Miracle-dz/UniFAR-FTUs huggingface dataset:
Proteome Analysis of Legionella Vacuoles Purified by Magnetic Immunoseparation Reveals Secretory and Endosomal GTPases

Legionella pneumophila, the causative agent of Legionnaires’ disease, replicates in macrophages and amoebae within ‘Legionella‐containing vacuoles’ (LCVs), which communicate with the early secretory pathway and the endoplasmic reticulum. Formation of LCVs requires the bacterial Icm/Dot type IV secretion system. The Icm/Dot‐translocated effector protein SidC selectively anchors to LCVs by binding the host lipid phosphatidylinositol‐4‐phosphate (PtdIns(4)P). Here, we describe a novel and simple approach to purify intact vacuoles formed by L. pneumophila within Dictyostelium discoideum by using magnetic immunoseparation with an antibody against SidC, followed by density gradient centrifugation. To monitor LCV purification by fluorescence microscopy, we used Dictyostelium producing the LCV marker calnexin‐GFP and L. pneumophila labeled with the red fluorescent protein DsRed. A proteome analysis of purified LCVs by liquid chromatography coupled to tandem mass spectrometry revealed 566 host proteins, including known LCV components, such as the small GTPases Arf1, Rab1 and Rab7. Rab8, an endosomal regulator of the late secretory pathway originating from the trans Golgi network, and the endosomal GTPase Rab14 were identified as novel LCV components, which were found to be present on vacuoles harboring wild‐type but not Icm/Dot‐deficient L. pneumophila. Thus, LCVs also communicate with the late secretory and endosomal pathways. Depletion of Rab8 or Arf1 by RNA interference reduced the amount of SidC on LCVs, indicating that the GTPases promote the recruitment of Legionella effectors by regulating the level of PtdIns(4)P.
Alnuheptanoid A: a new diarylheptanoid derivative from Alnus japonica

Extensive chromatographic investigation of the ethanolic extract of Alnus japonica Steud stem bark led to the isolation of a new diarylheptanoid named alnuheptanoid A [(5S)-7-(3,4-dihydroxyphenyl)-1-(4-hydroxyphenyl)-5-methoxyheptan-3-one] (8), together with seven known diarylheptanoid derivatives: platyphyllenone (5), (5S)-1,7-bis(4-hydroxyphenyl)-5-methoxyheptan-3-one (6), 1-(3,4-dihydroxyphenyl)-7-(4-hydroxyphenyl)-4-hepten-3-one (7), hirsutenone (9), (5R)-O-methylhirsutanonol (10), hirsutanonol (11) and oregonin (13), three triterpenes: α-amyrin (1), betulinaldehyde (3) and betulinic acid (4), and two sterols: β-sitosterol (2) and daucosterol (12). Compound 6 was isolated for the first time from natural source. The structures of the isolated compounds were determined on the basis of spectroscopic measurements (UV, IR, HR-ESI-MS, 1D and 2D NMR).
Young Children with Down's Syndrome

subject. For this reaison a more critical appraisal would have been helpful, and perhaps, the sections discussing the ininunodeficiency diseases, for example, could have been combined. Other chapteirs include "Complement in Immnunohaematology" by L. Petz and G. Garratty, "Autoimnunity and Lympphoid Malignancy in New Zealand Black Mice" by N. Talal, "Therapeutic Uses of Transfer Factor" by W. H. Hitzig and P. J. Grob, and "Carcinofetal Antigens" by M. Costanza and L. Nathanson. These should prove interesting to immunologists concerned with patients with cancer, thougth the account of transfer factor in the treatment of malignant diseases is somewhat less criitical than it might have been. The section on carcinofetal antigens is a useful review, appraising several controversial issues on the possible clinical applications of the detection of these antigens. The book should prove most valuable to clinical immunologists, clinicians with an interest in immunology or cancer medicine, and readers doing research in immunology who want to improve their knowledge in subjects which are not their direct concern.
Microtubule Binding and Trapping at the Tip of Neurites Regulate Tau Motion in Living Neurons

During the development of neurons, the microtubule‐associated tau proteins show a graded proximo‐distal distribution in axons. In tauopathies such as Alzheimer's disease, tau accumulates in the somatodendritic compartment. To scrutinize the determinants of tau's distribution and motion, we constructed photoactivatable green fluorescent protein (GFP)‐tagged tau fusion proteins and recorded their distribution after focal activation in living cells. Simulation showed that the motion of tau was compatible with diffusion/reaction as opposed to active transport/reaction. Effective diffusion constants of 0.7–0.8 μm2/second were calculated in neurites of PC12 cells and primary cortical neurons. Furthermore, tau's amino terminal projection domain mediated binding and enrichment of tau at distal neurites indicating that the tip of a neurite acts as an adsorber trapping tau protein. Treatment with taxol, incorporation of disease‐related tau modifications, experimentally induced hyperphosphorylation and addition of preaggregated amyloid β peptides (Aβ) increased the effective diffusion constant compatible with a decreased binding to microtubules. Distal enrichment was present after taxol treatment but was suppressed at disease‐relevant conditions. The data suggest that (i) dynamic binding of tau to microtubules and diffusion along microtubules and (ii) trapping at the tip of a neurite both contribute to its distribution during development and disease.
Rab10 Regulates Phagosome Maturation and Its Overexpression Rescues Mycobacterium‐Containing Phagosomes Maturation

Phagosome maturation follows a defined biochemical program and, in the vast majority of cases, the microbe inside the phagosome is killed and digested. Although, an important number of pathogens, including Mycobacterium tuberculosis, which kills around two million people every year, have acquired the ability to survive, and even replicate by arresting phagosomal maturation. To identify more of the machinery involved in phagocytosis and phagosomal maturation, we investigated the function of Rab10 in engulfment and maturation of inert particles and Mycobacterium bovis bacille Calmette‐Guérin (BCG). We showed that Rab10 association with phagosomes is transient and confocal microscopy revealed detectible levels of Rab10 on phagosomal membranes at very early time‐points, occurring even before Rab5 acquisition. Rab10 recruitment had strong functional consequence, as the knockdown of endogenous Rab10 by RNA interference or overexpression of Rab10 dominant‐negative mutant delayed maturation of phagosomes of IgG‐opsonized latex beads or heat killed‐mycobacteria. These results can be explained, at least in part, by the involvement of Rab10 in recycling of some phagosomal components. More importantly, overexpression of the constitutively active mutant of Rab10 partially rescued live‐Mycobacterium‐containing phagosomes maturation. Indeed, we found that the membrane harbouring Mycobacterium acquired early endosome antigen 1 (EEA‐1), a marker excluded from phagosomes in control cells. Altogether these results indicate that Rab10, acting upstream of Rab5, plays a prominent role in phagolysosome formation and can modulate Mycobacterium‐containing phagosomes maturation.
Rab27a and Rab27b Regulate Neutrophil Azurophilic Granule Exocytosis and NADPH oxidase Activity by Independent Mechanisms

Neutrophils rely on exocytosis to mobilize receptors and adhesion molecules and to release microbicidal factors. This process should be strictly regulated because uncontrolled release of toxic proteins would be injurious to the host. In vivo studies showed that the small GTPase Rab27a regulates azurophilic granule exocytosis. Using mouse neutrophils deficient in Rab27a (Rab27aash/ash), Rab27b [Rab27b knockout (KO)] or both [Rab27a/b double KO (DoKo)], we investigated the role of the Rab27 isoforms in neutrophils. We found that both Rab27a and Rab27b deficiencies impaired azurophilic granule exocytosis. Rab27aash/ash neutrophils showed upregulation of Rab27b expression which did not compensate for the secretory defects observed in Rab27a‐deficient cells, suggesting that Rab27 isoforms play independent roles in neutrophil exocytosis. Total internal reflection fluorescence microscopy analysis showed that Rab27aash/ash and Rab27b KO neutrophils have a decreased number of azurophilic granules near the plasma membrane. The effect was exacerbated in Rab27a/b DoKo neutrophils. Rab27‐deficient neutrophils showed impaired activation of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase at the plasma membrane although intraphagosomal reactive oxygen species (ROS) production was not affected. Exocytosis of secretory vesicles in Rab27‐deficient neutrophils was functional, suggesting that Rab27 GTPases selectively control the exocytosis of neutrophil granules.
Natural product extracts of the Canadian prairie plant, Thermopsis rhombifolia, have anti-cancer activity in phenotypic cell-based assays

Many plant species within the terrestrial ecological zones of Canada have not yet been investigated for anti-cancer activity. We examined the scientific literature describing the endemic flora from the prairie ecological zone and selected the species, Thermopsis rhombifolia, locally known as the buffalo bean, for investigation of its anti-cancer potential. We tested it in cell-based assays using phenotypic screens that feature some of the hallmarks of cancer. An ethanolic extract prepared from T. rhombifolia was cytotoxic to HT-29 (colon) and SH-SY5Y (brain) cancer cell lines, and showed little cytotoxicity to a normal human cell line (WI-38). In phenotypic assays, we identified activities in the extracts that target cell death, cell cycle and cell adhesion. These data highlight the anti-cancer potential of previously untested plants found in northern ecological zones and the feasibility of using pertinent phenotypic assays to examine the anti-cancer potential of natural product extracts.
Chemotyping of diverse Eucalyptus species grown in Egypt and antioxidant and antibacterial activities of its respective essential oils

The chemical composition of the essential oil from the leaves of Eucalyptus camaldulensis, Eucalyptus camaldulensis var. obtusa and Eucalyptus gomphocephala grown in northern Egypt was analysed by using GC-FID and GC–MS techniques. The antibacterial (agar disc diffusion and minimum inhibitory concentration methods) and antioxidant activities (2,2′-diphenypicrylhydrazyl) were examined. The main oils constituents were 1,8-cineole (21.75%), β-pinene (20.51%) and methyleugenol (6.10%) in E. camaldulensis; spathulenol (37.46%), p-cymene (17.20%) and crypton (8.88%) in E. gomphocephala; spathulenol (18.37%), p-cymene (19.38%) and crypton (16.91%) in E. camaldulensis var. obtusa. The essential oils from the leaves of Eucalyptus spp. exhibited considerable antibacterial activity against Gram-positive and Gram-negative bacteria. The values of total antioxidant activity were 70 ± 3.13%, 50 ± 3.34% and 84 ± 4.64% for E. camaldulensis, E. camaldulensis var. obtusa and E. gomphocephala, respectively. The highest antioxidant activity value of 84 ± 4.64% could be attributed to the high amount of spathulenol (37.46%).
Lipid Cosorting Mediated by Shiga Toxin Induced Tubulation

To maintain cell membrane homeostasis, lipids must be dynamically redistributed during the formation of transport intermediates, but the mechanisms driving lipid sorting are not yet fully understood. Lowering sphingolipid concentration can reduce the bending energy of a membrane, and this effect could account for sphingolipid depletion along the retrograde pathway. However, sphingolipids and cholesterol are enriched along the anterograde pathway, implying that other lipid sorting mechanisms, such as protein‐mediated sorting, can dominate. To characterize the influence of protein binding on the lipid composition of highly curved membranes, we studied the interactions of the B‐subunit of Shiga toxin (STxB) with giant unilamellar vesicles containing its glycosphingolipid receptor [globotriaosylceramide (Gb3)]. STxB binding induced the formation of tubular membrane invaginations, and fluorescence microscopy images of these highly curved membranes were consistent with co‐enrichment of Gb3 and sphingolipids. In agreement with theory, sorting was stronger for membrane compositions close to demixing. These results strongly support the hypothesis that proteins can indirectly mediate the sorting of lipids into highly curved transport intermediates via interactions between lipids and the membrane receptor of the protein.
Association of homocysteine level and vascular burden and cognitive function in middle‐aged and older adults with chronic kidney disease

Patients with chronic kidney disease (CKD) have been found to have cognitive impairment. However, the core features and clinical correlates of cognitive impairment are still unclear. Elevated homocysteine levels are present in CKD, and this is a risk factor for cognitive impairment and vascular diseases in the general population. Thus, this study investigated the core domains of cognitive impairment and investigated the associations of homocysteine level and vascular burden with cognitive function in patients with CKD.
Apoptogenic effects of β-sitosterol glucoside from Castanopsis indica leaves

β-Sitosterol glucoside (BSSG) is a natural biologically active substance isolated from the Castanopsis indica leaves. This study explored the apoptogenic mechanistic studies of BSSG against Ehrlich's ascites carcinoma (EAC) treated mice through morphological study, comet assay, flow cytometry (FACS) and Western blotting assay method. AO/EB staining and FACS analysis showed that BSSG possessed apoptosis induction activities on EAC cells. Dose dependent induction of DNA damage was observed after BSSG treatment. Increase the expression of apoptotic protein p53 and p21 in EAC, multiple downstream factors contributing to apoptosis pathway. The increase of caspase-9 and caspase-3 activities revealed that caspase was a key mediator of the apoptotic pathway induced by BSSG, and up-regulation of Bax and down-regulation of anti-apoptotic protein Bcl-2 resulted in the decrease of Bcl-2/Bax ratio. Owing to the combination of significant antitumour activity by inducing apoptosis, BSSG holds the promise of being an interesting chemo-preventive agent active in cancer therapy.
A Comparison of the Effects of 50 MeVd→Be Neutron and Cobalt-60 Irradiation of the Kidneys of Rhesus Monkeys

Twenty rhesus monkeys had one kidney irradiated (after undergoing unilateral nephrectomies) with one of four doses: 960 or 1080 rads of 50 MeVd→Be neutrons, or 2350 or 2700 rads of 60Co. Whereas animals treated with the lower dose of neutrons or 60Co are alive with relatively normal renal function, those treated with the higher dose of neutrons died of radiation nephritis. Animals treated with the higher dose of 60Co developed radiation nephritis but survived. The physiological and histopathological changes of radiation nephritis secondary to neutron irradiation are not qualitatively different from those reported for radiation nephritis secondary to photon irradiation.
In Vivo Roles of the Basic Domain of Dynactin p150 in Microtubule Plus‐End Tracking and Dynein Function

Microtubule (MT) plus‐end‐tracking proteins accumulate at MT plus ends for various cellular functions, but their targeting mechanisms are not fully understood (Akhmanova A and Steinmetz MO. Tracking the ends: a dynamic protein network controls the fate of microtubule tips. Nat Rev Mol Cell Biol 2008;9:309–322.). Here, we tested in the filamentous fungus Aspergillus nidulans the requirement for plus‐end localization of dynactin p150, a protein essential for dynein function. Deletion of the N‐terminal MT‐binding region of p150 significantly diminishes the MT plus‐end accumulation of both dynein heavy chain and p150, and causes a partial defect in nuclear distribution. Surprisingly, within the MT‐binding region, the basic domain is more critical than the CAP‐Gly (cytoskeleton‐associated protein glycine‐rich) domain for maintaining plus‐end tracking of p150, as well as for the functions of dynein in nuclear distribution and early endosome movement. Our results show that the basic domain of A. nidulans p150 is important for p150–MT interaction both in vivo and in vitro, and the basic amino acids within this domain are crucial for the plus‐end accumulation of p150 in the wild‐type background and for the p150–MT interaction in the ΔkinA (kinesin‐1) background. We suggest that the basic amino acids are required for the electrostatic interaction between p150 and MTs, which is important for kinesin‐1‐mediated plus‐end targeting of dynactin and dynein in A. nidulans.
Serum markers for the early detection of hepatocellular carcinoma in patients with chronic viral hepatitis C infection

Objective The aim of this study was to identify the serum markers and the use of abdominal ultrasound for early detection of hepatocellular carcinoma (HCC) in patients with chronic viral hepatitis C virus infection. Background HCC meets the criteria of a tumor that would benefit from a surveillance program, but the poor sensitivity and specificity of currently available tools have prevented widespread implementation of surveillance. Patients and methods This study included 110 patients, age from 23 to 70 years, from Menoufia University hospitals during the period from July 2011 to November 2013. They were classified into three groups: group I, non-HCC group (50 patients); group II, HCC group (40 patients with chronic hepatitis C virus infection); and group III, healthy controls (20 individuals). Members of the study were subjected to thorough history taking, complete physical examination, liver function testing (serum bilirubin, albumin, prothrombin time, serum transaminases), serum α-fetoprotein (α-FP), and transforming growth factor β1 (TGF-β1) level. Group I was subjected to serum TGF-β1 at 0-, 9-, and 18-month intervals. Results The mean age was 46.72 ± 9.03 years in the non-HCC group (group I), 58.70 ± 5.76 years in the HCC group (group II), and 42.15 ± 11.33 years in the control group (group III). The mean serum level of TGF-β1 was 232.25 ± 70.53 ng/ml in the HCC group, 42.16 ± 13.34 ng/ml in the non-HCC group, and 13.92 ± 7.73 ng/ml in the control group; there was a highly significant difference between all groups ( P Conclusion This study recommends TGF-β1 as being more accurate than α-FP in differentiating patients with HCC from those with nonmalignant chronic liver disease.
Botulinum Neurotoxin A Impairs Neurotransmission Following Retrograde Transynaptic Transport

The widely used botulinum neurotoxin A (BoNT/A) blocks neurotransmission via cleavage of the synaptic protein SNAP‐25 (synaptosomal‐associated protein of 25 kDa). Recent evidence demonstrating long‐distance propagation of SNAP‐25 proteolysis has challenged the idea that BoNT/A remains localized to the injection site. However, the extent to which distant neuronal networks are impacted by BoNT/A retrograde trafficking remains unknown. Importantly, no studies have addressed whether SNAP‐25 cleavage translates into structural and functional changes in distant intoxicated synapses. Here we show that the BoNT/A injections into the adult rat optic tectum result in SNAP‐25 cleavage in retinal neurons two synapses away from the injection site, such as rod bipolar cells and photoreceptors. Retinal endings displaying cleaved SNAP‐25 were enlarged and contained an abnormally high number of synaptic vesicles, indicating impaired exocytosis. Tectal injection of BoNT/A in rat pups resulted in appearance of truncated‐SNAP‐25 in cholinergic amacrine cells. Functional imaging with calcium indicators showed a clear reduction in cholinergic‐driven wave activity, demonstrating impairments in neurotransmission. These data provide the first evidence for functional effects of the retrograde trafficking of BoNT/A, and open the possibility of using BoNT/A fragments as drug delivery vehicles targeting the central nervous system.
Alzheimer's disease and Toxoplasma gondii infection; seromolecular assess the possible link among patients

We investigated the possible association between Parkinson's disease (PD), the second most common neurode-generative disorder and Toxoplasma gondii infection, the most common neurotropic protozoan parasitic infection, using serological and molecular techniques. One hundred and fifteen patients with confirmed PD and 115 healthy subjects in the same age and sex distribution were enrolled in this study. Blood samples were taken from each participant and the sera was screened for anti-Toxoplasma antibodies (IgG and IgM). PCR assay was performed in duplicate using the primer pair targeting the B1 gene of Toxoplasma. Amplicons were directly sequenced to conduct the phylogenetic analysis. The prevalence of Toxoplasma infection based on IgG titer was 53% in case and 55.6% in the control groups, revealing no statistically significant association between Toxoplasma seropositivity and PD (OR = 0.90; 95% CI = 0.54-1.51; P = 0.691). According to PCR assay, the prevalence of Toxoplasma infections was 19.3% in the case and 10.4% in control groups which the difference was statistically significant (OR = 3.02; 95% CI = 1.46-6.27; P = 0.002). Multiple sequence alignment of Toxoplasma gondii isolates manifested a common haplotype by the identity: 93.6-100% and divergence: 0-6.7%. We concluded that T. gondii infection not only could not be a risk factor to PD, but even it could be concluded that patients with PD are in more risk to acquisition of infection. These results provide fresh insights into the ambiguous association between T. gondii infection and PD. Keywords Author Keywords:Parkinson's disease; Toxoplasma gondii infection; Sero-molecular assessment KeyWords Plus:SCHIZOPHRENIA; METAANALYSIS; PREVALENCE; DOPAMINE
Protocol Development for Infants with Orthopedic Complications in the Neonatal Intensive Care Unit: Brachial Plexus Injuries and Clubfoot

Currently, there is limited information to guide occupational therapy or physical therapy treatment of high-risk neonates with secondary orthopedic conditions in the Neonatal Intensive Care unit (NICU). This article describes one master of occupational therapy student capstone project that aimed to develop intervention protocols and parent education materials for the treatment of brachial plexus injuries (BPI) and clubfoot for a hospital NICU. A literature review and needs assessment were conducted to determine whether clinical practices and standards existed and to inform the development of the intervention protocols. The critical need to document, define, and examine best practices was validated. Clinical treatment protocols and parent education materials were developed and are presented.