Beneficial effects of CCL8 inhibition at lipopolysaccharide-induced lung injury.

CCL8 (MCP-2) is a chemoattractive cytokine associated with various immune-related pathologies. Recent studies show that CCL8 is significantly stimulated during acute respiratory distress syndrome in severely ill patients with COVID-19, making the inhibition of CCL8 activity a promising treatment. Lipopolysaccharide (LPS)-induced lung injury was evaluated in mice using a neutralizing antibody (1G3E5) against human CCL8. Pharmacokinetic studies indicated that following IP administration, 1G3E5 was sustained at higher levels and for a longer period compared to IV administration. CCL8 expression in the lungs was not enhanced by LPS, but CCR2 and CCR5 receptors were significantly stimulated. 1G3E5-mediated inhibition of CCL8 was associated with the reduction of pulmonary inflammation and suppression of various pro-inflammatory cytokines. These results point to a previously unrecognized, permissive role for CCL8 in mediating cytokine induction and ultimately sustaining inflammation. Disruption of CCL8 activity may provide a strategy for mitigating pulmonary inflammation during lung injury when related to abnormal cytokine induction.

Methylation studies in Peromyscus: aging, altitude adaptation, and monogamy.

DNA methylation-based biomarkers of aging have been developed for humans and many other mammals and could be used to assess how stress factors impact aging. Deer mice (Peromyscus) are long-living rodents that have emerged as an informative model to study aging, adaptation to extreme environments, and monogamous behavior. In the present study, we have undertaken an exhaustive, genome-wide analysis of DNA methylation in Peromyscus, spanning different species, stocks, sexes, tissues, and age cohorts. We describe DNA methylation-based estimators of age for different species of deer mice based on novel DNA methylation data generated on highly conserved mammalian CpGs measured with a custom array. The multi-tissue epigenetic clock for deer mice was trained on 3 tissues (tail, liver, and brain). Two human-Peromyscus clocks accurately measure age and relative age, respectively. We present CpGs and enriched pathways that relate to different conditions such as chronological age, high altitude, and monogamous behavior. Overall, this study provides a first step towards studying the epigenetic correlates of monogamous behavior and adaptation to high altitude in Peromyscus. The human-Peromyscus epigenetic clocks are expected to provide a significant boost to the attractiveness of Peromyscus as a biological model.

Propensity to endoplasmic reticulum stress in deer mouse fibroblasts predicts skin inflammation and body weight gain.

The unfolded protein response (UPR) is involved in the pathogenesis of metabolic disorders, yet whether variations in the UPR among individuals influence the propensity for metabolic disease remains unexplored. Using outbred deer mice as a model, we show that the intensity of UPR in fibroblasts isolated early in life predicts the extent of body weight gain after high-fat diet (HFD) administration. Contrary to those with intense UPR, animals with moderate UPR in fibroblasts and therefore displaying compromised stress resolution did not gain body weight but developed inflammation, especially in the skin, after HFD administration. Fibroblasts emerged as potent modifiers of this differential responsiveness to HFD, as indicated by the comparison of the UPR profiles of fibroblasts responding to fatty acids in vitro, by correlation analyses between UPR and proinflammatory cytokine-associated transcriptomes, and by BiP (also known as HSPA5) immunolocalization in skin lesions from animals receiving HFD. These results suggest that the UPR operates as a modifier of an individual's propensity for body weight gain in a manner that, at least in part, involves the regulation of an inflammatory response by skin fibroblasts. This article has an associated First Person interview with the first author of the paper.

Persistent effects of pair bonding in lung cancer cell growth in monogamous Peromyscus californicus.

Epidemiological evidence suggests that social interactions and especially bonding between couples influence tumorigenesis, yet whether this is due to lifestyle changes, homogamy (likelihood of individuals to marry people of similar health), or directly associated with host-induced effects in tumors remains debatable. In the present study, we explored if tumorigenesis is associated with the bonding experience in monogamous rodents at which disruption of pair bonds is linked to anxiety and stress. Comparison of lung cancer cell spheroids that formed in the presence of sera from bonded and bond-disrupted deer mice showed that in monogamous Peromyscus polionotus and Peromyscus californicus, but not in polygamous Peromyscus maniculatus, the disruption of pair bonds altered the size and morphology of spheroids in a manner that is consistent with the acquisition of increased oncogenic potential. In vivo, consecutive transplantation of human lung cancer cells between P. californicus, differing in bonding experiences (n = 9 for bonded and n = 7 for bond-disrupted), and nude mice showed that bonding suppressed tumorigenicity in nude mice (p<0.05), suggesting that the protective effects of pair bonds persisted even after bonding ceased. Unsupervised hierarchical clustering indicated that the transcriptomes of lung cancer cells clustered according to the serum donors' bonding history while differential gene expression analysis pointed to changes in cell adhesion and migration. The results highlight the pro-oncogenic effects of pair-bond disruption, point to the acquisition of expression signatures in cancer cells that are relevant to the bonding experiences of serum donors, and question the ability of conventional mouse models to capture the whole spectrum of the impact of the host in tumorigenesis.

People's social interactions could influence their risk of developing various diseases, including cancer, according to population-level studies. In particular, studies have identified a so-called widowhood effect where a person's risk of disease increases following the loss of a spouse. However, the cause of the widowhood effect remains debatable, as it can be difficult to separate the impact of lifestyle changes from biological changes in the individual following bereavement. It is not possible to use laboratory mice to identify a causal biological mechanism, because they do not form long-term relationships with a single partner (pair bonds). However, several species of deer mouse form pair bonds, and suffer from anxiety and stress if these bonds are broken. Naderi et al. used these mice to study the widowhood effect on the risk of developing cancer. First, Naderi et al. grew human lung cancer cells in blood serum taken from mice that were either in a pair bond or had been separated from their partner. The cancer cells grown in the blood of mice with disrupted pair bonds changed size and shape, indicating that these mice were more likely to develop cancer. This effect was not observed when the cells were grown in the blood of bonded deer mice or of another deer mouse species that does not form pair bonds. Naderi et al. also found that the activity of genes involved in the cancer cells' ability to spread and to stick together was different in pair-bonded mice and in pair-separated mice. Next, Naderi et al. implanted lung cancer cells into the deer mice to study their effects on live animals. When cancer cells from the deer mice were transplanted into laboratory mice with a weakened immune system, the cells taken from pair-bonded deer mice were less likely to grow than the cells from deer mice with disrupted pair bonds. This suggests that the protective effects of pair bonding persist even after removal from the original mouse. These results provide evidence for a biological mechanism of the widowhood effect, where social experiences can alter gene activity relating to cancer growth. In the future, it will be important to determine whether the same applies to humans, and to find out if there are ways to mimic the effects of long-term bonds to improve cancer prognoses.

Neuroprotective Effect of Intravitreal Single-Dose Lithium Chloride after Optic Nerve Injury in Rats.

OBJECTIVE: Lithium is an old drug to control bipolar disorder. Moreover, it presents neuroprotective effects and supports neuronal plasticity. The aim of this study was to evaluate neuroprotective effect of intravitreal lithium after optic nerve injury. METHODS: Three dosages of lithium chloride, including 2 pmol, 200 pmol, and 2 nmol, were injected intravitreally after rat optic nerve injury. Proteins expression were assessed by western blot. Nitric oxide (NO) metabolites were measured by Griess test. Visual evoked potential (VEP) and optical coherence tomography (OCT) measurement were performed after trauma induction, in addition to H & E and TUJ1 staining of ganglion cells. RESULTS: Western blot depicted lithium can significantly increase antiapoptotic Bcl-2 protein level and reduce p-ERK, Toll-like receptor 4 (TLR4) and proapoptotic proteins such as Bax level in retinal tissue and Griess test reflected that NO metabolites level decreased in lithium treated eyes (P < .05). While, OCT showed no significant changes (P = .36 and P = .43 comparing treated group with trauma) in retinal ganglion cell layer thickness after lithium injection, VEP P2 wave amplitude increased significantly (P < .01) in lithium-treated eyes and its latency reduced (P < .05 for N1 wave and P < .01 for P2 wave). Tuj1 antibody-labeled retinal ganglion cells analyzing showed that the number of retinal ganglion cells were significantly higher in lithium treated eyes compared to untreated eyes with optic nerve injury. CONCLUSION: It seems intravitreally lithium has optic nerve neuroprotective effects by various mechanisms like overexpression of antiapoptotic proteins, suppressing proinflammatory molecules and proapoptotic factors, and decreasing nitric oxide.

Curcumin-silica nanocomplex preparation, hemoglobin and DNA interaction and photocytotoxicity against melanoma cancer cells.

Melanoma is a malignant cancer of the skin associated with a high mortality. Early medical diagnosis and surgical intervention are essential for the treatment of melanoma. The use of plant-based compounds is an important strategy for the prevention and treatment of different types of cancers. Curcumin is a promising natural anticancer compound used towards treatment for various kinds of cancers. Studies have shown that curcumin could be applied as a photosensitizer in cancer photodynamic therapy (PDT). PDT uses light and a photosensitizing agent which produce reactive oxygen species leading to cancer cell death. The main obstacle for using curcumin as photosensitizer is its low solubilization ability in an aqueous environment. To improve its application in cancer treatment, we synthetized curcumin-silica nanoparticles as photosensitizer for photodynamic treatment of human melanoma cancer cells. Scanning electron microscopy, Transmission electron microscopy, Powder X-ray diffraction and Thermo geometric analysis indicated that curcumin was loaded on silica. The solubility of curcumin in water increased by using silica nanoparticles which wasconfirmed by spectroscopy results. The spectroscopy study confirmed the interaction of curcumin-silica nanocomplex with double strand DNA and no interaction with hemoglobin. The curcumin-silica nanocomplex and curcumin photodynamic effect was investigated on human melanoma cancer cells (A375) and also human fibroblast cells. The cell toxicity experiments showed that the curcumin-silica nanocomplex had greater photodynamic effects on cancer cell death as compared to free curcumin. The apoptotic assay by acridine orange/ethidium bromide (AO/EB) dual staining and colony forming ability confirmed the MTT results. Therefore, these results suggest that the curcumin-silica nanocomplex has great potential to be employed in photodynamic treatment of melanoma cancer.

Role of Schwann Cells in Preservation of Retinal Tissue Through Reduction of Oxidative Stress.

The aim of this study was to evaluate the effect of subretinal injection of Schwann cells on preservation of retina by decreasing oxidative stress in Dystrophic Royal College of Surgeons (RCS) rats. Schwann cells were harvested from the sciatic nerve of postnatal day 5, RCS rats. Twenty-five RCS rats randomly assigned to cell and sham groups. Schwann cells injected in the sub-retinal space in one eye of the cell group and carrier medium was injected in one eye of the sham group. The proof for the appropriate site of injection of Schwann cells confirmed by the green fluorescent protein (GFP) positive cells. Electroretinogram (ERG) and enucleation for histopathology and enzymatic evaluation were performed 1, 2 and 3 months post-injection. The enzymatic evaluation included catalase, superoxide dismutase (SOD) and glutathione peroxidase 1 (GPx1) by enzyme-linked immunosorbent assay (ELISA) method. Three months after injection, histopathology assessments showed a complete absence of the outer nuclear layer (ONL), photoreceptors and obvious reduction of retinal pigment epithelium (RPE) in the sham group. Cell group showed marked preservation of RPE, choroidal congestion and mild presence of ONL. The green fluorescent protein positive Schwann cells remained in one integrated layer during the study under RPE. The enzymatic evaluation showed that in cell group expression of SOD and GPx1 until month 2 and catalase until month 1 were significantly more than the sham group. At the end of month 3, the amplitude of ERG waves significantly preserved in cell group in comparison to baseline waves and the sham group. We concluded that Schwan cells are able to preserve retinal in RCS rats by reducing oxidative stress.

Long term features of diabetic retinopathy in streptozotocin-induced diabetic Wistar rats.

Diabetic retinopathy is a complication of diabetes and a leading cause of vision loss among working-age adults. To assess whether the Wistar rat with Streptozotocin (STZ)-induced diabetes is a suitable animal model of human proliferative diabetic retinopathy we evaluated the vascular changes to assess the diabetic retinopathy (DR) stages in this model. After two weeks of intraperitoneal STZ (55 mg/kg) injection in male Wistar rats (270-300 g), they were considered diabetic with persistent blood glucose levels ≥ 16.65 mmol/L. The diabetic and control rats were investigated after 1, 3, 6 and 9 months by electroretinography, Evans blue assay, dextran fluorescence retinal angiography, and retinal histopathological studies. Retinal vascular permeability in the diabetic groups increased significantly in all diabetic groups. The amplitude of a- and b-waves decreased significantly in all diabetic groups compared with the age-matched control groups. The latent time of a-waves in the diabetic groups was delayed at 3 months of diabetes and this delay remained relatively constant till 9 months following the onset of diabetes. Although the latent time of b-wave in the diabetic groups increased slightly, a significant difference was found right at 9 months of diabetes. Vascular density and branching point numbers significantly decreased in the diabetic eyes at 3 and 6 months while they increased at 9 months, which was not significant. Intraretinal hemorrhage and ischemic changes were detected in the half of diabetic rats after 6 months and considered as preproliferative stage of diabetic retinopathy. Although preproliferative changes were detected in all diabetic rats at 9 months, half of them showed vitreous neovascularization attached to retina and retinal folds which can be considered as proliferative stage of DR. Intraretinal hemorrhage, extensive leakage of fluorescein, retinal folds, and vitreous neovascularization were the most prominent findings of severe and proliferative diabetic retinopathy in a fraction of the STZ-induced diabetic rats which were comparable to that of the human patients. STZ-induced diabetic rats can be considered to be a potentially useful model for studies on pathogenesis and treatment of diabetic retinopathy in human.

Erythropoietin in Treatment of Methanol Optic Neuropathy.

BACKGROUND: Methanol poisoning can cause an optic neuropathy that is usually severe and irreversible and often occurs after ingestion of illicit or homemade alcoholic beverages. In this study, we evaluated the potential neuroprotective effect of erythropoietin (EPO) on visual acuity (VA) in patients with methanol optic neuropathy. METHODS: In a prospective, noncomparative interventional case series, consecutive patients with methanol optic neuropathy after alcoholic beverage ingestion were included. All patients initially received systemic therapy including metabolic stabilization and detoxification. Treatment with intravenous recombinant human EPO consisted of 20,000 units/day for 3 successive days. Depending on clinical response, some patients received a second course of EPO. VA, funduscopy, and spectral domain optical coherence tomography were assessed during the study. Main outcome measure was VA. RESULTS: Thirty-two eyes of 16 patients with methanol optic neuropathy were included. Mean age was 34.2 years (±13.3 years). The mean time interval between methanol ingestion and treatment with intravenous EPO was 9.1 days (±5.56 days). Mean follow-up after treatment was 7.5 months (±5.88 months). Median VA in the better eye of each patient before treatment was light perception (range: 3.90-0.60 logMAR). Median last acuity after treatment in the best eye was 1.00 logMAR (range: 3.90-0.00 logMAR). VA significantly increased in the last follow-up examination (P < 0.0001). Age and time to EPO treatment after methanol ingestion were not significantly related to final VA. No ocular or systemic complications occurred in our patient cohort. CONCLUSIONS: Intravenous EPO appears to improve VA in patients with methanol optic neuropathy and may represent a promising treatment for this disorder.

Topical Tranexamic Acid Compared With Anterior Nasal Packing for Treatment of Epistaxis in Patients Taking Antiplatelet Drugs: Randomized Controlled Trial.

OBJECTIVE: We evaluated the efficacy of topical application of the injectable form of tranexamic acid (TXA) compared with anterior nasal packing (ANP) for the treatment of epistaxis in patients taking antiplatelet drugs (aspirin, clopidogrel, or both) who presented to the emergency department (ED). METHODS: A randomized, parallel-group clinical trial was conducted at two EDs. A total of 124 participants were randomized to receive topical TXA (500 mg in 5 mL) or ANP, 62 patients per group. The primary outcome was the proportion of patients in each group whose bleeding had stopped at 10 minutes. Secondary outcomes were the rebleeding rate at 24 hours and 1 week, ED length of stay (LOS), and patient satisfaction. RESULTS: Within 10 minutes of treatment, bleeding was stopped in 73% of the patients in the TXA group, compared with 29% in the ANP group (difference = 44%, 95% confidence interval, 26% to 57%; p < 0.001). Additionally, rebleeding was reported in 5 and 10% of patients during the first 24 hours in the TXA and the ANP groups, respectively. At 1 week, 5% of patients in the TXA group and 21% of patients in the ANP group had experienced recurrent bleeding (p = 0.007). Patients in the TXA group reported higher satisfaction scores (median [interquartile range {IQR}], 9 [8-9.25]) compared with the ANP group (median [IQR] = 4 [3-5]; p < 0.001). Discharge from the ED in <2 hours was achieved in 97% of patients in the TXA group versus 13% in the ANP group (p < 0.001). There were no adverse events reported in either group. CONCLUSIONS: In our study population, epistaxis treatment with topical application of TXA resulted in faster bleeding cessation, less rebleeding at 1 week, shorter ED LOS, and higher patient satisfaction compared with ANP.

Methionine synthase A2756G variation is associated with the risk of retinoblastoma in Iranian children.

Association of epigenetic modifications with cancer has been widely studied. Gene-specific hypermethylation and global DNA hypomethylation are the most frequently observed patterns in great number of tumors. The methionine synthase (MTR) gene plays key role in maintaining adequate intracellular folate, methionine and normal homocysteine concentrations and, its polymorphism have been associated with the risk of retinoblastoma and other neoplasms. We evaluated the association of MTR A2756G polymorphism with the risk of retinoblastoma in an Iranian population. Totally, 150 retinoblastoma patients and 300 individuals with no family history of cancer as control were included in this study. Genotyping of the A2756G polymorphism was performed by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) using the restriction enzymes HaeIII. Our results showed that the "G" was the minor allele with a frequency of 31.7% and 20.3% in both retinoblastoma and control groups, respectively. The frequency of the 2756GG genotype (P=0.023) and 2756G allele (P=0.0001) were significantly higher in the patients than control group, respectively. Individual with the 2756GG genotype had a 2.99 fold increased risk for retinoblastoma. According to our results, the MTR A2756G polymorphism was associated with the risk of retinoblastoma in Iranian patients.

Protection against brain tissues oxidative damage as a possible mechanism for the beneficial effects of Rosa damascena hydroalcoholic extract on scopolamine induced memory impairment in rats.

OBJECTIVES: Hypnotic, analgesic, anticonvulsant, and antioxidant effects of Rosa damascena have been reported. This study, investigated the effect of R. damascena hydroalcoholic extract on memory performance in a scopolamine-induced memory impairment model. METHODS: The rats were divided into control group received just saline; scopolamine group was treated by saline for 2 weeks, but was injected by scopolamine 30 minutes before each trial in Morris water maze test; treatment groups (scopolamine + extract 50; Sco + Ext 50) and (scopolamine + extract 250; Sco + Ext 250) were daily treated by 50 and 250 mg/kg of R. damascena extract (2 weeks) and were finally injected by scopolamine before each trial in Morris water maze. The brains were removed for biochemical measurements. RESULTS: Time latency and path length in the scopolamine group were higher than control (P < 0.01 to <0.001). Both treatment groups showed shorter traveled distance and time latency compared with scopolamine group (P < 0.05 to <0.001). Time spent in target quadrant by scopolamine group was lower than control (P < 0.05), while Sco + Ext 250 group spent longer time in target quadrant than scopolamine group (P < 0.05). Malondialdehyde concentrations in hippocampal and cortical tissues of scopolamine group were higher, while thiol concentrations were lower than control ones (P < 0.001). Treatment by both doses of the extract decreased the malondialdehyde concentration, while increased the thiol concentration (P < 0.05 to <0.001). DISCUSSION: The results of this study showed that the hydroalcoholic extract of R. damascena prevents scopolamine-induced memory deficits. This finding suggests that memory improvement may be in part due to the antioxidant effects.

Estradiol attenuates spinal cord injury-related central pain by decreasing glutamate levels in thalamic VPL nucleus in male rats.

Central neuropathic pain (CNP) is a complicated medical problem that involves both the spinal and supraspinal regions of the central nervous system. Estrogen, a neuroprotective agent, has been considered a possible candidate for CNP treatment. In this study, we examined the effects of a single dose of 17ß-estradiol on glutamate levels in the ventral posterolateral (VPL) nucleus of the rat thalamus. Furthermore, we determined whether there was a correlation between glutamate levels and neuropathic pain induced by unilateral electrolytic spinothalamic tract (STT) lesion. STT lesioning was performed in male Wistar rats at the T8-T9 vertebrae; rats were then administered 17ß-estradiol (4 mg/kg, i.p.) 30 min after injury. Glutamate samples were collected using a microdialysis probe and quantified by high performance liquid chromatography. Mechanical allodynia (MA) and thermal hyperalgesia (TH) thresholds were measured pre-injury and 7, 14, and 28 days post-injury. We found that STT lesion significantly increased glutamate levels in the ipsilateral VPL nucleus 14 and 28 days post-injury; this was accompanied by allodynia and hyperalgesia in the hind paws of the rats. Administering 17ß-estradiol to the rats decreased glutamate levels in the ipsilateral VPL nucleus and significantly increased MA and TH thresholds. These results suggest that glutamate in the VPL nucleus of the thalamus is involved in the pathology of neuropathic pain after STT injury; furthermore, 17ß-estradiol may attenuate this neuropathic pain by decreasing glutamate levels.

Treadmill exercise reduces self-administration of morphine in male rats.

Exercise can activate the same pathways as morphine. The aim of the present study was to clarify the effect of short-term and mid-term exercises on the self-administration of morphine in rats. Male Wistar rats were initially trained to receive small pellets of food by pressing the active lever in self-administration apparatus. Rats were divided into 4 groups: Saline, Morphine, Exercise 1 (11 days) and Exercise 2 (30 days). Their jugular vein was cannulated. The animals were placed in self-administration apparatus and allowed to self-administer morphine (0.5mg per infusion all test groups) or saline (Saline group) during consecutive days, for 2h/sessions. In the group 1 the rats were running before each session of self-administration and of group Exercise 2, 30 days before surgery as well as before each session. The pressing numbers of active and passive levers in each group and among different groups were compared. The number of active lever pressing of Morphine group was significantly higher than Saline group (p<0.001). In Exercise 1 and Exercise 2 groups, the number of active lever pressing was significantly lower than Morphine group (p<0.001). As exercise can activate many neurotransmitter systems involved in the addiction process and increase the release of endorphins, it is likely that could decrease the morphine self-administration in this experimental setup.