Humans For Therapeutic Purposes Research Articles

Comparison of colostrum and milk extracellular vesicles small RNA cargo in water buffalo

Recently, much interest has been raised for the characterization of signaling molecules carried by extracellular vesicles (EVs), which are particularly enriched in milk (mEVs). Such interest is linked to the capability of EVs to cross biological barriers, resist acidification in the gastric environment, and exert modulation of the immune system, mainly through their microRNA (miRNA) content. We characterized the small-RNA cargo of colostrum EVs (colosEVs) and mEVs from Italian Mediterranean buffalo through next generation sequencing. Colostrum (first milking after birth) and milk (day 50 of lactation) were sampled from seven subjects from five farms. ColosEVs and mEVs were subjected to morphological characterization, followed by high-depth sequencing of small RNA libraries produced from total RNA. The main difference was the amount of EV in the two samples, with colostrum showing 10 to 100-fold higher content than milk. For both matrices, miRNA was the most abundant RNA species (95% for colosEVs and 96% for mEVs) and three lists were identified: colosEV-specific, mEV-specific and shared most expressed. Gene ontology (GO) enrichment analysis on miRNA targets highlighted many terms related to the epigenetic, transcriptional and translational regulations across the three lists, with a higher number of enriched terms for colosEV-specific miRNAs. Terms specific to colosEVs were related to “cell differentiation” and “microvillus assembly”, while for mEV “cardiac and blood vessel development” and “mitochondria” emergerd. Immune modulation terms were found for both sample-specific miRNAs. Overall, both matrices carry a similar molecular message in terms of biological processes potentially modulated into receiving cells, but there is significant difference in the abundance, with colostrum containing much more EVs than milk. Moreover, colosEVs carry molecules involved in signal transduction, cell cycle and immune response, as for mEVs and EVs of other previously characterized species, but with a special enrichment for miRNAs with epigenetic regulation capacities. These beneficial characteristics of colosEVs and mEVs are essential for the calf and could also be exploited for the therapeutic purposes in humans, although further studies are necessary to measure the sanitization treatment impact on EV conservation, especially in buffalo where milk is consumed almost exclusively after processing.

Evaluation of Methylene Blue as An Effective Antiseptic for Medicinal Leeches (Hirudo verbana).

Medicinal leeches (Hirudo spp.) have been used for therapeutic purposes in humans since ancient times. Because of their growth conditions, leeches carry certain bacteria and endosymbionts (e.g., Aeromonas spp). In both leech farms and hirudotherapy clinics, there are no reliable antiseptics that can be used with leeches. This study aimed to determine whether methylene blue (MB) is a safe antiseptic for medicinal leeches and assess its safe usage. This study evaluated the efficacy of MB by determining lethal concentrations (LC), effective concentrations (EC), and lethal times (LT) for the medicinal leech Hirudo verbena Carena, 1820. A total of 570 H. verbana specimens obtained from a local farm were used in this study. Eighteen different concentrations of MB (between 1 ppm and 512 ppm) were tested. The LC50 and EC50 values for H. verbana were determined to be 60.381 (53.674-66.636) ppm and 2.013 (1.789-2.221) ppm, respectively. The LT50 durations for MB concentrations of 32 and 512 ppm were calculated as 212.92 h (138.43 h-1485.78 h) and 17.82 h (8.08 h-23.90 h), respectively. The results show that MB concentrations between 2 and 19 ppm can be safely used as antiseptics in hirudotherapy clinics and leech farms to address bacterial concerns caused by medicinal leeches.

Manipulating Myc for reparative regeneration.

The Myc family of proto-oncogenes is a key node for the signal transduction of external pro-proliferative signals to the cellular processes required for development, tissue homoeostasis maintenance, and regeneration across evolution. The tight regulation of Myc synthesis and activity is essential for restricting its oncogenic potential. In this review, we highlight the central role that Myc plays in regeneration across the animal kingdom (from Cnidaria to echinoderms to Chordata) and how Myc could be employed to unlock the regenerative potential of non-regenerative tissues in humans for therapeutic purposes. Mastering the fine balance of harnessing the ability of Myc to promote transcription without triggering oncogenesis may open the door to many exciting opportunities for therapeutic development across a wide array of diseases.

Antibacterial and antioxidant activities of plants consumed by western lowland gorilla (Gorilla gorilla gorilla) in Gabon.

Zoopharmacognosy is the study of the self-medication behaviors of non-human animals that use plant, animal or soil items as remedies. Recent studies have shown that some of the plants employed by animals may also be used for the same therapeutic purposes in humans. The aim of this study was to determine the antioxidant and antibacterial activity of Ceiba pentandra, Myrianthus arboreus, Ficus subspecies (ssp.) and Milicia excelsa bark crude extracts (BCE), plants consumed by western lowland gorillas (Gorilla gorilla gorilla) in Moukalaba-Doudou National Park (MDNP) and used in traditional medicine, and then to characterize their phytochemical compounds. DPPH (2,2-Diphenyl-1-Picrylhydrazyl), phosphomolybdenum complex and β-carotene bleaching methods were used to assess antioxidant activity. Antimicrobial susceptibility testing was performed using the diffusion method, while minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were assessed using the microdilution method. The highest level of total phenolics was found in Myrianthus arboreus aqueous extract [385.83 ± 3.99 mg [gallic acid equivalent (GAE)/g]. Total flavonoid (134.46 ± 3.39) mg quercetin equivalent (QE)/100 g of extract] were highest in Milicia excelsa, tannin [(272.44 ± 3.39) mg tannic acid equivalent (TAE)/100 g of extract] in Myrianthus arboreus and proanthocyanidin [(404.33 ± 3.39) mg apple procyanidins equivalent (APE)/100 g of extract] in Ceiba pentandra. Ficus ssp. (IC50 1.34 ±3.36 μg/mL; AAI 18.57 ± 0.203) ethanolic BCE and Milicia excelsa (IC50 2.07 ± 3.37 μg/mL; AAI 12.03 ± 0.711) showed the strongest antioxidant activity. Myrianthus arboreus ethanolic BCE (73.25 ± 5.29) and Milicia excelsa aqueous BCE (38.67 ± 0.27) showed the strongest percentage of total antioxidant capacity (TAC). Ceiba pentandra ethanolic BCE (152.06 ± 19.11 mg AAE/g) and Ficus ssp aqueous BCE (124.33 ± 39.05 mg AAE/g) showed strongest relative antioxidant activity (RAA). The plant BCE showed antimicrobial activity against multidrug resistant (MDR) E. coli (DECs) isolates, with MICs varying from 1.56 to 50 mg/mL and inhibition diameters ranging from 7.34 ± 0.57 to 13.67 ± 0.57mm. Several families of compounds were found, including total phenolic compounds, flavonoids, tannins and proanthocyanidins were found in the plant BCEs. The plant BCEs showed antioxidant activities with free radical scavenging and antimicrobial activities against 10 MDR E. coli (DECs) isolates, and could be a promising novel source for new drug discovery.

Features, management, and long-term outcome in dogs with pancreatitis and bile duct obstruction treated medically and surgically: 41 dogs (2015-2021).

Pancreatitis resulting in extrahepatic biliary obstruction (EHBO) can cause substantial morbidity and mortality. Endoscopic retrograde cholangiopancreatography is utilized for diagnostic and therapeutic purposes in humans; however, this is not available in veterinary medicine. Treatment options include medical management and biliary drainage procedures. The aim of this study was to describe the management of EHBO secondary to pancreatitis in dogs, treated medically and surgically and to determine whether the neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) differ between the treatment groups. 41 dogs treated for EHBO secondary to pancreatitis during the period of May 2015 to November 2021. Records from 41 dogs diagnosed with EHBO secondary to pancreatitis were reviewed, and information extracted included clinical signs, ultrasound findings, NLR, PLR, histopathology, treatment, and outcomes. 18 of 19 (95%) surgical patients survived, while 12 of 21 (57%) medical patients survived. There was no difference in the length of hospitalization or time to return to adequate function between the groups; however, there was a significant difference in the 2- and 12-month survival between those treated surgically and medically. There was no difference in the NLR or PLR between surgically versus medically treated dogs or between survivors and nonsurvivors. The mortality rate of surgery for EHBO secondary to pancreatitis may be lower than previously described, and in this cohort of dogs, those treated surgically had improved survival at 2 and 12 months compared to those treated medically.

Establishment of a method to detect human immunoglobulin G1 binding microorganisms

In this study, we developed an original and rapid anti-idiotypic antibody-based ELISA method, different from the techniques defined in the literature for this purpose so far, to detect immunoglobulin binding proteins (IBP) on the surface of bacteria. The test antibody used in our study to detect IBP is a recombinant human immunoglobulin G1 Kappa molecule, and has been used as a drug, Tocilizumab (Actemra®), in humans for therapeutic purposes. As a result, the test antibody in the supernatant after centrifugation is reduced compared to the initial moment due to antibody binding. Staphylococcus aureus cowan 1 strain used as positive control causes at least a 50% decrease in OD value in this respect. A similar observation at this level indicated that among a total of 189 microorganisms tested, 3 Staphylococcus aureus and 1 MRSA carrying high-affinity IgBP showed greater than 50% inhibition. This level of inhibition was not detected in the remaining microorganisms.

MELATONIN: ROLE IN THE PROPHYLAXIS OF MIGRAINE

Melatonin and migraine have been linked in most ways, from mechanisms to treatment. Migraine is one of the common neurological diseases and it is characterized by throbbing unilateral headache attacks. Migraine is an issue for both individuals and society. Migraine can lead to reduced work productivity and a flawed family and social life. It has been suggested that melatonin plays a key therapeutic role in treating migraines and several other types of headaches. Early and exact diagnosis is essential in enhancing treatment and in preventing progression. This review aimed to evaluate the available evidence related to the efficacy and safety of melatonin in the prophylaxis and prevention of migraine. Research in regard the administration of melatonin found it safe in migraine patients, with few or no side effects. This review concluded that Melatonin can be considered a safe integrative prophylactic treatment for children, adults and menstrual-related migraines, thus improving outcomes related to patient’s quality of life and patient-centered care. However, further studies and evaluation is required to confirm its effects.This review aimed to evaluate the available evidence related to the efficacy and safety of melatonin in migraine prophylaxis. Melatonin and migraine have been linked in a majority of ways, from mechanisms to treatment. Melatonin is an endogenous hormone and is secreted by the pineal gland. It plays an important role in regulating circadian rhythm and the sleep-wake cycle. It has been proposed for migraine prevention due to its anti-inflammatory effects against calcitonin gene-related peptides and other pro-inflammatory mediators in vitro, and its pro-regulatory effect on the circadian rhythm. Also, low melatonin levels have been reported in serum and urine due to hypothalamic dysfunction in patients with migraine. Hence, in this article, we review those papers describing the results of the administration of melatonin in humans for therapeutic purposes in treating migraines.This narrative literature review was based on studies on Melatonin's effectiveness, safety and tolerability for the prophylactic and preventive treatment of migraine in children and adolescents and menstruating women. The literature search was conducted using the Google Scholar and PubMed database and included articles published up to December 2021, utilizing the keywords: “melatonin”, “migraine”, “melatonin and migraine”, and “prophylaxis of migraine”. Observational studies, randomized controlled trials (RCTs), case reports and systematic reviews were included. The articles focusing on the latest medication for preventing migraines were also reviewed.

Hybrid Enzymatic/Photocatalytic Degradation of Antibiotics via Morphologically Programmable Light-Driven ZnO Microrobots.

Antibiotics are antimicrobial substances that can be used for preventive and therapeutic purposes in humans and animals. Their overdose usage has led to uncontrolled release to the environment, contributing significantly to the development of antimicrobial resistance phenomena. Here, enzyme-immobilized self-propelled zinc oxide (ZnO) microrobots are proposed to effectively target and degrade the released antibiotics in water bodies. Specifically, the morphology of the microrobots is tailored via the incorporation of Au during the synthetic process to lead the light-controlled motion into having on/off switching abilities. The microrobots are further modified with laccase enzyme by physical adsorption, and the immobilization process is confirmed by enzymatic activity measurements. Oxytetracycline (OTC) is used as a model of veterinary antibiotics to investigate the enzyme-immobilized microrobots for their removal capacities. The results demonstrate that the presence of laccase on the microrobot surfaces can enhance the removal of antibiotics via oxidation. This concept for immobilizing enzymes on self-propelled light-driven microrobots leads to the effective removal of the released antibiotics from water bodies with an environmentally friendly strategy.

Phytochemical constituents and assessment of crude extracts from Boerhavia diffusa L. and Lonchocarpus sericeus (Poir.) Kunth ex DC. leaves for antioxidant and antibacterial activities

Medicinal plants have been used by humans for therapeutic purposes since ancient times. Hence, this study investigated the human health benefits of aqueous and ethanolic extracts of Boerhavia diffusa and Lonchocarpus sericeus leaves after these solvents’ extraction. The ethanolic L. sericeus extract (ELS), aqueous L. sericeus extract (ALS), ethanolic B. diffusa extract (EBD) and aqueous B. diffusa extract (ABD), were respectively evaluated for their phytochemical constituents, antioxidant and antibacterial properties. Results showed that both plant species have various constituents such as flavonoid (7.11–21.05%), alkaloid (2.00–8.00%), oxalate (1.98- 3.51 mg/g), saponin (3.00–16.00%), tannin (9.70–18.10 mg/100 g), total phenol (7.36- 18.26 mg/100 g) and phytate (225.64–394.88 mg/100 g), respectively. The ethanolic extracts of the leaves showed higher phytochemicals values than the aqueous extracts, with the highest values in the L. sericeus species, which might be due to the less polarity nature of ethanol solvent when compared to the aqueous solvent. The antioxidant results showed that the ethanolic extracts of the L. sericeus species had higher DPPH (78.61%), ABTS (75.57%), Fe2+ chelation (72.13%), FRAP (2.89 mg AAE/g) and hydroxyl radical scavengers (79.81%) than the ethanolic extracts of the B. diffusa species (58.20%, 71.46%, 61.03%, 2.72 mg AAE/g, 66.58%), respectively. The leaves extracts possessed remarkable antibacterial effects against the selected bacterial strains such as Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and S. typhimurium. For instance, the ethanolic leaves extracts showed better inhibitory performances (2–14 mm) than the aqueous extracts (1–8 mm). Therefore, the study recorded the best activities from the ethanolic extracts of Lonchocarpus sericeus plant. Hence, the study substantiated the fact that the effective antibacterial performances of the ethanolic leaves extract is dependent on its high antioxidant capacities as well as the species of plants being used.

Mammalian Cell Culture as a Platform for Veterinary Vaccines.

For more than three decades, mammalian cells have been the host par excellence for the recombinant protein production for therapeutic purposes in humans. Due to the high cost of media and other supplies used for cell growth, initially this expression platform was only used for the production of proteins of pharmaceutical importance including antibodies. However, large biotechnological companies that used this platform continued research to improve its technical and economic feasibility. The main qualitative improvement was obtained when individual cells could be cultured in a liquid medium similar to bacteria and yeast cultures. Another important innovation for growing cells in suspension was the improvement in chemically defined media that does not contain macromolecules; they were cheaper to culture as any other microbial media. These scientific milestones have reduced the cost of mammalian cell culture and their use in obtaining proteins for veterinary use. The ease of working with mammalian cell culture has permitted the use of this expression platform to produce active pharmaceutic ingredients for veterinary vaccines. In this chapter, the protocol to obtain recombinant mammalian cell lines will be described.

The relevance of mitochondrial DNA variants fluctuation during reprogramming and neuronal differentiation of human iPSCs.

SummaryThe generation of inducible pluripotent stem cells (iPSCs) is a revolutionary technique allowing production of pluripotent patient-specific cell lines used for disease modeling, drug screening, and cell therapy. Integrity of nuclear DNA (nDNA) is mandatory to allow iPSCs utilization, while quality control of mitochondrial DNA (mtDNA) is rarely included in the iPSCs validation process. In this study, we performed mtDNA deep sequencing during the transition from parental fibroblasts to reprogrammed iPSC and to differentiated neuronal precursor cells (NPCs) obtained from controls and patients affected by mitochondrial disorders. At each step, mtDNA variants, including those potentially pathogenic, fluctuate between emerging and disappearing, and some having functional implications. We strongly recommend including mtDNA analysis as an unavoidable assay to obtain fully certified usable iPSCs and NPCs.

PD-L1 on macrophages protects from severe collagen induced arthritis

Abstract Immune related adverse events linked to severe inflammatory arthritis have been identified in 10–43% of patients receiving immune checkpoint inhibitor therapy, aCTLA-4, aPD-1 and aPD-L1, indicating the importance of immune inhibitory pathways in maintaining homeostasis in the synovium. We found that immune checkpoint, PD-L1, is primarily expressed in the joint compared to other immune checkpoints. Furthermore, blocking of PD-L1 during collagen induced arthritis (CIA) resulted in more severe arthritis. Mice synovial macrophages have the highest PD-L1 expression level at homeostasis and macrophage PD-L1 expression is further increased during CIA. Similarly, in human synovial fluid, macrophages have the highest frequency of PD-L1 expression. We hypothesize that the expression of PD-L1 by macrophages is essential for synovial homeostasis and regulation of joint inflammation. We selectively deleted PD-L1 in macrophages using the cre/flox system and induced CIA analyzing disease severity using flow cytometry. LyzcrePD-L1fl/fl showed more severe CIA with a greater change in paw thickness, higher disease and histology score and a higher frequency of CD4 and CD8 T cells in the synovium. Interestingly, female LyzcrePD-L1fl/fl mice have higher arthritis severity to LyzcrePD-L1fl/fl males. This indicates the importance of sex specific PD-L1 expression by synovial macrophages during CIA. Our project has high translational potential since joint specific PD-L1 expression could be modulated for therapeutic purposes in humans with rheumatoid arthritis or check point inhibitors induced arthritis.

In Vivo Validation of a Reversible Small Molecule-Based Switch for Synthetic Self-Amplifying mRNA Regulation

Synthetic mRNA therapeutics have the potential to revolutionize healthcare, as they enable patients to produce therapeutic proteins inside their own bodies. However, convenient methods that allow external control over the timing and magnitude of protein production after invivo delivery of synthetic mRNA are lacking. In this study, we validate the invivo utility of a synthetic self-amplifying mRNA (RNA replicon) whose expression can be turned off using a genetic switch that responds to oral administration of trimethoprim (TMP), a US Food and Drug Administration (FDA)-approved small-molecule drug. After intramuscular electroporation, the engineered RNA replicon exhibited dose-dependent and reversible expression of its encoded protein upon TMP administration. The TMP serum level needed for maximal downregulation of protein translation was approximately 45-fold below that used in humans for therapeutic purposes. To demonstrate the therapeutic potential of the technology, we injected mice with a TMP-responsive RNA replicon encoding erythropoietin (EPO) and successfully controlled the timing and magnitude of EPO production as well as changes in hematocrit. This work demonstrates the feasibility of controlling mRNA kinetics invivo, thereby broadly expanding the clinical versatility of mRNA therapeutics.

Zoopharmacology: A Way to Discover New Cancer Treatments.

Zoopharmacognosy is the multidisciplinary approach of the self-medication behavior of many kinds of animals. Recent studies showed the presence of antitumoral secondary metabolites in some of the plants employed by animals and their use for the same therapeutic purposes in humans. Other related and sometimes confused term is Zootherapy, which consists on the employment of animal parts and/or their by-products such as toxins, venoms, etc., to treat different human ailments. Therefore, the aim of this work is to provide a brief insight for the use of Zoopharmacology (comprising Zoopharmacognosy and Zootherapy) as new paths to discover drugs studying animal behavior and/or using compounds derived from animals. This work is focused on the approaches related to cancer, in order to propose a new promising line of research to overcome multidrug resistance (MDR). This novel subject will encourage the use of new alternative prospective ways to find new medicines.

PD-L1 on macrophages protects from severe collagen induced arthritis

Abstract Immune related adverse events linked to severe inflammatory arthritis have been identified in 10–43% of patients receiving immune checkpoint inhibitor therapy, aCTLA-4, aPD-1 and aPD-L1, indicating the importance of immune inhibitory pathways in maintaining homeostasis in the synovium. We found that immune checkpoint, PD-L1, is primarily expressed in the joint compared to other immune checkpoints. Furthermore, blocking of PD-L1 during collagen induced arthritis (CIA) resulted in more severe arthritis. Mice synovial macrophages have the highest PD-L1 expression level at homeostasis and macrophage PD-L1 expression is further increased during CIA. Similarly, in human synovial fluid, macrophages have the highest frequency of PD-L1 expression. We hypothesize that the expression of PD-L1 by macrophages is essential for synovial homeostasis and regulation of joint inflammation. We selectively deleted PD-L1 in macrophages using the cre/flox system and induced CIA analyzing disease severity using flow cytometry. LyzcrePD-L1fl/fl showed more severe CIA with a greater change in paw thickness, higher disease and histology score and a higher frequency of CD4 and CD8 T cells in the synovium. Interestingly, female LyzcrePD-L1fl/fl mice have higher arthritis severity to LyzcrePD-L1fl/fl males. This indicates the importance of sex specific PD-L1 expression by synovial macrophages during CIA. Our project has high translational potential since joint specific PD-L1 expression could be modulated for therapeutic purposes in humans with rheumatoid arthritis or check point inhibitors induced arthritis.

Overweight in the Elderly Induces a Switch in Energy Metabolism that Undermines Muscle Integrity.

Aging is characterized by a progressive loss of skeletal muscle mass and function (sarcopenia). Obesity exacerbates age-related decline and lead to frailty. Skeletal muscle fat infiltration increases with aging and seems to be crucial for the progression of sarcopenia. Additionally, skeletal muscle plasticity modulates metabolic adaptation to different pathophysiological situations. Thus, cellular bioenergetics and mitochondrial profile were studied in the skeletal muscle of overweight aged people without reaching obesity to prevent this extreme situation. Overweight aged muscle lacked ATP production, as indicated by defects in the phosphagen system, glycolysis and especially mostly by oxidative phosphorylation metabolic pathway. Overweight subjects exhibited an inhibition of mitophagy that was linked to an increase in mitochondrial biogenesis that underlies the accumulation of dysfunctional mitochondria and encourages the onset of sarcopenia. As a strategy to maintain cellular homeostasis, overweight subjects experienced a metabolic switch from oxidative to lactic acid fermentation metabolism, which allows continued ATP production under mitochondrial dysfunction, but without reaching physiological aged basal levels. This ATP depletion induced early signs of impaired contractile function and a decline in skeletal muscle structural integrity, evidenced by lower levels of filamin C. Our findings reveal the main effector pathways at an early stage of obesity and highlight the importance of mitochondrial metabolism in overweight and obese individuals. Exploiting mitochondrial profiles for therapeutic purposes in humans is an ambitious strategy for treating muscle impairment diseases.

Acute Effect of Whole-Body Vibration in a Female Dog with Metritis

Background: Whole-body vibration is a modality of exercise used in humans for therapeutic purposes or to increase physical performance. In veterinary medicine there are only a few reports on the use of this technology. The Whole-body vibration derivate from vibrating rhythmic movements caused by vibrating platforms. Vibrating platforms are used over 30 years in the treatment and prevention of injuries and other debilitating conditions in humans. This paper aims to describe the first report of a possible spontaneous opening of the cervix in a female dog with purulent metritis by Whole-body vibration using a platform vibration.Case: A sexually intact female American Pit Bull Terrier dog showed an acute effect after a single session of Whole- body vibration training. Physical examination and physiological parameters were within normal. Slight edema of the vulva was observed without signs of discharge. Complete blood cell count, serum chemistry and urinalysis yielded no significant abnormal findings. An enlarged uterus with content was observed during a transabdominal ultrasound. Despite this, a vibrating-platform session was performed during 15 min. A purulent vulvar discharge was observed 6 min. after Whole-body vibration exercise and remained continuous through session. After the Whole-body vibration exercise, the dog was treated with cephalexin for 15 days. Escherichia coli were isolated from vaginal discharge culture. Seven days after the Whole-body vibration session, no signs of vulvar edema or discharge were observed. A complete blood count, serum chemistry, urinalysis and uterus ultrasound showed no abnormalities. Ovariohysterectomy by minimally invasive technique was performed and was observed cysts in both ovaries. The patient was placed under general anesthesia withisoflurane/O2. The premedication used was carprofen, morphine sulphate, acepromazine and diazepam. The induction was with propofol. An open ventral midline celiotomy an ovariohysterectomy was performed. Ten days after the surgery the dog showed no abnormality. After recovered from anesthesia, the patient was discharged with meloxican and tramadol for postoperative inflammation and pain management. According to the owner, the immediate postoperative period was without any problems. The histopathology examination confirmed the presence of right ovary cystic and left uterine horn cystic, and uterine hyperplasia. Four months after the surgery, the dog returned for evaluation and the owner reported that patient had been doing well, with normal appetite, normal urination and defecation.Discussion: The Whole-body vibration is not yet fully understood and it is important that the patient is clinically evaluated before the Whole-body vibration session. Whole-body vibration is a modality of exercise used in humans for therapeutic purposes or to increase physical performance. Otherwise, in veterinary medicine there are only a few reports on the use of this technology. In the present report there were no clinical signs that indicated uterine infection. In human patients there are reports that showed any clinical signs of diseases before use the Whole-body vibration. The ultrasound exam had been done because was part of another study and showed an enlarged uterus. Therefore, the cervix opening after a single Whole- body vibration training may be considered a positive side effect of Whole-body vibration in female dogs since this event helped to diagnose the disease. Although, the effects of WBV on reproductive organs and endocrine system are not clear.

Drug repurposing for antivirulence therapy against opportunistic bacterial pathogens.

Antibiotic resistance is a serious public health concern at the global level. Available antibiotics have saved millions of lives, but are progressively losing their efficacy against many bacterial pathogens, and very few new antibiotics are being developed by the pharmaceutical industry. Over the last few decades, progress in understanding the pathogenic process of bacterial infections has led researchers to focus on bacterial virulence factors as potential targets for 'antivirulence' drugs, i.e. compounds which inhibit the ability of bacteria to cause damage to the host, as opposed to inhibition of bacterial growth which is typical of antibiotics. Hundreds of virulence inhibitors have been examined to date in vitro and/or in animal models, but only a few were entered into clinical trials and none were approved, thus hindering the clinical validation of antivirulence therapy. To breathe new life into antivirulence research and speed-up its transfer to the clinic, antivirulence activities have also been sought in drugs already approved for different therapeutic purposes in humans. If effective, these drugs could be repositioned for antivirulence therapy and have an easier and faster transfer to the clinic. In this work we summarize the approaches which have led to the identification of repurposing candidates with antivirulence activities, and discuss the challenges and opportunities related to antivirulence therapy and drug repurposing. While this approach undoubtedly holds promise for boosting antivirulence drug research, some important issues remain to be addressed in order to make antivirulence drugs viable alternatives to traditional antibacterials.

Unique Spatiotemporal Neuromodulation of the Lumbosacral Circuitry Shapes Locomotor Success after Spinal Cord Injury.

Spinal cord epidural stimulation has resulted in the initiation of voluntary leg movements and improvement in postural, bladder, and sexual function. However, one of the limitations in reaching the full potential of epidural stimulation for therapeutic purposes in humans has been the identification of optimal stimulation configurations that can neuromodulate the spinal cord for stepping. In the present work, we investigated the mechanisms underlying the specificity of interaction between the rostral and caudal spinal cord circuitries in enabling locomotion in spinal rats (n = 10) by epidural spinal cord stimulation. By using unique spatiotemporal epidural stimulation parameters of the lumbar and sacral spinal cords, a robust stepping pattern in spinal rats was observed with only six training sessions and as early as 3 weeks post-injury. Electrophysiological evidence reveals that in addition to frequency of stimulation pulses at the stimulation sites, the relative timing between stimulation pulses applied at the lumbar (L2) and sacral (S1) segments of the spinal cord heavily impacted stepping performance. Best stepping was established at a higher stimulation frequency (40 Hz vs. 5, 10, 15, and 20Hz) and at specific relative time-intervals between the stimulation pulses (L2 pulse applied at 18-25 msec after the onset of the S1 pulse; S1 pulse applied 0-7 msec after the L2 pulse). Our data suggest that controlling pulse-to-pulse timing at multiple stimulation sources provides a novel strategy to optimize spinal stepping by fine-tuning the physiological state of the locomotor networks. These findings hold direct relevance to the clinician who will incorporate electrical stimulation strategies for optimizing control of locomotion after complete paralysis.

CD1d-restricted peripheral T cell lymphoma in mice and humans.

Peripheral T cell lymphomas (PTCLs) are a heterogeneous entity of neoplasms with poor prognosis, lack of effective therapies, and a largely unknown pathophysiology. Identifying the mechanism of lymphomagenesis and cell-of-origin from which PTCLs arise is crucial for the development of efficient treatment strategies. In addition to the well-described thymic lymphomas, we found that p53-deficient mice also developed mature PTCLs that did not originate from conventional T cells but from CD1d-restricted NKT cells. PTCLs showed phenotypic features of activated NKT cells, such as PD-1 up-regulation and loss of NK1.1 expression. Injections of heat-killed Streptococcus pneumonia, known to express glycolipid antigens activating NKT cells, increased the incidence of these PTCLs, whereas Escherichia coli injection did not. Gene expression profile analyses indicated a significant down-regulation of genes in the TCR signaling pathway in PTCL, a common feature of chronically activated T cells. Targeting TCR signaling pathway in lymphoma cells, either with cyclosporine A or anti-CD1d blocking antibody, prolonged mice survival. Importantly, we identified human CD1d-restricted lymphoma cells within Vδ1 TCR-expressing PTCL. These results define a new subtype of PTCL and pave the way for the development of blocking anti-CD1d antibody for therapeutic purposes in humans.