Secretory Patterns in Colleters of Apocynaceae

Colleters of Apocynaceae are glands related to different types of protection of vegetative and floral meristems through the production of mucilage or a mixture of many different compounds. Although several anatomical papers have shown histological and histochemical aspects of colleters of the family, almost nothing is known about their secretory process. In this study, we analyzed two types of colleters in Apocynaceae: one produces mucilage and lipophilic compounds, while the other produces an exclusively mucilaginous secretion. The secretory epidermis of the colleters of Allamanda schottii and Blepharodon bicuspidatum has a dense cytoplasm with organelles responsible for the production of mucilage and lipids. This heterogeneous secretion is released through granulocrine and eccrine mechanisms and is temporarily stored in a subcuticular space before crossing the cuticle. Conversely, colleters of Mandevilla splendens and Peplonia axillaris produce only mucilage and have a very different secretory apparatus. The mechanism of secretion is granulocrine, and the exudate is firstly accumulated in a large periplasmic space and later in an intramural space before crossing the cuticle. Notably, the structure of the cuticle varies according to the secretion composition. Although the colleters of the family are histologically similar, this study demonstrates a metabolic and subcellular variability previously unknown for Apocynaceae.

Phosphoproteome Profiling of Mouse Liver During Normal Aging

BackgroundAging is a complex biological process accompanied by a time-dependent functional decline that affects most living organisms. Omics studies help to comprehensively understand the mechanism of aging and discover potential intervention methods. Old mice were frequently obese with a fatty liver. MethodsWe applied mass spectrometry-based phosphoproteomics to obtain a global phosphorylation profile of liver in mice aged 2 or 18 months. A total of 5,685 phosphosites in 2,335 proteins were filtered for quantitative analysis. Phosphoproteome weakly separated young and old mice. ResultsCombining kinase prediction, kinase-substrate interaction analysis, and KEGG functional enrichment analysis, we observed high phosphorylation of fatty acid biosynthesis, b-oxidation, and potential secretory process, together with low phosphorylation of Egfr-Sos1-Araf/Braf-Map2k1-Mapk1 pathway and Ctnnb1 during aging. Proteins with differentially expressed phosphosites seemed more directly related to aging-associated fatty liver phenotype compared to the differentially expressed transcripts. Phosphoproteome may observe distinctive biological functions lost in transcriptome and proteome. ConclusionsIn summary, we constructed a phosphorylation-associated network in the liver of mice during normal aging, which may help to discover novel anti-aging strategies.

Ultrastructural Alterations in Cells of Sunflower Linear Glandular Trichomes during Maturation

Sunflower and related taxa are known to possess a characteristic type of multicellular uniseriate trichome which produces sesquiterpenes and flavonoids of yet unknown function for this plant. Contrary to the metabolic profile, the cytological development and ultrastructural rearrangements during the biosynthetic activity of the trichome have not been studied in detail so far. Light, fluorescence and transmission electron microscopy were employed to investigate the functional structure of different trichome cells and their subcellular compartmentation in the pre-secretory, secretory and post-secretory phase. It was shown that the trichome was composed of four cell types, forming the trichome basis with a basal and a stalk cell, a variable number (mostly from five to eight) of barrel-shaped glandular cells and the tip consisting of a dome-shaped apical cell. Metabolic activity started at the trichome tip sometimes accompanied by the formation of small subcuticular cavities at the apical cell. Subsequently, metabolic activity progressed downwards in the upper glandular cells. Cells involved in the secretory process showed disintegration of the subcellular compartments and lost vitality in parallel to deposition of fluorescent and brownish metabolites. The subcuticular cavities usually collapsed in the early secretory stage, whereas the colored depositions remained in cells of senescent hairs.

RESIDUAL PHENOMENA OF THE TRANSFERRED COVID-19 IN THE MAXILLOFACIAL REGION OR ITS COMPLICATIONS

Today, the issues of the incidence of the Covid-19 virus and its complications are extremely relevant. Much attention in the scientific literature is paid to possible complications such as sufferings of the lungs, heart. Along with this, it became known that the coronavirus can affect the brain, nasopharynx, eyes, blood vessels, liver, kidneys and intestines. Interesting data from clinicians and morphologists has been received. Those who have had Covid-19 note the long-term effects of scarring of the lung tissue and kidney failure, inflammation of the heart muscle, arrhythmias, liver damage, cognitive impairment, psychosis, accompanied by a sharp change in mood. Interesting observations from the Department of Systems Biology at George Mason University were seen: 70% of patients who have had Covid-19 rarely observe pathology of internal organs. The purpose of our work was to highlight the complications from the large salivary glands in the patients who suffered from Covid-19. Objects and research methods. Our observations were carried out at the Poltava Center for Salivary Gland Pathology. There were only 17 patients. Three or four weeks ago, they reliably suffered from the disease. The function of the parotid salivary glands was studied using metal catheters, which were introduced into the mouth of the gland ducts for 10 minutes. The cellular composition of the secretion of the parotid glands was investigated according to the method generally accepted in cytology. 8 patients underwent ultrasound examination of salivary glands. Results. The antiviral and symptomatic treatment given to the patients eliminated the leading symptoms of the disease. At the time of completion of treatment, general somatic symptoms were eliminated and they were discharged healthy. 3 patients complained of dryness in the mouth, especially at night and in the morning. 4 patients complained of dry mouth, burning sensation in the eye area. In 5 patients, along with dryness in the mouth and nose, there was a complete lack of sense food taste. 5 patients complained of disorders of appearance. The parotid-masticatory areas were enlarged, we could observe dryness of the mucous membranes of the mouth and eyes. All patients had short-term or long-term headaches. Before the viral disease, none of the patients and their relatives had diseases of the salivary glands. Physical examination in 8 patients did not reveal any facial asymmetry. Palpation revealed slightly enlarged parotid salivary glands, slightly compacted, and slightly painful. The submandibular salivary glands were of normal consistency and were painless. These patients had a moderate amount of oral fluid in the mouth. Saliva of normal viscosity was released from the ducts of the parotid and submandibular glands. With sialometry of the parotid glands, there was a slight drop in secretion to 1.5 + 0.3 ml. Cytological examination of the secretion of the parotid glands determined a small number of cells of the columnar epithelium. Some of these cells had a tendency to necrosis; single squamous epithelial cells were found in the preparation. Conclusions. We evaluated the results of clinical examination data of 17 patients who had a viral Covid-19 infection and they were examined by us. In 3 - 4 weeks after recovery, there were complaints: dryness of the mucous membranes of the mouth, nose, conjunctiva of the eyes, loss food taste, enlargement of the parotid and submandibular salivary glands. It is possible to reliably state that patients have clinical phenomena of sialopathy. The results of additional studies of the secretion of the parotid glands, data of the cytology of their secretion and ultrasound examination confirm the secretory process in the glands of various degrees. The presence of elements of an inflammatory nature in the secret clearly defines the sialectatic process. The presence of headaches in the examined patients determines their probable vascular disorders in the components of the brain. Such changes can indirectly affect the vascular complex of the salivary glands and be the cause of the development of duct contractures and leads to sialadenosis. Perspectives. In order to examine this category of patients, the study of the features of blood circulation in the vessels of the brain and salivary glands should be considered to clarify the pathogenesis of sialadenosis.

Assembly and Cellular Exit of Coronaviruses: Hijacking an Unconventional Secretory Pathway from the Pre-Golgi Intermediate Compartment via the Golgi Ribbon to the Extracellular Space

Coronaviruses (CoVs) assemble by budding into the lumen of the intermediate compartment (IC) at the endoplasmic reticulum (ER)-Golgi interface. However, why CoVs have chosen the IC as their intracellular site of assembly and how progeny viruses are delivered from this compartment to the extracellular space has remained unclear. Here we address these enigmatic late events of the CoV life cycle in light of recently described properties of the IC. Of particular interest are the emerging spatial and functional connections between IC elements and recycling endosomes (REs), defined by the GTPases Rab1 and Rab11, respectively. The establishment of IC-RE links at the cell periphery, around the centrosome and evidently also at the noncompact zones of the Golgi ribbon indicates that—besides traditional ER-Golgi communication—the IC also promotes a secretory process that bypasses the Golgi stacks, but involves its direct connection with the endocytic recycling system. The initial confinement of CoVs to the lumen of IC-derived large transport carriers and their preferential absence from Golgi stacks is consistent with the idea that they exit cells following such an unconventional route. In fact, CoVs may share this pathway with other intracellularly budding viruses, lipoproteins, procollagen, and/or protein aggregates experimentally introduced into the IC lumen.

Nectary and elaiophore work together in flowers of Xylopia aromatica (Annonaceae): structure indicates a role in pollination

Secretory structures that produce floral rewards have rarely been reported for Annonaceae. We identified a glandular region in Xylopia aromatica (Lam.) Mart., which consisted of a nectary and an elaiophore. This study aimed to describe the structure and secretory process of these glandular structures, which are highly correlated with the reproductive biology of this species. Anatomical and ultrastructural studies were performed prior to and during anthesis, focusing on the channel and pollination chamber. The floral nectary is placed in the roof of the chamber. It has a secretory epidermis and subglandular parenchyma and is immediately contiguous with the elaiophore, a portion that delimits the pollination channel and produces lipids. The release of nectar begins in the pistillate phase, while the elaiophore starts secreting prior to anthesis, both of which finishing during the staminate phase. Lipids form a sticky layer covering the channel surface, which provides access to the chamber. The cell machinery of the epidermis for both nectary and elaiophore is highly correlated with the exudates, despite their highly similar structure. Nectar attracts pollinators to the pollination chamber, whereas lipids seem to act in pollen adhesion to the body of pollinators. Both of exudates appear to act in complementary ways during pollination.

A Comparison Between the Effect of Vaginal Sildenafil Versus Oral Estradiol Valerate on Ultrasound Parameters of Endometrial Receptivity and Follicular Growth in Iraqi Females Receiving Letrozole for Induction of Ovulation

Endometrial thickness (ET) is one of the best predictors of implantation rate and pregnancy success rate because endometrial growth depends on hormones. Estrogen regulates the proliferative phase, endometrial proliferation induced by estrogen after menstruation is largely dependent on blood flow to the basal endometrium that produces progesterone receptors which are required for progesterone-regulated endometrial growth in the secretory process. To compare the effect of estradiol valerate vs. sildenafil citrate on endometrial receptivity and subsequent pregnancy outcome in unexplained infertility patients receiving letrozole for ovulation induction. Herein, 21 patients with unexplained infertility underwent ovulation induction by letrozole (2.5 mg). Patients were divided into 2 groups; group A (10 patients) were given sildenafil tablets vaginally (25 mg) four times daily and group B (11 patients) were treated with estradiol valerate tablet 2 mg/12 hours. Patients were evaluated by transvaginal ultrasonography (TVS) to determine endometrial thickness, pattern, size, and the number of the dominant follicles, serum assessment of Vascular Endothelial Growth Factor (VEGF) level at trigger day, and chemical pregnancy outcome 14 days later. Results showed that the endometrial thickness, patterns of the endometrium, VEGF, size of the dominant follicle at the trigger day, and pregnancy rates were not significantly different between the two groups. However, the difference in the number of the dominant follicles at trigger day was statistically significant which is more in the sildenafil treated group. The present study confirmed the findings of both Sildenafil and E2 valerate to boost endometrial receptivity and pregnancy rate.

ON THE MATTER OF MILK SECRETION INTENSITY BY COW’S MAMMARY GLAND

The secretion of milk in the intervals between milkings has its own patterns, and therefore researchers have different opinions about the intensity of the secretory process. Some scientists believe that the secretion of milk is uneven. Immediately after emptying the udder, it is higher, and with increasing interval, the intensity of the secretory process decreases, i.e. it is curved. This issue is still not clarified scientifically. The aim of our research was to study the change in the intensity of milk secretions during lactation under the condition of "voluntary", physiologically motivated emptying of the mammary gland and its frequency. The study of changes in the intensity of milk secretion during lactation under conditions of "voluntary" emptying of mammary gland in Holstein cows of different ages and levels of milk productivity was evaluated by using automatic milking control system based on electronic meters "Fullflow", which is mounted and functions in the milking unit VMS - 2012. Analyzing the nature of the secretory activity of the mammary gland of the first-born cow during lactation, we found that even under the condition of "voluntary" emptying, the intensity of the secretory process decreases. The average value of the intensity of milk secretion per hour during the first five months was quite close (1192.1 ± 33.51… 1105.6 ± 24.63) and only starting from the sixth month, it decreases initially by 17.3%, in comparison to the first month of lactation, and for the ninth month of lactation - by as much as 31.0%. In addition, there were significant fluctuations in the intensity of milk secretion for individual milkings during each month of lactation. Thus, the difference between the minimum and maximum values of the intensity of milk secretion by the mammary gland was the largest in the first month of lactation – 4.65 times, and the smallest - in the sixth month – 2.22 times. This indicates that the intensity of milk secretion even in the condition of "voluntary" emptying of the mammary gland is influenced by both internal and external factors, and above all – the conditions in which cows are fed, kept and exploitated. All this did not allow the animal to realize the functionality of the secretory epithelium of the mammary gland to the maximum possible secretory process for each day of lactation. Thus, under the relatively optimal parameters of these conditions, the average daily milk yield in the first month of lactation should be more than 65.0 kg of milk, with the actual – 25.7 kg, or 2.56 times higher. A similar pattern is observed during all months of lactation, where this difference ranges from 1.67 to 1.93 times. No less important, in our opinion, is the question of the time of emptying the mammary gland during the day. We found out that during lactation, mammary gland emptying occurs at all hours of the day without exception, although with varying frequency. Thus, more than 5.0 % of mammary gland emptying occurred: from 0 to 1 o’clock, from 7 to 8, from 10 to 12, from 17 to 18 and from 21 to to 22 o'clock. The rarest (1.74 %) mammary gland emptying occurred from 9 to 10 o'clock. Also, less than 3.0 % of emptying were observed from 4 to 5 and from 19 to 20 o’clock. On average per day, the emptying of the mammary gland in first-born cows occurred 3.5 times, and depending on the month of lactation ranged from 4.2 to 2.7 times. So the assessment of the intensity of the secretory process in the mammary gland can be a criterion for establishing genetic capabilities to the productivity of each animal and the time of milking during the day practically has no effect on the secretory process in the cow's mammary glands.

Multi-gene signature of microcalcification and risk prediction among Taiwanese breast cancer

Microcalcification is one of the most common radiological and pathological features of breast ductal carcinoma in situ (DCIS), and to a lesser extent, invasive ductal carcinoma. We evaluated messenger RNA (mRNA) transcriptional profiles associated with ectopic mammary mineralization. A total of 109 breast cancers were assayed with oligonucleotide microarrays. The associations of mRNA abundance with microcalcifications and relevant clinical features were evaluated. Microcalcifications were present in 86 (79%) patients by pathological examination, and 81 (94%) were with coexistent DCIS, while only 13 (57%) of 23 patients without microcalcification, the invasive diseases were accompanied with DCIS (χ2-test, P < 0.001). There were 69 genes with differential mRNA abundance between breast cancers with and without microcalcifications, and 11 were associated with high-grade (comedo) type DCIS. Enriched Gene Ontology categories included glycosaminoglycan and aminoglycan metabolic processes and protein ubiquitination, indicating an active secretory process. The intersection (18 genes) of microcalcificaion-associated and DCIS-associated genes provided the best predictive accuracy of 82% with Bayesian compound covariate predictor. Ten genes were further selected for prognostic index score construction, and five-year relapse free survival was 91% for low-risk and 83% for high-risk group (log-rank test, P = 0.10). Our study suggested that microcalcification is not only the earliest detectable radiological sign for mammography screening but the phenomenon itself may reflect the underling events during mammary carcinogenesis. Future studies to evaluate the prognostic significance of microcalcifications are warranted.

Altered ISGylation drives aberrant macrophage-dependent immune responses during SARS-CoV-2 infection

Interferon stimulated gene 15 (ISG15) is a ubiquitin like modifier frequently induced during virus infections and involved in versatile host defense mechanisms. Not surprisingly, many viruses including SARS-CoV-2 have evolved de-ISGylating activities to antagonize its effect. In this study we compared ISG15-driven macrophage responses upon infection by influenza, Zika and SARS-CoV-2 viruses. ISG15 and its modifying enzymes were upregulated in human macrophages after infection with all three viruses. While influenza and Zika viruses induced cellular ISGylation, SARS-CoV-2 triggered hydrolysis of ISG15 modifications instead, to generate free, extracellular ISG15 from macrophages and dendritic cells, but not from bronchial epithelial cells. Extracellular ISG15 was released independent of the conventional secretory pathway or cell death, but instead, depended on a non-classical autophagy-related secretory process. Increase of extracellular ISG15 was also reflected in serum samples from COVID-19 patients. The high ratio of free versus conjugated ISG15 in SARS-CoV-2 infected cells triggered macrophage polarization towards a M1 phenotype, increased secretion of pro-inflammatory cytokines, e.g. MCP-1 (CCL2), IL-1, TNF and IL-6, and attenuated antigen presentation. Depleting ISG15 conjugating enzymes Ube1L and HERC5 further increased free ISG15 and exacerbated this effect. We could recapitulate this phenomenon by expressing the wild-type but not the catalytically inactive PLpro de-ISGylating enzyme of SARS-CoV-2. Proteomic analyses of the secretome from SARS-CoV-2 infected macrophages revealed that besides ISG15, it displayed significant enrichment in non-classical secretory proteins and inflammatory responses, which was further amplified by free ISG15. Collectively, our results indicate that increased proportions of free ISG15 dramatically alter macrophage responses and is likely a key feature of cytokine storms triggered by highly pathogenic respiratory viruses such as influenza and SARS-CoV-2.