Minimal Inflammatory Cell Infiltration Research Articles

The Different Therapeutic Effects of Traditional Chinese Medicine Shensong Yangxin Capsule and Salubrinal in High-intensity Exercise-induced Heart Failure in Rats with Acute Myocardial Infarction.

Currently, endoplasmic reticulum stress is studied utilizing a dephosphorylation inhibitor (Sal). The traditional Chinese patent medicine and simple formulation Shensong Yangxin Capsule is a commonly used medication for the treatment of arrhythmia. However, the efficacy and underlying mechanism of the capsule in treating post-ischemic heart failure in myocardial tissue have not yet been investigated. The therapeutic effects and the underlying mechanism of the Shensong Yangxin Capsule (SSYX) and the dephosphorylation inhibitor Salubrinal (Sal) on heart failure (HF) induced by high-intensity exercise in rats with acute myocardial infarction (AMI) were investigated. Male infants of 8 weeks Spragge-Dawley (SD) rats were randomly assigned to one of four groups: sham surgery group, AMI+placebo group, AMI+Shensong Yangxin Capsule group (AMI+SSYX), and AMI+Sal administration group. Rats' myocardial infarction was induced by left coronary artery ligation. Rats were subjected to a 3-week high-intensity exercise program to simulate heart failure after 7 days of postoperative rest. After the fourth postoperative week, echocardiography was applied to determine the left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), and left ventricular systolic volume (LVESV) in each group. HE and TUNEL labeling were employed to examine the morphology of cardiac cells and measure the percentage of apoptosis in each group; Western blotting was applied to detect the cardiomyocyte apoptosis-related proteins p-JNK, p-P38, and NOX2, while ELISA was used to detect glutathione(GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) in serum. Following a 4-week drug intervention:(1)LVFS and LVEF in the AMI+placebo group were statistically significantly reduced, while LVESV were significantly higher, compared to those in the sham surgery group (P<0.05); The AMI+SSYX group performed statistically significantly better than the AMI+placebo group(P<0.05). (2) The myocardial cells in the AMI+placebo group exhibited significant swelling and inflammatory cell infiltration; the myocardial cells in the AMI+SSYX group and AMI+Sal group displayed mild swelling and minimal inflammatory cell infiltration; the AMI+SSYX group's myocardial cell morphology was superior to that of the AMI+Sal group; (3) The apoptosis rate of the AMI+placebo group was around 95%, greater than that of the sham surgery group (2.55%). The apoptosis rate of the AMI+SSYX group is approximately 21%, while the apoptosis rate of the AMI+Sal group is about 43%. (4) In the AMI+placebo group, p-JNK, p-P38, and NOX2 protein expression dramatically increased compared to the sham surgery group. The expression of p-P38, NOX2, and p-JNK/t-JNK was considerably reduced in the AMI+Shensong group and AMI+Sal group, compared to the AMI+placebo group. (P<0.01)The AMI+SSYX group's result is superior to that of the AMI+Sal group. (5) Compared to the sham surgery group, the serum levels of SOD and GSH were significantly lower, and MDA was significantly higher in the AMI+placebo group. Compared to the AMI+placebo group, the serum levels of SOD and GSH were significantly higher, and MDA was significantly lower in the AMI+SSYX group and the AMI+Sal group. (P<0.05) Conclusion: In rats with acute myocardial infarction in high-intensity exercise-induced heart failure, Shensong Yangxin Capsule dramatically reduces myocardial cell death and cardiac dysfunction. SSYX has a shorter course of treatment and a better therapeutic effect than Sal.

Statin Myopathy: A Case series

Anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) immune-mediated necrotizing myopathy (anti-HMGCR IMNM) is a rare adverse effect associated with the use of statin medications [1]. While the investigation remains ongoing, statin exposure is thought to induce mitochondrial dysfunction due to inhibition of HMGCR, subsequently triggering an inflammatory cascade with oxidative stress and autoantibodies (aAbs) against HMGCR, ultimately leading to an immune-mediated necrotizing myopathy (IMNM) [2]. Clinical presentation may involve dermatomyositis-like features ranging from myalgias to severe and progressive proximal muscle weakness, often persisting after cessation of the offending statin [3]. Serum studies typically reveal significantly elevated Creatine Kinase (CK) levels, often ranging from 10 to 100 times the upper limit of normal [4]. Presumptive diagnosis is confirmed via muscle biopsy, which typically reveals necrotic fibers with minimal inflammatory cell infiltrates [5]. In addition to withdrawal of the offending agent, treatment may involve immunomodulatory medications (e.g. intravenous immunoglobulin (IVIG), steroids, and/or Rituximab). For most patients, resolution typically occurs over months, sometimes years. This paper highlights three cases of statin-induced myopathy and the clinical implications for providers in prompt recognition, diagnosis, and management of this rare condition. Further research is needed to better understand both the risk factors and underlying mechanisms in order to increase awareness, optimize care, and minimize the morbidity associated with this adverse drug reaction.

Lung Protection After Severe Thermal Burns With Adenosine, Lidocaine, and Magnesium (ALM) Resuscitation and Importance of Shams in a Rat Model.

The management of severe burns remains a complex challenge. Adenosine, lidocaine, and magnesium (ALM) resuscitation therapy has been shown to protect against hemorrhagic shock and traumatic injury. The aim of the present study was to investigate the early protective effects of small-volume ALM fluid resuscitation in a rat model of 30% total body surface area (TBSA) thermal injury. Male Sprague-Dawley rats (320-340 g; n = 25) were randomly assigned to: 1) Sham (surgical instrumentation and saline infusion, without burn, n = 5), 2) Saline resuscitation group (n = 10), or 3) ALM resuscitation group (n = 10). Treatments were initiated 15-min after burn trauma, including 0.7 mL/kg 3% NaCl ± ALM bolus and 0.25-0.5 mL/kg/h 0.9% NaCl ± ALM drip, with animals monitored to 8.25-hr post-burn. Hemodynamics, cardiac function, blood chemistry, hematology, endothelial injury markers and histopathology were assessed. Survival was 100% for Shams and 90% for both ALM and Saline groups. Shams underwent significant physiological, immune and hematological changes over time as a result of surgical traums. ALM significantly reduced malondialdehyde levels in the lungs compared to Saline (P = .023), and showed minimal alveolar destruction and inflammatory cell infiltration (P < .001). ALM also improved cardiac function and oxygen delivery (21%, P = .418 vs Saline), reduced gut injury (P < .001 vs Saline), and increased plasma adiponectin (P < .001 vs baseline). Circulating levels of the acute phase protein alpha 1-acid glycoprotein (AGP) increased 1.6-times (P < .001), which may have impacted ALM's therapeutic efficacy. We conclude that small-volume ALM therapy significantly reduced lung oxidative stress and preserved alveolar integrity following severe burn trauma. Further studies are required to assess higher ALM doses with longer monitoring periods.

Anti-HMGCR myopathy frequently coexists with other myositis-specific autoantibodies in Korean patients (P9-8.012)

<h3>Objective:</h3> To understand the characteristics of Korean patients with anti-HMGCR myopathy, we measured anti-HMGCR antibodies and analyzed clinical, radiological, and pathological features. <h3>Background:</h3> Immune-mediated necrotizing myopathy (IMNM) is an inflammatory myopathy subgroup characterized by subacute proximal weakness and prominent myofiber necrosis with minimal inflammatory cell infiltrate on muscle biopsy. Two autoantibodies have been reported in association with IMNM: antibodies against the signal recognition particle (SRP) and 3-hydroxy-3-methylglutaryl-coenxyme A reductase (HMGCR). To date, there only have been two case reports of Korea patients with anti-HMGCR myopathy. <h3>Design/Methods:</h3> We measured titers of anti-HMGCR antibodies in the sera of 99 patients with inflammatory myopathy using the enzyme-linked immunosorbent assay. We also tested 16 myositis-specific autoantibodies (MSAs) in all patients with anti-HMGCR myopathy. <h3>Results:</h3> Postivity for the anti-HMGCR antibody was observed in 17 (4 males and 13 females) of 99 patients with inflammatory myopathy. The median age at symptom onset was 60 years. Ten (59%) of the anti-HMGCR positive patients had taken statins. The most common symptoms were proximal muscle weakness in 15 (88%), followed by myalgia in 9 (53%), neck weakness in 4 (24%), dysphagia in 3 (18%), and skin lesions in 2 (12%). The median titer of anti-HMGCR antibody was 202 U/ml. We found eight different MSAs in nine (53%) patients. The median disease duration from symptom onset to diagnosis was significantly shorter in the MSA-positive group than in the MSA-negative group (p = 0.027). <h3>Conclusions:</h3> In conclusion, our study is the first to measure anti-HMGCR antibodies with inflammatory myopathy, and identify new findings including the coexistence of other MSAs in Korean patients. <b>Disclosure:</b> Miss Lee has nothing to disclose. Ms. Oh has nothing to disclose. Kee Duk Park has nothing to disclose. Young-Chul Choi has nothing to disclose. Hyung Jun Park has nothing to disclose.

Dual-effects of caffeinated hyalurosomes as a nano-cosmeceutical gel counteracting UV-induced skin ageing

Caffeine (CAF) is a challenging natural bioactive compound with proven antiaging efficacy. However, being hydrophilic hampers its permeation through the skin. Our aim is to develop a novel CAF-loaded nano-cosmeceutical tool counteracting skin photoaging via improving CAF skin permeation using a bioactive nanocarrier. Caffeinated hyalurosomes are novel biocompatible antiaging nanoplatforms designed by immobilization of phospholipid vesicles with a hyaluronan polymer. Physicochemical properties of the selected hyalurosomes formulation showed nano-sized vesicles (210.10 ± 1.87 nm), with high zeta potential (−31.30 ± 1.19 mv), and high encapsulation efficiency (84.60 ± 1.05%). In vitro release results showed outstanding sustained release profile from caffeinated hyalurosomes compared to the CAF-loaded in conventional gel over 24 h. The in-vivo study revealed a photoprotective effect of caffeinated hyalurosomes, reflected from the intact and wrinkling-free skin. Results of biochemical analyses of oxidative stress, pro-inflammatory mediators, and anti-wrinkling markers further confirmed the efficacy of the prepared hyalurosomes compared to the CAF conventional gel. Finally, histopathological examination demonstrated normal histological structures of epidermal layers with minimal inflammatory cell infiltrates in the caffeinated hyalurosomes group compared to the positive control group. Conclusively, caffeinated hyalurosomes successfully achieved enhanced CAF loading and penetration into the skin besides the hydration effect of hyaluronan. Consequently, the developed delivery system presents a promising skin protection nano-platforms via the double effects of both hyaluronan and CAF, hence it guards against skin photodamage.

Statins and muscle – friends or foes? Discussions based on a case report

Immune-mediated necrotizing myopathy (IMNM) is a type of autoimmune myopathy characterized by relatively severe proximal weakness, myofiber necrosis with minimal inflammatory cell infiltrate on muscle biopsy, and infrequent extra-muscular involvement Anti-3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) myopathy is a distinct subtype of INMN, often associated with statin exposure. Treatment of IMNM consists of immunosuppression with steroids, steroid-sparing agents, intravenous immune globulin and/or biologics. We present here a case of anti-HMCGR-associated IMNM and a brief review the pathophysiology, diagnosis and treatment to increase physician awareness of this rare and debilitating condition.

Substance P/NK1R Antagonistic Effect of 17-Trifluoromethyl Phenyl Trinor Prostaglandin F2α in Breast Cancer

17-Trifluoromethyl phenyl trinor prostaglandin F2α (17-TPGF2α) is extracted from a Zinc database, but its value for inhibiting breast cancer (BC) through the substance P (SP)/NK1R system remains unknown. This study was designed to investigate the potential antagonist effect of 17-TPGF2α through neurokinin 1 receptor (NK1R). The effect of 17-TPGF2α on the proliferation and apoptosis of BC cell lines was determined through in vitro cell lines. Based on in vitro results we planned to investigate anticancer activity in female Balb/c and used subcutaneous (SC) injection of DMBA for cancer induction. Oral administration of 17-TPGF2α significantly suppresses the tumor volume as compared with an untreated group. The serum parameters like ALP, AST, and ALT and hematological parameters were normalized in test treated group. Histological examination revealed normal histoarchitecture of the mammary gland and focal areas showed minimal inflammatory cell infiltration. There is no necrosis is seen in both test treated and standard treated groups when compared with the DMBA group. All these findings concluded that 17-TPGF2α may have potential as a novel antitumor candidate for BC.

The inflammatory pathology of dysferlinopathy is distinct from calpainopathy, Becker muscular dystrophy, and inflammatory myopathies

The descriptions of muscle pathology in dysferlinopathy patients have classically included an inflammatory infiltrate that can mimic inflammatory myopathies. Based on over 20 years of institutional experience in evaluating dystrophic and inflammatory myopathy muscle biopsies at the University of Iowa, we hypothesized the inflammatory histopathology of dysferlinopathy is more similar to limb-girdle pattern muscular dystrophies such as calpainopathy and Becker muscular dystrophy, and distinct from true inflammatory myopathies. Muscle biopsies from 32 dysferlinopathy, 30 calpainopathy, 30 Becker muscular dystrophy, and 30 inflammatory myopathies (15 each of dermatomyositis and inclusion body myositis) were analyzed through digital quantitation of CD3, CD4, CD8, CD20, and PU.1 immunostaining. The expression of MHC class I and deposition of complement C5b-9 was also evaluated. Dysferlinopathy, calpainopathy, and Becker muscular dystrophy muscle biopsies had similar numbers of inflammatory cell infiltrates and significantly fewer CD3+ T-lymphocytes than dermatomyositis (p = 0.05) and inclusion body myositis (p < 0.0001) biopsies. There was no statistically significant difference in the number of PU.1+ macrophages identified in any diagnostic group. MHC class I expression was significantly lower in the limb-girdle pattern muscular dystrophies compared to the inflammatory myopathies (p < 0.0001). In contrast, complement C5b-9 deposition was similar among dysferlinopathy, dermatomyositis, and inclusion body myositis biopsies but significantly greater than calpainopathy and Becker muscular dystrophy biopsies (p = 0.05). Compared to calpainopathy, Becker muscular dystrophy, and inflammatory myopathies, the unique profile of minimal inflammatory cell infiltrates, absent to focal MHC class I, and diffuse myofiber complement C5b-9 deposition is the pathologic signature of dysferlinopathy muscle biopsies.

Syzygium aqueum (Burm.f.) Alston Prevents Streptozotocin-Induced Pancreatic Beta Cells Damage via the TLR-4 Signaling Pathway.

Although several treatments are available for the treatment of type 2 diabetes mellitus, adverse effects and cost burden impose the search for safe, efficient, and cost-effective alternative herbal remedies. Syzygium aqueum (Burm.f.) Alston, a natural anti-inflammatory, antioxidant herb, may suppress diabetes-associated inflammation and pancreatic beta-cell death. Here, we tested the ability of the bioactive leaf extract (SA) to prevent streptozotocin (STZ)-induced oxidative stress and inflammation in pancreatic beta cells in rats and the involvement of the TLR-4 signaling pathway. Non-fasted rats pretreated with 100 or 200 mg kg−1 SA 2 days prior to the STZ challenge and for 14 days later had up to 52 and 39% reduction in the glucose levels, respectively, while glibenclamide, the reference standard drug (0.5 mg kg−1), results in 70% reduction. Treatment with SA extract was accompanied by increased insulin secretion, restoration of Langerhans islets morphology, and decreased collagen deposition as demonstrated from ELISA measurement, H and E, and Mallory staining. Both glibenclamide and SA extract significantly decreased levels of TLR-4, MYD88, pro-inflammatory cytokines TNF-α, and TRAF-6 in pancreatic tissue homogenates, which correlated well with minimal pancreatic inflammatory cell infiltration. Pre-treatment with SA or glibenclamide decreased malondialdehyde, a sensitive biomarker of ROS-induced lipid peroxidation, and restored depleted reduced glutathione in the pancreas. Altogether, these data indicate that S. aqueum is effective in improving STZ-induced pancreatic damage, which could be beneficial in treating type 2 diabetes mellitus.

Therapeutic Role of Platelet Rich Plasma in Formaldehyde-Induced Arthritis in Adult Male Albino Rats

Abstract Introduction Osteoarthritis (OA) is a common health problem. Platelet-rich plasma (PRP) has been recognized to enhance articular cartilage metabolism. Aim of the work the study was designed to investigate the influence of PRP on cartilage healing after induction of arthritis. Material and methods Forty two adult male albino rats were used in this study. The rats were randomly divided into three groups: Group I (n = 18): the control group (Ia, Ib &amp; Ic) Ib &amp; Ic were injected intra-articularly with saline and left for 3 and 6 weeks. Group II (n = 12): arthritic group, in which osteoarthritis was induced by injection of 0.02ml 5% formaldehyde once in the right knee joints, left without treatment, and were sacrificed after three weeks (IIa) or after six weeks (IIb). Group III(n = 12): arthritis was induced as group II, one week later, the rats were intra-articularly injected with single dose of 0.3ml PRP in the same joint then were sacrificed three weeks (IIIa) or six weeks (IIIb) after formaldehyde injection. At the end of the study the right knee joints were taken, decalcified then processed for paraffin sections to be examined by light microscope using H&amp;E, toluidine blue and Masson’s trichrome (MTC) stains. Immunohistochemistry for caspase-3 enzyme was done to demonstrate apoptotic chondrocytes. Morphometric study was conducted to measure the thickness of the non-calcified cartilage, count the chondrocytes and synovial membrane inflammatory cells and Mankin's score. Then statistical analysis was done. Results The arthritic groups revealed irregular surface of the articular cartilage, loss of the articular matrix and bone eburnation. Moreover, there was apparent hypocellularity and disorganization of the chondrocytes. Osteoclasts and osteoblasts were seen invading the osteochondral junction. MTC stained sections of the synovial membrane showed deposition of thick collagen bundles with heavy inflammatory cell infiltrate and numerous blood vessels. The affinity of the articular cartilage to toluidine blue stain was apparently decreased while caspase-3 immunoreactivity was apparent in many chondrocytes .Group IIIa demonstrated almost similar histological findings as the control group; regular articular cartilage surface with regularly arranged chondrocytes in the different cartilage zones. Synovial membrane illustrated minimal inflammatory cell infiltrate with thin collagen bundles and small blood vessels in MTC stained sections. There was high affinity of the articular cartilage to the toluidine blue stain and few chondrocytes showed positive caspase-3 immunoreactivity. Group IIIb revealed continuous surface of the articular cartilage, yet with minimal fibrillation in some areas. Osteoblasts and osteoclasts were seen invading the calcified cartilage. Synovial membrane showed deposition of dense collagen bundles with some inflammatory cell infiltrate. Toluidine blue sections revealed decreased articular cartilage affinity to the stain while caspase-3 immunoreactivity was evident in many chondrocytes. The morphometric results and statistical analysis confirmed the histological findings. Conclusion Intra-articular injection of PRP demonstrated advantageous role on articular cartilage healing, however, these effects appeared to be transient. So the need of multiple injections of PRP has to be considered in cases of OA.

Miopatía necrotizante autoinmune. Presentación de un caso clínico

Inflammatory myopathies (IM) or myositis are a heterogeneous group of muscle diseases of rare occurrence. Such diseases are characterized by inflammation of the different components of muscle tissue, which can occur either in isolation or, more commonly, as part of a systemic disorder. Immune-mediated necrotizing myopathies (IMNM) are a type of autoimmune myopathy characterized by proximal muscle weakness, myofiber necrosis with minimal inflammatory cell infiltrate on muscle biopsy and infrequent extramuscular involvement. Even though there are clinical and histopathological similarities. The spectrum of inflammatory myopathies is considerably variable. Therefore, the performance of complementary studies is essential for the proper identification of the IM subtype to contribute accurately on treatment so determine the better prognosis). The present article shows the case of a young 29 years old, with no personal and family history background of autoimmune disease and no relevant pathological back-ground. The patient consulted the medical ward of the Institution with pain, functional impairment of upper and lower extremities, muscle weakness mainly located in the pectoral girdle area and, although to a lesser degree, in the pelvic girdle area. It was also associated with asthenia, tendency to drowsiness and hyporeactivity.

Preclinical Safety Assessment of 2 Inhaled Single-Domain Antibodies in the Cynomolgus Monkey.

The safety of 2 single domain antibodies (dAbs) was evaluated by inhalation toxicology studies in the cynomolgus monkey. In the first case study, a 14-day repeat-dose study evaluating an anti-thymic stromal lymphopoietin (anti-TSLP) dAb resulted in minimal mononuclear inflammatory cell infiltrates in the lungs, increases in lymphocytes in bronchoalveolar lavage fluid, and development of antidrug antibodies (ADAs). In a 6-week inhalation study, there was an increase in incidence and/or severity of mononuclear cell infiltrates in the lung, increased cellularity in the tracheobronchial lymph node (TBLN), and development of ADA. The second case study evaluated a change in duration of inhalation dosing, a different route of exposure (intravenous or IV), and recovery following an off-dose period with an anti-tumor necrosis factor receptor 1 dAb. A 7-day repeat-dose inhalation study and a 14-day IV study produced no microscopic effects in the lung, whereas a 14-day inhalation study resulted in moderate increases in pulmonary perivascular/peribronchiolar/alveolar lymphocytic infiltrates and increased cellularity in the TBLN, with partial and full recovery, respectively, after 14 days. The lung and lymph node findings seen after inhalation of either dAb were considered secondary to the immunogenic response to a human protein and were considered nonadverse.

The "histological replacement growth pattern" represents aggressive invasive behavior in liver metastasis from pancreatic cancer.

BackgroundIn the case of liver metastasis (LM), tumors showing the replacement growth pattern (RGP), in which metastatic cells infiltrate and replace hepatocytes with minimal desmoplastic reaction and inflammatory cell infiltration, associate with a poor prognosis. The heterogeneity, frequency, and prognostic value of the RGP in LM from pancreatic cancer (PCa) are not well known.MethodsIn the circumference of treatment‐naïve resected LMs from patients with PCa, the heterogeneity of the GP was assessed. Next, the clinicopathological features of LMs showing the RGP in needle biopsy specimens were investigated in patients with treatment‐naïve advanced PCa.ResultsThirteen of the 14 (93%) in all resected LMs and 7 of the 9 (78%) in RGP component GP in resected LMs showed homogeneous GP. A RGP was found in 50% of the needle biopsy specimens of LMs obtained from 107 patients. The median overall survival times in the RGP group and non‐RGP group were 3.6 and 10.4 months. Multivariate analysis identified RGP as an independent poor prognostic factor. Median value of CD8 positive percentage in RGP was lower than that in non‐RGP (0.75 vs 1.46, P = .04). Median overall survival times in low CD8 groups tend to be shorter than those in high CD8 group (8.2 vs 4.2 months).ConclusionMost LMs from PCa show a homogeneous GP. The RGP was observed in about a half of the LMs from PCa patients, and was identified as a poor prognostic factor.

Formulation and evaluation of simvastatin polymeric nanoparticles loaded in hydrogel for optimum wound healing purpose.

PurposeThe aim of this study was to formulate a hydrogel loaded with polymeric nanoparticles (PoNPs) of simvastatin (SIM) for topical wound healing application.Materials and methodsThe SIM PoNPs were prepared by using the nanoprecipitation method to improve the drug solubility and skin permeation. Furthermore, drug content, solubility, particle size, surface charge, and transmission electron microscopy of the prepared PoNPs were evaluated. Then, the PoNPs were loaded on hydrogel, and physical characteristics, in vitro release, and ex vivo permeation of the hydrogel were evaluated. Finally, the prepared gel was applied on rat wounds, and a histopathological study was performed.ResultsThe results showed that the drug content in the PoNPs was 86.4%. The PoNPs were spherical in shape with a smooth surface and a uniform size distribution. The particle size was 268.4±2.6, polydispersity index was ≤0.302, and zeta potential was −33±1.67 mV. The hydrogel loaded with SIM PoNPs was homogenous, and the pH was accepted and compatible with the skin. Moreover, the viscosity and spreadability assured its ease of application. The drug content was 97.25±0.02%. Furthermore, about 81% of SIM was released within 24 hours, while in the ex vivo permeation study 69.19% of SIM passed through the skin after 24 hours. Finally, the histopathological studies confirmed the efficacy of the SIM PoNPs-loaded hydrogel in wound healing due to the formation of the normal epithelial layer on day 11 after wound creation.ConclusionThe hydrogel loaded with SIM PoNPs showed a good efficacy in accelerating the healing of the rat wound with complete epithelialization and minimal inflammatory cell infiltration.

Immune-Mediated Necrotizing Myopathy.

Immune-mediated necrotizing myopathy (IMNM) is a type of autoimmune myopathy characterized by relatively severe proximal weakness, myofiber necrosis with minimal inflammatory cell infiltrate on muscle biopsy, and infrequent extra-muscular involvement. Here, we will review the characteristics of patients with IMNM. Anti-signal recognition particle (SRP) and anti-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) autoantibodies are closely associated with IMNM and define unique subtypes of patients. Importantly, the new European Neuromuscular Centre criteria recognize anti-SRP myopathy, anti-HMGCR myopathy, and autoantibody-negative IMNM as three distinct subtypes of IMNM. Anti-SRP myopathy patients have more severe muscle involvement, have more common extra-muscular features, and may respond best to immunosuppressive regimens that include rituximab. In contrast, anti-HMGCR myopathy is often associated with statin exposure and intravenous immunoglobulin treatment may be an effective treatment, even as monotherapy. Both anti-SRP and anti-HMGCR myopathy tend to be most severe in younger patients. Furthermore, children with these forms of IMNM may present with dystrophy-like features which are potentially reversible with immunosuppressant treatment. IMNM patients with either autoantibody may experience fatty replacement of muscle soon after disease onset, suggesting that intense and early immunosuppressant therapy may provide the best chance to avoid long-term disability. IMNM is composed of anti-SRP myopathy, anti-HMGCR myopathy, and autoantibody-negative IMNM. Both anti-SRP and anti-HMGCR myopathy can cause severe weakness, especially in younger patients. Anti-SRP myopathy patients tend to have the most severe weakness and most prevalent extra-muscular features. Autoantibody-negative IMNM remains poorly described.

Inflammatory Cell Findings in the Female Rabbit Heart and Stress-associated Exacerbation with Handling and Procedures Used in Nonclinical Studies.

Despite the use of rabbits in biomedical research, including regulatory toxicology and cardiovascular studies, little data exist on heart findings in this species. This study was designed to document myocardial findings in female rabbits and the impact of study-related procedures typical for vaccine toxicology studies. One hundred and forty 6- to 8-month-old female New Zealand White rabbits were divided equally into 2 groups, high and low study procedure groups (group 1 and group 2, respectively). All animals received intramuscular (IM) injections of sterile saline every 2 weeks for 5 times and were necropsied 2 days after the final IM injection. Clinical chemistry, hematology, and urinalysis were evaluated. Blood for stress biomarkers (norepinephrine, epinephrine, cortisol, and corticosterone), C-reactive protein, cardiac troponin I, and creatine kinase were collected at time 0 (just before dose administration) and then at 4, 24, and 48 hr after dose administration in group 1 only. Hearts were assessed histologically. Focal to multifocal minimal inflammatory cell infiltrates were common (∼80%), particularly in the left ventricle and interventricular septum, and were similar to the types of infiltrates identified in other laboratory animal species. Additionally, study-related procedures elevated serum stress biomarkers and exacerbated the frequency and severity of myocardial inflammatory cell infiltrates.

Low-Initial-Modulus Biodegradable Polyurethane Elastomers for Soft Tissue Regeneration.

The mechanical match between synthetic scaffold and host tissue remains challenging in tissue regeneration. The elastic soft tissues exhibit low initial moduli with a J-shaped tensile curve. Suitable synthetic polymer scaffolds require low initial modulus and elasticity. To achieve these requirements, random copolymers poly(δ-valerolactone-co-ε-caprolactone) (PVCL) and hydrophilic poly(ethylene glycol) (PEG) were combined into a triblock copolymer, PVCL-PEG-PVCL, which was used as a soft segment to synthesize a family of biodegradable elastomeric polyurethanes (PU) with low initial moduli. The triblock copolymers were varied in chemical components, molecular weights, and hydrophilicities. The mechanical properties of polyurethanes in dry and wet states can be tuned by altering the molecular weights and hydrophilicities of the soft segments. Increasing the length of either PVCL or PEG in the soft segments reduced initial moduli of the polyurethane films and scaffolds in dry and wet states. The polymer films are found to have good cell compatibility and to support fibroblast growth in vitro. Selected polyurethanes were processed into porous scaffolds by a thermally induced phase-separation technique. The scaffold from PU-PEG1K-PVCL6K had an initial modulus of 0.60 ± 0.14 MPa, which is comparable with the initial modulus of human myocardium (0.02-0.50 MPa). In vivo mouse subcutaneous implantation of the porous scaffolds showed minimal chronic inflammatory response and intensive cell infiltration, which indicated good tissue compatibility of the scaffolds. Biodegradable polyurethane elastomers with low initial modulus and good biocompatibility and processability would be an attractive alternative scaffold material for soft tissue regeneration, especially for heart muscle.

Possible Protective Effect of Bone Marrow-Mesenchymal Stem Cells (BM-MSCs) Against the Remote Liver Injury Induced by Renal Ischemia Reperfusion in Male Albino Rats

Background: Renal ischemia-reperfusion injury (IRI) is a major cause of acute renal injury and leads to multiorgan dysfunction especially liver injury. Stem cell therapy has been used effectively in treatment of renal IRI. Aim of the work: To investigate the possible therapeutic effect of BM-MSC therapy on the remotely affected liver in a rat model of renal IRI and the mechanisms involved in this effect. Methods: Rats were divided into 3 groups; the sham-operated control group, the renal IRI group: bilateral renal IRI for 45 minutes followed by reperfusion, and BM-MSCs-injected group: rats were subjected to renal IRI and then received a single intravenous (iv) injection of BM-MSCs immediately after reperfusion. At the different time-points (days 1, 3, and 5) of the study, serum urea, creatinine, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were estimated. Livers were removed and were divided: some specimens were homogenized for assessment of oxidant and antioxidant parameters, and other specimens were used for paraffin embedding for histological and immunohistochemical study. Results: Renal IRI resulted in a significant increase in serum levels of urea, creatinine, AST, ALT, and hepatic tissue levels of malondialdehyde (MDA) which attenuated on days 3 and 5 after BM-MSCs injection. MDA showed an early significant decrease in the BM-MSCs-injected group at day 1. Reduced glutathione (GSH) showed a significant decrease in the IRI sub-groups while BM-MSCs-injected sub-groups showed significantly higher levels of GSH on days 1, 3 and 5. Tissue sections showed that labeled BM-MSCs were engrafted into liver tissue. The IRI subgroups showed marked destruction of liver tissue e.g. disrupted lobular architecture, marked hepatocellular ballooning and cytoplasmic vaculations, and marked recruitment of the inflammatory cells with vascular congestion. While in the BM-MSCs-injected subgroups, there were gradual regeneration and restoration of lobular architecture, reduced degenerated cells, decreased congestion, and minimal inflammatory cell infiltration. By day 5, most hepatocytes had more or less normal appearance, arranged in cords with preserved general architecture and lacking evidence of major morphological injury. In addition, BM-MSCs significantly attenuated the liver immunohistochemical expression of tumor necrosis factor-alpha (TNF-α). Conclusion: BM-MSCs protects against the remote liver injury induced by renal IRI through early inhibition of inflammatory cell recruitment, inflammatory reactions, suppressing oxidative stress and lipid peroxidation, and rapid restoration of cellular and architectural integrity of the liver. This provides another important basis for the therapeutic concept of BM-MSCs in treatment of renal IRI by adding a beneficial effect on decreasing the remote liver injury besides considering it as a promising treatment in renal IRI.

MA09.10 A NaPi2b Antibody-Drug Conjugate Induces Durable Complete Tumor Regressions in Patient-Derived Xenograft Models of NSCLC

The sodium-phosphate transporter NaPi2b is expressed at high levels in a majority of non-squamous non-small cell lung cancers (NSCLC), suggesting it may be an attractive therapeutic target for antibody-drug conjugate (ADC) development in this disease. However, NaPi2b is also expressed at high levels in type II alveolar cells, raising the potential for normal tissue toxicity with this approach. XMT-1536 is an ADC comprised of a humanized antibody against NaPi2b and approximately 15 auristatin-derived payload molecules per antibody conjugated via a multivalent hydrophilic polymer (Fleximer). The auristatin payload is enzymatically cleaved upon ADC trafficking to the endosome/lysosome compartment, releasing a cytotoxic auristatin-derivative that is capable of bystander effect killing. The anti-tumor activity of XMT-1536 was evaluated in seven patient-derived xenograft models of NSCLC adenocarcinoma, chosen for high NaPi2b-expression and representing a spectrum of oncogenic driver mutations prevalent in NSCLC adenocarcinoma (including tumors without oncogenic drivers). The standard dose of XMT-1536 used across models was 3 mg/kg administered intravenously once weekly for 3 weeks (last dose on Day 14). Experiments ran until tumor growth past a pre-specified endpoint or Day 60. XMT-1536 was also evaluated for tolerability in a cynomolgus monkey study. At the 3 mg/kg dose, XMT-1536 was active in 6/7 models: complete tumor regression in 3 models, partial tumor regression in 1 model, and significant tumor growth inhibition in 2 models. In 3 of the 4 models where XMT-1536 induced tumor regression, regressions were durable, with a majority of the animals maintaining partial or complete regression at Day 60. The antibody component of XMT-1536 is cross-reactive with cynomolgus NaPi2b with similar affinity as human NaPi2b. XMT-1536 was well tolerated up to 5 mg/kg (4294 mg/m2 auristatin payload equivalents), the highest dose tested. There was no body weight loss, no clinical observations attributable to XMT-1536, and no evidence of neutropenia. On pathology, there was minimal mixed inflammatory cell infiltrate in the lung in 1 high dose animal at each necropsy time point, but no evidence of significant lung toxicity. Exposure to XMT-1536 indicated good conjugate stability, low exposure to free drug payload in plasma (<1 ng/mL), and supported the 3 mg/kg dose level in mouse studies as a potentially clinically-relevant dose. These results indicate XMT-1536 can achieve durable tumor regressions in murine patient-derived NSCLC adenocarcinoma models at doses associated with good tolerability in cynomolgus monkey, and support evaluation of XMT-1536 in patients with NSCLC.

Genipin-crosslinked, immunogen-reduced decellularized porcine liver scaffold for bioengineered hepatic tissue

Liver disease affects millions of patients each year worldwide. Decellularized biologic matrices are plausible biomedical materials for bioengineered replacement hepatic tissue. However, one of the concerns for its safe medical application is the lack of objective assessment of the immunogen within the materials and in vivo immune responses to the matrices. The purpose of this study was to produce immunogen- reduced and biocompatible matrices from porcine liver. Whole porcine livers were perfusion decellularized and cross-linked with glutaraldehyde (GA) or genipin (GP). Proteins were extracted, and the migratory response of human leukocytes toward protein extracts was examined using an in vitro migration chamber. In addition, biopsy specimens of decellularized scaffolds were implanted subcutaneously into rodents to investigate scaffold immunogenicity. Histological staining confirmed cellular clearance from pig livers, with removal of nuclei and cytoskeletal components and widespread preservation of structural extracellular molecules. Polymerase chain reaction analysis showed that galactose-alpha-1,3-galactose-beta-1,4-N-acetylglucosamine (1,3 gal), swine leukocyte antigen, and porcine endogenous retrovirus were completely removed in the matrices. Decellularization significantly reduced the migration of monocytes compared with native porcine tissue. Although the proportion of transmigrating lymphocytes was much lower, repeating the cross-linking procedure reduced the migratory response. After implantation for 4 weeks, the decellularized and native samples were degraded, and the GA-treated group demonstrated a severe inflammatory reaction; however, minimal inflammatory cell infiltration was seen in the GPtreated group during the 8-week investigation period. In conclusion, our study provided evidence that GP crosslinking could significantly reduce the immunogenicity of decellularized liver biomaterials.