Cell crawling mediates collective cell migration to close undamaged epithelial gaps

Fundamental biological processes such as morphogenesis and wound healing involve the closure of epithelial gaps. Epithelial gap closure is commonly attributed either to the purse-string contraction of an intercellular actomyosin cable or to active cell migration, but the relative contribution of these two mechanisms remains unknown. Here we present a model experiment to systematically study epithelial closure in the absence of cell injury. We developed a pillar stencil approach to create well-defined gaps in terms of size and shape within an epithelial cell monolayer. Upon pillar removal, cells actively respond to the newly accessible free space by extending lamellipodia and migrating into the gap. The decrease of gap area over time is strikingly linear and shows two different regimes depending on the size of the gap. In large gaps, closure is dominated by lamellipodium-mediated cell migration. By contrast, closure of gaps smaller than 20 μm was affected by cell density and progressed independently of Rac, myosin light chain kinase, and Rho kinase, suggesting a passive physical mechanism. By changing the shape of the gap, we observed that low-curvature areas favored the appearance of lamellipodia, promoting faster closure. Altogether, our results reveal that the closure of epithelial gaps in the absence of cell injury is governed by the collective migration of cells through the activation of lamellipodium protrusion.     

Complete Denture Occlusion: An Evidence‐Based Approach

Purpose : This study involved an extensive search for randomized controlled clinical trials comparing bilateral balanced and canine‐guided dentures, and questioned whether a bilateral balanced occlusion is imperative for successful denture treatment. Materials and Methods : Studies were identified by searching electronic databases (PubMed/MEDLINE, ISI Web of Science, LILACS, and BBD). The keywords “denture” and “occlusion” were used. The minimum inclusion requirements were (1) randomized controlled trials with patients of any age wearing both maxillary and mandibular conventional complete dentures (CDs), (2) comparison between bilateral balanced and canine‐guided dentures, and (3) assessment of masticatory function and/or patients’ satisfaction. Results : The search resulted in the identification of 5166 articles. Subsequently, 5156 articles were excluded on the basis of title and abstract. By the end of the search phase, seven randomized controlled trials were considered eligible. Conclusions : Current scientific evidence suggests that bilateral balanced occlusion is not imperative for successful treatment with conventional CDs in average patients. More studies are necessary to identify if specific clinical conditions may benefit from a balanced occlusion.     

Characterization of endogenous betaine γ-amino-n-butyric acid cotransporter glycoform and its hyperosmotic regulation in MDCK cells

Increase in mRNA expression and transport activity of the betaine γ-amino-n-butyric acid cotransporter (BGAT) in response to hyperosmolality has been previously shown in MDCK cells. However, the hyperosmolality-induced response of endogenous BGAT protein expression was not investigated in detail. We show two forms of endogenous BGAT immunoreactivity that are expressed in MDCK II cells. Both are sensitive to Peptide N-Glycosidase F (PNGase F), suggesting that they are N-glycosylated proteins. One band, about 75 kDa, is resistant to Endo H, while the other 55 kDa band is sensitive to it, suggesting that they are fully N-glycosylated mature form in the post-Golgi compartment and core-glycosylated immature form in the endoplasmic reticulum (ER), respectively. When treated with hyperosmolality, they are significantly increased. But the rate of BGAT processing, as assessed by the ratio of mature to immature form, is not increased, suggesting that hyperosmolality does not facilitate the export of BGAT from the ER to the secretory pathway. Surface biotinylation and confocal microscopy show that hyperosmolality significantly increases the amount of the mature form of BGAT on the basolateral membrane with a very small fraction on the apical membrane. We conclude that BGAT is an N-glycosylated protein with two glycoforms and endogenous BGAT synthesis rather than processing is involved in the adaptation to the hyperosmotic stress. Xue-Mei Zhang and Xi-Tao Wang contributed equally to this work.     

Prediction of Human Brain Penetration of P-glycoprotein and Breast Cancer Resistance Protein Substrates Using In Vitro Transporter Studies and Animal Models

Four P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) substrates with human cerebrospinal fluid (CSF) concentrations and preclinical neuropharmacokinetics were used to assess in vitro–in vivo extrapolation of brain penetration in preclinical species and the ability to predict human brain penetration. Unbound brain (C_b,u), unbound plasma (C_p,u), and CSF compound concentrations (C_CSF) were measured in rats and nonhuman primates (NHPs), and the unbound partition coefficients (C_b,u/C_p,u and C_CSF/C_p,u) were used to assess brain penetration. The results indicated that for P-gp and BCRP dual substrates, brain penetration was severally impaired in all species. In comparison, for P-gp substrates that are weak or non-BCRP substrates, improved brain penetration was observed in NHPs and humans than in rats. Overall, NHP appears to be more predictive of human brain penetration for P-gp substrates with weak or no interaction with BCRP than rat. Although C_CSF does not quantitatively correspond to C_b,u for efflux transporter substrates, it is mostly within 3-fold higher of C_b,u in rat and NHP, suggesting that C_CSF can be used as a surrogate for C_b,u. Taken together, a holistic approach including both in vitro transporter and in vivo neuropharmacokinetics data enables a better estimation of human brain penetration of P-gp/BCRP substrates.     

Molecular Investigation of Canine Parvovirus-2 (CPV-2) Outbreak in Nevis Island: Analysis of the Nearly Complete Genomes of CPV-2 Strains from the Caribbean Region

To date, there is a dearth of information on canine parvovirus-2 (CPV-2) from the Caribbean region. During August–October 2020, the veterinary clinic on the Caribbean island of Nevis reported 64 household dogs with CPV-2-like clinical signs (hemorrhagic/non-hemorrhagic diarrhea and vomiting), of which 27 animals died. Rectal swabs/fecal samples were obtained from 43 dogs. A total of 39 of the 43 dogs tested positive for CPV-2 antigen and/or DNA, while 4 samples, negative for CPV-2 antigen, were not available for PCR. Among the 21 untested dogs, 15 had CPV-2 positive littermates. Analysis of the complete VP2 sequences of 32 strains identified new CPV-2a (CPV-2a with Ser297Ala in VP2) as the predominant CPV-2 on Nevis Island. Two nonsynonymous mutations, one rare (Asp373Asn) and the other uncommon (Ala262Thr), were observed in a few VP2 sequences. It was intriguing that new CPV-2a was associated with an outbreak of gastroenteritis on Nevis while found at low frequencies in sporadic cases of diarrhea on the neighboring island of St. Kitts. The nearly complete CPV-2 genomes (4 CPV-2 strains from St. Kitts and Nevis (SKN)) were reported for the first time from the Caribbean region. Eleven substitutions were found among the SKN genomes, which included nine synonymous substitutions, five of which have been rarely reported, and the two nonsynonymous substitutions. Phylogenetically, the SKN CPV-2 sequences formed a distinct cluster, with CPV-2b/USA/1998 strains constituting the nearest cluster. Our findings suggested that new CPV-2a is endemic in the region, with the potential to cause severe outbreaks, warranting further studies across the Caribbean Islands. Analysis of the SKN CPV-2 genomes corroborated the hypothesis that recurrent parallel evolution and reversion might play important roles in the evolution of CPV-2.     

Forced-eruption time for palatally impacted canines treated with and without ostectomy-decortication technique

Objectives:To compare forced-eruption times for palatally impacted canines treated with and without the ostectomy-decortication technique and to assess the influence of palatally impacted canine pretreatment position and angle on forced-eruption time.Materials and Methods:The sample was composed of 118 patient-subjects with 151 palatally impacted canines treated with the ostectomy-decortication technique (n = 72) and without (n = 79). The orthopantomogram radiographs (OPGs) were analyzed for palatally impacted canine angle and horizontal and vertical position. Recovery time was measured from the start of forced eruption until the canine was within ±1 mm of final dental arch position.Results:The time of forced canine eruption with ostectomy-decortication technique was significantly shorter than without (6.6 vs 21.0 months). Pretreatment canine position significantly increased forced-eruption time in the ostectomy-decortication group but not in the control sample.Conclusions:Forced-eruption time of palatally impacted canines using the ostectomy-decortication technique was 3.2 times more rapid than without. Forced-eruption time increased significantly as a function of pretreatment palatally impacted canine position severity in the ostectomy-decortication group but not in the control.     

Cone-beam computerized tomography evaluation of canine retraction using micro implant and conventional anchorage

ObjectiveVarious anchorage techniques have been designed for canine retraction. The aim was to measure and compare the rate of canine retraction with conventional method and micro implant using CBCT.Materials and methodsSample size comprising of 17 subjects were scheduled for extraction of all first premolars. After leveling and aligning, titanium micro implants were placed between the roots of the second premolar and the first molars on right side, in both the arches. Pre and post retraction CBCT scans were taken. Retraction was done using sliding mechanics using stainless steel arch wire.ResultsThe maxillary right canine (micro implant) retracted by 6.75 mm and tipped distally by 9.51° at a rate of 1.05 mm/month while the mandibular right canine retracted by 4.83 mm and tipped distally by 7.88° at a rate of 1.13 mm/month. On the left side (conventional) with molar as a source of anchorage, maxillary canine retracted by 6.03 mm and tipped distally by 6.51° at a rate of 1.46 mm/month while the mandibular canine retracted by 5.03 mm and tipped distally by 4.34° at a rate of 1.15 mm/month.ConclusionImplants can serve as a source of anchorage.