Apical End Research Articles

COMPARATIVE ANALYSIS OF STRESS-STRAIN STATE OF SINGLE-ROOTED TEETH DURING INDIRECT RESTORATION OF THE CROWN WITH STUMP INLAYS OF DIFFERENT DESIGNS

The aim of our study on the biomechanics of the tooth inlay is to select the most efficient design of stump pin inlay for indirect restoration of single-rooted teeth. Materials and methods. To implement the aim of the study, the biomechanics research was conducted in the following four designs of stump inlays: 1 - traditional, 2 - with a single-stage change in the diameter of the pin along its length, 3 - with a two-stage change in the pin along its length, and 4 - with a single-stage change in the diameter of the pin and a cap in the crown of the tooth, covering the end of the root. The mathematical modeling was performed using the common software MSC NASTRAN for finite element analysis. The package, with the help of which the elastic three-dimensional models of the dentition are built and analyzed based on the finite element procedure, determines the displacement of each node of the finite element along the axis system, with three coordinates, normal and tangential stresses, as well as equivalent Huber-Mises stresses, which are calculated according to the well-known formula of deformable solid mechanics Results. The minimum values of equivalent stresses in both the pin inlay and the dentin of the root of the tooth at the level of the apical end of the pin inlay occur when using a pin inlay with a one-stage change in the diameter of the pin and a cap on the crown of the premolar. At the same time, the values of equivalent stress in dentin at the level of the apical end of the inlay pin are closest to the maximum values of equivalent stresses observed on the surface of the root of the tooth (differences do not exceed 10%). The maximum values of equivalent stresses in both the pin inlay and the dentin of the root of the tooth at the level of the apical end of the pin inlay occur when using a pin inlay of conventional design. In this case, the difference in the values of equivalent stresses in the pin compared to the inlay with a one-stage change in the diameter of the pin and a cap is 9%, and in the dentin at the level of the apical end of the inlay pin, the difference in stresses is 10%. The maximum values of equivalent stresses in both the dentin of the root of the tooth to be restored and the pin of the stump inlay occur when applying a traditionally used pin inlay. Among the various pin inlays considered, a stump inlay featuring a single-stage change in the diameter of the pin and a cap on the crown of the tooth to be restored demonstrates the highest efficiency, yielding the lowest equivalent stresses in the dentin of the tooth's root.

Dentin Grafts: Navigating the Paradigm Shift in Regenerative Dentistry.

The biomaterial of dentin has emerged as a promising candidate for the tissue engineering of dental hard tissues. In bone tissue engineering, it may serve as either a scaffold or a reservoir of growth factors. The physical and chemical similarities between the dentin structure and bone have sparked scientific interest in using its features for the development of a new bone transplant material. Dentin, unlike hard and fragile enamel, is viscoelastic, making it a very effective bone replacement. The regeneration of pulp tissue has proven challenging due to its encasement in dentin, which lacks collateral blood flow except from the apical end of the root. Yet, the emergence of contemporary tissue engineering and the identification of dental stem cells have enabled experimentation with the regeneration of both pulp and dentin. This review will explain the different types of dentin grafts, their biocompatibility, safety, and effectiveness, along with difficulties. Additionally, the paper covers several strategies for creating autogenous dentin grafts and gives evidence-based insights into their clinical effectiveness. Overall, dentin grafts appear as a potential alternative to standard graft materials, stimulating tissue regeneration and enhancing patient outcomes in regenerative dentistry operations.

Toxoplasma FER1 is a versatile and dynamic mediator of differential microneme trafficking and microneme exocytosis

Toxoplasma gondii is a polarized cell concentrating several secretory organelles at the apical pole. The secretory micronemes come in two sub-populations differentiated by dependence on Rab5A/C in their biogenesis. Calcium-dependent exocytosis of micronemes occurs at the very apical tip and is critical for parasite egress from its host cell, adhesion and invasion of the next cell. Ferlins represent a protein family with roles in exocytosis containing multiple Ca2+-sensing C2 domains. We determined that T. gondii’s ferlin 1 (FER1) localized dynamically to the parasite’s secretory pathway. FER1 function was dissected by dominant negative overexpression strategies. We demonstrated that FER1 traffics microneme organelles along the following trajectories: (1) Along the cortex to the apical end; (2) To the apical tip for fusion with the plasma membrane; (3) Differential microneme sub-population traffic, and that FER1 could putatively be responsible for microneme protein trafficking. (4) From the trans-Golgi-endosomal network to the subpellicular cortex; (5) Retrograde transport allowing microneme recycling from mother to daughter. Finally, FER1 overexpression triggers a microneme exocytosis burst, supporting the notion that the radially organized micronemes at the apical tip comprise a readily-releasable microneme pool. In summary, FER1 is pivotal for dynamic microneme trafficking, acts differently on the two microneme subpopulations, and acts on the plasma membrane fusion step during microneme exocytosis.

THE FIRST CENOZOIC OCTOPOD: A LOWER EOCENE RECORD FROM BOLCA, NORTHEASTERN ITALY

To date, soft-tissue remains of extinct incirrate octopods have been described exclusively from UpperCretaceous deposits. Here, three specimens of an incirrate octopodid with well-preserved soft tissue imprints are described for the first time from Paleogene strata. This material originates from the upper Ypresian (lower Eocene) fish-bearing levels of the Bolca Konservat-Lagerstätte in the Pesciara of Bolca in north-eastern Italy. Previously, these specimens had tentatively been interpreted as teuthids. Based on a detailed study of their anatomical structure, a new genus and species of octopodid, Bolcaoctopus pesciaraensis, belonging to the extant family Octopodidae, are here erected. Bolcaoctopus gen. nov. may be distinguished from the Late Cretaceous genus Styletoctopus by the presence of long and thin arms and an elongated body shape with a narrowed apical end and covered by longitudinal wrinkles. Although finds of fossil octopuses in the Bolca Konservat-Lagerstätte remain extremely rare, it is worth noting that Cenozoic marine vertebrate localities constitute an important potential source of additional finds of coleoid cephalopods, the study of which may shed light on a poorly understood period in the evolutionary history of these molluscs.

The role of pressure and friction forces in automated insertion of cochlear implants.

Despite the success of cochlear implant (CI) surgery for hearing restoration, reducing CI electrode insertion forces is an ongoing challenge with the goal to further reduce post-implantation hearing loss. While research in this field shows that both friction and quasistatic pressure forces occur during CI insertion, there is a lack of studies distinguishing between these origins. The present study was conducted to analyze the contribution of both force phenomena during automated CI insertion. Five MED-EL FLEX28 CI electrode arrays were inserted into both a regular and uncoiled version of the same average scala tympani (ST). Both ST models had a pressure release hole at the apical end, which was kept open or closed to quantify pressure forces. ST models were filled with different sodium dodecyl sulfate (SDS) lubricants (1, 5, and 10% SDS, water). The viscosity of lubricants was determined using a rheometer. Insertions were conducted with velocities ranging from v= 0.125 mm/s to 2.0 mm/s. Viscosity of SDS lubricants at 20°C was 1.28, 1.96, and 2.51 mPas for 1, 5, and 10% SDS, respectively, which lies within the values reported for human perilymph. In the uncoiled ST model, forces remained within the noise floor (maximum: 0.049 × 10-3 N ± 1.5 × 10-3 N), indicating minimal contribution from quasistatic pressure. Conversely, forces using the regular, coiled ST model were at least an order of magnitude larger (minimum: Fmax = 28.95 × 10-3 N, v = 1 mm/s, 10% SDS), confirming that friction forces are the main contributor to total insertion forces. An N-way ANOVA revealed that both lubricant viscosity and insertion speed significantly reduce insertion forces (p < 0.001). For the first time, this study demonstrates that at realistic perilymph viscosities, quasistatic pressure forces minimally affect the total insertion force profile during insertion. Mixed friction is the main determinant, and significantly decreases with increaseing insertion speeds. This suggests that in clinical settings with similar ST geometries and surgical preparation, quasistatic pressure plays a subordinate role. Moreover, the findings indicate that managing the hydrodynamics of the cochlear environment, possibly through pre-surgical preparation or the use of specific lubricants, could effectively reduce insertion forces.

Accuracy of Orthodontic Anchor Screw Placement Using a 3D-Printed Surgical Guide.

Background Although radiographs and computed tomography (CT) images are reviewed before temporary anchorage device (TAD) implantation, implantation of TADs exactly as planned is difficult. This study aimed to evaluate the accuracy of TAD implantation using an original surgical guide fabricated using cone-beam CT data and computer-aided design software. Methodology The studyparticipants included six experienced orthodontists who had implanted ≥20 TADs, and six inexperienced orthodontists who had never implanted a TAD. Maxillary dental typodont models with radiopaque tooth crowns and roots were used. A total of four TADs were implanted on the buccal sides: between the second bicuspid and first molars and between the first and second molars bilaterally. The accuracy of TAD implantation was examined in two groups: in 12 dental typodont models, TAD implantation was performed using a surgical guide (guide group), and in 12 dental typodont models, TAD implantation was performed without a surgical guide (freehand group). All dental typodont models implanted a total of 96 TADs.The TAD position was evaluated using the CT coordinate system and 3D image measurement software. Using the long axis of the TAD as a reference, the distance between the coronal and apical ends of the implanted TAD and those of the planned TAD, i.e., the ideal implantation position, was measured in both groups along the x, y, and z axes. The medians of the values were compared between the groups. Additionally, the presence of root contact was compared between the experienced and inexperienced orthodontists. Results On the x-axis, the linear deviations (median) of the coronal and apical ends of the TAD in the freehand group were 1.06 mm and 1.36 mm, respectively. In contrast, in the guide group, the deviations were 0.65 mm and 0.90 mm, respectively, and the difference was statistically significant (p = 0.002 and p = 0.005, respectively). On the y-axis, the deviations in the freehand group were 1.13 mm and 1.08 mm, respectively. In contrast, the deviations in the guide group were 0.71 mm and 0.79 mm, respectively, and only the coronal deviations were significantly different between the groups (p = 0.006). On the z-axis, the deviations in the freehand group were 1.44 mm and 1.86 mm, respectively. In contrast, the deviations in the guide group were 0.75 mm and 1.16 mm, respectively, and the difference was statistically significant (p = 0.006 and p = 0.002, respectively). Conclusions The use of a surgical guide allowed for more accurate TAD implantation. Additionally, TAD implantation using a guide prevented root damage.

Metabolomics Simultaneously Derives Benchmark Dose Estimates and Discovers Metabolic Biotransformations in a Rat Bioassay.

Benchmark dose (BMD) modeling estimates the dose of a chemical that causes a perturbation from baseline. Transcriptional BMDs have been shown to be relatively consistent with apical end point BMDs, opening the door to using molecular BMDs to derive human health-based guidance values for chemical exposure. Metabolomics measures the responses of small-molecule endogenous metabolites to chemical exposure, complementing transcriptomics by characterizing downstream molecular phenotypes that are more closely associated with apical end points. The aim of this study was to apply BMD modeling to in vivo metabolomics data, to compare metabolic BMDs to both transcriptional and apical end point BMDs. This builds upon our previous application of transcriptomics and BMD modeling to a 5-day rat study of triphenyl phosphate (TPhP), applying metabolomics to the same archived tissues. Specifically, liver from rats exposed to five doses of TPhP was investigated using liquid chromatography-mass spectrometry and 1H nuclear magnetic resonance spectroscopy-based metabolomics. Following the application of BMDExpress2 software, 2903 endogenous metabolic features yielded viable dose-response models, confirming a perturbation to the liver metabolome. Metabolic BMD estimates were similarly sensitive to transcriptional BMDs, and more sensitive than both clinical chemistry and apical end point BMDs. Pathway analysis of the multiomics data sets revealed a major effect of TPhP exposure on cholesterol (and downstream) pathways, consistent with clinical chemistry measurements. Additionally, the transcriptomics data indicated that TPhP activated xenobiotic metabolism pathways, which was confirmed by using the underexploited capability of metabolomics to detect xenobiotic-related compounds. Eleven biotransformation products of TPhP were discovered, and their levels were highly correlated with multiple xenobiotic metabolism genes. This work provides a case study showing how metabolomics and transcriptomics can estimate mechanistically anchored points-of-departure. Furthermore, the study demonstrates how metabolomics can also discover biotransformation products, which could be of value within a regulatory setting, for example, as an enhancement of OECD Test Guideline 417 (toxicokinetics).

ANALYSIS OF THE FORM AND SURFACE OF DENTAL IMPLANTS

The aim of the work – to study the peculiarities of the shape and surface relief of the main types of dental implants. Materials and methods. 12 dental implants used in the medical practice of Ukrainian dental clinics were studied. The study of the shape and surface relief of the implants was carried out using a scanning electron microscope GSM-649 (Japan). Results. All implants in the apical part have the shape of a cone with threads and cutting grooves. The thread with a large profile is located in the apical part of the implant, has an average pitch of turns of 1.1±0.05 mm and a profile height of 0.3±0.02 mm. The thread with a small profile is located in the part of the implant near the neck, has a pitch of 0.4±0.02 mm and a profile height of 0.1±0.02 mm. The thread pitch of implants with a single profile is 0.8±0.05 mm, and the height of the profile is 0.2±0.01 mm. Threads with a triangular profile have a profile angle of 50.0±2.9 degrees. The highest roughness is the surface of the top of the fine profile thread with the height of micro-uniformities from 7.0±2.2 to 36.0±8.3 μm, the smallest roughness with the height of micro-uniformities from 3.0±1.1 to 14.0±6.1 μm have surface areas in the area of the implant neck. Conclusions. 50% of the examined implants were made with two thread profiles: a large one in the apical side and a small one closer to the neck, 50% – with the same thread profile from the apical end to the neck. In most implants, the thread with a small profile is made in the form of annular grooves. The large thread of the implants had a rectangular, rounded and triangular profile with one or two steps. In some implants, as they approach the neck, the shape changes and the height of the thread profile decreases. The shape of the profile of small threads in all implants was close to rounded. The implants had different surface roughness. The highest roughness was recorded on the surface of the top of the small profile thread, and the lowest – in the area without a thread in the area of the implant neck.

Pre-silking water deficit in maize induced kernel loss through impaired silk growth and ovary carbohydrate dynamics.

Both carbon limitation and developmentally driven kernel failure occur in the apical region of maize (Zea mays L.) ears. Failed kernel development in the basal and middle regions of the ear often is neglected because their spaces usually are occupied by adjacent ovaries at harvest. We tested the spatial distribution of kernel losses and potential underlying reasons, from perspectives of silk elongation and carbohydrate dynamics, when maize experienced water deficit during silk elongation. Kernel loss was distributed along the length of the ear regardless of water availability, with the highest kernel set in the middle region and a gradual reduction toward the apical and basal ends. Water deficit limited silk elongation in a manner inverse to the temporal pattern of silk initiation, more strongly in the apical and basal regions of the ear than in the middle region. The limited recovery of silk elongation, especially at the apical and basal regions following rescue irrigation was probably due to water potentials below the threshold for elongation and lower growth rates of the associated ovaries. While sugar concentrations increased or did not respond to water deficit in ovaries and silks, the calculated sugar flux into the developing ovaries was impaired and diverged among ovaries at different positions under water deficit. Water deficit resulted in 58% kernel loss, 68% of which was attributable to arrested silks within husks caused by lower water potentials and 32% to ovaries with emerged silks possibly due to impaired carbohydrate metabolism.

Timely Pulmonary Tuberculosis Diagnosis Based on the Epidemiological Disease Spectrum: Population-Based Prospective Cohort Study in the Republic of Korea.

Timely pulmonary tuberculosis (PTB) diagnosis is a global health priority for interrupting transmission and optimizing treatment outcomes. The traditional dichotomous time-divided approach for addressing time delays in diagnosis has limited clinical application because the time delay significantly varies depending on each community in question. We aimed to reevaluate the diagnosis time delay based on the PTB disease spectrum using a novel scoring system that was applied at the national level in the Republic of Korea. The Pulmonary Tuberculosis Spectrum Score (PTBSS) was developed based on previously published proposals related to the disease spectrum, and its validity was assessed by examining both all-cause and PTB-related mortality. In our analysis, we integrated the PTBSS into the Korea Tuberculosis Cohort Registry. We evaluated various time delays, including patient, health care, and overall delays, and their system-associated variables in line with each PTBSS. Furthermore, we reclassified the scores into distinct categories of mild (PTBSS=0-1), moderate (PBTBSS=2-3), and severe (PBTBSS=4-6) using a multivariate regression approach. Among the 14,031 Korean patients with active PTB whose data were analyzed from 2018 to 2020, 37% (n=5191), 38% (n=5328), and 25% (n=3512) were classified as having a mild, moderate, and severe disease status, respectively, according to the PTBSS. This classification can therefore reflect the disease spectrum of PTB by considering the correlation of the score with mortality. The time delay patterns differed according to the PTBSS. In health care delays according to the PTBSS, greater PTB disease progression was associated with a shorter diagnosis period, since the condition is microbiologically easy to diagnose. However, with respect to patient delays, the change in elapsed time showed a U-shaped pattern as PTB progressed. This means that a remarkable patient delay in the real-world setting might occur at both apical ends of the spectrum (ie, in both mild and severe cases of PTB). Independent risk factors for a severe PTB pattern were age (adjusted odds ratio 1.014) and male sex (adjusted odds ratio 1.422), whereas no significant risk factor was found for mild PTB. Timely PTB diagnosis should be accomplished. This can be improved with use of the PTBSS, a simple and intuitive scoring system, which can be more helpful in clinical and public health applications compared to the traditional dichotomous time-only approach.

Rhizome fragment weight and density of competing shoots determine belowground regrowth of Elymus repens

AbstractManagement of perennial weeds has become increasingly complex with the reduction of herbicide use and tillage. Their capacity to store nutrients into vegetative organs and to regenerate to establish new individuals makes them very persistent and hard to control. A key aspect for managing perennial weeds is to minimise storage reserves to reduce vegetative regeneration. For that prospect, better knowledge of the regrowth capacities from storage organs is needed. Our work focused on the mechanistic understanding of belowground regrowth dynamics from fragments of Elymus repens rhizomes by analysing the effect of source and sink factors. The effect of rhizome fragment weight (i.e., source) and the number of regrowing buds and belowground shoots (i.e., sinks) was assessed for four mechanisms: sprouting, belowground shoot elongation, belowground shoot biomass to length conversion and biomass allocation to belowground shoots versus roots. Heavier fragments contributed to faster sprouting, faster belowground shoot elongation and increased the proportion of remobilised biomass allocated to belowground shoots. Higher belowground shoot density increased the proportion of remobilised biomass allocated to belowground shoots but decreased individual elongation rate. In other words, larger reserves resulted in a better regeneration capacity and a higher number of growing belowground shoots increased the competition for reserves among them. The belowground shoot that sprouted first on a fragment grew faster and dominated the other shoots on the same fragment. Dominant belowground shoots were located toward the apical end of the fragment. These findings support the fact that tillage should aim at cutting storage organs into the smallest fragments possible to reduce regeneration capacity. This study also emphasised that belowground regrowth mechanisms are complex, and further studies should investigate other species with different types of storage organs.

A Unique Clinical Appearance of Upper Maxillary Premolar associated with Dentigerous Cyst: Case Report

During dental development, a variety of hereditary and environmental variables interact to cause dental abnormalities. The etiology may involve multi-tiered, multi-directional, and multi-factorial processes. Fusion is a dental anomaly that happens during tooth development, where two separated teeth bud fuse together to form a single tooth. The morpho-differentiation or histo-differentiation stages of tooth development, genetic and environmental variables can induce developmental dental abnormalities. A clinical resemblance of a "double-headed" maxillary second upper left premolar with two crowns, formed on the both coronal and apical root end or whether it is just a hypercementosis of apical root discovered after surgical extraction and enucleation of cyst. Investigations to confirm such unique findings were performed. This case report discusses the importance of clinical, radiographic and laboratory investigations to confirm the unique appearance of the maxillary second premolar.

Developing efficient strategies for localizing the enhanced yellow fluorescent protein subcellularly in transgenic Eimeria parasites

Eimeria species serve as promising eukaryotic vaccine vectors. And that the location of heterologous antigens in the subcellular components of genetically modified Eimeria may determine the magnitude and type of immune responses. Therefore, our study aimed to target a heterologous fluorescent protein to the cell surface or microneme, two locations where are more effective in inducing protective immunity, of Eimeria tenella and E. acervulina sporozoites. We used an enhanced yellow fluorescent protein (EYFP) as a tagging biomarker, fusing variously with some localization or whole sequences of compartmental proteins for targeting. After acquiring stable transgenic Eimeria populations, we observed EYFP expressing in expected locations with certain strategies. That is, EYFP successfully localized to the surface when it was fused between signal peptides and mature products of surface antigen 1 (SAG1). Furthermore, EYFP was efficiently targeted to the apical end, an optimal location for secretory organelle known as the microneme, when fused to the C terminus of microneme protein 2. Unexpectedly, EYFP exhibited dominantly in the apical end with only weak expression on the surface of the transgenic sporozoites when the parasites were transfected with plasmid with EYFP fused between signal peptides and mature products of E. tenella SAG 13. These strategies worked in both E. tenella and E. acervulina, laying a solid foundation for studying E. tenella and E. acervulina-based live vaccines that can be further tailored to the inclusion of cargo immunogens from other pathogens.

Comparison between Calcium Hydroxide with Mineral Trioxide Aggregate and Regenerative Endodontics in Inducing Root Apex Closure during Apexification – A Systematic Review and Meta-analysis

Abstract This study aimed to assess better effectiveness between regenerative endodontic procedures (REP) and apexification procedures (AP) with mineral trioxide aggregate (MTA) and calcium hydroxide for inducing root end apex closure. Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines were followed and registered in PROSPERO-CRD42023398997. Electronic databases were searched for studies evaluating effectiveness of REP and AP in terms of survival rate, success rate, increase in root length, root width and decrease in apical diameter. Cochrane risk of bias (RoB) -2 tool was used for quality assesssment using RevMan 5.3. The risk ratio (RR) and standardized mean difference (SMD) was used as summary statistic measure with random effect model (P &lt; 0.05). Nine studies were included in qualitative synthesis and eight studies for meta-analysis. Quality assessment revealed moderate to low risk of bias. The pooled estimate through RR and SMD favoured REP being superior to AP for better survival rate (RR = 1.01 (0.96 – 1.06)), success rate (RR = 1.09 (0.96 – 1.24)), increase in root length (SMD = 0.25 (-0.14 – 0.63)), root width (0.66 (0.22 – 1.10)) and decrease in apical diameter (SMD =0.66 (-0.51 – 1.83)). Funnel plot did not show any heterogeneity indicating absence of publication bias. REP significantly improved apical root end closure. AP are equally effective in forming calcific barrier, however it was concluded that regeneration procedures are comparably superior to apexification procedures with greater outcomes. Clinicians should consider employing the REP in cases when root development is severely deficient and where tooth’s prognosis is hopeless even with an apexification procedure.

Somatic Embryogenesis and Direct Shoot Bud Formation from in vitro Root Segments of Garlic (Allium sativum L.)

In vitro techniques are unconventionally used for garlic improvement. In garlic tissue culture, numerous roots are produced in vitro. Segments (1 cm) of those in vitro roots were cultivated on MS medium with 5.0 μM 2, 4-D for callus initiation followed by transfer to 0.5 μM 2, 4-D for somatic embryogenesis. Root segments had 51.67% callus and somatic embryo formation. Plantlet regeneration was obtained from 90% calli after being transferred to MS medium supplemented with 5.0 μM kinetin. Within 6 months, 54.8 plants per root segment and 42.2 plants per root tip explant were found. MS media supplemented with 1.0 or 5.0 μM NAA in combination with 10.0 or 50.0 μM BA were used for direct shoot bud induction from root segments. The highest percentage (23.33 %) of direct shoot bud regeneration was found in 5.0 μM NAA with 50.0 μM BA within 2 months. The number of shoots per explant was varied from 1-81 with the highest average of 15.75 shoots per explant. The shoots produced bulblets upon transfer to a medium containing higher amount of sucrose (12%). The largest bulblets weighed 356.57 g in the medium with 5.0 μM NAA and 50.0 μM BA. In both pathways, regeneration occurred from only the apical (distal) end of the root segments. The protocols are promising for continuous regeneration of propagules and genetic transformation study for improvement of garlic. Plant Tissue Cult. &amp; Biotech. 33(2): 135-142, 2023 (December)

The cilium like region of the Drosophila bifurca spermatocyte: Elongation of a giant axoneme without intraflagellar transport

AbstractThe growth of the ciliary axonemes mainly depends on the evolutionary conserved intraflagellar transport (IFT) machinery. However, insect spermatocytes are characterized by cilium‐like regions (CLRs) that elongate in the absence of IFT. It is generally believed that the dynamics of these structures relies on the free diffusion of soluble tubulin from the cytoplasm. However, this passive process could allow the elongation of short ciliary axonemes, but it is unclear whether simple diffusion of tubulin molecules can ensure the correct assembly of elongated ciliary structures. To decipher this point we analyzed the assembly of the CLRs held by the primary spermatocytes of Drosophila bifurca. These ciliary structures consist of a very elongated axoneme that grows without IFT and, therefore, could represent a good model in which to evaluate the role played by the free diffusion of soluble tubulin. The observation of wavy microtubules in the axonemal lumen of fully elongated CLRs of D. bifurca may be consistent with the diffusion of tubulin within the axonemal lumen. Progressive consumption of soluble tubulin used for axoneme growth at the apical tip of the CLRs could result in a gradient sufficient to move tubulin from the cytoplasm to the apical end of the forming ciliary structure. When the axoneme reaches its full length, tubulin molecules are not drawn to the tip of the CLRs and accumulate at the base of the axoneme, where its concentration may exceed the threshold need for microtubule polymerization. The presence of γ‐TuRCs at the proximal ends of the supernumerary microtubules could enhance their nucleation.

The role of partially consumed acorn remains in scatter hoards and their implications in oak colonization

Past studies have revealed the benefits of rodent participation in the colonization process of oak species. Certain rodent species (Apodemus sylvaticus and Mus spretus) partially consume acorns, beginning at the basal part and preserving the embryo. Perea et al. (2011) and Yang and Yi (2012) found that during periods of abundance, the remains left after partial consumption continue to be present on the surface and are not transported to caches, given that they are perceived as leftovers. These remains, produced after several visits by the cache owner or by thieving conspecifics, also appear in the caches. If they are perceived as offal, they will not be attacked and may remain in these stores for longer periods, serving as resources for the cache builder. Our objective is to determine whether these remnants are perceived as offal by the rodent generating them or if the remains left by other rodents are considered offal. This is relevant in cases of theft, a common behavior of this species, if the thieving animals reject the remains. The results suggest that foreign remains and the rodents’ own remains are not rejected, but rather, they are consumed in preference to intact acorns. The intact acorns remain in the cache for longer periods and have a greater opportunity to germinate and emerge. Rodents prefer to consume foreign remains first. This may be due to the fact that, in case of shortage, it is considered advantageous to finish the reserves of a potential competitor before depleting one’s own reserves.Significance statementRodents participate in the acorn dissemination process by constructing surface stores (caches). The rodent species studied here partially consumes acorns, beginning with the basal part and preserving the embryo located at the apical end. These partially consumed acorn remains are considered offal and remain in the caches for longer periods, serving as reserves for the rodent. Our objective is to examine whether these acorn remains are viewed as offal by the rodents. We have found that, to the contrary, they are consumed before intact acorns. Intact acorns remain in the caches for longer periods, assuming the role of reserves and taking on a greater capacity to germinate. This species of rodent differentiates between its own remains and those of others, first consuming the foreign offal. Therefore, their own offal remains in the stores for longer periods and may potentially germinate if the embryo is preserved. This behavior has been demonstrated by this rodent species.

Subcellular responses of fish cells to sewage effluents: Cell line-based and whole-animal based approaches

Acute toxicity determination is essential in the ecological risk assessment. Traditionally, acute toxicity testing requires substantial numbers of animals and uses death as an apical end point which requires large number of experimental animals and takes days to obtain the results. Application of fish cell lines can provide a possible alternative to traditional acute toxicity test. However, cell-based assay may show several orders of magnitude less sensitive than the animal-based results. Some changes in cellular organelles could have the sensitivity in responding to pollutants. For this reason, a cell-based fluorescent assay was developed using rabbitfish fin cells as model and fluorescent probes to visualize the subcellular responses. The subcellular responses under sewage effluents exposure were captured by confocal microscopy. These cellular responses were quantified and several subcellular indexes represented the toxicity. The optimized assay was then used to determine the toxicity of sewage effluents displaying toxicity to aquatic animals. Through visualization of cellular responses, we further screened several cellular indexes including lysosomal number and mitochondrial size which had a good linear relationship with sewage effluents content. Besides, these cellular indexes had a good agreement between in vivo and in vitro results, demonstrating the accuracy of cellular parameters in representing the acute toxicity of sewage effluents. The developed cell-based testing assay presented here has the characteristics of a faster and cheaper method, which does not require complex facilities and large amount of testing samples. The developed assay may be further applied in predicting the acute toxicity to sewage effluents.

Live Detection of Intracellular Chitin in Butterfly Wing Epithelial Cells In Vivo Using Fluorescent Brightener 28: Implications for the Development of Scales and Color Patterns.

Chitin is the major component of the extracellular cuticle and plays multiple roles in insects. In butterflies, chitin builds wing scales for structural colors. Here, we show that intracellular chitin in live cells can be detected in vivo with fluorescent brightener 28 (FB28), focusing on wing epithelial cells of the small lycaenid butterfly Zizeeria maha immediately after pupation. A relatively small number of cells at the apical surface of the epithelium were strongly FB28-positive in the cytosol and seemed to have extensive ER-Golgi networks, which may be specialized chitin-secreting cells. Some cells had FB28-positive tadpole-tail-like or rod-like structures relative to the nucleus. We detected FB28-positive hexagonal intracellular objects and their associated structures extending toward the apical end of the cell, which may be developing scale bases and shafts. We also observed FB28-positive fibrous intracellular structures extending toward the basal end. Many cells were FB28-negative in the cytosol, which contained FB28-positive dots or discs. The present data are crucial to understanding the differentiation of the butterfly wing epithelium, including scale formation and color pattern determination. The use of FB28 in probing intracellular chitin in live cells may be applicable to other insect systems.

The bulbils of Palaeonitella cranii (Charales, Charophyceae) revisited, with notes on other inflated cells of this alga that are not bulbils

Palaeonitella cranii is a charophyte alga (Charophyceae) from the Lower Devonian Rhynie chert of Scotland that consists of an axis of alternating internodes and nodes, with whorls of branchlets arising from the nodes. Conspicuous spherical to ovoid structures believed by most to be the bulbils of P. cranii are often found together with the axes and branchlets; however, they have so far only been insufficiently studied. Here, we revisit the bulbils, along with the rhizoids on which they were formed, based on new material from the Rhynie and Windyfield chert sites. The bulbils are up to 540 μm in diameter and occur singly or in groups on rhizoid nodes. A small node recognizable at the apical end in some specimens is evidence that the bulbils are modified branch rhizoids. There is also evidence of lower-order branch rhizoids extending from the apical node, as well as for sprouting. Palaeonitella cranii thrived in shallow pools and outflow channels of the Rhynie geothermal wetland. The bulbils might have enabled the plant to persist if photosynthesis was limited and to regenerate if the thallus was destroyed, but they might also have served as dispersal units. Other significantly inflated cells, which are not bulbils, may occur in both axes and branchlets of P. cranii, but are generally rare. Some of these cell alterations may be due to parasites. This study complements our understanding of P. cranii, the only fossil charophyte known from both above- and below-ground parts of the thallus, and from reproductive structures.