Effective reduction of primary dysmenorrheal symptoms through concurrent use of n-3 fatty acids and Rosa damascena extract (RDE)

Background: Primary dysmenorrhea (PD) is characterized by painful cramps of lower abdomen without abnormal pelvic pathology. PD begins some hours before or simultaneously with the onset of menstrual bleeding. Objectives: The purpose of this study was to investigate the effects of separate and concurrent supplementation of fish oils (FO) containing n-3 fatty acids and Rosa Damascena extract (RDE) on PD symptoms. Methods: In this double blind clinical trial in 2015, through convenience sampling, 105 university students with primary menstrual pain in most recent years, without abnormal pathology, and with moderate/severe dysmenorrheal symptoms according to visual analogue scale (VAS) were randomly assigned into one of four groups: 1. FO 1000 mg/day (n=26), 2. RDE 1000 mg/day (n=27), 3. FO and RDE concurrently, with the same dose (n=27), and 4. Control group (n=25). All measurements were performed three times, at the beginning, 30th day, and 60th day. Symptoms including nausea, vomiting, diarrhea, bloating, cramp, low back pain, headache, fatigue, anxiety, sweat, weakness, dizziness, drowsiness, and feeling cold were measured by VAS. Results: After 2-month treatment, supplementation with RDE significantly reduced severity of bloating (p<0.001) and sweat (p<0.001), but FO supplementation had no significant effect on PD symptoms. The concurrent use of FO and RDE significantly decreased severity of diarrhea (p=0.038), weakness (p<0.001), dizziness (p=0.003), and feeling cold (p=0.049). Conclusion: Our results suggest that the concurrent supplementation of omega-3 fatty acids and RDE could be more effective than their separate use in decreasing PD symptoms; however, larger trials are warranted to confirm these preliminary findings

Myelination generates aberrant ultrastructure that is resolved by microglia

To enable rapid propagation of action potentials, axons are ensheathed by myelin, a multilayered insulating membrane formed by oligodendrocytes. Most of the myelin is generated early in development, resulting in the generation of long-lasting stable membrane structures. Here, we explored structural and dynamic changes in central nervous system myelin during development. To achieve this, we performed an ultrastructural analysis of mouse optic nerves by serial block face scanning electron microscopy (SBF-SEM) and confocal time-lapse imaging in the zebrafish spinal cord. We found that myelin undergoes extensive ultrastructural changes during early postnatal development. Myelin degeneration profiles were engulfed and phagocytosed by microglia using exposed phosphatidylserine as one “eat me” signal. In contrast, retractions of entire myelin sheaths occurred independently of microglia and involved uptake of myelin by the oligodendrocyte itself. Our findings show that the generation of myelin early in development is an inaccurate process associated with aberrant ultrastructural features that require substantial refinement.</p

The anticancer activity of lytic peptides is inhibited by heparan sulfate on the surface of the tumor cells

<p>Abstract</p> <p>Background</p> <p>Cationic antimicrobial peptides (CAPs) with antitumor activity constitute a promising group of novel anticancer agents. These peptides induce lysis of cancer cells through interactions with the plasma membrane. It is not known which cancer cell membrane components influence their susceptibility to CAPs. We have previously shown that CAPs interact with the two glycosaminoglycans (GAGs), heparan sulfate (HS) and chondroitin sulfate (CS), which are present on the surface of most cells. The purpose of this study was to investigate the role of the two GAGs in the cytotoxic activity of CAPs.</p> <p>Methods</p> <p>Various cell lines, expressing different levels of cell surface GAGs, were exposed to bovine lactoferricin (LfcinB) and the designer peptide, KW5. The cytotoxic effect of the peptides was investigated by use of the colorimetric MTT viability assay. The cytotoxic effect on wild type CHO cells, expressing normal amounts of GAGs on the cell surface, and the mutant pgsA-745, that has no expression of GAGs on the cell surface, was also investigated.</p> <p>Results</p> <p>We show that cells not expressing HS were more susceptible to CAPs than cells expressing HS at the cell surface. Further, exogenously added heparin inhibited the cytotoxic effect of the peptides. Chondroitin sulfate had no effect on the cytotoxic activity of KW5 and only minor effects on LfcinB cytotoxicity.</p> <p>Conclusion</p> <p>Our results show for the first time that negatively charged molecules at the surface of cancer cells inhibit the cytotoxic activity of CAPs. Our results indicate that HS at the surface of cancer cells sequesters CAPs away from the phospholipid bilayer and thereby impede their ability to induce cytolysis.</p

Regeneration of myelin sheaths of normal length and thickness in the zebrafish CNS correlates with growth of axons in caliber

Demyelination is observed in numerous diseases of the central nervous system, including multiple sclerosis (MS). However, the endogenous regenerative process of remyelination can replace myelin lost in disease, and in various animal models. Unfortunately, the process of remyelination often fails, particularly with ageing. Even when remyelination occurs, it is characterised by the regeneration of myelin sheaths that are abnormally thin and short. This imperfect remyelination is likely to have implications for the restoration of normal circuit function and possibly the optimal metabolic support of axons. Here we describe a larval zebrafish model of demyelination and remyelination. We employ a drug-inducible cell ablation system with which we can consistently ablate 2/3rds of oligodendrocytes in the larval zebrafish spinal cord. This leads to a concomitant demyelination of 2/3rds of axons in the spinal cord, and an innate immune response over the same time period. We find restoration of the normal number of oligodendrocytes and robust remyelination approximately two weeks after induction of cell ablation, whereby myelinated axon number is restored to control levels. Remarkably, we find that myelin sheaths of normal length and thickness are regenerated during this time. Interestingly, we find that axons grow significantly in caliber during this period of remyelination. This suggests the possibility that the active growth of axons may stimulate the regeneration of myelin sheaths of normal dimensions

Glycosaminoglycans and Sialylated Glycans Sequentially Facilitate Merkel Cell Polyomavirus Infectious Entry

Merkel cell polyomavirus (MCV or MCPyV) appears to be a causal factor in the development of Merkel cell carcinoma, a rare but highly lethal form of skin cancer. Although recent reports indicate that MCV virions are commonly shed from apparently healthy human skin, the precise cellular tropism of the virus in healthy subjects remains unclear. To begin to explore this question, we set out to identify the cellular receptors or co-receptors required for the infectious entry of MCV. Although several previously studied polyomavirus species have been shown to bind to cell surface sialic acid residues associated with glycolipids or glycoproteins, we found that sialylated glycans are not required for initial attachment of MCV virions to cultured human cell lines. Instead, glycosaminoglycans (GAGs), such as heparan sulfate (HS) and chondroitin sulfate (CS), serve as initial attachment receptors during the MCV infectious entry process. Using cell lines deficient in GAG biosynthesis, we found that N-sulfated and/or 6-O-sulfated forms of HS mediate infectious entry of MCV reporter vectors, while CS appears to be dispensable. Intriguingly, although cell lines deficient in sialylated glycans readily bind MCV capsids, the cells are highly resistant to MCV reporter vector-mediated gene transduction. This suggests that sialylated glycans play a post-attachment role in the infectious entry process. Results observed using MCV reporter vectors were confirmed using a novel system for infectious propagation of native MCV virions. Taken together, the findings suggest a model in which MCV infectious entry occurs via initial cell binding mediated primarily by HS, followed by secondary interactions with a sialylated entry co-factor. The study should facilitate the development of inhibitors of MCV infection and help shed light on the infectious entry pathways and cellular tropism of the virus

Requirements for Receptor Engagement during Infection by Adenovirus Complexed with Blood Coagulation Factor X

Human adenoviruses from multiple species bind to coagulation factor X (FX), yet the importance of this interaction in adenovirus dissemination is unknown. Upon contact with blood, vectors based on adenovirus serotype 5 (Ad5) binds to FX via the hexon protein with nanomolar affinity, leading to selective uptake of the complex into the liver and spleen. The Ad5:FX complex putatively targets heparan sulfate proteoglycans (HSPGs). The aim of this study was to elucidate the specific requirements for Ad5:FX-mediated cellular uptake in this high-affinity pathway, specifically the HSPG receptor requirements as well as the role of penton base-mediated integrin engagement in subsequent internalisation. Removal of HS sidechains by enzymatic digestion or competition with highly-sulfated heparins/heparan sulfates significantly decreased FX-mediated Ad5 cell binding in vitro and ex vivo. Removal of N-linked and, in particular, O-linked sulfate groups significantly attenuated the inhibitory capabilities of heparin, while the chemical inhibition of endogenous HSPG sulfation dose-dependently reduced FX-mediated Ad5 cellular uptake. Unlike native heparin, modified heparins lacking O- or N-linked sulfate groups were unable to inhibit Ad5 accumulation in the liver 1h after intravascular administration of adenovirus. Similar results were observed in vitro using Ad5 vectors possessing mutations ablating CAR- and/or αv integrin binding, demonstrating that attachment of the Ad5:FX complex to the cell surface involves HSPG sulfation. Interestingly, Ad5 vectors ablated for αv integrin binding showed markedly delayed cell entry, highlighting the need for an efficient post-attachment internalisation signal for optimal Ad5 uptake and transport following surface binding mediated through FX. This study therefore integrates the established model of αv integrin-dependent adenoviral infection with the high-affinity FX-mediated pathway. This has important implications for mechanisms that define organ targeting following contact of human adenoviruses with blood

Molecular and Parasitological Study of Cryptosporidium Isolates From Cattle in Ilam, West of Iran

Background: Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP. Methods: Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN). Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes. Results: The prevalence of Cryptosporidium infection in cattle using both methods is 3.68. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum. Conclusion: C. parvum (bovine genotype) detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection

Biotransformacja mircenu z udziałem bakterii Pseudomonas putida PTCC 1694

Terpineol and linalool are sources of fragrances providing an unique volatile terpenoid alcohol of low toxicity, and thus are widely used in the perfumery industry. They are also being applied in folk medicine and in aromatherapy, as well as important chemical constituents of the essential oil of many plants. Previous studies have implicated the biotransformation of limonene by Pseudomonas putida. The objective of this research was to study the pathways involved in biotransformation of myrcene by Pseudomonas putida. The culture preparation was done by using such variables as different microbial methods and incubation periods to obtain maximum cells of P. putida for myrcene biotransformation. It was found that myrcene was converted to dihydrolinalool, cis-β-dihydroterpineol, linalool and cis-ocimene-8-oxo in high percentages. The biotransformation products were identified by theoretical study (TS), Fourier-transform infrared spectroscopy (FT-IR), ultraviolet visible (UV), gas chromatography (GC), nuclear magnetic resonance (NMR) and gas chromatography/ mass spectroscopy (GC-MS). Comparison of different incubation times showed that 120 h which was more effective, the major products were dihydrolinalool (4.1%), cis- β-dihydroterpineol (67.6%) and linalool (25.8%). The main compounds comprised 97.5%. The incubation period of 72 h yielded dihydrolinalool (16.7%), cis-ocimene-8-oxo (61.6%), Biotransformation of Myrcene by Pseudomonas putida PTCC 1694 trans-β-dihydroterpineol (8.4%) and β-cadinene (3.5%), with main compounds comprising 86.7%. Incubation for 30 h yielded dihydrolinalool (59.5%), cis-β-dihydroterpineol (25.0%), hexadecanoic acid (12.5%), and the main compounds comprising 97.0%.Terpineol i linalol są związkami zapachowymi należącymi do alkoholi terpenowych. Charakteryzują się wyjątkowo niską toksycznością, dlatego często są wykorzystywane w przemyśle perfumeryjnym, medycynie ludowej oraz aromaterapii. Są głównymi składnikami olejków eterycznych wielu roślin. Poprzednio prowadzone badania dotyczyły biotransformacji limonenu prowadzonej przez Pseudomonas putida. Celem niniejszej pracy było zbadanie możliwości biotransformacji mircenu prowadzonej przez bakterie Pseudomonas putida. W hodowli kultur wykorzystano różne metody mikrobiologiczne oraz okresy inkubacji w celu uzyskania maksymalnego stężenia komórek bakterii Pseudomonas putida zdolnych do biotransformacji mircenu. W trakcie badań stwierdzono wysokie stężenie takich metabolitów biotransformacji mircenu jak: dihydrolinalol, cis-β-dihydroterpineol, linalol oraz 82 A. Esmaeili, E. Hashemi, S. Safaiyan, A. Rustaiyan cis-ocymen-8-okso. Produkty metabolizmu mircenu zidentyfikowano, wykorzystując dane wynikające z rozważań teoretycznych (TS) oraz wykorzystując techniki spektroskopii w podczerwieni (FTIR), promieniowania ultrafioletowego (UV), chromatografii gazowej (GC), spektroskopii magnetycznego rezonansu jądrowego (NMR) i spektroskopii mas sprzężonej z chromatografią gazową (GC-MS). Na podstawie porównania różnych czasów inkubacji stwierdzono, że 120-godzinna inkubacja jest najbardziej wydajna. Dzięki niej otrzymano dihydrolinalol (16,7%), cis-β-dihydroterpineol (67.6%), linalol (25,8%), które stanowiły 97,5% całego substratu. W czasie 72-godzinnej inkubacji otrzymano 86,7% wszystkich związków: dihydrolinalol (16,7%), cis-ocymen-8-okso (61,6%), trans-β-dihydroterpineol (8,4%) oraz β-kadinen (3,5%). W wyniku inkubacji 30h otrzymano 97% wszystkich związków powstałych w procesie biotransformacji: dihydrolinalol (59,5%), cis-β-dihydroterpineol (25,0%) oraz kwas heksadekanowy (12,5%)

Surprises from Intravital Imaging of the Innate Immune Response

Successful immune responses depend on the spatiotemporal coordination of immune cell migration, interactions, and effector functions in lymphoid and parenchymal tissues. Real-time intravital microscopy has revolutionized our understanding of the dynamic behavior of many immune cell types in the living tissues of several species. Observing immune cells in their native environment has revealed many unanticipated facets of their biology, which were not expected from experiments outside a living organism. Here we highlight both classic and more recent examples of surprising discoveries that critically relied on the use of live in vivo imaging. In particular, we focus on five major cell types of the innate immune response (macrophages, microglia, neutrophils, dendritic cells, and mast cells), and how studying their dynamics in mouse tissues has helped us advance our current knowledge of immune cell–mediated tissue homeostasis, host defense, and inflammation

Age-related myelin degradation burdens the clearance function of microglia during aging

Myelin is synthesized as a multilamellar membrane, but the mechanisms of membrane turnover are unknown. We found that myelin pieces were gradually released from aging myelin sheaths and were subsequently cleared by microglia. Myelin fragmentation increased with age and led to the formation of insoluble, lipofuscin-like lysosomal inclusions in microglia. Thus, age-related myelin fragmentation is substantial, leading to lysosomal storage and contributing to microglial senescence and immune dysfunction in aging