Incorporation of Saqez essential oil into polyvinyl alcohol/chitosan bilayer hydrogel as a potent wound dressing material.

Nowadays, many studies are conducted on multilayer hydrogels for wound dressing. On the other hand, considering the emergence of bacterial resistance to common antibiotics, studies on the use of natural essential oils and their derivatives that have antibacterial and antioxidant activity can be useful. Herein, a novel bilayer hydrogel developed from polyvinyl alcohol and chitosan with the incorporation of Saqez essential oil (SEO) was synthesized. The results showed a gel-type structure with specific compression and flexibility, while the microscopic images confirmed the formation of a bilayer hydrogel. Further, the data showed that increasing the concentration of SEO reduces the swelling and water vapor permeability and increases the water retention and hydrophobicity of the hydrogel surface. The effects of the combination of SEO in the bilayer hydrogel led to a strong antioxidant property and increased antimicrobial activity. Also, the in vitro results demonstrated that the bilayer hydrogels are biocompatible, non-toxic, and blood compatible. Finally, the results of the in vivo tests showed that these bilayer hydrogels had good homeostatic efficiency. Overall, the obtained results indicate that these bilayer hydrogels are promising candidates for wound dressing.

Radiomics for differentiation of the posterior fossa pilocytic astrocytoma versus hemangioblastomas in adults. A pilot study.

PURPOSE: Both pilocytic astrocytoma (PA) and hemangioblastoma (HB) are common primary neoplasms of the posterior fossa with similar radiological manifestations. This study was conducted to evaluate the role of Radiomics in differentiating these two conditions in adults. MATERIALS AND METHODS: After a retrospective search of our institutional imaging archive, adult patients with a known diagnosis of PA or HB were included. We reviewed each patient's most recent preoperative brain magnetic resonance imaging (MRI). The solid enhancing nodule of each lesion on post-contrast T1 sequence was manually segmented. Multiple Radiomics features were then extracted from each nodule using the Pyradiomics library. Subsequently, the most predictive features were identified by feature selection models. Following this, different machine learning (ML) models were constructed based on these selected features to classify lesions as PA or HB. Finally, we evaluated the performance of each model by leave-one-out cross-validation. RESULTS: With inclusion and exclusion criteria, 34 enhancing PA nodules and 39 HB nodules were selected. A total of 115 features were extracted from each enhancing nodule. Twelve characteristics were detected as most predictive of histopathological diagnosis. Among various ML models, the neural network had the best performance in differentiating these two conditions with an AUC of 0.9 and an accuracy of 82%. CONCLUSIONS: In this retrospective study, Radiomics MRI techniques demonstrated high performance in distinguishing adult posterior fossa PA from HB. Future development of Radiomics models may advance presurgical diagnosis of these two conditions when added to routine clinical practice and thus improve patient management.

Higher dietary total antioxidant capacity (TAC) reduces the risk of cardio-metabolic risk factors among adults: An updated systematic review and meta-analysis.

Background: Numerous studies have revealed the protective role of dietary antioxidants against hypertension and diabetes. In the current systematic review and meta-analysis, we evaluated the possible role of dietary total antioxidant capacity (TAC) against metabolic parameters in the adult population. Methods: A literature search of authentic electronic resources including PubMed, Scopus, Web of Sciences, and Cochrane had been performed to retrieve the qualified observational studies that reported the mean plus/minus the SD for the parameter in subjects of the highest versus lowest dietary TAC categories up to July 2020. Results: Totally, the number of included studies was 13 for fasting blood sugar (FBS), 12 for systolic blood pressure (SBP), and 12 for diastolic blood pressure (DBP), with the participants' number of 26349. The results of the current study showed that being at highest category of TAC significantly reduces serum FBS (WMD: -2.381; CI: -3.445, -1.316; P<0.001), SBP (WMD: -1.080; CI: -1.458, -0.701; P<0.001) and DBP (WMD: -0.854; CI: -1.655, -0.052; P<0.001), while no significant difference in the serum insulin, HOMA-IR values, prevalence of type 2 diabetes (T2DM) (P=0.37) and hypertension (HTN) (P=0.09) was observed. Subgroup analysis revealed the geographical location, dietary assessment tool, sample size, and gender as possible sources of heterogeneity. Conclusion: Higher intake of dietary TAC was associated with reduced SBP, DBP, and FBS in the current observational studies of this meta-analysis. These findings further confirm the clinical importance of dietary antioxidants in the prevention of different metabolic disorders.

The Assessment of Marcaine Versus Meperidine for Spinal Anesthesia in Anorectal Surgery: A Randomized Clinical Trial.

Background: Spinal anesthesia (SA) for the surgical management of chronic anal fissures is favored by surgeons as it provides an early return to daily activities; however, the agents applied for SA to achieve the best outcomes with minimized adverse effects are a matter of debate. Objectives: This study aimed to assess the utility of Marcaine versus meperidine for SA induction of anoderm surgery. Methods: This randomized clinical trial (RCT) was conducted on 138 patients with chronic anal fissures who were candidates for surgical management in 2020. The patients were randomly assigned to two groups of SA using 2.5 mL of hyperbaric Marcaine 0.5% (n = 69) or 1 mg/kg of meperidine (n = 69). Pain severity (measured via Numerical Rating Scale (NRS)), anal sphincter tone manometry (measured at baseline and the end of the sphincterotomy), and drug-related adverse effects were compared between the groups. Results: Both agents led to significant pain relief within 24 hours after SA (P < 0.05); nevertheless, pain severity was remarkably lower in meperidine-treated patients in different measurements performed during the first 24 hours after SA (P < 0.05). The sphincteric tone significantly decreased in both groups (P < 0.001), while the postoperative tone was significantly less in the Marcaine-treated patients (65.22 ± 3.02 versus 46.04 ± 1.97, P < 0.001). The two groups did not differ regarding the adverse effects (P > 0.05). Conclusions: Meperidine for SA in anal fissure surgical management was relatively superior to Marcaine, as postoperative pain control was remarkably better achieved with meperidine. However, anal sphincter tone reached a normal range in Marcaine-treated cases, and the average tone in those anesthetized with meperidine was slightly above the normal limits.

Evaluation of the epidemiologic, clinical, radiologic, and treatment methods of patients with subacute and chronic meningitis.

BACKGROUND: Meningitis is known as a meningeal inflammation accompanied by pleocytosis in the cerebrospinal fluid (CSF), and can be classified into acute, subacute, and chronic meningitis based on symptoms duration of ≤ 5 days, ≥ 5 days and ≥ 4 weeks, respectively. Subacute and chronic meningitis are caused mainly by indolent infectious agents and noninfectious causes such as autoimmune, and neoplastic. In this study, we investigated the characteristics, diagnosis, and treatment of subacute and chronic meningitis. METHODS: We extracted the medical records of patients with chronic and subacute meningitis who were referred to three tertiary centers from Jun 2011 to Jun 2021. Initially, 2050 cases of meningitis were screened, and then 79 patients were included in the study. RESULTS: Headache (87.3%), nausea and vomiting (74.7%), fever (56.4%), and visual impairments (55.7%) were the most prevalent symptoms. The most common signs were nuchal rigidity (45.3%), altered mental status (26.9%), and papillary edema (37.5%). Brain computed tomography (CT) was normal in 68.6% of the patients while 22.9% of the cases had hydrocephalus. Brain magnetic resonance imaging (MRI) was normal in 60.0% of the patients. The most common abnormal MRI findings were leptomeningeal enhancement (16.0%) and hydrocephalus (16.0%). We had a 44.3% definite diagnosis with bacterial (n:25, 31.6%) and neoplastic (n:8, 10.1%) being the most prevalent etiologies. Mycobacterium tuberculosis (60%) and Brucella spp. (12%) were the most prevalent bacterial pathogens. CONCLUSIONS: The most common etiologies include infectious, neoplastic, and immunologic. Due to insidious presentation and uncommon etiologies, establishing a proper diagnosis, and providing timely targeted treatment for patients with subacute and chronic meningitis remains a challenge for clinicians.

Green extraction of nickel and valuable metals from pyrrhotite samples with different crystallographic structures through acidophilic bioleaching.

Nowadays, due to the strategic status of nickel in the global market, utilizing its disregarded resources like low-grade nickel containing pyrrhotite is of significant importance. A comprehensive set of experiments and analyses were performed to determine the bioleaching capability and mechanism for nickel extraction from hexagonal and monoclinic pyrrhotite. Over 95% Ni extraction was achieved from the hexagonal pyrrhotite sample. Ni extraction from the monoclinic sample reached its maximum value of 67% and 90% at 3% pulp density, with mixed mesophilic and moderately thermophilic cultures, respectively. Characterization analyses indicated that jarosite and elemental sulfur formation in mixed mesophilic bioleaching reduced the samples' bio-oxidation rate and metal dissolution. The kinetics study revealed that the controlling step in thermophilic bioleaching is the chemical reaction; however, the mixed control model was best fitted on mesophilic data. Electrochemistry studies confirmed bioleaching results and indicated that monoclinic pyrrhotite's oxidation rate under the operating conditions is faster than hexagonal pyrrhotite, and the temperature positively correlates with the oxidation rate. Toxicity assessment analysis showed that the final residues of both bioleached samples could be considered environmentally safe.

Discovery of Leptulipin, a New Anticancer Protein from theIranian Scorpion, Hemiscorpius lepturus.

Cancer is one of the leading causes of mortality in the world. Unfortunately, the present anticancer chemotherapeutics display high cytotoxicity. Accordingly, the discovery of new anticancer agents with lower side effects is highly necessitated. This study aimed to discover an anticancer compound from Hemiscorpius lepturus scorpion venom. Bioactivity-guided chromatography was performed to isolate an active compound against colon and breast cancer cell lines. 2D electrophoresis and MALDI-TOF were performed to identify the molecule. A partial protein sequence was obtained by mass spectrometry, while the full-length was deciphered using a cDNA library of the venom gland by bioinformatics analyses and was designated as leptulipin. The gene was cloned in pET-26b, expressed, and purified. The anticancer effect and mechanism action of leptulipin were evaluated by MTT, apoptosis, and cell cycle assays, as well as by gene expression analysis of apoptosis-related genes. The treated cells displayed inhibition of cell proliferation, altered morphology, DNA fragmentation, and cell cycle arrest. Furthermore, the treated cells showed a decrease in BCL-2 expression and an increase in Bax and Caspase 9 genes. In this study, we discovered a new anticancer protein from H. lepturus scorpion venom. Leptulipin showed significant anticancer activity against breast and colon cancer cell lines.

Artemisinins in Combating Viral Infections Like SARS-CoV-2, Inflammation and Cancers and Options to Meet Increased Global Demand.

Artemisinin is a natural bioactive sesquiterpene lactone containing an unusual endoperoxide 1, 2, 4-trioxane ring. It is derived from the herbal medicinal plant Artemisia annua and is best known for its use in treatment of malaria. However, recent studies also indicate the potential for artemisinin and related compounds, commonly referred to as artemisinins, in combating viral infections, inflammation and certain cancers. Moreover, the different potential modes of action of artemisinins make these compounds also potentially relevant to the challenges the world faces in the COVID-19 pandemic. Initial studies indicate positive effects of artemisinin or Artemisia spp. extracts to combat SARS-CoV-2 infection or COVID-19 related symptoms and WHO-supervised clinical studies on the potential of artemisinins to combat COVID-19 are now in progress. However, implementing multiple potential new uses of artemisinins will require effective solutions to boost production, either by enhancing synthesis in A. annua itself or through biotechnological engineering in alternative biosynthesis platforms. Because of this renewed interest in artemisinin and its derivatives, here we review its modes of action, its potential application in different diseases including COVID-19, its biosynthesis and future options to boost production.

Clinical applications of diffusion-weighted sequence in brain imaging: beyond stroke.

Diffusion-weighted imaging (DWI) is a well-established MRI sequence for diagnosing early stroke and provides therapeutic implications. However, DWI yields pertinent information in various other brain pathologies and helps establish a specific diagnosis and management of other central nervous system disorders. Some of these conditions can present with acute changes in neurological status and mimic stroke. This review will focus briefly on diffusion imaging techniques, followed by a more comprehensive description of the utility of DWI in common neurological entities beyond stroke.

The Role of Machine Learning and Radiomics for Treatment Response Prediction in Idiopathic Normal Pressure Hydrocephalus.

Introduction Ventricular shunting remains the standard of care for patients with idiopathic normal pressure hydrocephalus (iNPH); however, not all patients benefit from the shunting. Prediction of response in advance can result in improved patient selection for ventricular shunting. This study aims to develop a machine learning predictive model for treatment response after shunt placement using the clinical and radiomics features. Methods In this retrospective pilot study, the medical records of iNPH patients who underwent ventricular shunting were evaluated. In each patient, the "idiopathic normal pressure hydrocephalus grading scale" (iNPHGS) and a "Modified Rankin Scale" were calculated before and after surgery. The subsequent treatment response was calculated as the difference between the iNPHGS scores before and after surgery. iNPHGS score reduction of two or more than two were considered as treatment response. The presurgical MRI scans were evaluated by radiologists, the ventricular systems were segmented on the T2-weighted images, and the radiomics features were extracted from the segmented ventricular system. Using Orange data mining open-source platform, different machine learning models were then developed based on the presurgical clinical features and the selected radiomics features to predict treatment response after shunt placement. Results After the implementation of the inclusion criteria, 78 patients were included in this study. One hundred twenty radiomics features were extracted, and the 12 best predictive radiomics features were selected. Using only clinical data (iNPHGS and Modified Rankin Scale), the random forest model achieved the best performance in treatment prediction with an area under the curve (AUC) of 0.71. Adding the Radiomics analysis to the clinical data improved the prediction performance, with the support vector machine (SVM) achieving the highest rank in treatment prediction with an AUC of 0.8. Adding age and sex to the analysis did not improve the prediction. Conclusion Using machine learning models for treatment response prediction in patients with iNPH is feasible with acceptable accuracy. Adding the Radiomics analysis to the clinical features can further improve the predictive performance. SVM is likely the best model for this task.

Proteomic signature of the Dravet syndrome in the genetic Scn1a-A1783V mouse model.

BACKGROUND: Dravet syndrome is a rare, severe pediatric epileptic encephalopathy associated with intellectual and motor disabilities. Proteomic profiling in a mouse model of Dravet syndrome can provide information about the molecular consequences of the genetic deficiency and about pathophysiological mechanisms developing during the disease course. METHODS: A knock-in mouse model of Dravet syndrome with Scn1a haploinsufficiency was used for whole proteome, seizure, and behavioral analysis. Hippocampal tissue was dissected from two- (prior to epilepsy manifestation) and four- (following epilepsy manifestation) week-old male mice and analyzed using LC-MS/MS with label-free quantification. Proteomic data sets were subjected to bioinformatic analysis including pathway enrichment analysis. The differential expression of selected proteins was confirmed by immunohistochemical staining. RESULTS: The findings confirmed an increased susceptibility to hyperthermia-associated seizures, the development of spontaneous seizures, and behavioral alterations in the novel Scn1a-A1873V mouse model of Dravet syndrome. As expected, proteomic analysis demonstrated more pronounced alterations following epilepsy manifestation. In particular, proteins involved in neurotransmitter dynamics, receptor and ion channel function, synaptic plasticity, astrogliosis, neoangiogenesis, and nitric oxide signaling showed a pronounced regulation in Dravet mice. Pathway enrichment analysis identified several significantly regulated pathways at the later time point, with pathways linked to synaptic transmission and glutamatergic signaling dominating the list. CONCLUSION: In conclusion, the whole proteome analysis in a mouse model of Dravet syndrome demonstrated complex molecular alterations in the hippocampus. Some of these alterations may have an impact on excitability or may serve a compensatory function, which, however, needs to be further confirmed by future investigations. The proteomic data indicate that, due to the molecular consequences of the genetic deficiency, the pathophysiological mechanisms may become more complex during the course of the disease. As a result, the management of Dravet syndrome may need to consider further molecular and cellular alterations. Ensuing functional follow-up studies, this data set may provide valuable guidance for the future development of novel therapeutic approaches.

Emerging Applications of Radiomics in Neurological Disorders: A Review.

Radiomics has achieved significant momentum in radiology research and can reveal image information invisible to radiologists' eyes. Radiomics first evolved for oncologic imaging. Oncologic applications (histopathology, tumor grading, gene mutation analysis, patient survival, and treatment response prediction) of radiomics are widespread. However, it is not limited to oncologic analysis, and any digital medical images can benefit from radiomics analysis. This article reviews the current literature on radiomics in non-oncologic, neurological disorders including ischemic strokes, hemorrhagic stroke, cerebral aneurysms, and demyelinating disorders.

Mannose-binding Lectin Mediated Complement Pathway in Autoimmune Neurological Disorders.

Multiple sclerosis (MS) is a complex, demyelinating disease of the central nervous system (CNS) with variable phenotypic presentations, while Guillain-Barre Syndrome (GBS) is the prototypic acute inflammatory disorder that affects the peripheral nervous system. Myasthenia gravis (MG) is a T cell dependent and antibody mediated autoimmune disease. Although it has been shown that complement plays a critical role in the pathogenesis of MS, GBS, and MG, the role of mannose-binding lectin (MBL) as a biomarker of immunopathogensis of these diseases and also its association with the severity of them have been poorly investigated. Therefore, in this study we aimed to measure plasma levels of MBL in patients with MS, GBS, and MG. In a case-control study, plasma was obtained from healthy controls (n=100) and also patients with MS (n=120), GBS (n=30), and MG (n=30). Plasma level measurement of MBL was performed using enzyme-linked immunosorbent assay (ELISA). The mean serum level of MBL was significantly different between groups of patients and healthy controls (p<0.001). We also found a positive correlation between plasma levels of MBL and severity scores of MS, MG, and GBS patients including: expanded disability status scale (EDSS) (r=+0.60 and p=<0.001), quantitative myasthenia gravis score (QMGS) (r=+0.56 and p=0.01), and GBS disability scale (GDS) (r=+0.37 and p=0.04). Taken together, our findings suggest that complement activation mediated by MBL contributes to the pathogenesis and also severity of MS, MG, and GBS. However, because the lectin pathway can be involved in several phases of the immune response, further evidence will be required to elucidate the underlying mechanism.

A Novel Approach to Discriminate Subgroups in Multiple Sclerosis.

Multiple sclerosis (MS) is an autoimmune disease of central nervous system. Since different types of immune cells are involved in MS pathogenesis, in this study we aimed to evaluate serum levels of several immunological components including soluble CD4 (sCD4), sCD8, sCD163, and immunoglobulins as markers of activity of T-cells, macrophages, and B-cells in different types of MS. Serum levels of sCD4, sCD8, and sCD163 of patients with relapsing-remitting MS (RRMS, n=61), primary progressive MS (PRMS, n=31), secondary progressive MS (SPMS, n=31), clinical isolated syndrome (CIS, n=31) and neuromyelitis optica (NMO, n=31), and healthy controls (n=49) were measured using enzyme-linked immunosorbent assay (ELISA). Serum levels of Ig-G, Ig-M, and Ig-A were determined using nephelometric technique. Serum levels of sCD4, sCD8, sCD163, Ig-G, Ig-M, and Ig-A were significantly different in five groups of cases (p<0.05). Furthermore, application of stepwise method of discriminant analysis yielded 4 significant discriminant functions of classification due to the presence of six levels of categorical variables in the analysis. The most important function explained 85.5% of the total variance with the correlation value of 0.79. Taken together, our preliminary analysis suggests that although we found some functions to discriminate most of the patients, further studies will be required to individuate immunological markers characterizing the different type of MS including RRMS, PPMS, SPMS, CIS and NMO as proved by the data on sCD4, sCD163, Ig-M, and Ig-G in blood.

TNF-alpha Production by Peripheral Blood Monocytes in Multiple Sclerosis Patients and Healthy Controls.

BACKGROUND: Aberrant immune responses are evident in the pathogenesis of multiple sclerosis (MS) and it has been proposed that the spectrum of cytokines influence disease outcomes. Leptin and lipopolysaccharide (LPS) of Gram-negative bacteria are both potent cellular stimulators for production of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α). The aim of this study was to compare the TNF-α production by peripheral blood monocytes from MS patients with healthy controls. METHODS: Peripheral blood samples were stimulated with LPS or leptin. After blocking the Golgi apparatus, intracellular cytokine production was assessed using a monoclonal antibody against human TNF-α by the flow cytometry technique. Moreover, plasma level measurement of cytokines was performed using enzyme-linked immunosorbent assay (ELISA). RESULTS: Intracellular levels of TNF-α were 16.80 ± 8.21 and 16.52 ± 8.23in MS patients and healthy controls which showed no statistically significant difference between them (p = 0.850). Leptin-stimulated and LPS-stimulated TNF-α production showed no significant difference between MS patients and the control group (p = 0.263 and p = 0.191, respectively). However, after treatment with leptin, a weak significant difference was shown between cases and control group (p = 0.049). There were significant differences between cases and controls regarding serum levels of IL-6 and Toll-like receptor-4 (TLR-4) before and after stimulation with leptin and LPS, separately (p < 0.05). CONCLUSION: Taken together, we cannot definitely conclude that TNF-α does not play an important role in pathogenesis of MS. However, other characteristics of monocyte activation such as IL-6 or TLRs can elucidate implication of peripheral blood monocytes in MS pathogenesis.

Dynamic response of RNA editing to temperature in Drosophila.

BACKGROUND: Adenosine-to-inosine RNA editing is a highly conserved process that post-transcriptionally modifies mRNA, generating proteomic diversity, particularly within the nervous system of metazoans. Transcripts encoding proteins involved in neurotransmission predominate as targets of such modifications. Previous reports suggest that RNA editing is responsive to environmental inputs in the form of temperature alterations. However, the molecular determinants underlying temperature-dependent RNA editing responses are not well understood. RESULTS: Using the poikilotherm Drosophila, we show that acute temperature alterations within a normal physiological range result in substantial changes in RNA editing levels. Our examination of particular sites reveals diversity in the patterns with which editing responds to temperature, and these patterns are conserved across five species of Drosophilidae representing over 10 million years of divergence. In addition, we show that expression of the editing enzyme, ADAR (adenosine deaminase acting on RNA), is dramatically decreased at elevated temperatures, partially, but not fully, explaining some target responses to temperature. Interestingly, this reduction in editing enzyme levels at elevated temperature is only partially reversed by a return to lower temperatures. Lastly, we show that engineered structural variants of the most temperature-sensitive editing site, in a sodium channel transcript, perturb thermal responsiveness in RNA editing profile for a particular RNA structure. CONCLUSIONS: Our results suggest that the RNA editing process responds to temperature alterations via two distinct molecular mechanisms: through intrinsic thermo-sensitivity of the RNA structures that direct editing, and due to temperature sensitive expression or stability of the RNA editing enzyme. Environmental cues, in this case temperature, rapidly reprogram the Drosophila transcriptome through RNA editing, presumably resulting in altered proteomic ratios of edited and unedited proteins.

Multiple sclerosis and hyperprolactinemia: a case-control study.

UNLABELLED: Multiple sclerosis (MS) is an autoimmune disease of central nervous system which is characterized with demyelination. Prolactin, synthesized in the anterior pituitary cells, has a role in maturation of immune cells, suggesting its possible implication in autoreactivity. The aim of the current study is to investigate the role of hyperprolactinemia in MS. Twenty-two MS patients with hyperprolactinemia diagnosed with pituitary adenoma and 66 MS patients without hyperprolactinemia were enrolled in our case-control study. They were matched with regard to age, gender, and MS subtypes. Patients with other concomitant autoimmune diseases and pregnancy were excluded. Statistical analyses were carried out using SPSS (SPSS statistic package, version 21.0.0) statistical software. The Pearson Chi-square test and the t test were used to determine whether there were any significant differences. The level of significance was set at p < 0.05. Greater value of relapse rate among hyperprolactinemic MS patients in comparison to non-hyperprolactinemic MS patients was statistically significant (p < 0.001). Statistically significant difference between the two groups in terms of EDSS was observed (case group vs. CONTROL: 1.3 vs. 1.9; p = 0.007). The correlation between MS duration and duration of hyperprolactinemia was significant in the case group (p < 0.05, R = 0.752). No statistically significant difference was found between two groups regarding duration of MS. This study suggested a protective role of prolactin in demyelinating procedure of MS.

Co-administration of trientine and flaxseed oil on oxidative stress, serum lipids and heart structure in diabetic rats.

The administration of flaxseed oil or flaxseed oil plus trientine in diabetic rats reduced triglyceride, very low density lipoprotein, and total cholesterol. Furthermore, the combined treatment significantly increased superoxide dismutase activity and attenuated serum Cu2+. The results suggest that the administration of flaxseed oil plus trientine is useful in controlling serum lipid abnormalities, oxidative stress, restoring heart structure, and reducing serum Cu2+ in diabetic rats.