bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence and pathology of Cephalopina titillator infestation in Camelus bactrianus from Xinjiang, China
Authors: Yao Huaibing , Liu Mengli , Ma Wanpeng , Yue Haitao , Su Zhanqiang , Song Ruiqi , Ma Qiang , Li Ling , Wu Zhuangyuan , Ma Yingjun , Chen Gangliang , Chen Baojiang , Yang Jie
Published: September 2022

Background

In camels, nasopharyngeal myiasis is caused by the larvae of Cephalopina titillator, which parasitize the tissues of nasal and paranasal sinuses, pharynx, and larynx. C. titillator infestation adversely affects the health of camels and decreases milk and meat production and even death. However, the C. titillator infestation in Bactrian camels has not been widely studied.

Methods

The present study was conducted to determine the prevalence and risk factors of C. titillator in Bactrian camels of northwestern Xinjiang. Suspected larvae recovered from infested camels were evaluated for C. titillator by microscopy and polymerase chain reaction. Nucleotide sequences of the partial mitochondrial cytochrome c oxidase subunit I (COX1) and cytochrome b (CYTB) genes from the C. titillator of camels were aligned from the NCBI database. Furthermore, the gross and histopathological alterations associated with C. titillator infestation were evaluated via pathological examination.

Results

Of 1263 camels examined 685 (54.2%) camels were infested with suspected C. titillator larvae. Different larval stages were topically detected in the nasal passages and pharynx of the camel heads. Microscopy analysis of the pharyngeal mucosa tissue revealed necrotic tissue debris and some inflammatory cells. Molecular detection of the larval COX1 and CYTB genes indicated that pathogen collected in Bactrian camels was C. titillator. The epidemiological study demonstrated that the prevalence rate of C.titillator infestation was significantly higher in camels of Bestierek Town Pasture (67.2%) and Karamagai Town Pasture (63.6%) compared to Kitagel Town Pasture (38.7%) and Qibal Town Pasture (35.8%) (P < 0.05). No significant difference was observed between the prevalence rates in male (52.6%) and female (54.6%) camels (P > 0.05). The prevalence was higher in warm (64.2%) than that in cold (48.4%) seasons (P < 0.001). The prevalence in camels with non-nomadic method (67.2%) was significantly higher than in animals with nomadic method (47.5%) (P < 0.001). The prevalence of C.titillator infestation was significantly higher in animals of aged 5–10 (60.1%) and aged > 10 (61.1%) years old compared to those of aged < 5 (31.7%) years old camels (P < 0.001).

Conclusion

Our results confirm that there is a high prevalence of C. titillator in Bactrian camels from Xinjiang, closely related to age, season, pasture environment, and husbandry methods. Developing prevention, diagnosis, and control programs to prevent transmission is necessary.
Results of a prospective observational study of autologous peripheral blood mononuclear cell therapy for no-option critical limb-threatening ischemia and severe diabetic foot ulcers
Authors: Panunzi Andrea , Madotto Fabiana , Sangalli Elena , Riccio Federica , Sganzaroli Adriana Barbara , Galenda Paolo , Bertulessi Amelia , Barmina Maria Francesca , Ludovico Ornella , Fortunato Orazio , Setacci Francesco , Airoldi Flavio , Tavano Davide , Giurato Laura , Meloni Marco , Uccioli Luigi , Bruno Antonino , Spinetti Gaia , Caravaggi Carlo Maria Ferdinando
Published: September 2022

Background

Cell therapy with autologous peripheral blood mononuclear cells (PB-MNCs) may help restore limb perfusion in patients with diabetes mellitus and critical limb-threatening ischemia (CLTI) deemed not eligible for revascularization procedures and consequently at risk for major amputation (no-option). Fundamental is to establish its clinical value and to identify candidates with a greater benefit over time. Assessing the frequency of PB circulating angiogenic cells and extracellular vesicles (EVs) may help in guiding candidate selection.

Methods

We conducted a prospective, non-controlled, observational study on no-option CLTI diabetic patients that underwent intramuscular PB-MNCs therapy, which consisted of more cell treatments repeated a maximum of three times. The primary endpoint was amputation rate at 1 year following the first treatment with PB-MNCs. We evaluated ulcer healing, walking capability, and mortality during the follow-up period. We assessed angiogenic cells and EVs at baseline and after each cell treatment, according to primary outcome and tissue perfusion at the last treatment [measured as transcutaneous oxygen pressure (TcPO2)].

Results

50 patients were consecutively enrolled and the primary endpoint was 16%. TcPO2 increased after PB-MNCs therapy (17.2 ± 11.6 vs 39.1 ± 21.8 mmHg, p < .0001), and ulcers healed with back-to-walk were observed in 60% of the study population (88% of survivors) during follow-up (median 1.5 years). Patients with a high level of TcPO2 (≥ 40 mmHg) after the last treatment showed a high frequency of small EVs at enrollment.

Conclusions

In no-option CLTI diabetic patients, PB-MNCs therapy led to an improvement in tissue perfusion, a high rate of healing, and back-to-walk. Coupling circulating cellular markers of angiogenesis could help in the identification of patients with a better clinical benefit over time.
Interpretation of the role of germline and somatic non-coding mutations in cancer: expression and chromatin conformation informed analysis
Authors: Pudjihartono Michael , Perry Jo K. , Print Cris , O’Sullivan Justin M. , Schierding William
Published: September 2022

Background

There has been extensive scrutiny of cancer driving mutations within the exome (especially amino acid altering mutations) as these are more likely to have a clear impact on protein functions, and thus on cell biology. However, this has come at the neglect of systematic identification of regulatory (non-coding) variants, which have recently been identified as putative somatic drivers and key germline risk factors for cancer development. Comprehensive understanding of non-coding mutations requires understanding their role in the disruption of regulatory elements, which then disrupt key biological functions such as gene expression.

Main body

We describe how advancements in sequencing technologies have led to the identification of a large number of non-coding mutations with uncharacterized biological significance. We summarize the strategies that have been developed to interpret and prioritize the biological mechanisms impacted by non-coding mutations, focusing on recent annotation of cancer non-coding variants utilizing chromatin states, eQTLs, and chromatin conformation data.

Conclusion

We believe that a better understanding of how to apply different regulatory data types into the study of non-coding mutations will enhance the discovery of novel mechanisms driving cancer.
TUCAN: A molecular identifier and descriptor applicable to the whole periodic table from hydrogen to oganesson
Authors: Brammer Jan C. , Blanke Gerd , Kellner Claudia , Hoffmann Alexander , Herres-Pawlis Sonja , Schatzschneider Ulrich
Published: September 2022

TUCAN is a canonical serialization format that is independent of domain-specific concepts of structure and bonding. The atomic number is the only chemical feature that is used to derive the TUCAN format. Other than that, the format is solely based on the molecular topology. Validation is reported on a manually curated test set of molecules as well as a library of non-chemical graphs. The serialization procedure generates a canonical “tuple-style” output which is bidirectional, allowing the TUCAN string to serve as both identifier and descriptor. Use of the Python NetworkX graph library facilitated a compact and easily extensible implementation.

Graphical Abstract
Allosteric probe initiated triple signal recycles for sensitive analysis of adenosine triphosphate (ATP)
Authors: Chen Lin , Yu Lu
Published: September 2022

Adenosine triphosphate (ATP) is closely associated with a variety of pathophysiological process, providing energy for cell activities. However, it remains a huge challenge to develop a simple and sensitive ATP detection method. Herein, we describe an ultrasensitive approach for ATP detection by using an elegantly designed allosteric probe to bind with targets and to induce DNAzyme assisted triple signal recycles. To establish the approach, the allosteric probe is designed with three functional parts to bind with target ATP, to induce signal recycles and to assist chain extension. After recognition of ATP by allosteric probe, the triple signal recycles are initiated, including (i) released ATP-allosteric probe complex unfolds H1 probe, constituting the first signal recycle; (ii) DNAzyme based cleavage induces the second signal recycle; and (iii) chain extension induced release of ATP is the third signal recycle. Through a series of experiments, the approach exhibits a favorable ATP detection performance and shows a high anti-interference ability, indicating a potential application value in early diagnosis of diseases.
Investigation of the relationship between geomagnetic activity and solar wind parameters based on a novel neural network (potential learning)
Authors: Kitajima Ryozo , Nowada Motoharu , Kamimura Ryotaro
Published: September 2022

Predicting geomagnetic conditions based on in-situ solar wind observations allows us to evade disasters caused by large electromagnetic disturbances originating from the Sun to save lives and protect economic activity. In this study, we aimed to examine the relationship between the Kp index, representing global magnetospheric activity level, and solar wind conditions using an interpretable neural network known as potential learning (PL). Data analyses based on neural networks are often difficult to interpret; however, PL learns by focusing on the “potentiality of input neurons” and can identify which inputs are significantly utilized by the network. Using the full advantage of PL, we extracted the influential solar wind parameters that disturb the magnetosphere under southward Interplanetary magnetic field (IMF) conditions. The input parameters of PL were the three components of the IMF (Bx, By, Bz), solar wind flow speed (Vx), and proton number density (Np) in geocentric solar magnetospheric (GSM) coordinates obtained from the OMNI solar wind database between 1998 and 2019. Furthermore, we classified these input parameters into two groups (targets), depending on the Kp level: Kp = 6–9 (positive target) and Kp = 0 to 1 + (negative target). Negative target samples were randomly selected to ensure that numbers of positive and negative targets were equal. The PL results revealed that solar wind flow speed is an influential parameter for increasing Kp under southward IMF conditions, which was in good agreement with previous reports on the statistical relationship between the Kp index and solar wind velocity, and the Kp formulation based on the IMF and solar wind plasma parameters. Based on this new neural network, we aim to construct a more correct and parameter-dependent space weather forecasting model.

Graphical Abstract
Rare earth element identification and quantification in millimetre-sized Ryugu rock fragments from the Hayabusa2 space mission
Authors: Tack Pieter , De Pauw Ella , Tkalcec Beverley , Lindner Miles , Bazi Benjamin , Vekemans Bart , Brenker Frank , Di Michiel Marco , Uesugi Masayuki , Yurimoto Hisayoshi , Nakamura Tomoki , Amano Kana , Matsumoto Megumi , Fujioka Yuri , Enokido Yuma , Nakashima Daisuke , Noguchi Takaaki , Okazaki Ryuji , Yabuta Hikaru , Naraoka Hiroshi , Sakamoto Kanako , Tachibana Shogo , Yada Toru , Nishimura Masahiro , Nakato Aiko , Miyazaki Akiko , Yogata Kasumi , Abe Masanao , Okada Tatsuaki , Usui Tomohiro , Yoshikawa Makoto , Saiki Takanao , Tanaka Satoshi , Terui Fuyuto , Nakazawa Satoru , Watanabe Sei-Ichiro , Tsuda Yuichi , Vincze Laszlo
Published: September 2022

Millimetre-sized primordial rock fragments originating from asteroid Ryugu were investigated using high energy X-ray fluorescence spectroscopy, providing 2D and 3D elemental distribution and quantitative composition information on the microscopic level. Samples were collected in two phases from two sites on asteroid Ryugu and safely returned to Earth by JAXA’s asteroid explorer Hayabusa2, during which time the collected material was stored and maintained free from terrestrial influences, including exposure to Earth’s atmosphere. Several grains of interest were identified and further characterised to obtain quantitative information on the rare earth element (REE) content within said grains, following a reference-based and computed-tomography-assisted fundamental parameters quantification approach. Several orders of magnitude REE enrichments compared to the mean CI chondrite composition were found within grains that could be identified as apatite phase. Small enrichment of LREE was found for dolomite grains and slight enrichment or depletion for the general matrices within the Ryugu rock fragments A0055 and C0076, respectively.

Graphical Abstract
Efficacy of antifungal substances of three Streptomyces spp. against different plant pathogenic fungi
Authors: Ghanem Gamal A. M. , Gebily Doha A. S. , Ragab Mona M. , Ali Ayat M. , Soliman Nour El-Deen K. , El-Moity Tawfik H. Abd
Published: September 2022

Background

Soil-borne plant pathogenic fungi with a wide host range of crops cause a significant limitation on the global production of agronomic crops. Applications of synthetic pesticides are an important tool for managing plant diseases, but have deleterious influences on the environment as well as its incompatibility with organic agriculture. Recently, Streptomyces spp. became one of the best bio-control agents as a promising environmentally eco-friendly method for effective management of plant diseases.

Results

In a previous research, three species of Streptomyces spp., i.e., S. griseus (MT210913 “DG5”), S. rochei (MN700192 “DG4”) and S. sampsonii (MN700191 “DG1” strains) were identified, as exhibiting potent antifungal activities against plant pathogenic fungus, Sclerotinia sclerotiorum in vitro and greenhouse. GC–Mass analysis revealed the presence of 44, 47 and 54 substances of S. sampsonii DG1, S. griseus DG5 and S. rochei DG4, respectively. GC–MS revealed substances, with bio-control activity, were categorized as volatile organic compounds (VOCs), fatty acids and plant growth regulators, etc. GC–MS analysis exhibited the presence of 7, 13 and 20 volatile compounds produced by S. sampsonii, S. rochei and S. griseus, respectively. These substances exhibited potent antifungal activity against various plant pathogenic fungi, i.e., Botrytis cinerea, Macrophomina phaseolina, Rhizoctonia solani and S. sclerotiorum in vitro, by dual-culture assay. The three strains inhibited all the pathogenic fungi in dual-culture assay in the range of 30–73.67%. Also, the produced substances were applied in vivo (in the field) and supported their potential biocontrol agent against S. sclerotiorum as well as possessed significant biological properties for plant health and growth. Applying Streptomyces spp. culture broth in the field enhanced physiological responses of phenols, sugar, chlorophyll, protein contents and parameters as well as the yield of bean plants.

Conclusion

In field experiments, foliar application of Streptomyces spp. and their metabolites proved to be a great potential, as promising biocontrol agents, for controlling S. sclerotiorum and enhanced plant growth and yield. S. rochei and S. griseus proved to be strong antifungal, plant growth promoters and environmentally eco-friendly fungicides.
Shifts in therapeutic practices and decline of medicinal cannabis in Indian North-Eastern Frontier (1826–1925)
Authors: Bania Gita
Published: September 2022

Background

The emergence of colonial medicine in the North-Eastern Frontier witnessed different phases of consistent competition and resistance. Herbs such as cannabis provided native physicians with a coherent power to resist colonial medical intervention. Before British rule, cannabis assumed great significance in the socio-economic, cultural, and religious spheres. The colonizers’ bioprospection of cannabis shifted the production and use of cannabis from a medical and recreational plant to an industrial and commercial commodity. British policies on cannabis caused its ban leading to natives’ reliance on colonial cannabis products. As a result, the native medical practitioners resisted for reviving cannabis in the indigenous therapeutics. This paper mainly aims to investigate the decline of medicinal cannabis in indigenous therapeutics, causing subtle resistance of the native physicians of the North-Eastern Frontier.

Methods

This paper follows a nomadology method based on primary and secondary sources to understand the impact on native physicians after the ban on private use, cultivation, and sale of cannabis. The primary sources/data have been collected from the Directorate of Archives: Government of Assam and Directorate of State Archives and Research Centre, Kolkata, West Bengal. Secondary sources have been collected from books, articles, and theses accessed from various libraries and websites.

Results

Ban on cannabis led to dual responses from the indigenous population of the frontier. First is the interest of the native physicians resisting the revival of cannabis in indigenous therapeutics. The second is the interest of the frontier’s elites, who viewed cannabis as a “dangerous drug.” The British policies of control and restrictions on cannabis, the rift of response from the natives, and the over-powering of the indigenous therapeutics by the colonial medical system led to the decline of medicinal uses of cannabis in the North-Eastern Frontier.

Discussions

Various pre-colonial and colonial factors helped colonial medical practices to get the upper hand over indigenous therapeutics. Such a shift led to the decline of indigenous medicinal cannabis causing native resistance, which was patient and silent.

Conclusions

British ban on cannabis resulted in a rift of native responses, resistance, and decline of cannabis in the indigenous therapeutics of the North-Eastern Frontier.
High-risk breast lesions: role of multi-parametric DCE-MRI in detection and histopathological upgrade prediction
Authors: Hussein Samar Ahmed , EL-Dhurani Salwa , Abdelnaby Yasmine , Fahim Mohamed , Abdelazeem Hazem , Moustafa Amr Farouk
Published: September 2022

Background

High-risk breast lesions represent 3–21% of all breast lesions and are non-obligate precursors of malignancy. Various studies have evaluated the value of DCE-MRI including DWI and ADC mapping in the detection of high-risk breast lesions, differentiating them from malignant lesions and predicting upgrade risk after surgical excision reducing misdiagnosis and overtreatment. This study is a retrospective review of all image-guided breast biopsy procedures performed in 2021 at our institution, identified 68 patients with histopathologically proven high-risk breast and available MRI examinations with no concurrent ipsilateral malignancy. Image analysis of MRI examinations included morphological criteria, enhancement pattern, dynamic curves, DWI and ADC mapping assessment. Since our knowledge of high-risk breast lesions is still growing, this study is important to evaluate the merits of DCE-MRI in the assessment of high-risk breast lesions, to allow optimization of treatment, better limit it to those women at risk, and avoid overtreatment in women at low risk.

Results

The mean ADC value of high-risk breast lesions was not significantly different from that of malignant breast lesions (p value = 0.015). Non-mass enhancement and type III enhancing curve proved to be significant indicators of high-risk breast lesions upgrade in surgical pathology. Cut-off average ADC value for differentiating upgraded high-risk lesions from non-upgraded high-risk lesions proved 1.24 mm2/sec with sensitivity and specificity of 94 & 100%, respectively.

Discussion

Management of high-risk breast lesions is important in the screening setting, as they are non-obligate precursors of malignancy, and also function as risk indicators. Frequency and upgrade rates of high-risk lesions detected by MRI provide a reference for clinical management. DCE-MRI has a high negative predictive value in predicting the upgrade risk of high-risk lesions. In this study, non-mass enhancement and type III curve were proven to be indicators of high-risk lesion upgrade. Limitations of the study included small number of patients and limited follow-up period.

Conclusions

The use of multi-parametric DCE-MRI including DWI and ADC mapping provides complementary information to detect high-risk breast lesions and predict their upgrade rate.
Therapeutic manipulation of host cell death pathways to facilitate clearance of persistent viral infections
Authors: Cooney James , Allison Cody , Preston Simon , Pellegrini Marc
Published: September 2022

Most persistent viral infections can be controlled, but not cured, by current therapies. Abrogated antiviral immunity and stable latently infected cells represent major barriers to cure. This necessitates life-long suppressive antiviral therapy. Achieving a cure for HIV, hepatitis B virus, Epstein Barr-virus, and others, requires novel approaches to facilitate the clearance of infected cells from the host. One such approach is to target host cell death pathways, rather than the virus itself. Here, we summarize recent findings from studies that have utilized therapeutics to manipulate host cell death pathways as a means to treat and cure persistent viral infections.
Vascular endothelial cells provide T cells with costimulatory signals via the OX40/gp34 system
Authors: Kunitomi Akane , Hori Toshiyuki , Imura Akihiro , Uchiyama Takashi
Published: September 2022

We investigated whether gp34, the ligand of OX40, expressed on EC is involved in costimulation of T cells. Normal CD4+ T cells were stimulated with anti-CD3-coated beads, phytohemagglutinin (PHA), or concanavalin A (Con A) in the presence or absence of irradiated human umbilical vein endothelial cells (HUVEC). Stimulation of T cells with each of these mitogens results in significant T-cell proliferation only when HUVEC were present, and this proliferation was inhibited markedly by anti-OX40 or anti-gp34 monoclonal antibody (mAb). T cells cultured with HUVEC produced more interleukin (IL)-2 than those cultured without HUVEC. The addition of anti-IL-2R α chain and anti-IL-2R β chain mAbs abolished the costimulatory effects of HUVEC. Thus, the augmentation of T-cell proliferation appears to be attributable to increased IL-2 production. These results suggest that gp34 expressed on HUVEC plays a role in potentiation of T-cell immune response by providing OX40+ T cells with costimulatory signals.
The effects of NO synthase inhibitors on murine collagen-induced arthritis do not support a role of NO in the protective effect of IFN-γ
Authors: Vermeire Kurt , Thielemans Leen , Matthys Patrick , Billiau Alfons
Published: September 2022

DBA/1 mice deficient in expressing the interferon-γ (IFN-γ) membrane receptor (IFN-γR KO mice) are more susceptible to collagen-induced arthritis (CIA) than wild-type mice, indicating that endogenous IFN-γ plays a protective role in the pathogenesis of CIA. In IFN-γR KO mice, nitric oxide (NO) production during CIA is impaired. Because NO is known to exert immunosuppressive and anti-inflammatory effects in certain model systems, the protective effect of IFN-γ might be mediated by NO. Here, we tested in wild-type mice whether inhibition of NO production by metabolic inhibitors, aminoguanidine (AG) and L-N-(1-iminoethyl)lysine (L-NIL), could mimic the ablation of the IFN-γ receptor. A high-dose regimen of AG supplied in the drinking water inhibited NO production, disease development, and anticollagen antibody production but was also associated with transient body weight loss. At a dose and time regimen that still inhibited NO production but did not cause body weight loss, AG failed to affect disease scores. Treatment with L-NIL, which more specifically than AG affects inducible NO production, caused a slight increase in anticollagen antibody production although not significantly affecting disease occurrence. These data indicate that the diminished capacity of the IFN-γR KO mice to produce NO following immunization with collagen is unlikely to account for their higher susceptibility to CIA.
Altered caspase expression results in delayed neutrophil apoptosis in acute pancreatitis
Authors: O’Neill Sean , O’Neill Amanda J , Conroy Eimear , Brady Hugh R , Fitzpatrick John M , Watson R William G
Published: September 2022

Acute pancreatitis (AP) may lead to the development of multiple organ dysfunction syndrome (MODS), especially in severe cases. Resolution of such inflammatory responses is dependent on neutrophil apoptosis. Delays in this apoptotic response are associated with persistent inflammation and subsequent tissue damage. The aim of this study is to determine the effects of AP on neutrophil apoptosis and to investigate the underlying mechanisms involved. Neutrophils and serum were isolated from control (n=10) and from patients with AP (mild, n=35, and severe, n=5). Neutrophil apoptosis was assessed by propidium iodide DNA staining using flow cytometry. Caspase, glutathione-S-transferase (GST), and Mcl-1 protein expression were assessed by SDS-PAGE western blotting. Serum interleukin (IL)-1β and granulocyte-macrophage colony-stimulating factor (GM-CSF) levels were measured by ELISA. Neutrophils isolated from patients with AP show a significant delay in spontaneous neutrophil apoptosis. Serum factors contributed to this delay with increases in IL-1β and GM-CSF. Isolated neutrophils were resistant to Fas antibody-induced apoptosis. Caspases represent a central mechanism for spontaneous and Fas antibody-induced neutrophil apoptosis. Procaspase 3 expression was decreased in mild and severe cases, but this effect was independent of serum factors. Increases in GST expression may also contribute to the antiapoptotic effect. Altered caspase expression may represent an additional factor contributing to delayed neutrophil apoptosis. This may contribute to the development of AP and its related complications.
Age-associated increase of basal corticosterone levels decreases ED2high, NF-κBhigh activated macrophages
Authors: Kizaki Takako , Ookawara Tomomi , Iwabuchi Kazuya , Onoé Kazunori , Day Noorbibi K , Good Robert A , Maruyama Naoki , Haga Shukoh , Matsuura Nobuo , Ohira Yoshinobu , Ohno Hideki
Published: September 2022

The proportion of cells with a high density of ED2 (ED2high cells) in peritoneal cells from old rats was significantly lower than that from young rats. The expression of major histocompatibility complex class II (MHC class II) molecules, the antigen presentation, production of interleukin (IL)-1β and IL-6, and nuclear factor-κB activity in ED2high cells were markedly higher than those in cells with a low density of ED2 (ED2low cells), although no significant difference was observed in the expression of MHC class II molecules and the antigen presentation between ED2high cells from young and old rats. Meanwhile, basal corticosterone concentration in serum and glucocorticoid (GC) receptor mRNA expression in peritoneal cells increased significantly in old rats. The proportion of ED2high cells was increased by adrenalectomy in young rats. Furthermore, nuclear translocation of GC receptor was observed in ED2low cells, whereas GC receptor was detected in cytoplasmic extracts from ED2high cells. These results suggest that the decrease in functional ED2high macrophages with age results in the age-associated decline of immune responses, which is regulated, in part, by the basal GC concentration.
LFA-1 (CD11a/CD18) triggers hydrogen peroxide production by canine neutrophils
Authors: Lu Huifang , Ballantyne Christie , Smith C Wayne
Published: September 2022

The respiratory burst of neutrophils stimulated by chemotactic factors is markedly augmented by Mac-1-dependent adhesion such as the interaction of Mac-1 (CD11b/CD18) with intercellular adhesion molecule-1 (ICAM-1; CD54) expressed on the surface of parenchymal cells (e.g., cardiac myocytes). In the current study, we evaluate the hypothesis that lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18) can also trigger the respiratory burst in neutrophils. To isolate LFA-1/ICAM-1 interactions from Mac-1/ICAM-1 interactions, full-length chimeric ICAM-1 was developed and expressed in L cells with domains 1 and 2 from canine ICAM-1 and domains 3–5 from human ICAM-1 (C1,2;H3–5). We have shown that canine neutrophils do not bind to human ICAM-1. We demonstrated that chimeric ICAM-1 C1,2;H3–5 supported only LFA-1-dependent adhesion of canine neutrophils and that such adhesion triggered rapid onset of H2O2 production from canine neutrophils. The following seven experimental conditions distinguished LFA-1-dependent H2O2 production from Mac-1-dependent production: It did not require exogenous chemotactic stimulation; H2O2 release was more rapid, but the amount released was <40% of that mediated by Mac-1 adhesion; it was inhibited by anti-CD11a and anti-ICAM-1 antibodies; in contrast to that mediated by Mac-1, it was not inhibited by anti-CD11b antibody, neutrophil inhibitory factor (NIF), or cytochalasin B or H7. Thus, canine neutrophils seem to be able to utilize two members of the β2 integrin family to interact with ICAM-1 and signal H2O2 production, with LFA-1 at an early stage without prior chemotactic stimulation and Mac-1 at a later stage requiring chemotactic stimulation.
Human neutrophil defensins selectively chemoattract naive T and immature dendritic cells
Authors: Yang De , Chen Qian , Chertov Oleg , Oppenheim Joost J
Published: September 2022

Defensins, a family of cationic, structurally related, antimicrobial peptides, contribute to host defense by disrupting the cytoplasmic membrane of microbes. Here we show that human neutrophil defensins selectively induce the migration of human CD4+/CD45RA+ naive and CD8+, but not CD4+/CD45RO+ memory, T cells. Moreover, human neutrophil defensins are chemotactic for immature human dendritic cells derived from either CD34+ progenitors or peripheral blood monocytes. Upon maturation induced by treatment with tumor necrosis factor α (TNF-α), dendritic cells lose their responsiveness to human neutrophil defensins. The chemotactic effect of human neutrophil defensins on both T and dendritic cells is pertussis toxin-sensitive, suggesting that a Giα protein-coupled receptor is responsible. Human neutrophil defensins are also chemotactic for immature murine dendritic cells. These data suggest that, in addition to their antimicrobial role, human neutrophil defensins also contribute to adaptive immunity by mobilizing T cells and dendritic cells.
Suppression of TNF-α production in human mononuclear cells by an adenosine kinase inhibitor
Authors: Eigler Andreas , Matschke Verena , Hartmann Gunther , Erhardt Simon , Boyle David , Firestein Gary S , Endres Stefan
Published: September 2022

Adenosine exerts potent anti-inflammatory activities through inhibition of cytokine synthesis by activated monocytes. Adenosine is rapidly phosphorylated intracellularly by adenosine kinase. GP515, an adenosine kinase inhibitor, prevents the phosphorylation of adenosine to AMP and thereby locally enhances the adenosine concentration. GP515 has exhibited significant anti-inflammatory effects in several murine models of inflammation. In this study we investigated the effect of GP515 alone and in combination with exogenous adenosine or with rolipram, a phosphodiesterase inhibitor, on tumor necrosis factor α (TNF-α) synthesis in human peripheral blood mononuclear cells (PBMC) or whole blood. Lipopolysaccharide (LPS; 10 ng/mL)-stimulated PBMC were incubated in the absence or presence of these substances. GP515 alone showed a dose-dependent suppression of TNF-α production with an IC50 of 80 μM. The TNF-α-inhibiting effects of adenosine and GP515 were reversed in the presence of the cAMP antagonist (Rp)-cAMPS, supporting the hypothesis of a cAMP-mediated pathway. Combinations of GP515 with either adenosine or rolipram led to an additive inhibition of TNF-α synthesis. These experiments are the first to demonstrate efficacy of an adenosine kinase inhibitor in TNF-α suppression in cells of human origin. The findings form a basis to investigate these strategies in animal models of TNF-α-mediated chronic inflammatory diseases.
Induction of neutrophil death resembling neither apoptosis nor necrosis by ONO-AE-248, a selective agonist for PGE2 receptor subtype 3
Authors: Liu Jiajia , Akahoshi Tohru , Jiang Shixu , Namai Rie , Kitasato Hidero , Endo Hirahito , Kameya Toru , Kondo Hirobumi
Published: September 2022

An increase of intracellular cAMP mediated by prostaglandin E2 (PGE2) has been shown to delay spontaneous apoptosis of neutrophils. It has been demonstrated that a selective agonist for PGE2 receptor subtype 3 (the EP3 receptor) is capable of decreasing cAMP and stimulating phosphoinositide turnover in various types of cells. We investigated the effect of a selective EP3 receptor agonist, ONO-AE-248, on neutrophil viability. ONO-AE-248 rapidly caused a unique form of neutrophil death. The agonist primarily induced morphological changes of the nucleus, including fusion of the lobules, decreased compactness of the chromatin, and blebbing and rupture of the nuclear membrane. This was followed by an increase of plasma membrane permeability and cell lysis. During these processes, neither apoptotic changes such as nuclear condensation, DNA fragmentation, and expression of phospholipid phosphatidylserine on the plasma membrane nor necrotic changes such as chromatin clumping and organelle destruction were apparent in the treated neutrophils. The fatal effect of the agonist might be specific for neutrophils because it failed to promote the rapid death of other types of cells. Although activation of neutrophils by ONO-AE-248 was not evident, experiments using metabolic inhibitors demonstrated that the agonist caused neutrophil death via the activation of protein kinase C in the presence of intracellular ATP. These findings indicated that EP3 receptor-mediated signals might promote a novel form of neutrophil death, which differs from typical apoptosis or necrosis.
Human neutrophil cathepsin G down-regulates LPS-mediated monocyte activation through CD14 proteolysis
Authors: Le-Barillec Karine , Pidard Dominique , Balloy Viviane , Chignard Michel
Published: September 2022

A major property of monocytes/macrophages is to recognize and to be activated by bacterial wall components such as LPS, through membrane receptors including the key element CD14. We demonstrate that CD14 expression is down-regulated, as judged by flow cytometry analysis, upon incubation of human monocytes with purified cathepsin G (CG), a releasable neutrophil serine proteinase. The progressive decrease of CD14 expression due to increasing concentrations of CG highly correlates (P < 0.0001) with the decreased synthesis of tumor necrosis factor α (TNF-α) in response to lipopolysaccharide (LPS). This effect is dependent on the enzymatic activity of CG but is not exerted through an activation of monocytes. Immunoblot analysis reveals that CD14 (Mr = 57,000) is directly cleaved by CG and released into the extracellular medium as a high-Mr species (Mr = 54,000). In this context, incubation of monocytes with activated neutrophils leads to a down-regulation of CD14 expression, a process blocked by a serine proteinase inhibitor. These data suggest a paradoxical anti-inflammatory property for CG.
Expression of α4-integrins on human neutrophils
Authors: Kirveskari Juha , Bono Petri , Granfors Kaisa , Leirisalo-Repo Marjatta , Jalkanen Sirpa , Salmi Marko
Published: September 2022

α4 Integrins are important adhesion molecules mediating binding of lymphocytes, monocytes, and eosinophils to multiple cellular and extracellular ligands. Mature neutrophils have been recently suggested to express α4-integrins as well. We studied whether human neutrophils can synthesize α4-integrins upon activation in vitro or in vivo. Two anti-α4 mAbs, but not multiple subclass-matched non-binding controls, reacted with granulocytes in an inducer and time-dependent manner. Nevertheless, staining with Ig subclass-specific second-stage reagents surprisingly revealed that commercial anti-α4 mAbs contain two distinct Igs, the α4-specific IgG1 and an IgG2a of an unknown specificity. We showed that in vitro inductions used by us and others only induce the binding of nonspecific IgG2a from the commercial HP2/1 to activated neutrophils. By reverse-transcriptase polymerase chain reaction, α4 mRNA was not detectable in purified neutrophils. Our results show that α4 integrin protein and mRNA are absent from normal and stimulated human neutrophils.
β2 Integrin (CD11/CD18)-mediated signaling involves tyrosine phosphorylation of c-Cbl in human neutrophils
Authors: Willeke Thomas , Behrens Sandra , Scharffetter-Kochanek Karin , Gaehtgens Peter , Walzog Barbara
Published: September 2022

Leukocyte adhesion molecules of the β2 integrin (CD11/CD18) family mediate cell-cell and cell-substrate interactions of human polymorphonuclear neutrophils (PMN) during their recruitment to sites of inflammation. To elucidate the molecular events that follow extracellular ligand interactions of β2 integrins, protein tyrosine signaling was studied subsequent to integrin engagement by Western blotting technique. Upon adhesion to immobilized fibrinogen, a native ligand of the β2 integrins Mac-1 (CD11b/CD18) and gp150/95 (CD11c/CD18), tyrosine phosphorylation of several proteins including a 120-kDa protein was observed in human PMN. This effect was specific for β2 integrins because it was absent in PMN derived from CD18-deficient mice, which lack any β2 integrin expression. Moreover, no signaling was detectable upon engagement of CD29 and CD61, the β-subunits of the β1 and β3 integrins, respectively, revealing the unique function of the β2 integrins in PMN. By means of immunoprecipitation, the most prominent protein that became tyrosine phosphorylated upon β2 integrin engagement was identified as the 120-kDa protein c-Cbl. The observed signaling was independent of both pertussis toxin-sensitive heterotrimeric G-proteins as well as the small G-protein ras. Inhibition of β2 integrin-mediated signaling by herbimycin A prevented adhesion, shape change, and spreading of PMN to immobilized fibrinogen, demonstrating the biological significance of the observed effect. Together, the present data suggest that the β2 integrins fulfill a unique function among the leukocyte integrins in human PMN by activating an intracellular signal transduction cascade that leads to tyrosine phosphorylation of c-Cbl and allows subsequent adhesion, shape change, and spreading.
Permissive factors for HIV-1 infection of macrophages
Authors: Wahl Sharon M , Greenwell-Wild Teresa , Hale-Donze Hollie , Moutsopoulos Niki , Orenstein Jan M
Published: September 2022

Immunodeficiency, the consequence of HIV-1 infection, predisposes the host to opportunistic infections. In turn, opportunistic pathogens influence target cell susceptibility to HIV-1 infection and replication. Although the advent of highly active antiretroviral therapy (HAART) has altered these sequelae, co-infections may prevail in some parts of the world and in failed HAART regimens. Moreover, immune activation as occurs in tonsil and non-infectious mucosal inflammatory lesions may also be associated with proximal sites of viral replication. These connections between enhancement of HIV-1 infection and activation/inflammation warrant further elucidation of the factors promoting permissiveness to HIV-1 infection. Using the opportunistic pathogen Mycobacterium avium as an in vitro model, we demonstrated that co-infection facilitated HIV-1 infection of monocyte-macrophages by multiple pathways. M. avium activated NF-κB, the downstream consequences of which included augmented expression of tumor necrosis factor α and CCR5 receptors, both permissive for sustaining HIV-1 infection. Pronounced viral replication in lymph nodes co-infected with M. avium and HIV-1 paralleled these in vitro findings. Furthermore, reduction in viral burden is associated with treatment of infected or inflamed tissues, underscoring the link between immune activation and viral replication.
HIV-1 genotypes in peripheral blood monocytes
Authors: Zhu Tuofu
Published: September 2022

CD4+ T cells and tissue macrophages are well defined as the major targets for human immunodeficiency virus type 1 (HIV-1) infection and replication, and their infection accounts for many aspects of HIV-1 pathogenesis in vivo. HIV-1 genotype and phenotype in monocytes and their potential roles in pathogenicity in vivo remain unknown. Herein is an overview of our initial work on HIV-1 genotype in purified CD14+ monocytes isolated longitudinally during the course of infection starting from the time of infection. Our data provide evidence for HIV-1 evolution in monocytes and their role as a reservoir of HIV-1 in vivo. A better understanding of HIV-1 in monocytes may greatly help the development of new therapeutic strategies for HIV-1 infection.
Biological parameters of HIV-1 infection in primary intestinal lymphocytes and macrophages
Authors: Smith Phillip D , Meng Gang , Sellers Marty T , Rogers Tina S , Shaw George M
Published: September 2022

Mucosal surfaces are the portal of entry for most HIV-1 infections and play an important role in disease pathogenesis. To characterize the biological parameters of HIV-1 infection in mucosal cells, we used purified lamina propria lymphocytes and macrophages from normal human small intestine to determine the distribution of the HIV-1 receptor and coreceptors on intestinal mononuclear cells and the permissiveness of these cells to HIV-1 infection. Lamina propria lymphocytes expressed CD4, CCR5, and CXCR4. In contrast, lamina propria macrophages expressed CD4 but not CCR5 or CXCR4. Intestinal lymphocytes supported replication by R5 and X4 isolates of HIV-1, but lamina propria macrophages were permissive to neither. RANTES, macrophage inflammatory protein-1α (MIP-1α), and MIP-1β inhibited infection of intestinal lymphocytes by BaL, indicating that R5 infection of the intestinal lymphocytes was mediated by CCR5. Thus, resident lamina propria lymphocytes, not macrophages, are the target mononuclear cell for HIV-1 infection in the intestinal mucosa during early HIV-1 infection.
HIV-1 Vpr regulates expression of β chemokines in human primary lymphocytes and macrophages
Authors: Muthumani Karuppiah , Kudchodkar Sagar , Papasavvas Emmanouil , Montaner Luis J , Weiner David B , Ayyavoo Velpandi
Published: September 2022

The HIV-1 vpr gene encodes a 14-kDa virion-packaged protein that has been implicated in viral pathogenesis. Vpr exhibits profound effects on human primary cells influencing proliferation, differentiation, apoptosis, and cytokine production, in part through NF-κB-mediated transcription. NF-κB, a potent transcription factor, activates many proinflammatory cytokines/chemokines upon infection. Here, we analyzed the effect of extracellular Vpr as well as the virion-associated Vpr on β chemokines (MIP-1α, MIP-1β, and RANTES) production in human macrophages and primary lymphocytes (PBLs). Macrophages and PBLs exposed to HIV-1 vpr+ viruses or to recombinant Vpr protein produced significantly less β chemokines compared with cells infected with HIV-1 vpr− viruses or irrelevant control protein (Gag)-exposed cells. These results suggest that a Vpr-mediated increase in virus replication could be in part through down-regulation of chemokine production.
Mononuclear phagocytes mediate blood-brain barrier compromise and neuronal injury during HIV-1-associated dementia
Authors: Persidsky Yuri , Zheng Jialin , Miller Donald , Gendelman Howard E
Published: September 2022

The neuropathogenesis of HIV-1 infection revolves around the production of secretory factors from immune-activated brain mononuclear phagocytes (MP). MP-secreted chemokines may play several roles in HIV-1 encephalitis (HIVE). These can promote macrophage brain infiltration, blood-brain barrier (BBB) and neuronal dysfunction during HIV-1-associated dementia. We investigate how HIV-1-infected MP regulates the production of chemokines and how they influence HIV-1 neuropathogenesis. We demonstrate that HIV-1-infected and immune-activated MP (for example, microglia) and astrocytes produce β-chemokines in abundance, as shown in both laboratory assays and within infected brain tissue. HIV-1-infected microglia significantly modulate monocyte migration in a BBB model system and in brains of SCID mice with HIVE. HIV-1-infected MP down-regulate tight junction protein and special polarized transport systems on brain microvascular endothelial cells as shown in human autopsy brain tissue and in SCID mice with HIVE. Chemokines can damage neurons directly. Toxicity caused by binding of stromal-derived factor-1α to its receptor on neurons exemplifies such mechanism. In toto, these works underscore the diverse roles of chemokines in HIV-1 neuropathogenesis and lay the foundation for future therapeutic interventions.
Immunopharmacological activity ofEchinaceapreparations following simulated digestion on murine macrophages and human peripheral blood mononuclear cells
Authors: Rininger Joseph A , Kickner Shirley , Chigurupati Padmasree , McLean Anne , Franck Zsofia
Published: September 2022

We have investigated the immunostimulatory, anti-inflammatory, and antioxidant activities of variousEchinacea raw materials and commercially available products on murine macrophages and human peripheral blood mononuclear cells (PBMCs). To emulate oral dosing, a simulated digestion protocol was employed as a means of sample preparation.Echinacea-induced macrophage activation was used as a measure of immunostimulatory activity determined via quantitative assays for macrophage-derived factors including tumor necrosis factor α, interleukin (IL)-1α, IL-1β, IL-6, IL-10, and nitric oxide.Echinacea herb and root powders were found to stimulate murine macrophage cytokine secretion as well as to significantly enhance the viability and/or proliferation of human PBMCs in vitro. In contrast, Echinacea extracts chemically standardized to phenolic acid or echinocaside content and fresh pressed juice preparations were found to be inactive as immunostimulatory agents but did display, to varying degrees, anti-inflammatory and antioxidant properties.
Differential localization of protein kinase C isotypes in equine eosinophils and neutrophils
Authors: Greenaway E C , Cunningham F M , Goode N T
Published: September 2022

Phorbol esters, which activate protein kinase C (PKC), stimulate equine eosinophil superoxide production and adherence. After showing that superoxide production could be inhibited by the nonselective PKC inhibitors, staurosporine and bisindolymaleimide I, the PKC isotypes in equine eosinophils were characterized, because evidence suggests that individual isotypes may play distinct roles in regulating eosinophil function. Western blots demonstrated that equine eosinophils expressed PKC α, β, δ, ɛ, ι, and ζ. However, unlike the equine neutrophil, the majority of the PKC was detected in the particulate fraction of the cell. Despite this unusual location, the PKC in equine eosinophils was activatable, suggesting that it is functionally competent. The regulatory role of PKC in equine eosinophils may reflect the association of activity with the particulate fraction and the profile of isotype expression.