L. A. Orbeli Institute of Physiology NAS RA

Venoms have evolved >100 times in all major animal groups, and their components, known as toxins, have been fine-tuned over millions of years into highly effective biochemical weapons. There are many outstanding questions on the evolution of toxin arsenals, such as how venom genes originate, how venom contributes to the fitness of venomous species, and which modifications at the genomic, transcriptomic, and protein level drive their evolution. These questions have received particularly little attention outside of snakes, cone snails, spiders, and scorpions. Venom compounds have further become a source of inspiration for translational research using their diverse bioactivities for various applications. We highlight here recent advances and new strategies in modern venomics and discuss how recent technological innovations and multi-omic methods dramatically improve research on venomous animals. The study of genomes and their modifications through CRISPR and knockdown technologies will increase our understanding of how toxins evolve and which functions they have in the different ontogenetic stages during the development of venomous animals. Mass spectrometry imaging combined with spatial transcriptomics, in situ hybridization techniques, and modern computer tomography gives us further insights into the spatial distribution of toxins in the venom system and the function of the venom apparatus. All these evolutionary and biological insights contribute to more efficiently identify venom compounds, which can then be synthesized or produced in adapted expression systems to test their bioactivity. Finally, we critically discuss recent agrochemical, pharmaceutical, therapeutic, and diagnostic (so-called translational) aspects of venoms from which humans benefit.

INTRODUCTION: Orthopaedic surgery is among the least diverse fields in all of medicine. To promote the recruitment of minorities, a commonly proposed strategy is to increase the exposure of minority medical students to orthopaedic surgeons and residents who are minorities themselves. This study examines the degree to which the racial/ethnic diversity of the orthopaedic faculty and residency program influences underrepresented in medicine (URM) medical students at that institution to pursue a career in orthopaedics. METHODS: Using data provided by the Association of American Medical Colleges, we identified all US medical schools that were affiliated with an orthopaedic department and an orthopaedic residency program (n = 110). For each institution, data were collected on URM representation among the orthopaedic faculty and residents (2013 to 2017), as well as the proportion of URM medical students who applied to an orthopaedic residency program (2014 to 2018). The association between institutional factors and the URM medical student orthopaedic application rate was then assessed. RESULTS: Of 11,887 URM students who graduated from medical school during the 5-year study period, 647 applied to an orthopaedic residency program (5.4%). URM students who attended medical school at institutions with high URM representation on the orthopaedic faculty were more likely to apply in orthopaedics (odds ratio 1.27, 95% confidence interval 1.04 to 1.55, P = 0.020), as were URM students at institutions with high URM representation in the residency program (odds ratio 1.45, 95% confidence interval 1.17 to 1.79, P < 0.001). DISCUSSION: The benefits of a diverse orthopaedic workforce are widely acknowledged. In this study, we found that increased URM representation among the orthopaedic faculty and residents was associated with a greater likelihood that URM medical students at that institution would apply in orthopaedics. We also suggest a set of strategies to break the cycle and promote the recruitment of minorities into the field of orthopaedic surgery.

Hydrogen gas production was observed to occur during ATP-driven H+/K+ exchange in anaerobically grown E. coli. Neither process was found in aerobically grown cells or anaerobic cells grown on nitrate medium or when the osmotic pressure was decreased or K+ removed, or finally when DCCD, arsenate or CCCP was applied. Dithiothreitol restored the process even in the presence of CCCP but not in other cases of inhibition. A model of a multienzyme transport super-complex is proposed. The supercomplex consists of three genetically independent mechanisms: F0F1 H+-ATPase to provide energy, the K+-transporting Trk system as energy sink and formate-hydrogen lyase as donor of reducing equivalents. Within this supercomplex direct transduction of energy is accomplished via oxidation of 2 SH to S-S.

Autism spectrum disorders (ASD) are neurodevelopmental disorders, that are characterized by core symptoms, such as alterations of social communication and restrictive or repetitive behavior. The etiology and pathophysiology of disease is still unknown, however, there is a strong interaction between genetic and environmental factors. An intriguing point in autism research is identification the vulnerable time periods of brain development that lack compensatory homeostatic corrections. Valproic acid (VPA) is an antiepileptic drug with a pronounced teratogenic effect associated with a high risk of ASD, and its administration to rats during the gestation is used for autism modeling. It has been hypothesized that valproate induced damage and functional alterations of autism target structures may occur and evolve during early postnatal life. Here, we used prenatal and postnatal administrations of VPA to investigate the main behavioral features which are associated with autism spectrum disorders core symptoms were tested in early juvenile and adult rats. Neuroanatomical lesion of autism target structures and electrophysiological studies in specific neural circuits. Our results showed that prenatal and early postnatal administration of valproate led to the behavioral alterations that were similar to ASD. Postnatally treated group showed tendency to normalize in adulthood. We found pronounced structural changes in the brain target regions of prenatally VPA-treated groups, and an absence of abnormalities in postnatally VPA-treated groups, which confirmed the different severity of VPA across different stages of brain development. The results of this study clearly show time dependent effect of VPA on neurodevelopment, which might be explained by temporal differences of brain regions' development process. Presumably, postnatal administration of valproate leads to the dysfunction of synaptic networks that is recovered during the lifespan, due to the brain plasticity and compensatory ability of circuit refinement. Therefore, investigations of compensatory homeostatic mechanisms activated after VPA administration and directed to eliminate the defects in postnatal brain, may elucidate strategies to improve the course of disease.

The progressive disappearance of the postural and oculomotor syndrome triggered by unilateral labyrinthectomy (vestibular compensation) is a model of plasticity in the adult central nervous system. This recovery may involve modifications of the pharmacological profile of central vestibular neurones, in particular their sensitivity to inhibitory amino acids. We therefore compared the sensitivity of medial vestibular nucleus neurones to glycine and muscimol in slices taken either from control animals, or from guinea-pigs labyrinthectomized 3 days before. We demonstrate that the loss of excitatory inputs experienced by the ipsilesional vestibular neurones induces a decrease in their sensitivity to inhibitory amino acids. These pharmacological changes should facilitate the recovery of a normal balance between the average resting discharge of neurones in both vestibular nuclei.

Armenuhi Moghrovyan, Lilya Parseghyan, Gohar Sevoyan, Anna Darbinyan, Naira Sahakyan, Monica Gaboyan, Zaruhi Karabekian, and Armen Voskanyan. Korean J Pain 2022;35:140-51. https://doi.org/10.3344/kjp.2022.35.2.140

Abstract —Intraperitoneal injections of GABA (5 mg/kg) to rats lowered the level of norepinephrine in brain, heart and spleen but not suprarenals and raised that of serotonin in brain. Changes of these monoamines were most pronounced in the hypothalamic region after 20min. A reduction of hypothalamic norepinephrine was also observed 15min following the intracarotid administration of 0·5 mg/kg of GABA. In these experiments there was a concomitant increase in the level of free GABA in the anterior portion of the ventral hypothalamus. Brain dopamine level and 5‐hydroxytryptophan decarboxylase, dihydroxyphenylalanine decarboxylase and monoamine oxidase activities were not affected. The 20 per cent increase of endogenous GABA observed in the midbrain 30 min following the administration of amino‐oxyacetic acid was accompanied by a sharp fall in norepinephrine level (39 per cent) and an increase in serotonin (20 per cent). In in vitro studies 10–300 μg/ml of GABA were shown to release norepinephrine from cortical and hypothalamic slices, and to inhibit serotonin release without affecting 5‐hydroxytryptophan uptake and to have no effect on the release of dopamine from slices of the region of the corpus striatum nor on the activity of the enzymes mentioned. Subcellular studies showed that the particulate:supernatant ratio for norepinephrine was reduced from a control value of 2·04 to 1·75 and that of serotonin was raised from 2·8 to 3·5. Following pretreatment with iproniazid, GABA reduced the raised level of brain norepinephrine to a greater extent than reserpine but not as intensively as amphetamine. The results obtained suggest that these monoamines may be involved in the mechanisms underlying the action of GABA in brain and that the effect of GABA on brain monoamines may be of certain significance in synaptic events.

In the last decade, there has been remarkable progress in research toward understanding and refining the hallmarks of cancer. In this review, we propose a new hallmark - "pro-survival autophagy." The importance of pro-survival autophagy is well established in tumorigenesis, as it is related to multiple steps in cancer progression and vital for some cancers. Autophagy is a potential anti-cancer therapeutic target. For this reason, autophagy is a good candidate as a new hallmark of cancer. We describe two enabling characteristics that play a major role in enabling cells to acquire the hallmarks of cancer - "tumor-promoting microenvironment and macroenvironment" and "cancer epigenetics, genome instability and mutation." We also discuss the recent updates, therapeutic and prognostic implications of the eight hallmarks of cancer described by Hanahan et al. in 2011. Understanding these hallmarks and enabling characteristics is key not only to developing new ways to treat cancer efficiently but also to exploring options to overcome cancer resistance to treatment.

Excess dietary fructose intake associated with metabolic syndrome and insulin resistance and increased risk of developing type 2 diabetes. Previous animal studies have reported that diabetic animals have significantly impaired behavioural and cognitive functions, pathological synaptic function and impaired expression of glutamate receptors. Correction of the antioxidant status of laboratory rodents largely prevents the development of fructose-induced plurimetabolic changes in the nervous system. We suggest a novel concept of efficiency of Stevia leaves for treatment of central diabetic neuropathy. By in vivo extracellular studies induced spike activity of hippocampal neurons during high frequency stimulation of entorhinal cortex, as well as neurons of basolateral amygdala to high-frequency stimulation of the hippocampus effects of Stevia rebaudiana Bertoni plant evaluated in synaptic activity in the brain of fructose-enriched diet rats. In the conditions of metabolic disorders caused by fructose, antioxidant activity of Stevia rebaudiana was assessed by measuring the NOX activity of the hippocampus, amygdala and spinal cord. In this study, the characteristic features of the metabolic effects of dietary fructose on synaptic plasticity in hippocampal neurons and basolateral amygdala and the state of the NADPH oxidase (NOX) oxidative system of these brain formations are revealed, as well as the prospects for development of multitarget and polyfunctional phytopreparations (with adaptogenic, antioxidant, antidiabetic, nootropic activity) from native raw material of Stevia rebaudiana. Stevia modulates degree of expressiveness of potentiation/depression (approaches but fails to achieve the norm) by shifting the percentage balance in favor of depressor type of responses during high-frequency stimulation, indicating its adaptogenic role in plasticity of neural networks. Under the action of fructose an increase (3–5 times) in specific quantity of total fraction of NOX isoforms isolated from the central nervous system tissue (amygdala, hippocampus, spinal cord) was revealed. Stevia exhibits an antistress, membrane-stabilizing role reducing the level of total fractions of NOX isoforms from central nervous system tissues and regulates NADPH-dependent O2 − −producing activity. Generally, in condition of metabolic disorders caused by intensive consumption of dietary fructose Stevia leaves contributes to the control of neuronal synaptic plasticity possibly influencing the conjugated NOX-specific targets.

It has been the goal of this review to describe the functional interrelations between Deiters' vestibular nucleus and numerous brain structures. Emphasis is placed on dynamic and integrative properties of linkages between the neurons of Deiters' nucleus and many other brain structures in order to begin considering the capabilities of the loops in the light of motor control and coordination of movement. The problem of somatotopy within the loops is also considered. Putting this information together, the possible roles of Deiters' nucleus in the control of movements are described. It is suggested that Deiters' nucleus in co-operation with cerebral cortex, cerebellum, subcortical and brainstem structures are responsible for the integration and realization of different movements.

Experiments were performed on 48 albino rats. Part of the experimental animals were initially trained to a balancing instrumental conditioned reflex (ICR). Unilateral bulbar pyramidotomy performed in all rats caused contralateral hemiparesis. On the next day following the operation 24 rats were injected intramuscularly with bacterial melanin solution. 12 of these rats were initially trained to ICR. Recovery periods of ICR and paralyzed hindlimb movements were registered for melanin injected rats (n=24) and for operated rats, not treated with melanin (n=24). In rats injected with bacterial melanin the posttraumatic recovery is shorter than in animals not treated with melanin. The fastest and complete recovery was registered in rats initially trained to ICR and injected after the operation with bacterial melanin. Electrophysiological experiments were performed in transected animals treated with melanin, transected animals without melanin treatment and intact animals. Spiking activity of motoneurons was registered in lumbar motoneurons of rats in response to high frequency stimulation above the corticospinal tract transection. Spiking activity was very similar in motoneurons of melanin injected and intact or non operated animals. In animals, not dosed with bacterial melanin after the operation, areactivity or no change in firing rate was registered in response to stimulus. Stimulation of the corticospinal tract of melanin injected rats produced potentiation of the motoneuronal firing rate and is an evidence of regeneration in corticospinal tract. Similarity in spiking activity of intact and melanin injected rats shows the recovery of conductance in pyramidal tract. Morphohistochemical examination was carried out to confirm the results of behavioral and electrophysiological experiments. Medulla slices were prepared to trace the regeneration of nerve fibers. Examination of transection area revealed that bacterial melanin increases vascularization, dilates the capillaries in nervous tissue and stimulates the process of sprouting.

In recent years, molecularly imprinted polymer nanoparticles (nanoMIPs) have proven to be an attractive alternative to antibodies in diagnostic and therapeutic applications. However, several key questions remain: how suitable are intracellular epitopes as targets for nanoMIP binding? And to what extent can protein function be modulated via targeting specific epitopes? To investigate this, three extracellular and three intracellular epitopes of epidermal growth factor receptor (EGFR) were used as templates for the synthesis of nanoMIPs which were then used to treat cancer cells with different expression levels of EGFR. It was observed that nanoMIPs imprinted with epitopes from the intracellular kinase domain and the extracellular ligand binding domain of EGFR caused cells to form large foci of EGFR sequestered away from the cell surface, caused a reduction in autophosphorylation, and demonstrated effects on cell viability. Collectively, this suggests that intracellular domain-targeting nanoMIPs can be a potential new tool for cancer therapy.

INTRODUCTION: As of the 2022 to 2023 match cycle, orthopaedic residency programs began offering applicants 30 signals as part of a preference signaling program. Many have assumed that signals would become powerful tools in the match process, yet no objective data currently exist analyzing their effect. This study aims to provide such analysis while also offering comparisons with other factors affecting match success. METHODS: Self-reported survey data on applicants and applications from 2017 to 2023 from the Texas Seeking Transparency in Application to Residency database were queried. Variables associated with receiving interviews and match success were analyzed using two-sided Student t -tests, chi-squared tests, variance ratio testing, and receiver operating characteristic analysis. RESULTS: Compared with 2017 to 2022, 2023 applicants submitted fewer applications (61.8 versus 78; P < 0.001), received fewer interview offers (11.6 versus 13.8; P < 0.001), and interview offers were spread more evenly among applicants (SD, 6.82 versus 9.10; P < 0.001). For 2023 applications, odds of securing an interview were increased most by away rotations (odds ratios [OR] 61.8; P < 0.001), use of a signal (OR, 9.61; P < 0.001), and geographic connection (OR, 4.70; P < 0.001). Female applicants received more interview offers from signaled programs than their male counterparts (11.2 versus 8.94; P = 0.003). Applicant variables most predictive of match success in 2023 were interview offers (area under the receiver operating characteristic curve [AUC] = 0.802), step 2 CK score (AUC = 0.673), and step 1 score (AUC = 0.648). DISCUSSION: The preference signaling program seems to be accomplishing its goals of reducing applications and more evenly distributing interviews. Away rotations, signals, and geographic connections represent the strongest predictors of applications resulting in a successful match. Applicants must use their signals carefully to maximize their chance of success. LEVEL OF EVIDENCE: Level III.

Parkinson's disease (PD) is the most common neurodegenerative disease of unknown etiology and characterized by motor symptoms of tremor, rigidity, bradykinesia, and postural instability. Interactions between the dopaminergic systems and the hippocampus in synaptic plasticity and behavior are found. The rotenone-induced animal model is commonly used in research studies involved in PD. Administration of rotenone causes alterations of electrical neuronal activity. Rotenone (2.5 mg/kg/day) was administered intraperitoneally for 21 days to adult rats, and rotenone effects on rearing activity and electrophysiology were examined. Dose-dependent reduction of evoked neural activity and a reduction in firing strength were found in the hippocampus. Behaviorally, Rotenone rats showed a more consistent decrease in rearing across the 3 weeks, compared with animals in the control group. Thus, rotenone causes changes in hippocampal electrical activity and behavioral changes.

After intraperitoneal injection of 20 mg/kg lead acetate, rats received 8 weeks of treadmill exercise (15-22 m/min, 25-64 minutes) and/or treadmill exercise at 1.6 km/h until exhaustion. The markers related to neurotoxicity were measured by enzyme-linked immunosorbent assay method. 8 weeks of treadmill exercise significantly increased brain-derived neurotrophic factor level in the hippocampus (P = 0.04) and plasma level of total antioxidant capacity of rats exposed to lead acetate (P < 0.001), and significantly decreased plasma level of malondialdehyde (P < 0.001). Acute exercise only decreased the hippocampal malondialdehyde level (P = 0.09) and increased brain-derived neurotrophic factor level in the hippocampus (P = 0.66). Acute exercise also enhanced the total antioxidant capacity in rats exposed to lead acetate, insignificantly (P = 0.99). These findings suggest that chronic treadmill exercise can significantly decrease neurotoxicity and alleviate oxidative stress in rats exposed to lead acetate. However, acute endurance exercise was not associated with these beneficial effects.

The widespread use of silver nanoparticles (AgNPs) requires a study of their safety. The aim of the present study was to assess the levels of oxidative stress markers and histopathological changes in the experimental model of sarcoma S-180 of outbred mice caused by biogenic AgNPs. AgNPs were synthesized using 50% ethanol extract of Ocimum araratum leaves that was standardized for rosmarinic acid content. The effects of AgNPs were tested on chemiluminescence (ChL), malonic dialdehyde (MDA) content and activity of superoxide dismutase (SOD) in healthy and experimental model of sarcoma S-180 mice. It was shown that, under the influence of AgNPs, the intensity of ChL decreased, in contrast with control groups (with the exception of the hepatocytes of animals with transplanted sarcoma). The presence of AgNPs leads to the decrease of MDA in the tissues of healthy mice and to a slight increase of MDA content in the tumour and kidney tissues. AgNPs neutralize the activity of SOD in kidney tissue samples in animals with transplanted sarcoma, and in tumour tissue, they reduce SOD activity by three times. The results of the histological analysis indicate that AgNPs not only cause the destruction of tumour tissue but also lead to structural changes in hepatocytes and nephrons, which can affect the function of these organs. AgNPs are potential agents for antitumor therapy. Future studies are needed using biocompatible non-toxic NPs that meet the requirement for these drugs.

OBJECTIVES: To measure baseline bilateral tibial torsion in a cohort of uninjured patients to assess for a difference in torsion between sides. METHODS: Consecutive bilateral lower extremity CT angiography scans from 229 patients without tibial or fibular pathology were collected and reviewed. Torsion of each tibia was measured by two independent reviewers, and individual differences in torsion were calculated. RESULTS: On average, patients have a 6.0° difference in tibial torsion between sides. A difference of greater than 10° was present in 18% of patients. Across the cohort of patients, the right tibia was on average 4.4° more externally rotated than the left. In patients with a greater than 5° difference, the right tibia was more externally rotated than the left in 85% of cases. Tibial torsion did not correlate with age or sex. DISCUSSION: Differences in tibial torsion are common and should be considered during intramedullary nailing of tibial fractures. When a difference in torsion is present, external torsion of the right tibia when compared with the left occurs predominantly. LEVEL OF EVIDENCE: Prognostic level IV.

The hippocampus is a target of ovarian hormones, and is necessary for memory. Ovarian hormone loss is associated with a progressive reduction in synaptic strength and dendritic spine. Teucrium polium has beneficial effects on learning and memory. However, it remains unknown whether Teucrium polium ameliorates hippocampal cells spike activity and morphological impairments induced by estrogen deficiency. In the present study, we investigated the effects of hydroponic Teucrium polium on hippocampal neuronal activity and morpho-histochemistry of bilateral ovariectomized (OVX) rats. Tetanic potentiation or depression with posttetanic potentiation and depression was recorded extracellularly in response to ipsilateral entorhinal cortex high frequency stimulation. In morpho-histochemical study revealing of the activity of Ca2+-dependent acid phosphatase was observed. In all groups (sham-operated, sham + Teucrium polium, OVX, OVX + Teucrium polium), most recorded hippocampal neurons at HFS of entorhinal cortex showed TD-PTP responses. After 8 weeks in OVX group an anomalous evoked spike activity was detected (a high percentage of typical areactive units). In OVX + Teucrium polium group a synaptic activity was revealed, indicating prevention OVX-induced degenerative alterations: balance of types of responses was close to norm and areactive units were not recorded. All recorded neurons in sham + Teucrium polium group were characterized by the highest mean frequency background and poststimulus activity. In OVX+ Teucrium polium group the hippocampal cells had recovered their size and shape in CA1 and CA3 field compared with OVX group where hippocampal cells were characterized by a sharp drop in phosphatase activity and there was a complete lack of processes reaction. Thus, Teucrium polium reduced OVX-induce neurodegenerative alterations in entorhinal cortex-hippocamp circuitry and facilitated neuronal survival by modulating activity of neurotransmitters and network plasticity.

INTRODUCTION: Effective pain management after joint arthroplasty is essential for optimal participation in rehabilitation. However, this needs to be balanced with potential risks associated with opioid use and community exposure. The aim of this study was to evaluate opioid use and appropriateness of supply on discharge after total knee arthroplasty or total hip arthroplasty at a major Australian health service. METHODS: A prospective observational study was undertaken at an Australian 980-bed metropolitan health service. Patient interviews were conducted 3 weeks after hospital discharge to evaluate analgesic management and functional outcomes. The primary end point was the number of hospital-supplied opioid pills remaining 3 weeks postdischarge. Secondary end points included (1) factors associated with opioid use 3 weeks postdischarge, (2) opioid use in patients with poor functional outcomes, and (3) proportion of opioid naive patients who became chronic opioid users. RESULTS: One hundred forty patients were included, and 137 were supplied opioids on discharge. At 3 weeks postdischarge, the median number of opioid pills remaining was 0 (interquartile range 0 to 8). There were 77 patients (56.2%) still taking opioids; surgery type, opioid use before admission, and the number of "as required" doses used 24 hours before discharge were independent predictors of opioid continuation. Patients with poor functional outcomes were supplied with more opioids on discharge, often not satisfied with the quantity supplied and more likely to be taking opioids 3 weeks postdischarge. There were 5 of 93 opioid naive patients (5.3%) who developed chronic opioid usage. DISCUSSION: More than half of the patients undergoing total knee arthroplasty or total hip arthroplasty were still using opioids at 3 weeks postdischarge. Most patients were not supplied with excessive quantities at discharge. Future research should focus on identifying patients at risk of prolonged opioid use and improving the transition of these patients into the community. LEVEL OF EVIDENCE: Level II-Prognostic study = prospective observational study.

Thirty five percent of the American population is considered obese (body mass index [BMI] > 30). Obesity disproportionately affects African Americans, Hispanics, and women. Obesity is associated with postoperative complications, including wound complications, infections, and revision total joint arthroplasty (including total hip arthroplasty and total knee arthroplasty). Current BMI benchmarks (many institutions rely on a BMI of 40) selectively preclude patients from having surgery. Patients in these underserved populations can be optimized through the lens of shared decision making through the assessment of food security (eg, food deserts and food swamps), ability to afford healthy food, knowledge of social safety net and community resources to access healthy food, nutrition and weight loss referrals to programs that accept all forms of insurance, weight loss measurements as a percentage of body weight lost instead of BMI cutoffs, pharmacologic modalities, and bariatric surgery.