bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of bone sialoprotein coated three‐dimensional printed calcium phosphate scaffolds on primary human osteoblasts
Authors: Klein Anja , Baranowski Andreas , Ritz Ulrike , Götz Hermann , Heinemann Sascha , Mattyasovszky Stefan , Rommens Pol M. , Hofmann Alexander
Published: January 2018

The combination of the two techniques of rapid prototyping 3D‐plotting and bioactive surface functionalization is presented, with emphasis on thein vitroeffect of Bone Sialoprotein (BSP) on primary human osteoblasts (hOBs). Our primary objective was to demonstrate the BSP influence on the expression of distinctive osteoblast markers in hOBs. Secondary objectives included examinations of the scaffolds' surface and the stability of BSP‐coating as well as investigations of cell viability and proliferation. 3D‐plotted calcium phosphate cement (CPC) scaffolds were coated with BSP via physisorption. hOBs were seeded on the coated scaffolds, followed by cell viability measurements, gene expression analysis and visualization. Physisorption is an effective method for BSP‐coating. Coating with higher BSP concentrations leads to enhanced BSP release. Two BSP concentrations (50 and 200 μg/mL) were examined in this study. The lower BSP concentration (50 µg/mL) decreased ALP and SPARC expression, whereas the higher BSP concentration (200 μg/mL) did not change gene marker expression. Enhanced cell viability was observed on BSP‐coated scaffolds on day 3. hOBs developed a polygonal shape and connected in an intercellular network under BSP influence. Quantitative cell morphology analyses demonstrated for BSP‐coated CPCs an enhanced cell area and reduced circularity. The strength of the above‐mentioned effects of BSP‐coated scaffoldsin vivois unknown, and future work is focusing on bone ingrowth and vascularizationin vivo. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2565–2575, 2018.
Treatment-Related Changes in Bone Turnover and Fracture Risk Reduction in Clinical Trials of Anti-Resorptive Drugs: A Meta-Regression
Authors: Bauer Douglas C , Black Dennis M , Bouxsein Mary L , Lui Li-Yung , Cauley Jane A , de Papp Anne E , Grauer Andreas , Khosla Sundeep , McCulloch Charles E , Eastell Richard ,   for the Foundation for the National Institutes of Health (FNIH) Bone Quality Project
Published: January 2018

Few pooled analyses of antiresorptive (AR) treatment trials relate short-term changes in bone turnover markers (BTMs) to subsequent fracture reduction. Such information would be useful to assess new ARs or novel dosing regimens. In the Foundation for the National Institutes of Health (FNIH) Bone Quality project, we analyzed individual-level data from 28,000 participants enrolled in 11 bisphosphonate (BP) and three selective estrogen receptor modulator (SERM) placebo-controlled fracture endpoint trials. Using BTM results for two bone formation markers (bone-specific alkaline phosphatase [bone ALP] and pro-collagen I N-propeptide [PINP]) and two bone resorption markers (N-terminal and C-terminal telopeptide of type I collagen) and incident fracture outcome data, we performed a meta-regression relating the mean net effect of treatment on change in bone turnover (active minus placebo % difference after 3 to 12 months) to the log of study-wide fracture risk reduction, and used linear regression to plot the best fitting line. Separate analyses were performed for incident morphometric vertebral, nonvertebral, and hip fractures over 1 to 4 years of follow-up. Change in bone ALP and PINP were available for over 16,000 and 10,000 participants, respectively. For vertebral fracture, the results showed a strong relationship between treatment-related bone ALP or PINP changes and vertebral fracture risk reduction (r2 = 0.82 [p < 0.001] and r2 = 0.75 [p = 0.011], respectively) Relationships were weaker and no longer statistically significant for nonvertebral (r2 = 0.33 [p = 0.053] and r2 = 0.53 [p = 0.065], respectively) and hip fracture (r2 = 0.17 [p = 0.24] and r2 = 0.43 [p = 0.11], respectively) outcomes. Analyses limited to BP trials gave similar results. For all fracture types, relationships were weaker and nonsignificant for bone resorption markers. We conclude that short-term AR treatment-related changes in bone ALP and PINP strongly predict vertebral fracture treatment efficacy, but not nonvertebral or hip fracture treatment efficacy. Change in bone formation markers might be useful to predict the anti-vertebral fracture efficacy of new AR compounds or novel dosing regiments with approved AR drugs. © 2017 American Society for Bone and Mineral Research.
Synthesis, in vitro Antimicrobial, and Cytotoxic Activities of New 1,3,4‐Oxadiazin‐5(6H)‐one Derivatives from Dehydroabietic Acid
Authors: Jin Xiao‐Yan , Zhang Kang‐Ping , Chen Hao , Miao Ting‐Ting , Wang Shi‐Fa , Gu Wen
Published: January 2018

A series of new 1,3,4‐oxadiazin‐5(6H)‐one derivatives (6a–n) of dehydroabietic acid were designed and synthesized as potential antimicrobial and antitumor agents. Their structures were characterized by IR, 1H NMR, 13C NMR, MS, and elemental analyses. All the title compounds were evaluated for their antimicrobial activity against four bacterial and three fungal strains using the serial dilution method. Among them, compound 6e showed the highest antibacterial activity against Bacillus subtilis with a minimum inhibitory concentration (MIC) value of 1.9 μg/mL. In addition, the in vitro cytotoxic activities of the title compounds were also assayed against three human carcinoma cell lines (MCF‐7, SMMC‐7721, and HeLa) through the MTT colorimetric method. As a result, compounds 6b, 6g, 6k, and 6m exhibited significant inhibition against at least one cell line with IC50 values below 10 μM. Compound 6m was especially found to be the most potent derivative with IC50 values of 2.26 ± 0.23, 0.97 ± 0.11, and 1.89 ± 0.31 μM against MCF‐7, SMMC‐7721, and HeLa cells, respectively, comparable to positive control etoposide.
Church and family support networks and depressive symptoms among African Americans: Findings from The National Survey of American Life
Authors: Chatters Linda M. , Nguyen Ann W. , Taylor Robert Joseph , Hope Meredith O.
Published: January 2018

Aims:

We examined the associations between informal social support from church members and social support from extended family members and depressive symptoms within a national probability sample of African American adults ranging in age from 18 to 93.

Methods:

This analysis used data from the National Survey of American Life and accounted for religious service attendance and various demographic variables that have known associations with and are consequential for mental health.

Results:

Frequency of contact with church and family members and emotional support from family were inversely associated with depressive symptoms, and negative church and family interactions were positively associated with depressive symptoms. Emotional support from church members, however, was unrelated to depressive symptoms.

Conclusion:

This study underscores the important contributions of church relationships to depressive symptoms among African Americans across the adult life span, and confirms that these associations are independent of family relationship factors and religious service attendance.
A five‐gene signature may predict sunitinib sensitivity and serve as prognostic biomarkers for renal cell carcinoma
Authors: Chen Yuan‐Lei , Ge Guang‐Ju , Qi Chao , Wang Huan , Wang Huai‐Lan , Li Li‐Yang , Li Gong‐Hui , Xia Li‐Qun
Published: January 2018

Sunitinib resistance is, nowadays, the major challenge for advanced renal cell carcinoma patients. Illuminating the potential mechanisms and exploring effective strategies to overcome sunitinib resistance are highly desired. We constructed a reliable gene signature which may function as biomarkers for prediction of sunitinib sensitivity and clinical prognosis. The gene expression profiles were obtained from The Cancer Genome Atlas database. By performing GEO2R analysis, numerous differentially expressed genes (DEGs) were found to be associated with sunitinib resistance. To acquire more precise DEGs, we integrated three different microarray datasets. Functional analysis revealed that these DEGs were mainly involved in Rap1 signaling pathway, p53 signaling pathway and Ras signaling pathway. Then, top five hub genes, BIRC5, CD44, MUC1, TF, CCL5, were identified from protein–protein interaction (PPI) network. Sub‐network analysis carried out by MCODE plugin revealed that key DEGs were related with PI3K‐Akt signaling pathway, Rap1 signaling pathway and VEGF signaling pathway. Next, we established sunitinib‐resistant OS‐RC‐2 and 786‐O cell lines and validated the expression of five hub genes in cell lines. To further evaluate the potentials of five‐gene signature for predicting clinical prognosis, we analyzed RCC patients with gene expressions and overall survival information from two independent patient datasets. The Kaplan–Meier estimated the OS of RCC patients in the low‐ and high‐risk groups according to gene expression signature. Multivariate Cox regression analysis indicated that the prognostic power of five‐gene signature was independent of clinical features. In conclusion, we developed a five‐gene signature which can predict sunitinib sensitivity and OS for advanced RCC patients, providing novel insights into understanding of sunitinib‐resistant mechanisms and identification of RCC patients with poor prognosis.
No Increased Risk of Fracture in Patients Receiving Antimuscarinics for Overactive Bladder Syndrome: A Retrospective Cohort Study
Authors: Kao Li‐Ting , Huang Chao‐Yuan , Lin Herng‐Ching , Chu Chi‐Ming
Published: January 2018

To date, the actual relationship between antimuscarinics for overactive bladder (OAB) syndrome and the subsequent risk of a fracture remains unclear. The aim of this retrospective cohort study was to demonstrate the association between antimuscarinics for OAB and fractures using a large population‐based data set. Data used in this study were taken from the Taiwan Longitudinal Health Insurance Database 2005. We identified 2927 OAB patients who received antimuscarinics as the study cohort and 11 708 matched OAB patients who did not receive antimuscarinics as the comparison cohort. Each individual was tracked for a 3‐year period to define those who received a diagnosis of a fracture. Stratified Cox proportional hazard regression analyses were conducted to estimate the association between antimuscarinics for OAB and a subsequent fracture. Results revealed that the incidence rate of fracture per 100 person‐years within the 3‐year follow‐up period were 3.01 (95% confidence interval [CI], 2.65–3.40) for OAB patients who received antimuscarinics and 2.77 (95%CI, 2.60–2.95) for those OAB patients who did not receive antimuscarinics. The adjusted hazard ratio (HR) for a subsequent fracture for OAB patients who received antimuscarinics was 1.11 (95%CI, 0.97–1.28) compared with those OAB patients who did not receive antimuscarinics. Consequently, we concluded that antimuscarinics for OAB was not significantly predictive of fracture.
Temporal and spatial expression of insulin‐like peptide (insl5a and insl5b) paralog genes during the embryogenesis of Danio rerio
Authors: Venditti Massimo , Donizetti Aldo , Fiengo Marcella , Fasano Chiara , Santillo Alessandra , Aniello Francesco , Minucci Sergio
Published: January 2018

Relaxin (RLN) and insulin (INSL)‐like peptides are member of the INSL/RLN superfamily, which are encoded by seven genes in humans and can activate the G‐protein coupled receptor RXFP 1–4. These peptides evolved from a common ancestor, RLN3‐like gene. Two rounds of whole genome duplication (WGD) in early vertebrate evolution, together with an additional WGD in the teleost lineage, caused an expansion of RLN genes set in the genome of Danio rerio. In particular, six RLN genes are present: a single copy of rln and insl3 genes, and two paralogs for the rln3 gene (rln3a and rln3b), and the insl5 gene (insl5a and insl5b). We have already reported the presence of rln3a and rln3b genes in the developing zebrafish brain, as well as the expression of rln gene in the developing zebrafish brain and extraneural territories, such as thyroid gland and pancreas. Here, we report for the first time the expression of the two parologs genes for insl5, insl5a, and insl5b in D. rerio embryonic development. The corresponding transcripts of both the paralogs are present in all embryonic stages analyzed by RT‐qPCR. In situ hybridization analyses showed a restricted signal in intestinal cells and the pancreatic region at 72 hpf for insl5a, while at 96 hpf both genes are expressed in specific intestinal cells. Furthermore, in adult zebrafish intestine tissue, in situ hybridation experiments showed that insl5a transcript is specifically localized in the goblet cells, while insl5b transcript is in enteroendocrine cells. These data revealed a high degree of gene expression pattern conservation for such genes in vertebrate evolution.
Enatioselective Synthesis of Tetrahydrocarbazoles via Chiral Phosphoric Acid Promoted Domino Friedel–Crafts‐type Reaction of Indole‐3‐butanal with Indoles
Authors: Yu Shuowen , Chen Hui , Xu Xiaoying , Yuan Weicheng , Zhang Xiaomei
Published: January 2018

By employing a chiral phosphoric acid as a catalyst, enantioselective domino Friedel–Crafts‐type reaction of indole‐3‐butanal with indoles was realized. This transformation allowed for the synthesis of a wide variety of enantioenriched 1‐indolyl‐tetrahydrocarbazoles with good yields (up to 99%) in moderate enantioselectivities (up to 81% ee).
Detecting deception through small talk and comparable truth baselines
Authors: Palena Nicola , Caso Letizia , Vrij Aldert , Orthey Robin
Published: January 2018

The present experiment investigates similarities in participants' nonverbal and verbal behaviours when responding to baseline and investigative questions, comparing two different types of baselines. Police literature suggests to obtain a baseline through small talk, whereas academic literature underlines the importance of baseline and investigative themes to be comparable. First, a baseline was obtained (either small talk or comparable), then the investigative questioning started. During the investigative questioning, participants either truthfully reported a set of actions they had actually performed or lied about them. Findings revealed that truth tellers and liars in the small talk condition did not differ in their level of similarity when responding to the baseline and investigative questions. In the comparable truth condition, levels of verbal similarity between the baseline and investigative questions were higher for truth tellers than for liars, but only for one variable: spatial detail. Results therefore showed that a small talk baseline should not be used to assess interviewees' credibility, and that a comparable truth baseline, although better than a small talk baseline, is still problematic.
In Vitro Release of Lysozyme From Monocytes and Granulocytes
Authors: Haneberg Bjørn , Glette Johan , Talstad Ingebrigt , Sørnes Steinar , Solberg Claus Ola
Published: January 2018

When exposed to zymosan or latex particles or heat-inactivated staphylococci, freshly prepared human blood monocytes and granulocytes rapidly released a large fraction of their lysozyme content. Within 24 hours the total lysozyme activity in the monocyte suspensions tripled, while it doubled in the granulocyte suspensions, indicating synthesis of the enzyme following release. The monocytes in particular seemed to release and synthesize lysozyme without any other stimulus than contact with lymphocytes and the tube walls. Potassium caseinate in solution did not influence the lysozyme release. Myeloperoxidase and β-glucuronidase, which in the granulocytes are kept in lysosomal fractions separate from most of the lysozyme, were neither released nor synthesized to a significant degree. Moreover, the minute amount of lactate dehydrogenase released indicated that the lysozyme release was not the result of cell lysis. Accordingly, the monocytes, which are not already stimulated by adherence to nonphagocytosable surfaces, are capable of selective enzyme release similar to that of the granulocytes.
Increased Transglutaminase Activity in Elicited and Activated Macrophages: Relationship to Production of Superoxide Anion
Authors: Harris Lesley K , Pabst Michael J , Johnston Richard B
Published: January 2018

To evaluate whether transglutaminase (TG) might be involved in production of oxygen metabolites, TG activity was measured in resident, in activated, and in elicited murine peritoneal macrophages. These various types of macrophages show a wide diversity in their ability to generate oxygen metabolites. Like other transglutaminases, the macrophage enzyme was found to be a calcium-dependent enzyme that was stabilized by dithiothreitol and inhibited by dansylcadav- erine. The TG activity was approximately six-times higher in lysates of LPS- elicited macrophages compared with lysates of resident macrophages; this paralleled the six-fold increase in phorbol myristate acetate-stimulated release of superoxide anion (O2). From mixing experiments, the difference in TG activity was not due to an endogenous cellular inhibitor in the resident cells. The apparent Km of TG for putrescine was similar in lysates of resident and LPS-elicited macrophages, but the Vmax was significantly greater in lysates of LPS-elicited cells. Macrophages obtained from mice injected with either viable bacillus Calmette-Guérin, muramyl dipeptide, or killed Corynebacterium parvum released three- to six- times more O2 than did resident macrophages. The TG activity in these macrophages was also two- to six- times higher than in resident cells. However, macrophages that were primed by exposure to LPS in vitro exhibited increased production of O2 but no increase in TG activity. We conclude that enhanced TG activity is not a prerequisite for the enhanced O2 production observed in activated macrophages.
Modulation of Endotoxin-Induced Neutrophil Alveolitis by Captopril and by Hyperoxia
Authors: Rinaldo J E , Dauber J H
Published: January 2018

We compared survival and the intensity of bronchoalveolar inflammation reflected by lung lavage after the intraperitoneal injection of endotoxin from Escherichia coli serotype 055B5 in rats breathing air and those breathing 60% oxygen for six days after endotoxin injection. Survival following 7.5 mg/kg of endotoxin was comparable in air-breathing rats (50%) and in oxygen-breathing rats (63%). Endotoxin caused a dose-dependent increase in the recovery of polymorphonuclear leukocytes from the lung. Oxygen breathing reduced the percentage of neutrophils recovered by lavage 24 hr after endotoxin from 17% to 9% after 2.5 mg/kg of endotoxin and from 34% to 12% after 7.5 mg/kg of endotoxin. The absolute number of neutrophils recovered was also significantly decreased in oxygen-breathing rats. The activity of pulmonary angiotensin- converting enzyme (ACE) has been reported to be affected by oxygen tension, and ACE degrades bradykinin, a proinflammatory mediator. Therefore, we questioned whether the salutary effect of increased inspired oxygen tension on the magnitude of neutrophil influx into the airspaces could be related to changes in ACE activity. We found that after 48 hr of peroral pretreatment of the rats with captopril, a specific ACE inhibitor, there was increased recovery of neutrophils by lavage 24 hr after injection of endotoxin in air-breathing rats. Captopril pretreatment also increased the chemotactic activity of bronchoalveolar lavage fluid (BALF). There was no concomitant alteration in the accumulation of 125l albumin in the lung following captopril pretreatment either in endotoxin-treated rats or in controls. Thus, breathing 60% oxygen decreased the accumulation of neotrophils in airspaces after intraperitoneal endotoxin injection and pharmacologic inhibition of ACE had the opposite effect. Alterations in the activity of pulmonary angiotensin-converting enzyme related to alveolar oxygen tension is a potential speculative mechanism for modulation of alveolar inflammation by the inspired oxygen concentration in this model.
A Lectinlike Receptor on Murine Macrophage Cell Line Cells, Mm1: Involvement of Sialic Acid-Binding Sites in Opsonin-Independent Phagocytosis for Xenogeneic Red Cells
Authors: Kyoizumi Seishi , Masuda Tohru
Published: January 2018

The recognition mechanism of xenogenic red cells by mouse macrophages was studied by using established cell lines. Approximately 30% of cell line cells Mm1, which lack la antigen, as well as of thioglycollate-induced peritoneal macrophages from SL/Am mice (TGC-Mφ) could ingest unopsonized quail red cells (QRC). In contrast, an undifferentiated type of cell line, M1″, and another type of macrophage cell line, Mk1C, possessing accessory cell activity in association with the expression of la antigen, had no phagocytic activity for QRC. Approximately 80% of Mm1, cells, as well as TGC-Mφ formed rosettes with QRC, whereas M1 and Mk1 C cells did not; indicating that specific binding sites for QRC are expressed on a large portion of Mm1 and TGC-Mφ but not on M1 - and Mk1,-C cells. No requirement of divalent cation (Mg++, Ca++) and metabolic energy was observed for rosette formation between Mm1 cells and QRC. Protease treatment of Mm1 cells eliminated the rosetting activity, whereas periodate oxidation or glycosidase treatment slightly enhanced this activity, suggesting the involvement of surface protein in binding sites of Mm1, cells. In contrast to these findings on Mm1 cells, binding components of QRC were sensitive to periodate oxidation or neuraminidase treatment but resistant to protease, suggesting that the terminal sialic acid residues of carbohydrate of QRC are recognized by Mm1, cells. Furthermore, N-acetylneuraminic acid (NeuNAc) inhibited the rosette formation and promoted the dissociation of rosettes already formed. N-Acetylneuramin lactose (NeuNAc-Lact) was more efficient in rosette inhibition than NeuNAc. These sugars also blocked the phagocytosis of QRC by Mm1 cells but had no effect on either Fc-mediated phagocytosis or latex ingestion. These results suggest that phagocytosis of QRC by murine macrophages is mediated by protease-sensitive binding sites recognizing terminal sialic acid residues of QRC in conjunction with additional carbohydrates.
Antileukocyte Activity II. Induction of Tolerance to Systemic Anti-Inflammation Associated With Local Irritation and Major Surgery
Authors: Normann S J , Schardt M , Sorkin E
Published: January 2018

Anergy associated with cancer or major surgery may derive from the systemic antileukocyte effect induced by local inflammatory reactions (counter-irritation). Since the mechanism of the latter phenomenon is unknown, we approached the problem by asking if tolerance develops to repeated local irritant injections. Our results demonstrate that both tolerance and cross-desensitization occur rapidly to inflammatory agents (inflammagens) such as proteose peptone, thioglycollate, and carrageenan but not to the mitogens Con A, PHA-P, or LPS which also induce local inflammation. We interpret this data as supporting the notion that a common mechanism underlies the counter-irritant action of inflammagens but that injection of mitogens induces an additional mechanism of anti-inflammation distinguished from the former by its lack of tolerance induction. Based upon cross-desensitization experiments, we show that the anti-inflammatory effect of surgical amputation is analogous to that induced by inflammagens. In contrast, the systemic anti-inflammatory effect of tumor bearing, like that induced by mitogens, resists cross-desensitization suggesting that its mediation is not caused solely by the mechanism common to the counter-irritant action of inflammagens or major surgery.
Enhancement of Hamster Alveolar Macrophage Phagocytic Activity by Lymphokines
Authors: Widen Raymond H , Paradise Lois J
Published: January 2018

Modulation of phagocytic activity of resident hamster pulmonary alveolar macrophages was accomplished by incubation of the cells in lymphokines prepared by stimulation of hamster splenocytes with concanavalin A or alloantigens in mixed lymphocyte cultures. Alveolar macrophages preincubated in either of these lymphokine preparations possessed significantly greater ability to ingest IgG or IgM plus complement-coated sheep erythrocytes, via their Fc or complement receptors, respectively, than macrophages exposed to control preparations. Ingestion of yeast particles also was enhanced with macrophages incubated in supernatants from cultures of stimulated splenocytes. Supernatant fluids from either mitogen- or alloantigen-stimulated splenocytes possessed migration inhibitory activity with characteristics similar to MIF from other animals; the phagocytosis-enhancing activity shared some of these characteristics.
Effects of N-Nitrosodimethylamine on Tumor Susceptibility
Authors: Duke Scherer S , Schook Lawrence B , Holsapple Michael P
Published: January 2018

Dimethylnitrosamine (DMN) exposure altered the activity of the macrophage and natural killer (NK) cell defense mechanisms against the B16F10 melanoma In B6C3F1 adult female mice as assessed by several immunologic assays. Following 14 daily exposures (i.p.) to 1.5, 3.0, or 5.0 mg DMN/kg, mice were injected (i.v.) with B16F10 melanoma. The incidence and number of lung nodules, determined 18 days after challenge, were decreased in the DMN-exposed animals. The initial observation indicated the mice exposed to 3 mg/kg DMN were afforded the greatest protection. However, when mice exposed to the highest dose of DMN were divided into subgroups of mice with or without ascites, there was a degree protection seen in the 5-mg/kg-treated mice without ascites that was comparable to that of the 3-mg/kg group. The development of ascites is an overt toxic effect reflecting damage to the liver and was frequently associated with exposure to 5 mg/kg DMN. Exposure to DMN produced only slight changes in the activity of splenic NK cells as determined by the cytotoxicity of 51Cr-labelled YAC-1 cells. The activity was significantly increased only in mice exposed to 3 mg/kg DMN and only at effectortarget (E:T) ratios of 30:1 and 10:1. Interestingly, the activity of the NK cells was significantly decreased at all E:T ratios in mice exposed to 5 mg/kg that developed ascites. The number of peritoneal exudate cells was decreased, albeit nonsignificantly, in a dose-related fashion. The phagocytic activity, as measured by the uptake of fluorescent latex beads, was increased in a dose- related fashion with significance noted at the highest dose of DMN. The role of the macrophage in the increased resistance to the tumor challenge was assessed with bone marrow derived macrophages. The cytostatic activity versus B16F10 tumor cells, as measured by the uptake of 3H-thymidine, was markedly increased in the bone marrow derived macrophages from DMN (5 mg/kg) mice when compared to vehicle controls. These results suggest that exposure to DMN alters bone marrow, particularly the differentiation of effector tumoricidal cells, which renders the host more resistant to metastatic tumor formation.
The Effect of Local Antibody Responses on In Vivo and In Vitro Phagocytosis by Pulmonary Alveolar Macrophages
Authors: Harmsen Allen G , Bice David E , Muggenburg Bruce A
Published: January 2018

This study used both in vitro and in vivo techniques to determine if local antigen deposition in the lung has a localized effect on immune phagocytosis by pulmonary alveolar macrophages (PAM). Using a fiberoptic bronchoscope, dogs were immunized in the left cardiac and left diaphragmatic lobes with sheep red blood cells (sRBC). The right cardiac and right diaphragmatic lobes of the same animals received saline as controls. Unimmunized dogs served as additional controls. On days 2, 6, 9, 13, and 16 after immunization, the left and right diaphragmatic lobes were lavaged, and the cells and fluids were analyzed in vitro. Opsonizing antibody in lavage fluids was first detectable at 6 days, peaked at 9–13 days, and was significantly higher in the immunized lobe than in the control lobe. Phagocytosis of sRBC caused by cytophilic antibody on PAM also peaked at 9 to 13 days. Significantly more cytophilic antibody activity was detected on day 9 in the immunized lobes, than in the control lobes. In vivo phagocytosis of sRBC was evaluated in the alveoli of immunized and control lobes of immunized dogs and a control lobe of unimmunized dogs. Phagocytosis of sRBC by PAM in the immunized lobes was about four times greater than that of the control lobes and about 40 times greater than that of a control lobe of an unimmunized dog. These results indicate that the local deposition of a particulate antigen in the lung had a localized effect on immune phagocytosis. These data suggest that the accumulation of antibody-secreting cells in the alveolus may play a critical role in pulmonary defense mechanisms.
Reactivity of Anti-Asialo GM1 Serum With Tumoricidal and Non-Tumoricidal Mouse Macrophages
Authors: Wiltrout Robert H , Santoni Angela , Peterson Erik S , Knott Denise C , Overton W Roy , Herberman Ronald B , Holden Howard T
Published: January 2018

All peritoneal macrophage (pMφ) populations studied exhibited some binding of the anti-asGM1 serum as assessed by flow cytometry. The levels of reactivity varied quantitatively among populations, depending on the combination of eliciting and activating agents employed prior to the harvest of pMφ. Resident pMφ contained a very small percentage (4%) of cells that were strongly asGM1 +. Any treatment of these cells that induced them to become stimulated or activated increased the percentage of highly asGM1 + cells. Treatments that enhanced anti-asGMi binding including eliciting pMφ with proteose peptone (16% asGM1+) or Brewer's thioglycollate medium (66% asGM1+), treatment with the activating biological response modifiers (BRMs) MVE-2 (12% asGM1 +) and P acnes (18% asGM1 +), or treatment with both peptone + MVE-2 (37% asGM1 +) or peptone + poly IC/LC (33%). Increased expression of anti-asGM1 was accompanied by some increase in the reactivity of the various pMφ populations to treatment with anti-asGMj serum. This conclusion was based on the reduced viabilities of cells treated with both an eliciting agent and an activating agent prior to in vitro treatment with anti-asGM1 + C, as well as by reductions in cytolytic activity of pMφ elicited with peptone and activated by MVE-2, following anti-asGM1 treatment in vitro or administration in vivo. Conversely, the cytolytic activity of resident pMφ activated in vivo by MVE-2 or heat-killed P acnes, agents that induced relatively small increases in the percentage of asGM1φ cells, was resistant to the effects of in vivo and/or in vitro treatment with doses of anti-asGM1 serum that inhibit NK activity. These results indicate that stimulation of pMφ by eliciting or activating agents can increase the level of expression of asGM1 This increased expression of asGM1may be a useful marker for some aspects of macrophage heterogeneity, but increased expression is not necessarily directly related to expression of tumor- icidal activity. In fact, the results of this study demonstrate that anti-asGM1 serum can be used for specific depletion of NK activity in vivo in normal mice and in mice treated with at least some BRMs. However, the results also demonstrate that the use of eliciting agents, particularly thioglycollate, or eliciting agents in conjunction with activating agents can cause pMφ to become reactive with anti-asGM1 serum.
Effect of Prostaglandins on Complement Production by Tissue Macrophages
Authors: Rediske J J , Pickering R J
Published: January 2018

Tissue macrophages produce several proteins of the complement system. The mechanisms that regulate this process are poorly understood. The established ability of certain prostaglandins to influence macrophage secretory activity suggests that these lipid mediators may also modulate complement production (CP). Using the guinea pig peritoneal macrophages, we determined the effects of selected prostaglandins on in vitro CP and found that 1) PGE2 inhibited production of complement proteins but not lysozyme; 2) the response of elicited and resident peritoneal cells to PGE2 was identical; and 3) PGE1 PGF2 and PGI2 had no detectable effect. PGE2 may contribute to regulation of CP in vivo.
Interferon Produced Endogenously in Response to CSF-1 Augments the Functional Differentiation of Progeny Macrophages
Authors: Moore Robert N , Pitruzzello Frank J , Robinson Richard M , Rouse Barry T
Published: January 2018

The role of endogenously produced interferon α/β in the functional maturation of newly derived mononuclear phagocytes was investigated. Addition of highly specific anti-interferon α+ βantiserum to murine marrow cultures stimulated with colony-stimulating factor-1 (macrophage growth factor) markedly suppressed the capacity of resulting progeny mononuclear phagocytes to ingest opsonized sheep erythrocytes (EAlgG). This impairment was corrected either by direct addition of interferon α+ βat a concentration in excess of that neutralized by the antiserum or by the addition of lesser amounts of interferon (33 U/ml) following removal of the anti-interferon from the cultures. Conditioned media from control colony-stimulating factor-stimulated cultures similarly reversed the impairment of maturation resulting from 5 days of growth in the presence of anti-interferon. This enhancement of EA|gG ingestion reflected upon the interferon activity in the conditioned media and was neutralized by anti-interferon. Lastly, the endogenous interferon was found to enhance EAlgG ingestion by a majority of the mononuclear phagocyte progeny and not by a limited subpopulation.
Reactivation of Tritonated Models of Human Polymorphonuclear Leukocytes (PMNs): A Computer-Assisted Analysis
Authors: Cheung Anthony T W , Miller Michael E , Donovan Richard M , Goldstein Elliot , Kimura Gregory M
Published: January 2018

The orientation (chemotaxis) and locomotion (chemokinesis) of human polymorphonuclear leukocytes (PMNs) are generated by an internal movement mechanism that involves active cytoplasmic movement; they are influenced by external environmental and ionic conditions. We have studied the degree to which the orientation and movement mechanisms of PMNs are self-contained within the cell and the degree to which they are under membrane control.

PMNs were partially and selectively demembranated by treatment with the non-ionic detergent, octyl-phenoxyl-polyethoxyethanol (commercially known as Triton X-100) under controlled conditions. The tritonated PMNs (referred to in the literature as models) were non-motile and non-locomotory. Addition of ATP/Mg+ + with a trace amount of Ca+ + to the medium was followed by reactivation of the tritonated PMN models to move again as motile cells. Although these reactivated PMN models actively locomoted, they could no longer orient to chemoattractants. Thus, the reactivation process restored the physical self-contained movement parameters but could not reestablish the orientation capacity (chemotactic responsiveness) that was characteristic of live PMNs. The demembranation process apparently destroyed the chemotactic receptors and/or eradicated the coordination function of the membrane.

Videotapes of normal (control) as well as reactivated PMN movement were analyzed for movement characteristics. These characteristics were objectively analyzed with a newly designed computer-assisted micro-image-processing technique whereby the videotapes were digitized and quantified and the actual PMN movement printed out in computer-graphics and tracings (Freeman codes) for confirmation of orientation and movement arising as a result of reactivation.
Isolation of Canine Neutrophil Plasma Membranes
Authors: O'Donnell Robert T , Andersen Burton R
Published: January 2018

This paper describes the isolation of plasma membrane vesicles formed by nitrogen cavitation of canine neutrophils. Plasma membranes from disrupted cells were separated from other membranes and organelles by Percoll-density gradient centrifugation. Transmission electron microscopic examination of membrane preparations chromatographed on either Sephacryl S-1000 or Sepharose 4B revealed that two populations of plasma membrane vesicles were formed: large (176 ± 22nm), and small (119 ± 11 nm). Purified large vesicles were separated from Percoll and contaminating cytosol by Sephacryl S-1000 chromatography. Small vesicles were obtained free of Percoll by recavitating purified large vesicles. Problems encountered due to the presence of a soluble 5′-nucleotidase inhibitor also are discussed. Large and small membrane vesicles were separated into adherent and non-adherent populations by affinity chromatography on either concanavalin A-Sepharose or lentil lectin-Sepharose columns.
Accessory Cells Provide More Than One Signal for Lectin Mitogen-Stimulated Proliferation of Human Lymphocytes
Authors: Kern Jeffrey A , Daniele R P , Nowell P C
Published: January 2018

A culture system was developed in which human peripheral blood mononuclear cells (PBMC) depleted of la-expressing cells did not proliferate in response to the lectin mitogen phytohemagglutinin (PHA). These cells were able to respond to mitogen if purified autologous accessory cells were added back to the culture, thus showing an absolute requirement for la-expressing accessory cells in mitogen-driven T-cell proliferation. The identity of these accessory cells was shown to be not only monocytes but also la-expressing B cells and possibly other unidentified la-bearing cell types.

Human interleukin-1 (IL-1) in the lectin mitogen-unresponsive culture system was found unable to reconstitute normal human T-cell proliferation. This suggests that those cells with la surface antigen acting as accessory cells must deliver more than an IL-1 signal for T-cell proliferation.

The phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) was tested for its ability to replace necessary la-expressing accessory cells. TPA was able to replace accessory cells in culture, thus mimicking the la-expressing accessory cells and all their delivered signals in the triggering of human lymphocytes.
Continuous Expression of I-A Antigen by Peritoneal Macrophages From Mice Resistant to Mycobacterium bovis (Strain BCG)
Authors: Johnson Sarah C , Zwilling Bruce S
Published: January 2018

Monoclonal antibodies were used to monitor the expression of I-A antigen on the surface of macrophages obtained from mice immunized to Mycobacterium bovis (strain BCG). Unlike the transient nature of I-A expression by macrophages from Listeria-injected mice, peritoneal macrophages from mice injected 28 days previously with 104 BCG expressed I-A continuously. The continued expression was not due to the presence of antigen or of contaminating lymphocytes. When we compared the kinetics of I-A expression from different strains of mice, the continuous expression of I-A correlated with the genetic resistance of the mice to BCG. Macrophages from mice that were resistant to BCG expressed I-A continuously, while macrophages from BCG susceptible mice expressed I-A transiently. Injection of resistant mice with Salmonella typhimurium did not result in the induction of a population of macrophages that expressed I-A continuously. This suggests that the Bcg gene may not be the same as that responsible for resistance to Salmonella (Ity) or Leishmania (Lsh).
Truncation of neurokinin-1 receptor—Negative regulation of substance P signaling
Authors: Spitsin Sergei , Pappa Vasiliki , Douglas Steven D
Published: January 2018

Substance P (SP) is a tachykinin peptide, which triggers intracellular signaling in the nervous and immune systems, as well as, other local and systemic events. The interaction between SP and its receptor, neurokinin-1 receptor (NK1R), results in major downstream cellular actions, which include changes in calcium fluxes, ERK, and p21-activated kinase phosphorylation and NFκB activation. Two naturally occurring variants of the NK1R, the full-length, 407 aa receptor (NK1R-F) and the truncated, 311 aa isoform (NK1R-T), mediate the actions of SP. Receptor truncation partially disrupts signaling motifs of the carboxyl tail, a critical site for mediating NK1R signaling, resulting in a “less-efficient” receptor. Although NK1R-F is the predominant isoform in the central and peripheral nervous systems, NK1R-T is expressed in several tissues and cells, which include monocytes, NK cells, and T-cells. The SP binding domain is not affected by truncation and this site is identical in both NK1R receptor isoforms. However, while cells expressing NK1R-F respond to nanomolar concentrations of SP, monocyte and macrophage activation, mediated through NK1R-T, requires micromolar concentrations of SP in order to elicit signaling responses. Elevated plasma levels of SP are associated with increased inflammatory responses and NK1R antagonists reduce inflammation and cytokine production in vivo. This mini review presents and discusses the novel hypothesis that the expression of NK1R-T on immune system cells prevents immune activation in a milieu, which usually contains low concentrations of SP and, thus, maintains immune homeostasis. In contrast, in the activated neuronal microenvironment, when SP levels reach the threshold at tissue sites, SP promotes immune activation and modulates monocyte/macrophage polarization.
Partial Characterization of a Mitomycin-C Resistant T Cell
Authors: Anderson Robert E , Standefer Jimmy C , Troup Gary M , Pogue Laura E
Published: January 2018

A cell resistant to concentrations of mitomycin-C associated with the inactivation of many in vitro immune responses is described. This cell is responsive to concanavalin A (Con A), poorly adherent to nylon wool, present in relatively greater numbers in lymph node than other lymphoid tissues, and is relatively radiosensitive. In part, the resistance of this T cell appears to relate to a reduced activity of a microsomal reductase responsible for converting mitomycin-C to its more active form. Discrepancies in reductase activity may represent a way to differentiate among subsets of lymphocytes.
In Vitro Control of Neutrophilic Phagocytosis of IgG-Coated SRBC by Macromolecules Involved in Released Products From Platelets
Authors: Sakamoto Haruhiko , Yokoya Yukihiro , Ooshima Akira
Published: January 2018

Analysis of macromolecular fraction of platelet release products (PRPr) was undertaken after we observed that pretreatment of human neutrophils with macromolecules (>105 daltons) involved in PRPr enhanced phagocytosis of IgG-sensitized sheep red blood cells (IgG-SRBC). Sephadex G-200 gel filtration of PRPr revealed two macromolecular activators and two suppressors of neutrophilic phagocytosis of IgG-SRBC: MAPP(s), macromolecular activator(s) of phagocytosis from platelets; MSPP(s) macromolecular suppressor(s) of phagocytosis from platelets. The estimated molecular weights of the respective MAPPs were 100 kdaltons to 140 kdaltons (kd) (s-MAPP) and 200 kd to 290 kd (I-MAPP) and those of the respective MSPPs were 60 kd to 100 kd (s-MSPP) and 400 kd to 500 kd (I-MSPP). It is suggested that an l-MAPP molecule has at least an s-MAPP molecule as a functional unit and that MAPPs and larger MSPP (I-MSPP) might be glycoproteins, while smaller MSPP (s-MSPP) seems to contain neither carbohydrates nor lipids. As far as the effects of serum and plasma on phagocytosis of IgG-SRBC by neutrophils are concerned, the serum gave twice as much phagocytic activity as the plasma. Substantial MAPP-like activities were observed In the serum and MSPP-like activities were observed in both the serum and plasma.
Nitroblue Tetrazolium Reduction in Lymphocytes
Authors: Melinn Mary , McLaughlin Hugh
Published: January 2018

Certain populations of lymphocytes have been shown to reduce tetrazolium salts, indicating that superoxide anion may be present in lymphocytes. These experiments were done to determine if nitroblue tetrazolium reduction by lymphocytes was due to the presence of superoxide anion. Mitogen-activated lymphocytes showed increased nitroblue tetrazolium reduction compared to unstimulated cells, and the cells reducing nitroblue tetrazolium were both T-cells and non-T-cells. Release of superoxide anion or hydrogen peroxide by either resting or stimulated lymphocytes was not detected. There was no difference between resting and stimulated lymphocytes in the amount of chemiluminescence produced in the presence of lucigenin, an agent which appears to be sensitive to intracellular and extracellular superoxide anion. The results, then, indicate superoxide anion is not present in lymphocytes.
Low-Dose X-Irradiation and 4-Hydroperoxycyclophosphamide Distinguishes Three LYT-1+ Lymph Node T-Cell Subtypes That Respond to Specific Antigen In Vitro
Authors: Clark Connie , LaSota Irene , Borch Richard F
Published: January 2018

We investigated whether preculture exposure to low-dose X-irradiation and culture treatment with 4-hydroperoxycyclophosphamide (4-HPCY), a synthetic derivative of cyclophosphamide (CY), might distinguish subtypes of thymic-dependent (T) lymphocytes that respond to specific antigen in vitro. Lymph node (LN) cells were obtained from mice pretreated with CY and immunized with aggregated (A) human IgG (HGG) in Freund's complete adjuvant (CFA), and proliferation was assessed by incorporation of tritiated thymidine. Primed LN cells were untreated or exposed to low-dose irradiation before being cultured in medium alone and in medium containing 4-HPCY. The results show that these agents (irradiation and 4-HPCY) distinguished, in a dose-dependent manner, subtypes of T-cells which contribute to the specific antigen-stimulated proliferative response in vitro. For LN T-cells and LN Lyt-1 + T-cells, 20-25 rads and 1.0 μM 4-HPCY inactivated non-overlapping cell subtypes that respectively accounted for 26% and 28% of the response to HGG. The remaining 46% of HGG-responding cells were not affected by either agent. Although similar cell subtypes were discerned in unseparated LN cells, it required use of higher agent-doses. Cell cycle analysis revealed that treatment with irradiation, 4-HPCY, and the combination (both agents) caused S-phase arrest of 29%, 30%, and 55% of HGG-responding cells, respectively. Thus, identification of these cell subtypes could not be attributed to agent-mediated Inactivation of HGG-responding cells that might be in exclusively different phases of the cell cycle.
Detection of the Production of Oxygen-Centered Free Radicals by Human Neutrophils Using Spin Trapping Techniques: A Critical Perspective
Authors: Britigan Bradley E , Cohen Myron S , Rosen Gerald M
Published: January 2018

Human neutrophils generate oxygen reduction products as a consequence of membrane interactions with a number of stimuli. One oxygen-centered free radical (superoxide) has been uneqivocally shown to result from this “respiratory burst,” and some experimental evidence for another (hydroxyl radical) has been published, although debate remains as to its significance. The role of phagocyte-derived free radicals in microbicidal and tumoricidal activity as well as tissue damage at sites of inflammation has been the focus of extensive investigation in recent years. Of the techniques available to study free radical generation in biological systems, spin trapping has emerged as a powerful tool for detection and identification of these reactive species, owing in part to its ability to measure production of free radicals inside the phagosome. However, Interpretation of resulting spectra is extremely complex and filled with pitfalls and limitations. In this communication we review spin-trapping techniques and discuss the application of this system to the identification of free radicals resulting from stimulation of human neutrophils. Criteria are established which are necessary for definitive identification of superoxide and hydroxyl radical when employing this technology. In this context a critical perspective of previous studies of neutrophil-derived free radicals is offered.