A narrative review of the phenomenon of predatory journals to create awareness among researchers in veterinary medicine.

In recent years, especially since the COVID-19 pandemic, the number of predatory journals has increased significantly. Predatory journals exploit the "open-access model" by engaging in deceptive practices such as charging high publication fees without providing the expected quality and performing insufficient or no peer review. Such behaviors undermine the integrity of scientific research and can result in researchers having trouble identifying reputable publication opportunities, particularly early-career researchers who struggle to understand and establish the correct criteria for publication in reputable journals. Publishing in journals that do not fully cover the criteria for scientific publication is also an ethical issue. This review aimed to describe the characteristics of predatory journals, differentiate between reliable and predatory journals, investigate the reasons that lead researchers to publish in predatory journals, evaluate the negative impact of predatory publications on the scientific community, and explore future perspectives. The authors also provide some considerations for researchers (particularly early-career researchers) when selecting journals for publication, explaining the role of metrics, databases, and artificial intelligence in manuscript preparation, with a specific focus on and relevance to publication in veterinary medicine.

Does Nitrofurantoin Improve the Portfolio of Vets against Resistant Bacteria in Companion Animals?

In clinical practice in dogs and cats, antimicrobials are frequently used, sometimes overused or misused, increasing antimicrobial resistance (AMR). In order to limit the phenomenon, laws have been enacted and guidelines for prudent and rational use of antibiotics have been developed. Interestingly, old molecules such as nitrofurantoin could be used to achieve therapeutic success and overcome AMR. To better understand the suitability of this molecule in veterinary medicine, the authors performed a revision of the literature, searching on PubMed and entering the following keywords: nitrofurantoin, veterinary medicine, dog, and cat connected by the Boolean operator "and", without restrictions on the date of publication. Thirty papers were finally selected. It is possible to appreciate that papers dealing with nitrofurantoin have been written from the early 1960s to the middle of the 1970s, and then a long period passed without publications. Only at the beginning of the new century, nitrofurantoin was included or was sometimes the focus of papers dealing with its efficacy in veterinary medicine, mainly in the treatment of urinary tract infections. One recent paper dealt with pharmacokinetic features, and none was dedicated to pharmacokinetic/pharmacodynamic integration or modeling. Nitrofurantoin appears to be still effective against several pathogens that rarely develop resistance to this molecule.

Doxorubicin-Loaded Lipid Nanoparticles Coated with Calcium Phosphate as a Potential Tool in Human and Canine Osteosarcoma Therapy.

Osteosarcoma (OSA) is the most frequently diagnosed primary malignant bone tumor in humans and dogs. In both species, standard chemotherapy can be limited by multidrug resistance of neoplastic cells, which prevents intracellular accumulation of cytotoxic drugs, resulting in chemotherapy failure. In this study, a lipophilic ester of doxorubicin (C12DOXO) was loaded into nanoparticles (NPs) using the "cold microemulsion dilution" method. The resulting NPs were then coated with calcium phosphate (CaP) in two different ways to have calcium or phosphate ions externally exposed on the surface. These systems were characterized by determining mean diameter, zeta potential, and drug entrapment efficiency; afterward, they were tested on human and canine OSA cells to study the role that the coating might play in increasing both drug uptake into tumor cells and cytotoxicity. Mean diameter of the developed NPs was in the 200-300 nm range, zeta potential depended on the coating type, and C12DOXO entrapment efficiency was in the 60-75% range. Results of studies on human and canine OSA cells were very similar and showed an increase in drug uptake and cytotoxicity for CaP-coated NPs, especially when calcium ions were externally exposed. Therefore, applications in both human and veterinary medicine can be planned in the near future.

Implications of Veterinary Medicine in the comprehension and stewardship of antimicrobial resistance phenomenon. From the origin till nowadays.

Antimicrobial resistance (AMR) is defined by the entire scientific community as the major threat for human health and it is responsible for an increase in morbidity and mortality rates. The reasons behind this phenomenon are complex and the solution is achievable only considering the One Health approach, that encompasses the integration and implementation of human health, veterinary medicine and environmental status. Authors aimed to write this review to summarize to readers the three milestones of One-Health, underlying the most important topics in which veterinary medicine is mostly involved. Therefore, a short introduction about the history of AMR in veterinary medicine is provided, then more detailed aspects about the impact of AMR related to pets, food producing animals, wild animals and environment are discussed. Finally, some critical aspects about current and future issues are considered.

TRPV1 Receptor Identification in Bovine and Canine Mitral Valvular Interstitial Cells.

Myxomatous mitral valve degeneration (MMVD) is the most common acquired cardiac disease in canine species, and valvular interstitial cells (VICs) are considered the main responsible for the development of this pathology. The scientific interest is focused on isolating and characterizing these cells. The aims of the present study were to verify a novel VICs mechanical isolation method and to characterize isolated cells using immunocytochemistry and immunofluorescence, with parallel histological and immunohistochemistry assays on bovine and canine healthy and MMVD mitral valves. Antibodies against vimentin (VIM), smooth muscle actin (SMA), von Willebrand (vW) factor, Transforming Growth Factor (TGF) ß1, and Transient Receptor Potential Vanilloid 1 (TRPV1) were used. The isolation method was considered reliable and able to isolate only VICs. The different assays demonstrated a different expression of SMA in healthy and MMVD mitral valves, and TRPV1 was isolated for the first time from bovine and canine VICs and the correspondent mitral valve leaflets. The novelties of the present study are the new isolation method, that may allow correlations between laboratory and clinical conditions, and the identification of TRPV1, which will lead to further investigations to understand its function and possible role in the etiology of MMVD and to the design of new therapeutic strategies.

In vitro and in vivo evaluation of a new phytotherapic blend to treat acute externa otitis in dogs.

Canine otitis externa is frequently encountered in veterinary practice, caused by primary factors with bacteria and yeast overgrowth acting as secondary and perpetuating factors. The pharmacological support includes anti-inflammatory, antimicrobials, and antimycotic drugs, but therapeutic failure and antimicrobial resistance are leading to alternative strategies based on phytotherapic products. This study aimed to evaluate an essential oil blend (Otogen® ) to treat otitis externa in dogs. The experimental design was divided in: (a) an in vitro approach, based on the European Normative UNI EN 1275:2006, to assess the efficacy of the product against the most frequently isolated microorganisms during otitis externa. (b) an in vivo part, 12 owned dogs presenting with acute otitis externa were enrolled. A significant growth reduction (>99.9%) of Malassezia pachydermatis and Candida albicans after 15 min of contact and Pseudomonas aeruginosa after 1 h of incubation was recorded. For Staphylococcus pseudintermedius, 50% of growth reduction were appreciated after 15 min. Results obtained in vivo after 7 days of blend administration, noted a significant improvement of all the considered parameters (most important were head shaking, erythema, and scraping). The results obtained may support the usefulness of the tested phytotherapic blend to manage acute otitis externa in dogs.

Antibiotic Stewardship for Canine and Feline Acute Urinary Tract Infection: An Observational Study in a Small Animal Hospital in Northwest Italy.

Antimicrobial stewardship programs (ASPs) have been suggested to reduce antimicrobial resistance phenomena in veterinary medicine, as antibiotics are commonly used without microbiological confirmation. The aim of the present study is to design a specific working flow for a tailored antimicrobial treatment in the case of canine and feline urinary tract infections (UTIs). Urine samples were collected by cystocentesis from 16 dogs and 12 cats presenting acute signs of UTI. The therapy was decided according to the minimal inhibitory concentration, and it was possible to monitor 14 dogs and 11 cats. Rescue therapy (amoxicillin and clavulanic acid) was included in emergency cases. Escherichia coli, Proteus mirabilis, and Streptococcus canis were isolated in dogs, and Escherichia coli, Staphylococcus pseudintermedius, and Staphylococcus aureus were isolated in cats. No multidrug-resistant strains were detected, but all Staphylococci were methicillin resistant. Only one cat received rescue therapy, and only one dog was recruited. Dogs were treated with tetracycline (1/14), fluoroquinolones (6/14), beta-lactams (6/14), and gentamicin (1/14), while cats received fluoroquinolones (3/11), nitrofurans (1/11), clindamycin (1/11), and beta-lactams (6/11). The success rate was very high. Our findings are interesting because this is the first ASP in Italy, and it may be used as a model to develop ASPs for other pathologies.

Diagnostic and therapeutic management of Cryptococcosis in a kitten with practical considerations to veterinary pediatric therapeutic approach.

A 3-months-old male domestic kitten was referred for repeated seizures. Analysis revealed Cryptococcus neoformans. Levetiracetam and fluconazole were administered without significant clinical improvements and without negativization. Hypothesizing resistance, therapy was switched to amphotericin B. Seizure disappeared. Haematological controls highlighted transitory increasing of CK, BUN, ALP and cholesterol. PCR repeated two weeks after the treatment was negative for Cryptococcus neoformans. Nowadays the cat is 5 years old, and no seizures occurred since the age of 5 months.

Direct in situ rt-PCR.

In situ polymerase chain reaction (PCR) is a histological technique that exploits the advantages of PCR for detection of mRNA directly in tissue sections. It somehow conjugates together PCR and in situ hybridization that is more traditionally employed for mRNA localization in cell organelles, intact cells, or tissue sections. This chapter describes the application of in situ PCR for neuropeptide mRNA localization. We provide here a detailed protocol for direct in situ reverse transcription (RT) PCR (RT-PCR) with nonradioactive probes after fixation and paraffin embedding or cryosectioning. Digoxigenin-labeled nucleotides (digoxigenin-11-dUTP) are incorporated in the PCR product after RT and subsequently detected with an anti-digoxigenin antibody conjugated with alkaline phosphatase. The procedure can be modified for use with fluorescent probes and employed in combination with enzyme/fluorescence immunocytochemical labeling.

Modulation of inhibitory neurotransmission by the vanilloid receptor type 1 (TRPV1) in organotypically cultured mouse substantia gelatinosa neurons.

The vanilloid receptor type 1 (TRPV1) plays a pivotal role in modulating thermal, chemical, and inflammatory pain. TRPV1s are expressed in some dorsal horn (DH) neurons, but their contribution, if any, to central pain processing still remains unclear. We studied the effects of 2microM capsaicin-induced TRPV1 activation in organotypically cultured substantia gelatinosa neurons from post-natal (8-12) mice. Capsaicin affected sIPSC frequency (272+/-60% of control, n=14, P<0.02), but not amplitude (131+/-12% of control, n=14, P>0.05) in patch clamp recordings, also in the presence of 50microM AP-5 (frequency: 265+/-69% of control; n=8, P<0.05; amplitude: 156+/-28% of control; n=8, P>0.05). The frequency increase was reduced by TTX (181+/-21% of control; n=12, P<0.05). Pre-administration of I-RTX (1microM), a TRPV1 antagonist, prevented the capsaicin effect (frequency: 149+/-28% of control, P>0.05, n=12; amplitude: 97+/-4% of control, P>0.05, n=12). NADA (1microM), an endovanilloid/endocannabinoid agonist of TRPV1, induced a significant increase of sISPC frequency (191+/-40% of control; n=8, P<0.05) without affecting the amplitude (102+/-6% of control; n=8, P>0.05), and the co-application of two naturally occurring N-acyldopamines, PALDA (5microM) and STEARDA (5microM) that facilitate the effect of TRPV1 agonists, also induced a significant increase of sIPSC frequency (278+/-67% of control, n=6, P<0.05). The presence of TRPV1 protein and mRNA in DH neurons was confirmed by histological (immunocytochemistry, in situ PCR) and biochemical (Western blotting, PCR) procedures. These data show that TRPV1 modulates inhibitory neurotransmission in cultured substantia gelatinosa neurons, and suggest that endogenous agonists can activate the spinal receptors in vivo.

Autophagy regulates the post-translational cleavage of BCL-2 and promotes neuronal survival.

B-cell lymphoma 2 protein (BCL-2) is one of the more widely investigated anti-apoptotic protein in mammals, and its levels are critical for protecting from programmed cell death. We report here that the cellular content of BCL-2 is regulated at post-translational level along the autophagy/lysosome pathways in organotypic cultures of post-natal mouse cerebellar cortex. Specifically this mechanism appears to be effective in the cerebellar granule cells (CGCs) that are known to undergo massive programmed cell death (apoptosis) during post-natal maturation. By the use of specific agonists/antagonist of calcium channels at the endoplasmic reticulum it was possible to understand the pivotal role of calcium release from intracellular stores in CGC neuroprotection. The more general significance of these findings is supported by a very recent study Niemann-Pick transgenic mice.

Posttranslational regulation of BCL2 levels in cerebellar granule cells: A mechanism of neuronal survival.

Apoptosis can be modulated by K(+) and Ca(2+) inside the cell and/or in the extracellular milieu. In murine organotypic cultures, membrane potential-regulated Ca(2+) signaling through calcineurin phosphatase has a pivotal role in development and maturation of cerebellar granule cells (CGCs). P8 cultures were used to analyze the levels of expression of B cell lymphoma 2 (BCL2) protein, and, after particle-mediated gene transfer in CGCs, to study the posttranslational modifications of BCL2 fused to a fluorescent tag in response to a perturbation of K(+)/Ca(2+) homeostasis. There are no changes in Bcl2 mRNA after real time PCR, whereas the levels of the fusion protein (monitored by calculating the density of transfected CGCs under the fluorescence microscope) and of BCL2 (inWestern blotting) are increased. After using a series of agonists/antagonists for ion channels at the cell membrane or the endoplasmic reticulum (ER), and drugs affecting protein synthesis/degradation, accumulation of BCL2 was related to a reduction in posttranslational cleavage by macroautophagy. The ER functionally links the [K(+)](e) and [Ca(2+)](i) to the BCL2 content in CGCs along two different pathways. The first, triggered by elevated [K(+)](e) under conditions of immaturity, is independent of extracellular Ca(2+) and operates via IP3 channels. The second leads to influx of extracellular Ca(2+) following activation of ryanodine channels in the presence of physiological [K(+)](e), when CGCs are maintained in mature status. This study identifies novel mechanisms of neuroprotection in immature and mature CGCs involving the posttranslational regulation of BCL2.

Apoptosis of the cerebellar neurons.

Naturally occurring neuronal death (NOND) is an essential phenomenon during the course of normal development of the nervous system. Studies in vivo and on organotypic cultures have helped to elucidate the basic histological and ultrastructural features, as well as the main cellular mechanisms of NOND in several areas of the brain. This review examines the existing evidence about the two waves of apoptotic cell death that affect the different types of cerebellar neurons in normal development and certain pathological conditions. The first wave regards neuronal progenitors and pre-migratory neuroblasts, the second post-migratory neuroblasts and mature neurons. The underlying cellular and molecular mechanisms are discussed critically also in the light of their relevance to neurodegenerative diseases.

In vivo analysis reveals different apoptotic pathways in pre- and postmigratory cerebellar granule cells of rabbit.

Naturally occurring neuronal death (NOND) has been described in the postnatal cerebellum of several species, mainly affecting the cerebellar granule cells (CGCs) by an apoptotic mechanism. However, little is known about the cellular pathway(s) of CGC apoptosis in vivo. By immunocytochemistry, in situ detection of fragmented DNA, electron microscopy, and Western blotting, we demonstrate here the existence of two different molecular mechanisms of apoptosis in the rabbit postnatal cerebellum. These two mechanisms affect CGCs at different stages of their maturation and migration. In the external granular layer, premigratory CGCs undergo apoptosis upon phosphorylation of checkpoint kinase 1 (Chk1), and hyperphosphorylation of retinoblastoma protein. In postmigratory CGCs within the internal granular layer, caspase 3 and to a lesser extent 7 and 9 are activated, eventually leading to poly-ADP-ribose polymerase-1 (PARP-1) cleavage and programmed cell death. We conclude that NOND of premigratory CGCs is linked to activation of DNA checkpoint and alteration of normal cell cycle, whereas in postmigratory CGCs apoptosis is, more classically, dependent upon caspase 3 activation.