Retinoic acid induces the osteogenic differentiation of cat adipose tissue-derived stromal cells from distinct anatomical sites.

Mesenchymal stromal cells-based regenerative orthopedic therapies have been used in cats as a promising and innovative therapeutic approach to enhance the repair of bone defects. Adipose tissue-derived stromal cells (ADSCs) can be obtained from two main sites-subcutaneous and visceral-with established differences regarding structure, composition, cell content, and functionality. However, in cats, to the best of the authors' knowledge, no studies have been conducted to compare the functional activity of the ADSCs isolated from the two sites, and the impact of these differences on the induced osteogenic potential. Additionally, retinoic acid has been recently regarded as a new osteogenic inducer within cells of distinct species, with undisclosed functionality on cat-derived cell populations. Thus, the present study aimed to evaluate the functional activity of ADSCs isolated from the subcutaneous and visceral adipose sites (SCAT and VAT, respectively) of the cat, as well as the effects of two osteogenic-inducing conditions-the classic dexamethasone, ß-glycerophosphate and ascorbic acid-supplemented media (Dex + ß + AAM), and Retinoic Acid-supplemented media (RAM). The adipose tissue of subcutaneous and visceral origin was isolated, characterized, and ADSCs were isolated and grown in the presence of the two osteogenic-inducing conditions, and characterized in terms of proliferation, metabolic activity, morphology, and osteogenic activity. Our results demonstrated a distinct biological profile of the two adipose tissue sites regarding cell size, vascularization, and morphology. Further, osteogenic-induced ADSCs from both sites presented an increased expression of alkaline phosphatase activity (ALP) and cytochemical staining, as compared with control. Overall, RAM induced higher levels of ALP activity than Dex + ß + AAM, supporting an increased osteogenic activation. Additionally, VAT was the tissue with the best osteogenic potential, showing higher levels of ALP expression, particularly with RAM. In conclusion, different characteristics were found between the two adipose tissue sites-SCAT and VAT, which probably reflect the differences found in the functionality of isolated ADSCs from both tissues. Furthermore, for cat, VAT shows a greater osteogenic-inductive capacity than SCAT, particularly with RAM, which can be of therapeutic relevance for regenerative medicine applications.

Long-Term Exposure to Supraphysiological Levels of Testosterone Impacts Rat Submandibular Gland Proteome.

The salivary glands play a central role in the secretion of saliva, whose composition and volume affect oral and overall health. A lesser-explored dimension encompasses the possible changes in salivary gland proteomes in response to fluctuations in sex hormone levels. This study aimed to examine the effects of chronic exposure to testosterone on salivary gland remodeling, particularly focusing on proteomic adaptations. Therefore, male Wistar rats were implanted with subcutaneous testosterone-releasing devices at 14 weeks of age. Their submandibular glands were histologically and molecularly analyzed 47 weeks later. The results underscored a significant increase in gland mass after testosterone exposure, further supported by histologic evidence of granular duct enlargement. Despite increased circulating sex hormones, there was no detectable shift in the tissue levels of estrogen alpha and androgen receptors. GeLC-MS/MS and subsequent bioinformatics identified 308 proteins in the submandibular glands, 12 of which were modulated by testosterone. Of note was the pronounced upregulation of Klk3 and the downregulation of Klk6 and Klk7 after testosterone exposure. Protein-protein interaction analysis with the androgen receptor suggests that Klk3 is a potential target of androgenic signaling, paralleling previous findings in the prostate. This exploratory analysis sheds light on the response of salivary glands to testosterone exposure, providing proteome-level insights into the associated weight and histological changes.

TLR7 and TLR8 evolution in lagomorphs: different patterns in the different lineages.

Toll-like receptors (TLRs) are one of the most ancient and widely studied innate immune receptors responsible for host defense against invading pathogens. Among the known TLRs, TLR7 and TLR8 sense and recognize single-stranded (ss) RNAs with a dynamic evolutionary history. While TLR8 was lost in birds and duplicated in turtles and crocodiles, TLR7 is duplicated in some birds, but in other tetrapods, there is only one copy. In mammals, with the exception of lagomorphs, TLR7 and TLR8 are highly conserved. Here, we aim to study the evolution of TLR7 and TLR8 in mammals, with a special focus in the order Lagomorpha. By searching public sequence databases, conducting evolutionary analysis, and evaluating gene expression, we were able to confirm that TLR8 is absent in hares but widely expressed in the European rabbit. In contrast, TLR7 is absent in the European rabbit and quite divergent in hares. Our results suggest that, in lagomorphs, more in particular in leporids, TLR7 and TLR8 genes have evolved faster than in any other mammalian group. The long history of interaction with viruses and their location in highly dynamic telomeric regions might explain the pattern observed.

Comparison of dorsoventral and ventrodorsal approaches for ultrasound-guided quadratus lumborum block in cats: a cadaver study.

OBJECTIVE: To assess the feasibility of an ultrasound (US)-guided quadratus lumborum (QL)-psoas muscle (Pm) interfascial plane injection using a dorsoventral (DV) approach, and to compare needle visualization and distribution pattern between DV and ventrodorsal (VD) approaches. STUDY DESIGN: Experimental anatomical study. ANIMALS: A total of 14 cat cadavers. METHODS: Bilateral in-plane US-guided injections in the QL-Pm plane were performed at the level of the second lumbar (L2) transverse process. A spinal needle was inserted using either a VD or DV approach. A total volume of 0.4 mL kg-1 of a solution of ropivacaine 0.18%, tissue dye and iohexol (350 mg mL-1), in a proportion of 3:1:1, respectively, was injected bilaterally. Spread of injectate was compared and evaluated using US, computed tomography (CT) scanning and anatomical dissection. Presence of dye on the sympathetic trunk was compared between methods using Kappa (κ) coefficient of agreement. RESULTS: The QL-Pm plane was visualized, and dye was present in the target plane using both approaches. Needle visualization in the VD approach was enhanced compared with the DV approach (p = 0.0005). Mean distribution along the sympathetic trunk was 4.8 and 4.1 segments in the VD and DV approaches, respectively, showing non-statistical differences. The ventral branches of L1, L2 and L3 were successfully stained in 11/14 and 6/14 using the VD and DV approaches, respectively. Substantial agreement (κ = 0.61) of sympathetic trunk staining was observed between CT scan and anatomical dissection. Dye was observed cranial to the diaphragm in 7/8 cases on CT with both approaches. Epidural and abdominal cavity dye distribution was not observed. CONCLUSIONS AND CLINICAL RELEVANCE: The DV approach to the QL-Pm plane could be an alternative to the VD approach. Both approaches seem capable of reaching the spinal nerves, the sympathetic trunk and both splanchnic and coeliac nerves in feline cadavers.

HPV16 is sufficient to induce squamous cell carcinoma specifically in the tongue base in transgenic mice.

Head and neck squamous cell carcinomas (HNSCCs) associated with human papillomavirus (HPV) occur specifically in the tonsils and the tongue base, but the reasons for this specificity remain unknown. We studied the distribution of oral and pharyngeal lesions in HPV16-transgenic mice where the expression of all the HPV16 early genes is targeted to keratinising squamous epithelia by the cytokeratin 14 (Krt14) gene promoter. At 30 weeks of age, 100% of mice developed low- and high-grade intraepithelial dysplasia at multiple sites. Twenty per cent of animals developed invasive cancers that remarkably were restricted to the tongue base, in association with the circumvallate papilla. The lesions maintained expression of CK14 (KRT14) and the HPV16 E6 and E7 oncogenes, and displayed deregulated cell proliferation and up-regulation of p16INK4A . Malignant lesions were poorly differentiated and destroyed the tongue musculature. We hypothesised that the tongue base area might contain a transformation zone similar to those observed in the cervix and anus, explaining why HPV-positive cancers target that area specifically. Immunohistochemistry for two transformation zone markers, CK7 (KRT7) and p63 (TP63), revealed a squamocolumnar junction in the terminal duct of von Ebner's gland, composed of CK7+ luminal cells and p63+ basal cells. Dysplastic and invasive lesions retained diffuse p63 expression but only scattered positivity for CK7. Site-specific HPV-induced carcinogenesis in the tongue base may be explained by the presence of a transformation zone in the circumvallate papilla. This mouse model reproduces key morphological and molecular features of HPV-positive HNSCC, providing a unique in vivo tool for basic and translational research. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

HPV16 induces penile intraepithelial neoplasia and squamous cell carcinoma in transgenic mice: first mouse model for HPV-related penile cancer.

Penile cancer is an under-studied disease that occurs more commonly in developing countries and 30-50% of cases show high-risk human papillomavirus (HPV) infection. Therapeutic advances are slow, largely due to the absence of animal models for translational research. Here, we report the first mouse model for HPV-related penile cancer. Ten-week-old mice expressing all the HPV16 early genes under control of the cytokeratin 14 (Krt14) gene promoter and matched wild-type controls were exposed topically to dimethylbenz(a)anthracene (DMBA) or vehicle for 16 weeks. At 30 weeks of age, mice were sacrificed for histological analysis. Expression of Ki67, cytokeratin 14, and of the HPV16 oncogenes E6 and E7 was confirmed using immunohistochemistry and quantitative PCR, respectively. HPV16-transgenic mice developed intraepithelial lesions including condylomas and penile intraepithelial neoplasia (PeIN). Lesions expressed cytokeratin 14 and the HPV16 oncogenes E6 and E7 and showed deregulated cell proliferation, demonstrated by Ki67-positive supra-basal cells. HPV16-transgenic mice exposed to DMBA showed increased PeIN incidence and squamous cell carcinoma. Malignant lesions showed varied histological features closely resembling those of HPV-associated human penile cancers. Wild-type mice showed no malignant or pre-malignant lesions even when exposed to DMBA. These observations provide the first experimental evidence to support the etiological role of HPV16 in penile carcinogenesis. Importantly, this is the first mouse model to recapitulate key steps of HPV-related penile carcinogenesis and to reproduce morphological and molecular features of human penile cancer, providing a unique in vivo tool for studying its biology and advancing basic and translational research. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

A description of an ultrasound-guided technique for a quadratus lumborum block in the cat: a cadaver study.

OBJECTIVES: To describe the ultrasonographic (US) anatomy of the sublumbar region at the level of second lumbar vertebra (L2), to assess the feasibility of US-guided quadratus lumborum (QL) injections and to evaluate the dye distribution pattern in feline cadavers. STUDY DESIGN: Descriptive feline cadaver study. ANIMALS: A total of eight cat cadavers. METHODS: Using a linear probe (6-14 MHz), with the cadavers in lateral recumbency, the QL and psoas muscles (Pm) were identified at the level of L2 and landmarks recorded. Using an in-plane technique, a spinal needle was inserted in a ventrodorsal direction to reach the interfascial plane between the QL and Pm. Using a ropivacaine, dye and iohexol solution, a total volume of 0.4 mL kg-1 was injected. Computed tomography (CT) scanning and anatomic dissection were performed to evaluate the spread of injectate. Presence of dye on the sympathetic trunk was compared between assessment methods using kappa coefficient of agreement (κ). RESULTS: Using US guidance, the QL-Pm interfascial plane was identified and dye solution was present in the target fascial plane in all animals. Injectate was distributed on the ventral roots of the spinal nerves between the first and third lumbar vertebrae in 6/8 cats, and on the sympathetic trunk from the thirteenth thoracic to the third lumbar vertebrae in 7/8. Dye was found on the major splanchnic nerve in 7/8 cats and on the minor splanchnic nerve and coeliac ganglion pathways in all animals. Agreement between dissection and CT images dye distribution on the sympathetic trunk was κ = 0.72. CONCLUSIONS AND CLINICAL RELEVANCE: The US-guided QL injection was feasible and injectate was present in the QL-Pm interfascial plane in all feline cadavers. In vivo studies are warranted to evaluate the analgesic efficacy of this technique for abdominal surgical procedures.

Encapsulated bacteriophages in alginate-nanohydroxyapatite hydrogel as a novel delivery system to prevent orthopedic implant-associated infections.

An innovative delivery system based on bacteriophages-loaded alginate-nanohydroxyapatite hydrogel was developed as a multifunctional approach for local tissue regeneration and infection prevention and control. Bacteriophages were efficiently encapsulated, without jeopardizing phage viability and functionality, nor affecting hydrogel morphology and chemical composition. Bacteriophage delivery occurred by swelling-disintegration-degradation process of the alginate structure and was influenced by environmental pH. Good tissue response was observed following the implantation of bacteriophages-loaded hydrogels, sustaining their biosafety profile. Bacteriophages-loaded hydrogels did not affect osteoblastic cells' proliferation and morphology. A strong osteogenic and mineralization response was promoted through the implantation of hydrogels system with nanohydroxyapatite. Lastly, bacteriophages-loaded hydrogel showed excellent antimicrobial activity inhibiting the attachment and colonization of multidrug-resistant E. faecalis surrounding and within femoral tissues. This new local delivery approach could be a promising approach to prevent and control bacterial contamination during implantation and bone integration.

Association of lifestyle factors with semen quality: A pilot study conducted in men from the Portuguese Trás-os-Montes and Alto Douro region followed in fertility support consultations.

A cross-sectional pilot study was conducted in men followed in fertility consultations, from the portuguese Trás-os-Montes and Alto Douro region, in order to associate several lifestyle factors with the spermatic parameters. Of a total of 522 men, 373 were compared based on the occupational exposure to harmful factors, smoking habits and practice of physical exercise per week, and the other 149 men according to their body mass index (normal weight vs. overweight vs. obesity). In the absence of harmful occupational factors, physical exercise seems to be associated with sperm quality improvement, whether individuals smoke or not. When exposed to harmful environments, non-smokers that practice physical exercise more than two times per week tended to present the best vitality, normal morphology and sperm concentration (p > .05). However, if they smoke, physical exercise seems not enough to enhance the spermatic parameters. The BMI correlated negatively with the spermatic quality, especially with sperm concentration (p < .05). Concerning men that did not present lifestyle risks associated, the motility, midpiece and tail abnormalities, and teratozoospermia index were significantly worse on obese individuals comparing to overweight men (p < .05). Thus, patients should also be recommended to control their weight and to have a BMI under 30 kg/m2 .

An Overview of Current Alternative Models for Use in the Context of Prostate Cancer Research.

Prostate cancer is one of the most commonly diagnosed cancers worldwide, particularly in elderly populations. To mitigate the expected increase in prostate cancer-related morbidity and mortality as a result of an expanding aged population, safer and more effective therapeutics are required. To this end, plenty of research is focusing on the mechanisms underlying cancer initiation and development, the metastatic process and on the discovery of new therapies. While animal models are used (mainly rats and mice) for the study of prostate cancer, alternative models and methods are increasingly being considered to replace, or at least reduce, the number of animals used in this particular field of research. In this review, we cover some of the alternative models that are currently available for use in the study of prostate cancer, including: mathematical models; 2-D and 3-D cell cultures; microfluidic devices; the chicken egg chorioallantoic membrane-based model; and zebrafish embryo-based models. The main advantages and limitations, as well as some examples of applications, are given for each type of model. According to our analysis, immortalised cell lines are still the most commonly used models in the field of prostate cancer research. However, the use of alternative models for prostate cancer research will likely become more prevalent in the coming years partly because of the increasing societal pressure to reduce the numbers of laboratory animals. In this context, the development and dissemination of effective non-animal alternative models assumes particular relevance and will be instrumental in leveraging their success. Taking these perspectives into account, we believe that technological advances will lead to more effective cell culture systems, namely 3-D cultures or organ-on-a-chip devices, which can be used to replace animal-based models in prostate cancer research.

Assessment of the exposure to heavy metals and arsenic in captive and free-living black kites (Milvus migrans) nesting in Portugal.

Due to their high trophic level, raptor species may serve as important indicators of environmental contamination by heavy metals. This study was conducted to determine if the habitat of the black kite (Milvus migrans) is contaminated by heavy metals and arsenic and to assess the degree and type of exposure that may be present. For this purpose, this study was conducted on a group of captive birds (n = 12) and on a group of free-living birds admitted to two wildlife rehabilitation centers (n = 31). Blood samples were taken for analysis of arsenic (As), mercury (Hg) and lead (Pb) concentrations by inductively coupled plasma mass spectrometry (ICP-MS). Captive birds had the lowest blood concentrations for all toxic elements examined, but significant differences from the concentrations found in free-living birds were only observed for Hg and Pb (p < 0.01). Arsenic concentrations were almost three times higher in free-living birds (4.521 ±â€¯5.695 µg/dl) then in captive birds (1.566 ±â€¯0.753 µg/dl). In all the samples of captive birds' mercury was not detected, while in free-living birds we observed a concentration of 7.493 ±â€¯8.464 µg/dl (p < 0.01). Regarding lead, we observed a concentration almost four-fold higher in free-living birds (19.430 ±â€¯29.294 µg/dl) then in captive birds (4.449 ±â€¯1.987 µg/dl) (p < 0.01). Therefore, available sources of Pb and Hg seem to be present in the habitat of the black kite.

Potential effects of sulforaphane to fight obesity.

Obesity is linked to the onset of many diseases such as diabetes mellitus, cardiovascular diseases and cancer, among others. The prevalence of obesity nearly doubled worldwide between 1980 and 2014. Simultaneously, in the last decade, the effects of sulforaphane as a potential treatment for obesity have been investigated, with promising results. Fruits and vegetables and their processed agri-food co-products are good sources of natural health-promoting compounds. Brassica crops are among the most produced crops in the world and are a good source of glucoraphanin, which, following hydrolysis, releases sulforaphane. The Brassicaceae family generates large amounts of co-products with no intended use, causing negative economic and environmental impact. Valorization of these co-products could be achieved through their exploitation for the extraction of bioactive compounds such as sulforaphane. However, the extraction process still needs further improvement for its economic feasibility. This article reviews the potential effects of sulforaphane in the treatment of obesity, linked to the relevance of giving Brassica co-products added value, which is of key importance for the competitiveness of farmers and the agri-food industry. © 2018 Society of Chemical Industry.

Effects of exercise training on breast cancer metastasis in a rat model.

Exercise training is thought to play a protective role against cancer development and metastasis, either by reducing hormonal stimulation of hormone-dependent cancers or by reducing the permeability of vascular walls towards invading metastatic cells. The purpose of this work was to evaluate the role of long-term exercise training in the development and metastasis of breast cancer, in an immune-competent 1-methyl-1-nitrosourea (MNU) induced rat model. A single MNU dose was administered to Sprague-Dawley rats at 50 days of age and the rats were subjected to exercise training on a treadmill at 20 m/min, 60 min/day, 5 days/week for 35 weeks. Exercised animals developed slightly less (2.30 ± 1.42) tumours per animal than sedentary animals (2.55 ± 1.44) and did not develop any metastasis, while two pulmonary metastases were observed in the sedentary group. All primary neoplasms and their metastases were positive for oestrogen (ER) α and progesterone (PR) receptors, indicating high hormonal sensitivity. Interestingly, exercise training increased circulating oestrogen levels, thus suggesting that the mechanism might involve either or both of a protective hormone-independent effect and modulation of tumoural vascularization.

Radiographic assessment of humeroulnar congruity in a medium and a large breed of dog.

Elbow joint incongruity is recognized as an important factor in the development, treatment, and prognosis of canine elbow dysplasia. Elbow incongruity has been measured based on radiographic joint space widths, however these values can be affected by the degree of elbow joint flexion. Recent studies have reported radiographic curvature radii as more precise measures of humeroulnar congruity in dogs. The aim of this prospective observational study was to describe radiographic curvature radii measured from flexed and extended elbow radiographs for a sample of dogs representing a medium breed (Portuguese Pointing Dog) and a large breed (Estrela Mountain Dog). The curvature radii from the ulnar trochlear notch and humeral trochlea were measured in 114 mediolateral elbow extended radiographic views (30 Portuguese Pointing Dog and 27 Estrela Mountain Dog), and 84 mediolateral flexed views (22 Portuguese Pointing Dog and 20 Estrela Mountain Dog). The sampled animals' ages ranged from 12 to 84 months (34.6 ± 17.8 months). Good agreement was observed between curvature radii measurements for flexed vs. extended views in both breed groups. Ulnar trochlear notch curvature radii measurements were greater than humeral trochlea curvature radii measurements in both breed groups. Both curvature radii were greater in the large-breed dog group vs. the medium-breed dog group. Both breed groups had ulnar and humeral curves with similar typology. However, the large breed group had greater intermediate differences between the humeroulnar surface curvature radii. Results from this study supported the use of curvature radii as measures of humeroulnar congruity in mediolateral flexed elbow radiographs of medium and large breed dogs.

Lead Poisoning Due to Lead-Pellet Ingestion in Griffon Vultures ( Gyps fulvus ) From the Iberian Peninsula.

Avian scavengers that typically include game birds and mammals in their diets are at risk of lead poisoning from ingestion of carcasses with fragmented or residual lead ammunition that is used in hunting. Thus, lead may be one of the threats that the griffon vulture ( Gyps fulvus ) faces in the Iberian Peninsula and particularly in Portugal, where their conservation status is considered to be near-threatened. This is the first report that details 3 cases of lead poisoning, associated with the ingestion of lead shot, in adult female griffon vultures found in the Iberian Peninsula. The birds were found prostrate and immediately transferred to a wildlife rehabilitation center, where they died within 24 hours after supportive treatment. Necropsy and histopathologic examinations were done in 2 birds and metal analyses were done in all birds to determine the birds' causes of death. In one vulture, 9 uneroded lead pellets were recovered from the stomach, and moderate to severe hemosiderosis was seen histologically in the liver, lungs, and kidneys. Diagnosis of lead poisoning was confirmed by results of metal analyses, which revealed extremely high lead concentrations in blood (969-1384 µg/dL), liver (309-1077 µg/g dry weight), and kidneys (36-100 µg/g dry weight) for all 3 vultures. To prevent lead poisoning in vultures and preserve their populations in the Iberian Peninsula, more resources are needed for diagnosis and treatment of wildlife in rehabilitation centers, new regulations enabling the abandonment of fallen stock in the field must be approved, and lead ammunition must be prohibited in big-game hunting.

Cross-species comparison of mammalian saliva using an LC-MALDI based proteomic approach.

Despite the importance of saliva in the regulation of oral cavity homeostasis, few studies have been conducted to quantitatively compare the saliva of different mammal species. Aiming to define a proteome signature of mammals' saliva, an in-depth SDS-PAGE-LC coupled to MS/MS (GeLC-MS/MS) approach was used to characterize the saliva from primates (human), carnivores (dog), glires (rat and rabbit), and ungulates (sheep, cattle, horse). Despite the high variability in the number of distinct proteins identified per species, most protein families were shared by the mammals studied with the exception of cattle and horse. Alpha-amylase is an example that seems to reflect the natural selection related to digestion efficacy and food recognition. Casein protein family was identified in all species but human, suggesting an alternative to statherin in the protection of hard tissues. Overall, data suggest that different proteins might assure a similar role in the regulation of oral cavity homeostasis, potentially explaining the specific mammals' salivary proteome signature. Moreover, some protein families were identified for the first time in the saliva of some species, the presence of proline-rich proteins in rabbit's saliva being a good example.

Molecular insights into mitochondrial dysfunction in cancer-related muscle wasting.

Alterations in muscle mitochondrial bioenergetics during cancer cachexia were previously suggested; however, the underlying mechanisms are not known. So, the goal of this study was to evaluate mitochondrial phospholipid remodeling in cancer-related muscle wasting and its repercussions to respiratory chain activity and fiber susceptibility to apoptosis. An animal model of urothelial carcinoma induced by exposition to N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) and characterized by significant body weight loss due to skeletal muscle mass decrease was used. Morphological evidences of muscle atrophy were associated to decreased respiratory chain activity and increased expression of mitochondrial UCP3, which altogether highlight the lower ability of wasted muscle to produce ATP. Lipidomic analysis of isolated mitochondria revealed a significant decrease of phosphatidic acid, phosphatidylglycerol and cardiolipin in BBN mitochondria, counteracted by increased phosphatidylcholine levels. Besides the impact on membrane fluidity, this phospholipid remodeling seems to justify, at least in part, the lower oxidative phosphorylation activity observed in mitochondria from wasted muscle and their increased susceptibility to apoptosis. Curiously, no evidences of lipid peroxidation were observed but proteins from BBN mitochondria, particularly the metabolic ones, seem more prone to carbonylation with the consequent implications in mitochondria functionality. Overall, data suggest that bladder cancer negatively impacts skeletal muscle activity specifically by affecting mitochondrial phospholipid dynamics and its interaction with proteins, ultimately leading to the dysfunction of this organelle. The regulation of phospholipid biosynthetic pathways might be seen as potential therapeutic targets for the management of cancer-related muscle wasting.

Assessment of the exposure to heavy metals in Griffon vultures (Gyps fulvus) from the Iberian Peninsula.

Griffon vultures (Gyps fulvus), by virtue of their position at the top of the food chain and as obligate scavengers, are at risk of accumulating and concentrating heavy metals in their tissues and may be more predisposed to their toxic effects. The aim of this study is to investigate heavy metal concentrations in Griffon vultures in Portugal and Catalonia, Spain and to determine if heavy metal concentrations in the blood of weak and/or injured Griffon vultures admitted to wildlife rehabilitation centres (WRC) reflect contamination profiles in the local, free-living and outwardly healthy population. Whole-blood samples taken from 121 Griffon vultures caught in the wild or admitted to WRC in Portugal and Catalonia, Spain were examined for cadmium (Cd), mercury (Hg) and lead (Pb) by inductively coupled plasma mass spectrometry (ICP-MS). Cd and Hg were not detected in most samples (98.3% and 95%, respectively), while Pb was detected in all birds in concentrations ranging between 4.97 and 300.23 µg/dl. Birds admitted to WRC had significantly lower Pb concentrations (24.15 ± 15.07 and 25.98 ± 18.04 µg/dl in Portugal and Catalonia, Spain, respectively) than animals caught in the wild (29.67 ± 13.19 and 42.22 ± 50.08 µg/dl in Portugal and Catalonia, Spain, respectively) (p<0.05). This may be explained by the fact that malnutrition was the main cause of admission of Griffon vultures to WRC, as ingestion has been described as the most significant pathway for Pb exposure in raptors. Therefore Griffon vultures admitted to WRC do not seem to be representative of the local, free-flying populations, so it remains necessary to continue catching when one intends to monitor Pb exposure in this species. The population of vultures captured in Catalonia, Spain showed the highest mean blood Pb concentration, perhaps due to the municipal rubbish dump located near the feeding station, with rubbish providing a significant fraction of their trophic needs. The ingestion of game meat with bullet fragments in carcasses or with Pb shots embedded in their flesh could also be the cause of the high blood Pb concentrations found in some vultures. The potential risk of Pb exposure in Griffon vulture populations must be given consideration, since most individuals evaluated had Pb concentrations between 20 and 100µg/dl, which is considered to be subclinical exposure to Pb, and which is above the threshold level at which Pb can affect antioxidant system in this species (15 µg/dl).

Bisphosphonates induce the osteogenic gene expression in co-cultured human endothelial and mesenchymal stem cells.

Bisphosphonates (BPs) are known to affect bone homeostasis and also to have anti-angiogenic properties. Because of the intimate relationship between angiogenesis and osteogenesis, this study analysed the effects of Alendronate (AL) and Zoledronate (ZL) in the expression of endothelial and osteogenic genes on interacting endothelial and mesenchymal stem cells, an issue that was not previously addressed. Alendronate and ZL, 10(-12) -10(-6) M, were evaluated in a direct co-culture system of human dermal microvascular endothelial cells (HDMEC) and human bone marrow mesenchymal stem cells (HMSC), over a period of 14 days. Experiments with the respective monocultures were run in parallel. Alendronate and ZL caused an initial dose-dependent stimulation in the cell proliferation in the monocultures and co-cultures, and did not interfere with their cellular organization. In HDMEC monocultures, the expression of the endothelial genes CD31, VE-cadherin and VEGFR2 was down-regulated by AL and ZL. In HMSC monocultures, the BPs inhibited VEGF expression, but up-regulated the expression of the osteogenic genes alkaline phosphatase (ALP), bone morphogenic protein-2 (BMP-2) and osteocalcin (OC) and, to a greater extent, osteoprotegerin (OPG), a negative regulator of the osteoclastic differentiation, and increased ALP activity. In co-cultured HDMEC/HMSC, AL and ZL decreased the expression of endothelial genes but elicited an earlier and sustained overexpression of ALP, BMP-2, OC and OPG, compared with the monocultured cells; they also induced ALP activity. This study showed for the first time that AL and ZL greatly induced the osteogenic gene expression on interacting endothelial and mesenchymal stem cells.

Biomonitoring of heavy metals (Cd, Hg, and Pb) and metalloid (As) with the Portuguese common buzzard (Buteo buteo).

The accumulation of heavy metals in the environment may have a wide range of health effects on animals and humans. Thus, in this study, the concentrations of arsenic (As), cadmium (Cd), lead (Pb), and mercury (Hg) in the blood and tissues (liver and kidney) of Portuguese common buzzards (Buteo buteo) were determined by inductively coupled plasma-mass spectrometer (ICP-MS) in order to monitor environmental pollution to these elements. In general, Hg and As were the elements which appeared in the highest and lowest concentrations, respectively. A highest percentage of non-detected concentration was found for blood Cd (94.6 %) but, in turn, it was the only metal that was detected in all kidney samples. The kidney was the analyzed sample which showed the highest concentrations of each element evaluated. Statistically, significant differences among blood, liver, and kidney samples were observed for As and Cd (P < 0.05). Cd concentrations in kidney and liver varied significantly with age: Adults showed higher hepatic and renal Cd concentrations than juveniles. Blood Pb concentration seems to show an association with the hunting season. Although raptors are at the top of the food chain and are thus potentially exposed to any biomagnification processes that may occur in a food web, the individuals evaluated in this study generally had low levels of heavy metals in blood and tissues. However, chronic exposure to these metals was verified. The results presented here lend weight to arguments in favor of continuous biomonitoring of metals and metalloids, since heavy metals may accumulate to levels that will pose a risk to both human health and the environment.