additional roles of WAPO1 on wheat floral development. Previously, we found three widespread haplotypes in the QTL region (H1, H2 and H3), each associated with particular WAPO-A1 alleles. Results from this and our previous study, show that the WAPO-A1 allele in the H1 haplotype (115-bp deletion in the promoter) is expressed at significantly lower levels in the developing spikes than the alleles in the H2 and H3 haplotypes, resulting in reduced SNS. Field experiments also showed that the H2 haplotype is associated with the strongest effects in increasing SNS and GNS (H2>H3>H1). The H2 haplotype is already present in most modern common wheat varieties but is rare in durum wheat, where it might be particularly useful to improve grain yield.

Comprehensive assessment of Mahaim accessory pathways’ anatomic distribution

Objective To present the authors’ experience of Mahaim-type accessory pathways (MAPs), focusing on anatomic localizations. Methods Data from consecutive patients who underwent electrophysiological study (EPS) for MAP ablation in two tertiary centres, between January 1998 and June 2020, were retrospectively analysed. Results Of the 55 included patients, 27 (49.1%) were male, and the overall mean age was 29.5 ± 11.6 years (range, 12–66 years). MAPs were ablated at the tricuspid annulus in 43 patients (78.2%), mitral annulus in four patients (7.3%), paraseptal region in three patients (5.5%), and right ventricle mid-apical region in five patients (9.1%). Among 49 patients who planned for ablation therapy, the success rate was 91.8% (45 patients). Conclusion MAPs were most often ablated at the lateral aspect of the tricuspid annuli, sometimes at other sides of the tricuspid and mitral annuli, and infrequently in the right ventricle. The M potential mapping technique is likely to be a useful target for ablation of MAPs.

In vitro evaluation of the efficacy of different protocols on calcium hydroxide dressing removal

Objective: To compare in vitro irrigation systems’ efficacy in removing calcium hydroxide paste (Ca(OH)2) in single-root teeth. Methods: Sixty permanent teeth were prepared using ProTaper® rotatory instruments, up to F3. Specimens were split longitudinally into two halves, and two grooves located in the apical and coronal zones were created. Ca(OH)2 application into the grooves was assessed by radiographic and photographic methods. After the halves were reassembled in a silicone mold, teeth were randomly divided into experimental groups (n=15), testing Ca(OH)2 removal: 1 – CanalBrush™; 2 – E1 – Irrisonic tip; 3 – manual dynamic irrigation (MDI); 4 – XP-endo Finisher® (XP). A total volume of 10 mL of 2.5% sodium hypochlorite was used in each tooth. Finally, all groups were scored regarding the Ca(OH)2 remnants using the same methods as before. Statistical analysis was conducted using the Kruskal-Wallis test followed by Mann-Whitney U tests (with Bonferroni correction). A value of p<0.05 was considered statistically significant. Results: None of the techniques demonstrated 100% efficacy in both regions. CanalBrush™ was more effective in the apical region. Overall, E1 – Irrisonic tip and CanalBrush™ removed significantly more Ca(OH)2 than MDI and XP-endo Finisher® (p<0.05). Conclusion: Globally, E1 – Irrisonic tip and CanalBrush™ were more effective, although without statistically significant differences between them.

BT-1 An autopsy case of multicentric malignant glioma, H3K27M mutant

Introduction: In 2016, the World Health Organization (WHO) defined diffuse midline glioma, H3K27M mutant (WHO Grade 4) as a tumor with K27M mutation in histone H3.3 (H3F3A) or H3.1 (HIST1H3B/C) that develops mainly in the midline regions of the central nervous system. Here, we report a rare case of the abovementioned disease with remote multiple lesions in addition to the midline regions that was diagnosed on the initial visit. Case: The 52-year-old man, suffered from dysarthria, dysphagia, gait disturbance, and headache that gradually worsened over several months. Non-contrast-enhanced lesions were noted in the pons(swelling and involvement of the basilar artery trunk), cerebellum, thalamus, fornix, periventricular area, hippocampus, medial aspect of bilateral frontal lobes, and distally in the right frontal cortex and apical region of the left temporal lobe. The open biopsy was performed for left cerebellar surface lesion, and the pathological and genetic diagnosis was diffuse midline glioma, H3K27M mutant. Extended focal radiation at 50Gy/25fr and corpus callosal/cerebellar boost at 10Gy/5fr were performed. The lesions were markedly reduced, and neurological symptoms were also alleviated. However, 20 months after the initial visit, neurological symptoms had worsened and cerebrospinal fluid dissemination occurred, after that died at 29 months. An autopsy revealed tumor invasion mainly in the midline regions of the cerebrum and in the cerebellum, brain stem, pituitary gland, entire spinal cord, and cauda equina. Immunostaining of the distally cerebral cortex lesions showed that with a negative result for H3G34V. Discussion/Conclusion: It was suggested that caution is required for primary differential diagnosis may be presented at multiple lesions such as remote cerebral cortex of diffuse midline glioma, H3K27M-mutant.

Actin Crosslinking Family Protein 7 Deficiency Does Not Impair Hearing in Young Mice

To enable hearing, the sensory hair cell contains specialized subcellular structures at its apical region, including the actin-rich cuticular plate and circumferential band. ACF7 (actin crosslinking family protein 7), encoded by the gene Macf1 (microtubule and actin crosslinking factor 1), is a large cytoskeletal crosslinking protein that interacts with microtubules and filamentous actin to shape cells. ACF7 localizes to the cuticular plate and the circumferential band in the hair cells of vertebrates. The compelling expression pattern of ACF7 in hair cells, combined with conserved roles of this protein in the cytoskeleton of various cell types in invertebrates and vertebrates, led to the hypothesis that ACF7 performs a key function in the subcellular architecture of hair cells. To test the hypothesis, we conditionally target Macf1 in the inner ears of mice. Surprisingly, our data show that in young, but mature, conditional knockout mice cochlear hair cell survival, planar cell polarity, organization of the hair cells within the organ of Corti, and capacity to hear are not significantly impacted. Overall, these results fail to support the hypothesis that ACF7 is an essential hair cell protein in young mice, and the purpose of ACF7 expression in the hair cell remains to be understood.

Influence of Bone Quality on the Use of Implant Prostheses with Intermediate Pontic: Three-dimensional Finite Element Method

Objective The present study aimed to observe the differences in the dissipation of the main minimum stresses with the use of a fixed pontic partial prosthesis supported by two regular length implants in cortical and medullary bone tissues of different qualities. Materials and Methods Experimental groups were as follows: QI (two regular length implants with fixed pontic partial prosthesis and bony qualities consistent with type I), QII (identical to QI, with bony qualities consistent with type II), and QIII (identical to QI, with bony qualities consistent with type III). All the groups were developed and analyzed in virtual simulation environment using AnsysWorkbench software. Results The results showed highest stress concentrations in the region of the turns of the implants, especially in the apical region surrounding the implants and most notably in those positioned in the posterior region, supporting the molars. In addition, comparing the cortical bone among the groups, the results revealed increasing levels of stress in the order of QI, QII, and QIII. Comparing the medullary bone among the groups, the results revealed increasing levels of stress in the order of QIII, QII, and QI. Conclusion It was concluded that greater stress disparity occurred in the comparison between groups QI and QIII. There was a higher TMiP in QI in the cortical bone, but considering the literature values, it would not pose risks to its physiological limits. The use of a pontic fixed partial prosthesis on two regular implants of type III bone quality may cause unfavorable physiological repercussions for the posterior implant apical medullary bone.

Dynamic Hormone Gradients Regulate Wound-Induced de novo Organ Formation in Tomato Hypocotyl Explants

Plants have a remarkable regenerative capacity, which allows them to survive tissue damage after biotic and abiotic stresses. In this study, we use Solanum lycopersicum ‘Micro-Tom’ explants as a model to investigate wound-induced de novo organ formation, as these explants can regenerate the missing structures without the exogenous application of plant hormones. Here, we performed simultaneous targeted profiling of 22 phytohormone-related metabolites during de novo organ formation and found that endogenous hormone levels dynamically changed after root and shoot excision, according to region-specific patterns. Our results indicate that a defined temporal window of high auxin-to-cytokinin accumulation in the basal region of the explants was required for adventitious root formation and that was dependent on a concerted regulation of polar auxin transport through the hypocotyl, of local induction of auxin biosynthesis, and of local inhibition of auxin degradation. In the apical region, though, a minimum of auxin-to-cytokinin ratio is established shortly after wounding both by decreasing active auxin levels and by draining auxin via its basipetal transport and internalization. Cross-validation with transcriptomic data highlighted the main hormonal gradients involved in wound-induced de novo organ formation in tomato hypocotyl explants.

The antibacterial efficacy of photon-initiated photoacoustic streaming in root canals with different diameters or tapers

Background In recent years, the concept of minimally invasive endodontics has been proposed, which could be described as preventing or treating diseases by preserving more dental tissue and creating minimal damage. In the process of root canal preparation, it was recommended to use instruments with a smaller taper to preserve more tooth tissue and improve the preservation rate of the affected teeth. Photon-induced photoacoustic streaming (PIPS) was a new type of laser-activated irrigation technology, which was now widely used in endodontic treatment. The purpose of this article was to evaluate the bactericidal effect of PIPS with NaOCl in root canals with different widths or tapers. Methods Twenty-three maxillary first molars with three independent root canals were included in this study. The mesiobuccal (MB), distobuccal (DB), and palatal (P) root canals were prepared at sizes of #10/.02, #25/.02, and #25/.06, respectively. After being incubated with a bacterial suspension for 4 weeks, the specimen were irrigated with 2% NaOCl activated by conventional needle irrigation (CNI) (n = 10) or PIPS (n = 10). Three specimen were not treated (control group). Before and after irrigation, the presence of bacteria was assessed with an adenosine 5'-triphosphate (ATP) assay kit and biofilms were assessed using confocal laser scanning microscopy and scanning electron microscopy. Results In specimen prepared using PIPS irrigation, the ATP was reduced by more than 98%. When the root canal taper was 0.02, the size #25 root canals had a higher percentage of dead bacteria than the size #10 root canals in all regions (P < 0.05) in the PIPS group. When the root canal width was #25, the 0.02 taper group had a higher percentage of dead bacteria than the 0.06 taper group in the apical region (P < 0.05), except coronal and middle regions (P > 0.05). PIPS irrigation results in a greater percentage of dead bacteria and reduction of ATP in size #10/.02 root canals than CNI in size #25/.06 root canals in three regions (P < 0.05). Conclusion Increasing the width from #10 to #25 improves the bactericidal effect of PIPS in the root canal. Increasing the taper of the root canal from 0.02 to 0.06 at size #25 did not affect the bactericidal effects of PIPS. PIPS resulted in more dead bacteria in specimen with smaller tapers and root canal widths than CNI. PIPS can be used to clean the smear layer in the coronal region and open the dentin tubules. Clinical significance: Activation of irrigants with PIPS brought about significant bacterial reduction smaller tapers and width root canals compared to CNI, which was beneficial to prevent excessive loss of tooth tissue and conserve the structural integrity of teeth.

OAC-39 is an O-acyltransferase required for the synthesis of maradolipids in the dauer larva of C. elegans

Upon overcrowding or low food availability, the nematode C. elegans enters a specialized diapause stage for survival, called the dauer larva. The growth-arrested, non-feeding dauer larva undergoes a profound metabolic and physiologic switch underlying its extraordinary stress resistance and longevity. One of the metabolic signatures of dauer larvae is the accumulation of the disaccharide trehalose, which lowers the sensitivity of worms to desiccation and hyperosmotic shock. Previously, we have found that trehalose is incorporated as a headgroup into dauer-specific 6,6′-di-O-acyltrehalose lipids, named maradolipids. Despite comprising a bulk fraction of the polar lipids in dauer larvae, little is known about the physiological function of maradolipds because the enzyme(s) involved in their synthesis has not yet been identified. Here, we report that the dauer-upregulated O-acyltransferase homolog OAC-39 is essential for the synthesis of maradolipids. This enzyme is enriched at the apical region of the intestinal cells of dauer larvae, where it might participate in the structuring of the gut lumen. As OAC-39 is most probably responsible for the last step of maradolipid synthesis, its identification will pave the way for the elucidation of the function of this obscure class of lipids.