Increased Macrophage Activation Research Articles

P311 Influences Macrophage Polarization

Obesity is a pandemic disease that has become a serious concern worldwide. In United States, recently self-reported obesity rates of adults were escalated to 40%. In the early phase of obesity, excessive macrophage infiltration and disequilibrium of macrophage phenotypes (M1 vs. M2 or vice versa) play a critical role in the microenvironment of adipose tissue. It often leads to chronic inflammation and is a major cause for metabolic disorders including insulin resistance, type-2 diabetes, and cardiovascular diseases. P311 is an intrinsically disordered protein, contributes to adipogenesis as transcription regulator through activation of PPARγ2, master regulator of adipogenesis. However, the P311 role on the macrophage polarization was not known. Here we studied the macrophage polarization in presence of P311 using Raw 264.7 cells. Raw cells were treated with either interleukin-4 (IL4) or lipopolysaccharide (LPS) for macrophage polarization towards M2 and M1 phenotype respectively in presence and absence of P311 overexpression. Macrophage phenotypic switch was assessed using CD206 (M2 marker) and CD86 (M1 marker) expression. The dose and time dependent effects of IL4 and LPS on the expression of P311 and macrophage markers was also studied. A significant increase in endogenous P311 expression observed with IL4 treatment whereas LPS treatment showed the opposite effect. Our findings showed that the P311 overexpression promotes anti-inflammatory macrophage phenotype (M2) as indicated by the significant increase in mRNA and protein expression of CD206 when compared to empty vector (EV, pCMV) transfected control group. Further, P311 overexpression in LPS treated RAW cells significantly increased the expression of CD206, on contrary to the classical M1 polarization of LPS, compared to EV transfected cells. In addition, P311 suppressed the expression of CD86 there by facilitating macrophage switch towards anti-inflammatory phenotype in presence of LPS treatment. IL4 treatment alone increased macrophage activation towards M2 phenotype however IL4 treated P311 overexpressed cells showed further increase in CD206 expression compared to EV transfected cells. Our studies indicate that the P311 expression plays an important role in promoting macrophage polarization toward M2 phenotype. This study was supported by research funding to KB, from NIH/NIGMS #SC1GM141937. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Empowering macrophages: the cancer fighters within the tumour microenvironment in mantle cell lymphoma.

In Mantle Cell Lymphoma (MCL), the role of macrophages within the tumour microenvironment (TME) has recently gained attention due to their impact on prognosis and response to therapy. Despite their low absolute number in MCL tumour tissue, recent findings reveal an association between the levels of macrophages and prognosis, consistent with trends observed in other lymphoma subtypes. M2-like macrophages, identified by markers such as CD163, contribute to angiogenesis and suppression of the immune response. Clinical trials with MCL patients treated with chemoimmunotherapy and targeted treatments underscore the adverse impact of high levels of M2-like macrophages. Immunomodulatory drugs like lenalidomide reduce the levels of MCL-associated CD163+ macrophages and enhance macrophage phagocytic activity. Similarly, clinical approaches targeting the CD47 "don't eat me" signalling, in combination with the anti-CD20-antibody rituximab, demonstrate increased macrophage activity and phagocytosis of MCL tumour cells. Cell-based therapies such as chimeric antigen receptor (CAR) T-cell have shown promise but various challenges persist, leading to a potential interest in CAR-macrophages (CAR-M). When macrophages are recruited to the TME, they offer advantages including phagocytic function and responsiveness to microenvironment alterations, suggesting their potential as a manipulable and inducible alternative when CAR T-cell therapies fails in the complex landscape of MCL treatment.

Pengaruh Mucoadhesive Plester Ekstrak Daun Ciplukan (Physalis angulata L.) terhadap Jumlah Limfosit dalam Penyembuhan Ulkus Traumatikus Tikus Putih (Rattus norvegicus)

Traumatic ulcer is an open wound in the oral cavity caused by trauma. Flavonoids in ciplukan leaf have an antiinflammatory effect. Drug delivery using mucoadhesive plasters can increase the effectiveness, absorption, and bioavailability of drugs because directly attached to the wound. Lymphocytes can increase macrophage activation that can accelerate wound healing. The purpose of this research is to determine the effect of mucoadhesive plaster of ciplukan leaf extract (Physalis angulata L.) on the number of lymphocytes in healing traumatic ulcers of white rats (Rattus norvegicus). 30 rats were divided into 5 groups; the negative control group, the positive control group and the treatment group with mucoadhesive plaster ciplukan leaf extract doses of 100, 200, 400 mg/kg BW. Conducting heat induction on the labial mucosa of white rats using an amalgam stopper and then given treatment according to groups. The rats were dissected on day 5. The number of lymphocytes was counted on the preparate with Hematoxylin Eosin staining using an Olympus digital microscope with 400x magnification. The treatment group with mucoadhesive plaster of ciplukan leaf extract dose 100 mg/kg BB had the highest number of lymphocytes, while the positive control group had the lowest number of lymphocytes. The conclusion of this research is mucoadhesive plaster of ciplukan leaf extract (Physalis angulata L.) affects the number of lymphocytes in healing traumatic ulcer of white rats (Rattus norvegicus).

Differences peripheral of blood in HIV patients with and without cognitive disorders at “Prof. Dr. IGNG Ngoerah” Hospital

Background and objective. The prevalence of mild neurocognitive Disorder (MND) has increased after using antiretroviral therapy (ARV), which is around 51.5%. Inflammation plays an essential role in the neurodegenerative state of HIV patients. Peripheral blood images can assess increased macrophage activation by evaluating the number of leukocytes, neutrophils, total lymphocytes, hemoglobin, platelets, platelets, and neutrophil-lymphocyte ratio (NLR) to determine the condition of systemic inflammation in HIV patients with cognitive impairment. This study to know differences in the appearance of peripheral blood in HIV patients with and without cognitive impairment. Method. Observational analytic study with a cross-sectional design on 44 HIV patients who were outpatients at the polyclinic Voluntary Counseling and Testing (VCT) at Prof Dr. IGNG Ngoerah Denpasar Hospital. Subjects were divided into 2 groups based on cognitive status assessed using the MoCA-INA questionnaire. The peripheral blood images considered were the average levels of leukocytes, neutrophils, lymphocytes, hemoglobin, hematocrit, platelets, and the ratio of neutrophils to lymphocytes. Results. 31-40 years old, male, with >9 years of education, has a CD4 count >200, and has received ARV therapy >1 year. The Shapiro-Wilk normality test performed on both groups showed that the data were normally distributed with p>0.05. Bivariate analysis using an independent t-test with a 95% confidence level (α=0.05) found significant differences in mean leukocyte, neutrophil, lymphocyte, and neutrophil-lymphocyte ratios (p<0.05) between groups of HIV patients with cognitive impairment and no cognitive impairment. Conclusion. Inflammatory markers from the peripheral blood picture are increased in HIV-infected persons.

FKN secreted by kidney epithelial cells regulates macrophage activation in lupus nephritis via the Hippo signaling pathway.

Lupus nephritis (LN) is a serious complication of systemic lupus erythematosus (SLE), and its pathogenesis is not fully understood. Previously, we showed that fractalkine (FKN) expression was positively correlated with the severity of LN. Here, we aimed to study the role of the Hippo signaling pathway (HSP) and its interaction with FKN in LN in an attempt to provide novel strategies for LN treatment. In this study, lipopolysaccharide (LPS)/interferon-γ (IFN-γ)-stimulated THP-1 cells were co-cultured with FKN up-regulated or down-regulated kidney epithelial cells Hkb20. FKN-knockout (KO-FKN) mice were used to construct LN model. Flow cytometric analysis, quantitative real-time polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), pathological staining, Western blot, and immunofluorescence (IF) staining were employed to investigate the role of FKN and its interaction with the Hippo signaling pathway (HSP) in LN. Up-regulation of FKN in kidney epithelial cells was associated with increased macrophage activation. FKN overexpression in kidney epithelial cells suppressed apoptosis, inflammation levels, and M1 polarization of THP-1 cells and inhibited the HSP. Oppositely, FKN knockdown in kidney epithelial cells increased apoptosis, inflammation, and M1 polarization and activated the HSP. HSP inhibitor reversed the effect of FKN knockdown on THP-1 cells. In LN mice, FKN knockout and YAP inhibitor decreased the levels of renal function markers, alleviated kidney injury induced by LN, and inhibited macrophage activation in LN mice. FKN down-regulation reduced the activation of macrophages in renal tissue and alleviated kidney damage by activating HSP. The regulatory effect of FKN on HSP should be confirmed in patients with LN, and the mechanism of FKN in LN should be further explored.

The Stria Vascularis in Mice and Humans Is an Early Site of Age-Related Cochlear Degeneration, Macrophage Dysfunction, and Inflammation.

Age-related hearing loss, or presbyacusis, is a common degenerative disorder affecting communication and quality of life for millions of older adults. Multiple pathophysiologic manifestations, along with many cellular and molecular alterations, have been linked to presbyacusis; however, the initial events and causal factors have not been clearly established. Comparisons of the transcriptome in the lateral wall (LW) with other cochlear regions in a mouse model (of both sexes) of "normal" age-related hearing loss revealed that early pathophysiological alterations in the stria vascularis (SV) are associated with increased macrophage activation and a molecular signature indicative of inflammaging, a common form of immune dysfunction. Structure-function correlation analyses in mice across the lifespan showed that the age-dependent increase in macrophage activation in the stria vascularis is associated with a decline in auditory sensitivity. High-resolution imaging analysis of macrophage activation in middle-aged and aged mouse and human cochleas, along with transcriptomic analysis of age-dependent changes in mouse cochlear macrophage gene expression, support the hypothesis that aberrant macrophage activity is an important contributor to age-dependent strial dysfunction, cochlear pathology, and hearing loss. Thus, this study highlights the SV as a primary site of age-related cochlear degeneration and aberrant macrophage activity and dysregulation of the immune system as early indicators of age-related cochlear pathology and hearing loss. Importantly, novel new imaging methods described here now provide a means to analyze human temporal bones in a way that had not previously been feasible and thereby represent a significant new tool for otopathological evaluation.SIGNIFICANCE STATEMENT Age-related hearing loss is a common neurodegenerative disorder affecting communication and quality of life. Current interventions (primarily hearing aids and cochlear implants) offer imperfect and often unsuccessful therapeutic outcomes. Identification of early pathology and causal factors is crucial for the development of new treatments and early diagnostic tests. Here, we find that the SV, a nonsensory component of the cochlea, is an early site of structural and functional pathology in mice and humans that is characterized by aberrant immune cell activity. We also establish a new technique for evaluating cochleas from human temporal bones, an important but understudied area of research because of a lack of well-preserved human specimens and difficult tissue preparation and processing approaches.

Activation of estrogen receptor α inhibits TLR4 signaling in macrophages and alleviates the instability of atherosclerotic plaques in the postmenopausal stage

Acute cardiovascular events increase significantly in postmenopausal women. The relationship between estrogen receptor (ER) and plaque stability in the postmenopausal stage remains to be elucidated. We aimed to explore whether ERα activation improves plaque instability in the postmenopausal stage. Here, we report that postmenopausal women showed increased macrophage activation and plaque instability with increased MCP-1, MMP9, TLR4, MYD88 and NF-κB p65 and decreased ERα and TIMP1 expression in the vascular endothelium. Moreover, ovariectomy in LDLR-/- mice resulted in a significant increase in plaque area and necrotic core area, as well as a significant decrease in collagen content and an increase in macrophage accumulation in the artery. Ovariectomy also reduced serum estrogen levels and ERα expression and upregulated TLR4 and MMP9 expression in arteries in LDLR-/- mice. Estrogen or phytoestrogen therapy upregulated the expression level of ERα in ovariectomized mice and increased plaque stability by inhibiting macrophage accumulation and TLR4 signaling. In vitro, LPS incubation of RAW264.7 cells resulted in a significant decrease in ERα and TIMP1 expression and an increase in TLR4 activation, and estrogen or phytoestrogen treatment increased ERα and TIMP1 expression and inhibited TLR4 activation and MMP9 expression in LPS-treated RAW264.7 cells. Compared to control siRNA transfected RAW264.7 cells, TLR4 siRNA promoted TIMP1 expression in RAW264.7 cells with LPS incubation, but did not affect ERα expression in RAW264.7 cells with or without LPS treatment. The ERα inhibitor MPP abolished the regulatory effect of estrogen or phytoestrogen on LPS-induced RAW264.7 cells. In conclusion, the present study demonstrates that decreased ERα expression promotes macrophage infiltration and plaque instability in the postmenopausal stage, and activation of ERα in the postmenopausal stage alleviates atherosclerotic plaque instability by inhibiting TLR4 signaling and macrophage-related inflammation.

Living with chronic infection: Persistent immunomodulation during avirulent haemoparasitic infection in a wild rodent.

Apicomplexans are a protozoan phylum of obligate parasites which may be highly virulent during acute infections, but may also persist as chronic infections which appear to have little fitness cost. Babesia microti is an apicomplexan haemoparasite that, in immunocompromised individuals, can cause severe, potentially fatal disease. However, in its natural host, wild field voles (Microtus agrestis), it exhibits chronic infections that have no detectable impact on survival or female fecundity. How is damage minimized, and what is the impact on the host's immune state and health? We examine the differences in immune state (here represented by expression of immune-related genes in multiple tissues) associated with several common chronic infections in a population of wild field voles. While some infections show little impact on immune state, we find strong associations between immune state and B. microti. These include indications of clearance of infected erythrocytes (increased macrophage activity in the spleen) and activity likely associated with minimizing damage from the infection (anti-inflammatory and antioxidant activity in the blood). By analysing gene expression from the same individuals at multiple time points, we show that the observed changes are a response to infection, rather than a risk factor. Our results point towards continual investment to minimize the damage caused by the infection. Thus, we shed light on how wild animals can tolerate some chronic infections, but emphasize that this tolerance does not come without a cost.

Abstract 15356: Serial Evaluation of Chemotherapy-Induced Vascular Toxicity in a Rat Model Using 18 F-FDG PET/CT Imaging

Introduction: Cancer treatment with chemotherapeutics can lead to off-target effects on the vasculature. Fluorine-18 ( 18 F)-fluorodeoxyglucose (FDG) PET/CT imaging is standard of care for staging and monitoring of numerous cancers and may offer a tool for assessing vascular toxicity. We hypothesized that PET/CT imaging could quantify changes in chemotherapy-induced arterial inflammation. Thus, we sought to validate PET/CT imaging as a tool for assessing vascular toxicity in a rat model using in vivo imaging and post-mortem analyses. Methods: Male Wistar rats (n = 11 total) were divided into a control group that received twice weekly IV saline (n=5; 1 ml/kg) or a group that received twice weekly IV doxorubicin (n=6; 1 mg/kg) for 2.5 weeks (i.e., 7 mg/kg cumulative dose). Rats underwent 18 F-FDG microPET/CT imaging of the abdominal aorta (AA) at 3 time points (baseline, after 4 mg/kg dosing, and after 7 mg/kg cumulative dose) to quantify serial changes in arterial inflammation, expressed as changes in the maximum standard uptake value (SUV max ). Following terminal imaging, AA tissue was harvested and cut into 3 mm sections for gamma counting of 18 F-FDG uptake, hematoxylin and eosin morphological staining, and immunohistochemistry of macrophage expression. Results: In vivo image analyses demonstrated that 18 F-FDG uptake remained unchanged for the control group during the treatment period, but significantly increased from baseline levels in the doxorubicin-treated group after 4 mg/kg (p=0.0007) and 7 mg/kg (p=0.001) cumulative dosing. In vivo image analyses were confirmed by post-mortem gamma counting of 18 F-FDG uptake in anatomically-matched AA tissue samples. Histological analyses revealed increased levels of macrophage expression in sections with increased 18 F-FDG uptake identified by in vivo and ex vivo analyses. Conclusions: 18 F-FDG PET/CT imaging non-invasively quantifies serial changes in chemotherapy-induced arterial toxicity that are at least partially mediated by increased macrophage activity. Routine assessment of 18 F-FDG uptake on PET/CT imaging studies during cancer treatment could provide a translatable approach for characterizing the time course of vascular toxicity associated with various chemotherapeutics.

Peripheral Auditory Nerve Impairment in a Mouse Model of Syndromic Autism.

Dysfunction of the peripheral auditory nerve (AN) contributes to dynamic changes throughout the central auditory system, resulting in abnormal auditory processing, including hypersensitivity. Altered sound sensitivity is frequently observed in autism spectrum disorder (ASD), suggesting that AN deficits and changes in auditory information processing may contribute to ASD-associated symptoms, including social communication deficits and hyperacusis. The MEF2C transcription factor is associated with risk for several neurodevelopmental disorders, and mutations or deletions of MEF2C produce a haploinsufficiency syndrome characterized by ASD, language, and cognitive deficits. A mouse model of this syndromic ASD (Mef2c-Het) recapitulates many of the MEF2C haploinsufficiency syndrome-linked behaviors, including communication deficits. We show here that Mef2c-Het mice of both sexes exhibit functional impairment of the peripheral AN and a modest reduction in hearing sensitivity. We find that MEF2C is expressed during development in multiple AN and cochlear cell types; and in Mef2c-Het mice, we observe multiple cellular and molecular alterations associated with the AN, including abnormal myelination, neuronal degeneration, neuronal mitochondria dysfunction, and increased macrophage activation and cochlear inflammation. These results reveal the importance of MEF2C function in inner ear development and function and the engagement of immune cells and other non-neuronal cells, which suggests that microglia/macrophages and other non-neuronal cells might contribute, directly or indirectly, to AN dysfunction and ASD-related phenotypes. Finally, our study establishes a comprehensive approach for characterizing AN function at the physiological, cellular, and molecular levels in mice, which can be applied to animal models with a wide range of human auditory processing impairments.SIGNIFICANCE STATEMENT This is the first report of peripheral auditory nerve (AN) impairment in a mouse model of human MEF2C haploinsufficiency syndrome that has well-characterized ASD-related behaviors, including communication deficits, hyperactivity, repetitive behavior, and social deficits. We identify multiple underlying cellular, subcellular, and molecular abnormalities that may contribute to peripheral AN impairment. Our findings also highlight the important roles of immune cells (e.g., cochlear macrophages) and other non-neuronal elements (e.g., glial cells and cells in the stria vascularis) in auditory impairment in ASD. The methodological significance of the study is the establishment of a comprehensive approach for evaluating peripheral AN function and impact of peripheral AN deficits with minimal hearing loss.

Integrated metabolism and epigenetic modifications in the macrophages of mice in responses to cold stress.

The negative effects of low temperature can readily induce a variety of diseases. We sought to understand the reasons why cold stress induces disease by studying the mechanisms of fine-tuning in macrophages following cold exposure. We found that cold stress triggers increased macrophage activation accompanied by metabolic reprogramming of aerobic glycolysis. The discovery, by genome-wide RNA sequencing, of defective mitochondria in mice macrophages following cold exposure indicated that mitochondrial defects may contribute to this process. In addition, changes in metabolism drive the differentiation of macrophages by affecting histone modifications. Finally, we showed that histone acetylation and lactylation are modulators of macrophage differentiation following cold exposure. Collectively, metabolism-related epigenetic modifications are essential for the differentiation of macrophages in cold-stressed mice, and the regulation of metabolism may be crucial for alleviating the harm induced by cold stress.

IL‐25 Supplementation Induces M2 Macrophage Polarization, Reduces Blood Pressure, and Improves Fetal Weight in Placental Ischemic Rats

Preeclampsia (PE) is characterized by maternal hypertension, intrauterine growth restriction (IUGR), and increased macrophage activation. Macrophages can be broadly categorized into pro‐inflammatory M1 macrophages (M1s) and anti‐inflammatory M2 macrophages (M2s). The M1:M2 ratio is significantly increased in PE women. However, the precise contribution of increased M1s to PE pathophysiology remains unclear. Using the reduced uterine perfusion pressure (RUPP) rat model of placental ischemia, we tested the hypothesis that supplementation of RUPP rats with IL‐25, a cytokine that drives macrophage polarization to anti‐inflammatory M2s, will increase M2s, decrease mean arterial pressure (MAP), and improve fetal growth. On gestation day (GD) 14, timed pregnant 10‐12 week old Sprague Dawley rats underwent the RUPP or Sham procedure and a mini‐osmotic pump infusing IL‐25 (0.5µg/day) or vehicle was implanted i.p., in both groups. Carotid catheters were placed on GD18. On GD19, MAP was measured, animals were euthanized, and blood and tissues were collected for analysis. Flow cytometry analysis was performed on placental and renal tissues. M2 macrophages (CD3‐/CD68+/CD163+) were markedly reduced in placentas of RUPP rats compared to Sham (0.2% vs. 4.2% of total macrophages) and in kidneys of RUPP rats vs. Sham (0.19% vs. 1.445% total of macrophages). IL‐25 supplementation elevated M2 populations in placental (3.985%) and renal (5.985%) tissues of RUPP. Likewise, IL‐25 treatment increased M2 macrophages in Sham placental (18.56%) and renal (12.99%) tissues. Increased MAP was observed in RUPP rats compared with Sham (123.5 mmHg vs. 106.5 mmHg, respectively) and was reduced in RUPP + IL‐25 (107.5 mmHg). No changes were observed in MAP of Sham rats treated with IL‐25. Fetal weights were reduced in RUPP vs. Sham (2.035g vs. 2.405g, respectively) and were normalized in RUPP after IL‐25 supplementation (2.425g). IL‐25 supplementation had no effect on fetal weight of Sham rats. Additional analyses will include assessment of blood and tissue cytokines and reactive oxygen species, and vascular function in all groups. In conclusion, IL‐25 supplementation increased placental and renal anti‐inflammatory M2 macrophage populations, reduced MAP, and improved fetal weights in RUPP recipients. These data suggest that macrophage activation is a key contributor to PE pathophysiology and may represent a therapeutic target to improve maternal and fetal outcomes in PE.

Archetypal Analysis of Injury in Kidney Transplant Biopsies Identifies Two Classes of Early AKI.

All transplanted kidneys are subjected to some degree of injury as a result of the donation-implantation process and various post-transplant stresses such as rejection. Because transplants are frequently biopsied, they present an opportunity to explore the full spectrum of kidney response-to-wounding from all causes. Defining parenchymal damage in transplanted organs is important for clinical management because it determines function and survival. In this study, we classified the scenarios associated with parenchymal injury in genome-wide microarray results from 1,526 kidney transplant indication biopsies collected during the INTERCOMEX study. We defined injury groups by using archetypal analysis (AA) of scores for gene sets and classifiers previously identified in various injury states. Six groups and their characteristics were defined in this population: No injury, minor injury, two classes of acute kidney injury (“AKI,” AKI1, and AKI2), chronic kidney disease (CKD), and CKD combined with AKI. We compared the two classes of AKI, namely, AKI1 and AKI2. AKI1 had a poor function and increased parenchymal dedifferentiation but minimal response-to-injury and inflammation, instead having increased expression of PARD3, a gene previously characterized as being related to epithelial polarity and adherens junctions. In contrast, AKI2 had a poor function and increased response-to-injury, significant inflammation, and increased macrophage activity. In random forest analysis, the most important predictors of function (estimated glomerular filtration rate) and graft loss were injury-based molecular scores, not rejection scores. AKI1 and AKI2 differed in 3-year graft survival, with better survival in the AKI2 group. Thus, injury archetype analysis of injury-induced gene expression shows new heterogeneity in kidney response-to-wounding, revealing AKI1, a class of early transplants with a poor function but minimal inflammation or response to injury, a deviant response characterized as PC3, and an increased risk of failure. Given the relationship between parenchymal injury and kidney survival, further characterization of the injury phenotypes in kidney transplants will be important for an improved understanding that could have implications for understanding native kidney diseases (ClinicalTrials.gov #NCT01299168).

“Disseminated histoplasmosis and erythrophagocytosis in an immunocompromised host: the role of bone marrow evaluation for prompt diagnosis of invasive fungal infections”

“Disseminated histoplasmosis and erythrophagocytosis in an immunocompromised host: the role of bone marrow evaluation for prompt diagnosis of invasive fungal infections”

Formulation of Black Mulberry Fruit (Morus nigra l.) Ethanol Extract as Anti-Acne Cream Against Propionibacterium acne

Various parts of mulberry have been identified consisting of a large number of phenols and flavonoids, which are effective for immunosuppression, antioxidants, and are antibacterial, and anticancer agents. A polysaccharide and an alkaloid related to increased macrophage activity from mulberry fruit extracts. This study aims to formulate black mulberry fruit into cream dosage form and determine the antibacterial activity against Propionibacterium acnes. This is an experimental research which varying extract into 4 different concentrations: 6.25%, 12.5%, 25%, and 50%. There is two control group which is positive using vitacid cream and the negative control is cream base without extract. The cream was made and examined for its physical characteristic and the cream was tested for antibacterial activity using the Kirby-Bauer disk diffusion method. The result showed that the cream met the requirements of homogeneity, pH as its requirement, stable during storage, not irritating, and the antibacterial activity of 6,25% concentration has an average inhibition zone diameter of 10.15mm, a concentration of 12,5% is 12.36mm, a concentration of 25% is 13.16mm, and a concentration of 50% is 15.5 mm. From this study can be concluded that the cream preparation from an ethanol extract of black mulberry has good antibacterial activity against P.acne bacteria with a very strong inhibition zone.

Effect of oral intake of royal jelly on endothelium function in hemodialysis patients: study protocol for multicenter, double-blind, randomized control trial

BackgroundHemodialysis (HD) is a common renal replacement therapy for patients with renal failure. Cardiovascular and cerebrovascular diseases are known to shorten survival periods and worsen the quality of life of HD patients. Atherosclerosis is a major cause of vascular diseases, and various factors such as abnormality of lipid metabolism and increased macrophage activity, oxidative stress, and endothelial dysfunction are associated with its pathogenesis and progression. Further, endothelial stem cells (ESCs) have been reported to play important roles in endothelial functions. Royal jelly (RJ) affects atherosclerosis- and endothelial function-related factors. The main aim of this trial is to investigate whether oral intake of RJ can maintain endothelial function in HD patients. In addition, the effects of RJ intake on atherosclerosis, ESC count, inflammation, and oxidative stress will be analyzed.MethodsThis will be a multicenter, prospective, double-blind, randomized controlled trial. We will enroll 270 participants at Nagasaki Jin Hospital, Shinzato Clinic Urakami, and Maeda Clinic, Japan. The participants will be randomized into RJ and placebo groups. The trial will be conducted according to the principles of the Declaration of Helsinki, and all participants will be required to provide written informed consent. The RJ group will be treated with 3600 mg/day of RJ for 24 months, and the placebo group will be treated with starch for 24 months. The primary endpoint will be the change in flow-mediated dilation (FMD), a parameter of endothelium function, from the time before treatment initiation to 24 months after treatment initiation. The secondary and other endpoints will be changes in FMD; ESC count; serum levels of vascular endothelial cell growth factor, macrophage colony-stimulating factor, 8-hydroxydeoxyguanosine, and malondialdehyde; the incidence of cardiovascular diseases, cerebrovascular diseases, and stenosis of blood access; and safety.DiscussionThis trial will clarify whether oral intake of RJ can maintain endothelial function and suppress the progression of atherosclerosis in HD patients. In addition, it will clarify the effects of RJ on ESCs, oxidative stress, and angiogenic activity in blood samples.Trial registrationThe Japan Registry of Clinical Trials jRCTs071200031. Registered on 7 December 2020.

Are Serum Ion Levels Elevated in Pediatric Patients With Metal Implants?

Previous studies report elevated serum titanium (Ti) levels in children with spinal implants. To provide additional data on this topic, we sought to assess serum ion levels at multiple timepoints in pediatric patients with growing spine devices, spinal fusion instrumentation, and extremity implants placed for fracture treatment. We hypothesized that serum Ti, cobalt (Co), and chromium (Cr) levels would be elevated in pediatric patients with growing spine devices compared with patients with extremity implants. Pediatric patients undergoing any primary spine implant placement, those with spine implant revision or removal surgery and patients with other appendicular implant removal had serum Ti, Co, and Cr ion levels drawn at the time of surgery. Fifty-one patients (12 growing spine devices, 13 fusions, and 26 extremity implants) had one set of labs, 31 of whom had labs drawn both preoperatively and postoperatively. Biopsies obtained from tissue specimens at the time of implant revision were analyzed histologically for the presence of metal debris and macrophage activity. Patients with growing spine implants had elevated serum Ti (3.3 vs. 1.9 ng/mL, P=0.01) and Cr levels (1.2 vs. 0.27 ng/mL, P=0.01) in comparison to patients with fusion rods or extremity implants. With respect to patients with extremity implants, patients with growing spine devices had elevated serum Ti (3.3 vs. 0.98 ng/mL, P=0.013), Co (0.63 vs. 0.26 ng/mL, P=0.017), and Cr levels (1.18 vs. 0.26 ng/mL, P=0.005). On matched pairs analysis, patients who had labs drawn before and after spine implantation had significant increase in serum Ti levels (0.57 vs. 3.3 ng/mL, P=0.02). Histology of tissue biopsies adjacent to growing spine implants showed presence of metal debris and increased macrophage activity compared with patients with extremity implants. Serum Ti, Co, and Cr levels are elevated in children with spinal implants compared with those with extremity implants, particularly in those with growing spine devices. However, the clinical significance of these findings remains to be determined. Level II-prospective comparative study.

Spinal cord injury in mice affects central and peripheral pathology in a severity-dependent manner.

Chronic pain is a common medical complication experienced by those living with spinal cord injury (SCI) and leads to worsened quality of life. The pathophysiology of SCI pain is poorly understood, hampering the development of safe and efficacious therapeutics. We therefore sought to develop a clinically relevant model of SCI with a strong pain phenotype and characterize the central and peripheral pathology after injury. A contusion (50 kdyn) injury, with and without sustained compression (60 seconds) of the spinal cord, was performed on female C57BL/6J mice. Mice with compression of the spinal cord exhibited significantly greater heat and mechanical hypersensitivity starting at 7 days postinjury, concomitant with reduced locomotor function, compared with those without compression. Immunohistochemical analysis of spinal cord tissue revealed significantly less myelin sparing and increased macrophage activation in mice with compression compared with those without. As measured by flow cytometry, immune cell infiltration and activation were significantly greater in the spinal cord (phagocytic myeloid cells and microglia) and dorsal root ganglia (Ly6C+ monocytes) after compression injury. We also decided to investigate the gastrointestinal microbiome, as it has been shown to be altered in patients with SCI and has recently been shown to play a role in immune system maturation and pain. We found increased dysbiosis of the gastrointestinal microbiome in an injury severity-dependent manner. The use of this contusion-compression model of SCI may help advance the preclinical assessment of acute and chronic SCI pain and lead to a better understanding of mechanisms contributing to this pain.

Pharmacological Activities for Morus alba L., Focusing on the Immunostimulatory Property from the Fruit Aqueous Extract.

Depending on the extraction method, numerous compounds that have specific pharmacological effects can be obtained from M. alba L. There is a growing scientific interest in health problems related to aging. Efforts to develop safe immune-enhancing pharmaceuticals are increasing. This review aims to summarize and critically discuss the immunity enhancement effects and pharmaceutical efficacy of M. alba L. extracts. The scientific database search was conducted using Google Scholar, Web of Science, and PubMed until May 2021. Additional articles were identified and obtained from references in the retrieved articles. Ethanol or methanol extraction of various parts of M. alba L. identified a large amount of phenols and flavonoids, which are effective for immunosuppression, antioxidants, and cardiovascular diseases, and are antibacterial, and anticancer. Water extraction of M. alba L. enhanced the innate immune response based on immune cell activation. A polysaccharide and an alkaloid related to increased macrophage activity were isolated from M. alba L. fruit extracts. M. alba L. fruit water extracts primarily induced the production of pro-inflammatory substances, in model organisms, via TLR4 in immune cells. Water extracts have been shown to be effective in pathogen defense and tumor suppression by enhancing macrophage activity. Based on our literature review on the bioactivity of M. alba L. fruit extracts, particularly in relation to their immunity enhancement activity, we anticipate that M. alba-derived pharmaceuticals will have excellent potential in future medical research.

Low Intensity Shockwave Treatment Modulates Macrophage Functions Beneficial to Healing Chronic Wounds.

Extracorporeal Shock Wave Therapy (ESWT) is used clinically in various disorders including chronic wounds for its pro-angiogenic, proliferative, and anti-inflammatory effects. However, the underlying cellular and molecular mechanisms driving therapeutic effects are not well characterized. Macrophages play a key role in all aspects of healing and their dysfunction results in failure to resolve chronic wounds. We investigated the role of ESWT on macrophage activity in chronic wound punch biopsies from patients with non-healing venous ulcers prior to, and two weeks post-ESWT, and in macrophage cultures treated with clinical shockwave intensities (150–500 impulses, 5 Hz, 0.1 mJ/mm2). Using wound area measurements and histological/immunohistochemical analysis of wound biopsies, we show ESWT enhanced healing of chronic ulcers associated with improved wound angiogenesis (CD31 staining), significantly decreased CD68-positive macrophages per biopsy area and generally increased macrophage activation. Shockwave treatment of macrophages in culture significantly boosted uptake of apoptotic cells, healing-associated cytokine and growth factor gene expressions and modulated macrophage morphology suggestive of macrophage activation, all of which contribute to wound resolution. Macrophage ERK activity was enhanced, suggesting one mechanotransduction pathway driving events. Collectively, these in vitro and in vivo findings reveal shockwaves as important regulators of macrophage functions linked with wound healing. This immunomodulation represents an underappreciated role of clinically applied shockwaves, which could be exploited for other macrophage-mediated disorders.