Loss of Zona Pellucida Binding Proteins in the Acrosomal Matrix Disrupts Acrosome Biogenesis and Sperm Morphogenesis | ScienceGate (2025)

Author(s):

Yi-Nan Lin ◽

Angshumoy Roy ◽

Wei Yan ◽

Kathleen H. Burns ◽

Martin M. Matzuk

Keyword(s):

Eggs have long been a favorite material for studying the mechanism of karyokinesis in-vivo and in-vitro. They can be obtained in great numbers and, when fertilized, divide synchronously over many cell cycles. However, they are not considered to be a practical system for ultrastructural studies on the mitotic apparatus (MA) for several reasons, the most obvious of which is that sectioning them is a formidable task: over 1000 ultra-thin sections need to be cut from a single 80-100 μm diameter egg and of these sections only a small percentage will contain the area or structure of interest. Thus it is difficult and time consuming to obtain reliable ultrastructural data concerning the MA of eggs; and when it is obtained it is necessarily based on a small sample size.We have recently developed a procedure which will facilitate many studies concerned with the ultrastructure of the MA in eggs. It is based on the availability of biological HVEM's and on the observation that 0.25 μm thick serial sections can be screened at high resolution for content (after mounting on slot grids and staining with uranyl and lead) by phase contrast light microscopy (LM; Figs 1-2).

Background: Many drugs are delivered intranasally for local or systemic effect, typically in the form ofdroplets or aerosols. Because of the high cost of in vivo studies, drug developers and researchers often turn to in vitro or insilico testing when first evaluating the behavior and properties of intranasal drug delivery devices and formulations.Recent advances in manufacturing and computer technologies have allowed for increasingly realistic and sophisticated invitro and in silico reconstructions of the human nasal airways.Objective:To perform a summary of advances in understanding of intranasal drug delivery based on recent in vitro and insilico studies.Conclusion: The turbinates are a common target for local drug delivery applications, and while nasal sprays are able toreach this region, there is currently no broad consensus across the in vitro and in silico literature concerning optimalparameters for device design, formulation properties and patient technique which would maximize turbinate deposition.Nebulizers are able to more easily target the turbinates, but come with the disadvantage of significant lung deposition.Targeting of the olfactory region of the nasal cavity has been explored for potential treatment of central nervous systemconditions. Conventional intranasal devices, such as nasal sprays and nebulizers, deliver very little dose to the olfactoryregion. Recent progress in our understanding of intranasal delivery will be useful in the development of the nextgeneration of intranasal drug delivery devices.

Aim and Objective: Lipoxygenase (LOX) enzymes play an important role in thepathophysiology of several inflammatory and allergic diseases including bronchial asthma, allergicrhinitis, atopic dermatitis, allergic conjunctivitis, rheumatoid arthritis and chronic obstructivepulmonary disease. Inhibitors of the LOX are believed to be an ideal approach in the treatment ofdiseases caused by its over-expression. In this regard, several synthetic and natural agents are underinvestigation worldwide. Alkaloids are the most thoroughly investigated class of natural compoundswith outstanding past in clinically useful drugs. In this article, we have discussed various alkaloids ofplant origin that have already shown lipoxygenase inhibition in-vitro with possible correlation in insilico studies.Materials and Methods: Molecular docking studies were performed using MOE (MolecularOperating Environment) software. Among the ten reported LOX alkaloids inhibitors, derived fromplant, compounds 4, 2, 3 and 1 showed excellent docking scores and receptor sensitivity.Result and Conclusion: These compounds already exhibited in vitro lipoxygenase inhibition and theMOE results strongly correlated with the experimental results. On the basis of these in vitro assaysand computer aided results, we suggest that these compounds need further detail in vivo studies andclinical trial for the discovery of new more effective and safe lipoxygenase inhibitors. In conclusion,these results might be useful in the design of new and potential lipoxygenase (LOX) inhibitors.

One of the main goals of in silico Caco-2 cell permeability models is to identify those drugsubstances with high intestinal absorption in human (HIA). For more than a decade, several in silicoCaco-2 models have been made, applying a wide range of modeling techniques; nevertheless, their capacityfor intestinal absorption extrapolation is still doubtful. There are three main problems related tothe modest capacity of obtained models, including the existence of inter- and/or intra-laboratory variabilityof recollected data, the influence of the metabolism mechanism, and the inconsistent in vitro-invivo correlation (IVIVC) of Caco-2 cell permeability. This review paper intends to sum up the recentadvances and limitations of current modeling approaches, and revealed some possible solutions to improvethe applicability of in silico Caco-2 permeability models for absorption property profiling, takinginto account the above-mentioned issues.

Background:Chronic Myeloid Leukaemia (CML) is associated with the BCRABL1gene, which plays a central role in the pathogenesis of CML. Thus, it is crucial tosuppress the expression of BCR-ABL1 in the treatment of CML. MicroRNA is known tobe a gene expression regulator and is thus a good candidate for molecularly targeted therapyfor CML.Objective: This study aims to identify the microRNAs from edible plants targeting the 3’Untranslated Region (3’UTR) of BCR-ABL1.Methods:In this in silico analysis, the sequence of 3’UTR of BCR-ABL1 was obtainedfrom Ensembl Genome Browser. PsRNATarget Analysis Server and MicroRNA TargetPrediction (miRTar) Server were used to identify miRNAs that have binding conformitywith 3’UTR of BCR-ABL1. The MiRBase database was used to validate the species ofplants expressing the miRNAs. The RNAfold web server and RNA COMPOSER wereused for secondary and tertiary structure prediction, respectively.Results:In silico analyses revealed that cpa-miR8154, csi-miR3952, gma-miR4414-5p,mdm-miR482c, osa-miR1858a and osa-miR1858b show binding conformity with strongmolecular interaction towards 3’UTR region of BCR-ABL1. However, only cpa-miR-8154, osa-miR-1858a and osa-miR-1858b showed good target site accessibility.Conclusion:It is predicted that these microRNAs post-transcriptionally inhibit the BCRABL1gene and thus could be a potential molecular targeted therapy for CML. However,further studies involving in vitro, in vivo and functional analyses need to be carried out todetermine the ability of these miRNAs to form the basis for targeted therapy for CML.

It has been previously demonstrated that synthetic antibody-derived peptides could exert a significant activity in vitro, ex vivo, and/or in vivo against microorganisms and viruses, as well as immunomodulatory effects through the activation of immune cells. Based on the sequence of previously described antibody-derived peptides with recognized antifungal activity, an in silico analysis was conducted to identify novel antifungal candidates. The present study analyzed the candidacidal and structural properties of in silico designed peptides (ISDPs) derived by amino acid substitutions of the parent peptide KKVTMTCSAS. ISDPs proved to be more active in vitro than the parent peptide and all proved to be therapeutic in Galleria mellonella candidal infection, without showing toxic effects on mammalian cells. ISDPs were studied by circular dichroism spectroscopy, demonstrating different structural organization. These results allowed to validate a consensus sequence for the parent peptide KKVTMTCSAS that may be useful in the development of novel antimicrobial molecules.

Present study was aimed to synthesize and characterize Chitosan-Catechol conjugates and to design and develop mucoadhesive pellets loaded with lafutidine. SEM images indicated the presence of fibrous structures responsible for enhanced mucoadhesive potential of Chitosan-Catechol conjugates. Thermodynamic stability and amorphous nature of conjugates was confirmed by DSC and XRD studies respectively. Rheological studies were used to evaluate polymer mucin interactions wherein strong interactions between Chitosan-Catechol conjugate and mucin was observed in comparison to pristine chitosan and mucin. The mucoadhesion potential of Chitosan-Catechol (Cht-C) versus Chitosan (Cht) was assessed in silico using molecular mechanics simulations and the results obtained were compared with the in vitro and ex vivo results. Cht-C/mucin demonstrated much higher energy stabilization (∆E ≈ −65 kcal/mol) as compared to Cht/mucin molecular complex. Lafutidine-loaded pellets were prepared from Chitosan (LPC) and Chitosan-Catechol conjugates (LPCC) and were evaluated for various physical properties viz. flow, circularity, roundness, friability, drug content, particle size and percent mucoadhesion. In vitro drug release studies on LPC and LPCC pellets were performed for computing t50%, t90% and mean dissolution time. The values of release exponent from Korsmeyer-Peppas model was reported to be 0.443 and 0.759 for LPC and LPCC pellets suggesting Fickian and non-Fickian mechanism representing drug release, respectively. In vivo results depicted significant controlled release and enhanced residence of the drug after being released from the chitosan-catechol coated pellets. Chitosan-Catechol conjugates were found to be a promising biooadhesive polymer for the development of various mucoadhesive formulations.