Beam narrowing test: a motor index of post-stroke motor evaluation in an aged rat model of cerebral ischemia.

Each year, 15 million people worldwide suffer from strokes. Consequently, researchers face increasing pressure to develop reliable behavioural tests for assessing functional recovery after a stroke. Our aim was to establish a new motor performance index that can be used to evaluate post-stroke recovery in both young and aged animals. Furthermore, we validate the proposed procedure and recommend the necessary number of animals for experimental stroke studies. Young (n = 20) and aged (n = 27) Sprague-Dawley rats were randomly assigned to receive either sham or stroke surgery. The newly proposed performance index was calculated for the post-stroke acute, subacute and chronic phases. The advantage of using our test over current tests lies in the fact that the newly proposed motor index test evaluates not only the performance of the unaffected side in comparison to the affected one but also assesses overall performance by taking into account speed and coordination. Moreover, it reduces the number of animals needed to achieve a statistical power of 80%. This aspect is particularly crucial when studying aged rodents. Our approach can be used to monitor and assess the effectiveness of stroke therapies in experimental models using aged animals.

Investigating the Neuroprotective and Neuroregenerative Effect of Trazodone Regarding Behavioral Recovery in a BL6C57 Mice Stroke Model.

Stroke is a major cause of death and disability worldwide. Between 1990 and 2010, its global burden increased notably with reference to the absolute number of incident events, number of deaths, and disability-adjusted life-years lost. Trazodone is a triazolopyridine derivative that was approved for more than 40 years as monotherapy or in combination with other antidepressant drugs for the treatment of major depressive disorder in adult patients. The aim was investigated if trazodone can improve behavioural outcome after stroke in a mice model of middle cerebral artery occlusion (MCAo) due to the potential neuroprotective and neurodegenerative effects by using three behavioural tests: adhesive tape test, beam test and hole board test. Trazodone administration show modest improvements regarding the motor-sensorial function after stroke especially in the acute post-stroke phase in aged and young animals. The antidepressant effect of the drug was observed in the post-stroke period in aged animals and to a lesser extent in young animals. Future research is needed to evaluate the effects of trazodone at the cellular level to be sure that it has no benefit in stroke patients who do not suffer from depression.

Molecular prognostic factors in colorectal cancer: 5-year follow-up.

Colorectal cancer (CRC) is a frequently diagnosed and lethal disease. The risk of developing CRC is determined by environmental and genetic factors. Surgical treatment is the main curative modality for patients with CRC up to stage III. In recent years, a special place has been given to biological agents used as targeted therapy following the genetic analysis of the tumor: Bevacizumab (Avastin), Cetuximab (Erbitux), Ziv-aflibercept (Zaltrap). We present a study based on 46 colorectal tumor resection specimens from patients operated for CRC in the Surgery Departments of the Emergency County Clinical Hospital of Craiova, Romania. Histopathological examination and immunohistochemistry staining of tissue sections were performed to determine the degree of aggressiveness. Using the Kaplan-Meier test, we calculated the correlation coefficient between survival time and immunohistochemical prognostic factors. The patients were followed for 60 months postoperatively.

From Molecule to Patient Rehabilitation: The Impact of Transcranial Direct Current Stimulation and Magnetic Stimulation on Stroke-A Narrative Review.

Stroke is a major health problem worldwide, with numerous health, social, and economic implications for survivors and their families. One simple answer to this problem would be to ensure the best rehabilitation with full social reintegration. As such, a plethora of rehabilitation programs was developed and used by healthcare professionals. Among them, modern techniques such as transcranial magnetic stimulation and transcranial direct current stimulation are being used and seem to bring improvements to poststroke rehabilitation. This success is attributed to their capacity to enhance cellular neuromodulation. This modulation includes the reduction of the inflammatory response, autophagy suppression, antiapoptotic effects, angiogenesis enhancement, alterations in the blood-brain barrier permeability, attenuation of oxidative stress, influence on neurotransmitter metabolism, neurogenesis, and enhanced structural neuroplasticity. The favorable effects have been demonstrated at the cellular level in animal models and are supported by clinical studies. Thus, these methods proved to reduce infarct volumes and to improve motor performance, deglutition, functional independence, and high-order cerebral functions (i.e., aphasia and heminegligence). However, as with every therapeutic method, these techniques can also have limitations. Their regimen of administration, the phase of the stroke at which they are applied, and the patients' characteristics (i.e., genotype and corticospinal integrity) seem to influence the outcome. Thus, no response or even worsening effects were obtained under certain circumstances both in animal stroke model studies and in clinical trials. Overall, weighing up risks and benefits, the new transcranial electrical and magnetic stimulation techniques can represent effective tools with which to improve the patients' recovery after stroke, with minimal to no adverse effects. Here, we discuss their effects and the molecular and cellular events underlying their effects as well as their clinical implications.

Immunoexpression of Mismatch Repair Proteins in a Cohort of Colorectal Cancer Patients.

One of the molecular routes of colorectal carcinogenesis is the lack of mismatch repair (MMR) proteins, which may have substantial clinical consequences in predicting therapy success. This study aimed to analyze the expression of the MutL homolog 1 (MLH1), MutS homolog 2 (MSH2), and MutS homolog 6 (MSH6) in a cohort of 91 colorectal cancer (CRC) patients, and to evaluate the relationship between patient clinicopathological characteristics and immunoexpression of these biomarkers. In this study, we obtained the highest scores of the MLH1 immunoexpression in non-mucinous tumors, moderately differentiated lesions, and in stage IV. The highest values of the MSH2 and MSH6 scores were observed in mucinous tumors, and poorly differentiated lesions, in stages II-III, and stages III-IV, respectively. To improve the stratification criteria for targeted oncological therapy and to predict patient outcomes, markers used may help evaluate the aggressiveness of lesions.

Zinc-Boron Complex-Based Dietary Supplements for Longevity and Healthy Life.

The nutritional role of zinc (Zn) and boron (B) in the plant-animal-human food chain is highly topical worldwide research. Our data indicated that Zn-B complex (ZBC)-based dietary supplements can be used as stable non-toxic formulations, with high intestinal absorption rate, inducing alpha2-macroglobulin (A2M) expression for longevity and healthy life. ZBC is metabolized by hydrolysis, mainly at the absorption site (intestinal level), and most of it is excreted in the urine. Within seven hours from the administration in mice, almost the entire amount of orally absorbed ZBC is eliminated in a metabolized form. The highest amount of A2M protein in mouse liver was determined by immunoabsorbance assay in the chronic experiment (1000mg/kg of ZBC), followed by the subchronic experiment (at the same dose of ZBC), and by the acute experiment (5000mg/kg of ZBC).

Retrospective Study of a Patient with Multiple de Novo Skin Tumor Formations.

In Romania, the incidence of malignant melanocytic tumors is continuously increasing. According to the World Health Organization, the incidence of melanocytic and non-melanocytic skin neoplasms has increased considerably and globally, in the last decade. We present the case of a 49-year-old patient who, over the course of 7 years, came in the Plastic Surgery Clinic of the Emergency County Hospital of Craiova for the excision of a number of 25 skin tumor formations, located on the face, cervical region, trunk and upper limbs. Treatment included complete microsurgical excision and supervision. In the end, the patient's treatment compliance decreased significantly.

Testosterone and quality of life in patients with dilated cardiomyopathy.

BACKGROUND: Testosterone is an important factor that influences the quality of life in men. The purpose of this study is to evaluate how testosterone level impacts the quality of life in patients with dilated cardiomyopathy. METHODS: This cross-sectional single-center included 97 male patients with dilated cardiomyopathy, in whom serum testosterone was measured. Health-related quality of life was measured using the translated validated version of the Kansas City Cardiomyopathy Questionnaire (KCCQ-12). We used correlation and multivariable regression to assess the association between KCCQ-12 score, serum testosterone level, and clinical and paraclinical variables. RESULTS: The mean age of study participants was 58 (range 29-88). The mean LVEF was 25 ±8.61%. The average total serum testosterone level was 3.13 ±2.72 (range 0.19-13.5 ng/ml). The median global KCCQ-12 score was 44.8 (6.2-90.6) representing a poor to fair impairment in quality of life. There was an inverse correlation between the KCCQ-12 score and NYHA class (Pearson coefficient r = 0.847 p<0.001) and a direct correlation with LVEF (r=0.445, p<0.001). Also, the KCCQ-12 score correlated with hemoglobin level (r=0.214, p=0.037) and plasmatic creatinine level (r=-0.296 p= 0.004). In multivariable regression, the independent predictors of health-related quality of life were testosterone, LVEF, and NYHA class. CONCLUSIONS: The results of this study showed for the first time a significant direct relationship between serum testosterone levels and quality of life in patients with dilated cardiomyopathy.

Osteoarthritis induces gender-related changes in the knee range of motion.

In time, osteoarthritis (OA) generates the misalignment of the affected joint structures. However, due to the nature of bipedal gait, OA in the lower limb can also cause pathological gait patterns, which can generate instability and falls, with great consequence, especially in the aged population. With goniometry used to evaluate the range of motion (ROM) of joints, we wanted to evaluate how gender impacts gait dynamics in OA patients. For this study, we have compared 106 OA patients (74 females and 32 males) to age matched controls. All participants had their right leg as dominant. Video recording of normal gait was analyzed with a digital goniometry tool phone application, and the knee's ROM was measured in midstance and midswing moment of the gait. During midstance, significant extension and flexion of the knee excursion have been observed in both males and females. During midswing, knee OA presents more differences, whereas subjects with hip and knee OA present changes on the dominant knee. Midstance changes suggest that the knee's joint degenerative changes, such as synovitis, can be linked to hip OA secondary changes. Midswing changes in lower limb OA suggest a connection to the activities of daily life. Gender differences generated by OA must furthermore be studied in both lower limbs so that the best therapeutic approach can be chosen.

New Insights into Boron Essentiality in Humans and Animals.

Boron (B) is considered a prebiotic chemical element with a role in both the origin and evolution of life, as well as an essential micronutrient for some bacteria, plants, fungi, and algae. B has beneficial effects on the biological functions of humans and animals, such as reproduction, growth, calcium metabolism, bone formation, energy metabolism, immunity, and brain function. Naturally organic B (NOB) species may become promising novel prebiotic candidates. NOB-containing compounds have been shown to be essential for the symbiosis between organisms from different kingdoms. New insights into the key role of NOB species in the symbiosis between human/animal hosts and their microbiota will influence the use of natural B-based colon-targeting nutraceuticals. The mechanism of action (MoA) of NOB species is related to the B signaling molecule (autoinducer-2-borate (AI-2B)) as well as the fortification of the colonic mucus gel layer with NOB species from B-rich prebiotic diets. Both the microbiota and the colonic mucus gel layer can become NOB targets. This paper reviews the evidence supporting the essentiality of the NOB species in the symbiosis between the microbiota and the human/animal hosts, with the stated aim of highlighting the MoA and targets of these species.

Metabolic Alterations in Acute Cerebral Ischemia.

Cerebral ischemia is a major health problem worldwide, that affects millions of people, leaving a major percentage of them with major disabilities, therefore becoming one of the most resource consuming pathology. Beside the blockage of blood supply of the brain that leads to loss of cellular function and neuronal necrosis, metabolic processes are modified in the whole body through mechanisms that are not fully explained yet. The results in the analysis of the 2 groups, one with 70 patients with stroke and another with 68 patients with no cerebral infarction, revealed that brain ischemia is more often found in patients with atrial fibrillation and higher blood pressure values. The metabolic changes, found in the stroke group, are represented by increased values of blood glucose, serum urea and lower levels of creatinine levels. Also, the value of leucocytes count and the erythrocyte sedimentation rate were shown to be increased in stroke patients, indicating that inflammation is highly present in cerebral infarction. In the regard of these findings, cerebral ischemia is associated with major systemic disruptions that could be significant pathogenic factors and also effects of the complex processes that take place in the affected brain region, but further investigation should be done in order to explain all the mechanisms involved and also the possible impact in prophylaxis and acute management of stroke.

Hericium erinaceus (Bull.) Pers. Ethanolic Extract with Antioxidant Properties on Scopolamine-Induced Memory Deficits in a Zebrafish Model of Cognitive Impairment.

Hericium erinaceus (H. erinaceus) is a rare and appreciated fungal species belonging to the division Basidiomycota used for centuries in traditional Chinese medicine for its medicinal value. This species of mushrooms brings the most diverse benefits for the human body, and can have beneficial effects for treating Alzheimer's disease (AD). This study investigated whether ethanolic extract from the fungal biomass of H. erinaceus enhances cognitive function via the action on cholinergic neurons using the scopolamine (SCOP)-induced zebrafish (Danio rerio) model of memory impairment. The ethanolic extract from the fungal biomass of H. erinaceus was previously obtained using an ultrasonic extraction method (UE). The administration of H. erinaceus extract to zebrafish, with a pattern of AD induced by scopolamine, showed an improvement in memory evaluated by behavioral and biochemical tests on brain tissue. These results suggest that H. erinaceus has preventive and therapeutic potentials in managing memory deficits and brain oxidative stress in zebrafish with AD.

Effects of the Hydroethanolic Extract of Lycopodium selago L. on Scopolamine-Induced Memory Deficits in Zebrafish.

This scientific research focused on the production of hydroethanolic extract of the plant species Lycopodium selago L. (L. selago) by the ultrasound-assisted extraction (USAE) and the identification of biocompounds with high antioxidant activity is of interest for possible phytotherapeutic treatment against Alzheimer's disease (AD). The extract was phytochemically analyzed to investigate polyphenols, flavonoids, and identify the sesquiterpenoid alkaloid huperzine A (HupA), which is known in the literature for its great relevance in AD. Evaluation and comparison of the antioxidant activity of the extract were performed by four complementary spectrophotometric methods (DPPH, FRAP, ABTS, ORAC). In vitro tests of the extract showed an excellent reciprocal link between the concentration of polyphenols and the measurement of the antioxidant activity of the extract with the sesquiterpenoid HupA. To confirm the antioxidant activity, L. selago hydroethanolic extract was administered in vivo to zebrafish (Danio rerio) with a pattern of scopolamine-induced cognitive impairment. Moreover, this study explored a possible correlation between the expression of oxidative stress markers in the brain tissue with the behavior of the scopolamine zebrafish model. In vivo tests showed that this fern could be used as a nutritional supply and as a phytotherapeutic method to prevent or treat various neurodegenerative diseases that call for high-nutritive-value medications.

Testosterone, cardiomyopathies, and heart failure: a narrative review.

Testosterone exerts an important regulation of cardiovascular function through genomic and nongenomic pathways. It produces several changes in cardiomyocytes, the main actor of cardiomyopathies, which are characterized by pathological remodeling, eventually leading to heart failure. Testosterone is involved in contractility, in the energy metabolism of myocardial cells, apoptosis, and the remodeling process. In myocarditis, testosterone directly promotes the type of inflammation that leads to fibrosis, and influences viremia with virus localization. At the same time, testosterone exerts cardioprotective effects that have been observed in different studies. There is increasing evidence that low endogenous levels of testosterone have a negative impact in some cardiomyopathies and a protective impact in others. This review focuses on the interrelationships between testosterone and cardiomyopathies and heart failure.

Hip Joint Abnormalities During Midstance in Osteoarthritic Patients.

Osteoarthritis is one of the most debilitating diseases in Europe affecting the lower limb joints, especially the hip and knee, having a bad influence on gait in the long run as well. Rehabilitation physicians use gait in order for the whole body to be seen in ensemble, and through midstance as moment of gait to also take predilection to falls into consideration. Goniometry is the quantifiable measure of a rehabilitation treatment by measuring the range of motion of each treated joint and studied during time. The patients that volunteered to be part of this study have been divided into four groups, depending on the level of osteoarthritis present at the lower limb joints: hip, knee, both hip and knee osteoarthritis or control group with no osteoarthritis, have been asked to walk for a few times and the video recordings were uploaded into the Angles App where we measured the lower limb joint angles during midstance. Patients with knee osteoarthritis present a more extended hip on both dominant and non-dominant sides compared to the ones with hip osteoarthritis, hip and knee osteoarthritis or control group. The results can be explained through the body's kinematic chains that link the knee and hip, hip and pelvis during the midstance phase in the sagittal plane. A physician can use a video goniometry app in order for him to thoroughly evaluate an osteoarthritic patient as well as follow him or her during the entire course of treatment.

Complexity of Gait Angle Measurements at the Ankle Joint During Midstance in Patients with Osteoarthritis.

The evolution of rehabilitation treatments can be quantified through goniometric measurements. Thus, a video goniometer, and an app-based goniometry program can be both useful and a reliable method of obtaining a data base through which we can see if a certain rehabilitation treatment works out for our patients and during times such as the Covid-19 pandemic, a telemedicine approach can be done. Midstance is a sub-moment of the gait pattern, important in the stability of the lower limb, but that can also direct us towards a patient prone to falls. Osteoarthritis is a disease that causes high disability because of the cellular degradation that also affects normal gait. Four groups of subjects: subjects suffering from hip osteoarthritis, knee osteoarthritis, hip and knee osteoarthritis and control group, have been filmed and recorded their midstance joint range of motion in the Angles App. The dominant limb has been proven to have a more extended ankle in the hip osteoarthritis group, compared to knee osteoarthritis, hip and knee osteoarthritis or control group. Females have presented a more extended ankle, wearing high heels for a long period of time can be the cause of that. Subjects with knee osteoarthritis have presented a more flexed ankle in the dominant limb compared to the ones suffering from hip and knee osteoarthritis or control group. The ankle joint can also have its range of motion measured with a video goniometer, helping us compare results in between sessions of rehabilitation in osteoarthritic patients.

Inhibition of Aquaporin 4 Decreases Amyloid Aß40 Drainage Around Cerebral Vessels.

Aquaporin-4 (AQP4) is located mainly in the astrocytic end-feet around cerebral blood vessels and regulates ion and water homeostasis in the brain. While deletion of AQP4 is shown to reduce amyloid-ß (Aß) clearance and exacerbate Aß peptide accumulation in plaques and vessels of Alzheimer's disease mouse models, the mechanism and clearing pathways involved are debated. Here, we investigated how inhibiting the function of AQP4 in healthy male C57BL/6 J mice impacts clearance of Aß40, the more soluble Aß isoform. Using two-photon in vivo imaging and visualizing vessels with Sulfurodamine 101 (SR101), we first showed that Aß40 injected as a ≤ 0.5-µl volume in the cerebral cortex diffused rapidly in parenchyma and accumulated around blood vessels. In animals treated with the AQP4 inhibitor TGN-020, the perivascular Aß40 accumulation was significantly (P < 0.001) intensified by involving four times more vessels, thus suggesting a generalized clearance defect associated with vessels. Increasing the injecting volume to ≥ 0.5 ≤ 1 µl decreased the difference of Aß40-positive vessels observed in non-treated and AQP4 inhibitor-treated animals, although the difference was still significant (P = 0.001), suggesting that larger injection volumes could overwhelm intramural vascular clearance mechanisms. While both small and large vessels accumulated Aß40, for the ≤ 0.5-µl volume group, the average diameter of the Aß40-positive vessels tended to be larger in control animals compared with TGN-020-treated animals, although the difference was non-significant (P = 0.066). Using histopathology and ultrastructural microscopy, no vascular structural change was observed after a single massive dose of TGN-020. These data suggest that AQP4 deficiency is directly involved in impaired Aß brain clearance via the peri-/para-vascular routes, and AQP4-mediated vascular clearance might counteract blood-brain barrier abnormalities and age-related vascular amyloidopathy.

In vitro and in vivo studies of novel fabricated bioactive dressings based on collagen and zinc oxide 3D scaffolds.

The paper reports the synthesis and physico-chemical and biological characterization of novel wound dressings based on collagen and essential oil functionalized ZnO nanoparticles intended to improve the treatment of burns and to reduce the risk for developing wound sepsis in patients with burns or chronic wounds. The prepared wound dressings were physico-chemical characterized by Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). In vitro biocompatibility and cytotoxicity was proved on human fibroblast cells, antimicrobial potential was assessed on Gram positive and Gram negative bacteria models (Staphylococcus aureus and Escherichia coli, respectively), while in vivo studies were performed on a rat burn wound experimental model. Functionalized ZnO nanoparticles (NPs) proved to range 15-20 nm in size and contain about 1% orange essential oil (EO), which was utilized as a natural antimicrobial agent. NPs are grain-shapped and have a low tendency to form aggregates. No toxicity was noticed in vitro, as human fibroblasts maintained a normal growth and their membrane integrity in the presence of EO functionalized NPs. The capacity of the prepared wound dressings to act as implantable bioresorbable scaffolds that accelerates wounds healing along with an excellent biocompatibility, lack of cytotoxicity and a good antibacterial activity, make these materials promising and safe candidates for wound dressing, especially in burn patients.

Microglial morphology determined with confocal and two-photon laser scanning microscopy.

Microglia are the first and main form of active immune defense in the nervous system. The immune status of microglia is directly correlated to their morphology. Therefore, microglia morphology is used to distinguish between active and surveilling microglia. For the actual paper, we used confocal laser scanning microscopy (cLSM) and two-photon laser scanning microscopy (2P-LSM), to investigate microglia morphology of 14-16 weeks old male, transgenic mice (n=6). After obtaining, in vivo and fixed tissue, single cells images, we manually tracked individually branch segments of normal microglia. The total number of branches and their overall length were analyzed. Additionally, the number and mean length of each branch order were measured. The overall microglia branching morphology was not different between the two acquisition methods. However, a higher number of fifth branches was observed using cLSM and 2P-LSM, in both fixed and in vivo tissue. Although results from the two methods are mainly comparable, small differences between them should be taken in consideration when formulating an activating∕surveilling conclusion that is purely based on pure microscopic findings. Furthermore, in our opinion, due to their highly dynamic nature, microglia should be carefully labeled as resting or active, taking also into consideration the imaging method used to obtain the data.

Microglia morphology in the physiological and diseased brain - from fixed tissue to in vivo conditions.

First mentioned almost 100 years ago, neuroresearch has linked microglia to the initiation and/or maintenance of most central nervous system pathologies. Since their discovery, we learned that the microglia immune status is often correlated directly to their morphology. However, only recently, have we realized that, in vivo microglia are extremely dynamic cells capable to respond within minutes. Therefore, the name "resting" microglia is replaced slowly with surveilling microglia. Even more recent, it has been shown that microglia change their morphology and/or immune status depending on the way the tissue is obtained, processed and imaged, making labeling microglia, based on their morphology alone, as active or surveying even more difficult. All these observations correlated with a better understanding of cellular and subcellular process that microglia undergo in pathological conditions, could have profound implications on the time window and/or targets used to ensure the best outcome, when treating brain disease.