Clinical utility of urine kidney injury molecule-1 (KIM-1) and gamma-glutamyl transferase (GGT) in the diagnosis of canine acute kidney injury

The aim of the present study was to evaluate the sensitivity and specificity of urine KIM-1 and urine GGT for the detection of naturally-occurring AKI, compared to healthy control dogs, dogs with stable chronic kidney disease (CKD), and dogs with lower urinary tract disorders (LUTD). The study included AKI grade 1 (n = 21), AKI grade 2 to 5 (n = 11), stable CKD (n = 11), LUTD (n = 15), and healthy dogs (n = 37). Urine KIM-1 (ng/mg) and GGT (U/l) were normalized to urine creatinine (uCr). Statistically significant difference in KIM/uCr (p = 0.0007) and GGT/uCr (p < 0.0001) was found among the study groups. Area under the curve (AUC) for KIM-1/uCr and GGT/uCr as predictors of AKI was 0.81 and 0.91 respectively. Values of KIM-1/uCr of 0.73 ng/mg and of GGT/uCr of 54.33 showed the best combination of sensitivity and specificity (75% and 75.6%; 85.7% and 89.1% respectively). A significant positive correlation (p < 0.0001) between KIM-1/uCr and GGT/uCr was found. Both urine KIM-1/uCr and GGT/uCr seemed to be potentially good markers for the diagnosis of AKI. Dogs with AKI showed significantly higher levels of urine KIM-1/uCr and urine GGT/uCr, compared with healthy dogs. Caution should be used in the evaluation of elevated urine KIM-1/uCr and GGT/uCr in dogs with pre-existing CKD and/or LUTD. Urine KIM-1/uCr and GGT/uCr might have a significant clinical utility, as complementary test, particularly in diagnosis early, non-azotemic stages of AKI

Social Preference Tests in Zebrafish: A Systematic Review

The use of animal models in biology research continues to be necessary for the development of new technologies and medicines, and therefore crucial for enhancing human and animal health. In this context, the need to ensure the compliance of research with the principles Replacement, Reduction and Refinement (the 3 Rs), which underpin the ethical and human approach to husbandry and experimental design, has become a central issue. The zebrafish (Danio rerio) is becoming a widely used model in the field of behavioral neuroscience. In particular, studying zebrafish social preference, by observing how an individual fish interacts with conspecifics, may offer insights into several neuropsychiatric and neurodevelopmental disorders. The main aim of this review is to summarize principal factors affecting zebrafish behavior during social preference tests. We identified three categories of social research using zebrafish: studies carried out in untreated wild-type zebrafish, in pharmacologically treated wild-type zebrafish, and in genetically engineered fish. We suggest guidelines for standardizing social preference testing in the zebrafish model. The main advances gleaned from zebrafish social behavior testing are discussed, together with the relevance of this method to scientific research, including the study of behavioral disorders in humans. The authors stress the importance of adopting an ethical approach that considers the welfare of animals involved in experimental procedures. Ensuring a high standard of animal welfare is not only good for the animals, but also enhances the quality of our science

Deregulation of MiR-34b/Sox2 Predicts Prostate Cancer Progression.

Most men diagnosed with prostate cancer will have an indolent and curable disease, whereas approximately 15% of these patients will rapidly progress to a castrate-resistant and metastatic stage with high morbidity and mortality. Therefore, the identification of molecular signature(s) that detect men at risk of progressing disease remains a pressing and still unmet need for these patients. Here, we used an integrated discovery platform combining prostate cancer cell lines, a Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) model and clinically-annotated human tissue samples to identify loss of expression of microRNA-34b as consistently associated with prostate cancer relapse. Mechanistically, this was associated with epigenetics silencing of the MIR34B/C locus and increased DNA copy number loss, selectively in androgen-dependent prostate cancer. In turn, loss of miR-34b resulted in downstream deregulation and overexpression of the stemness marker, Sox2. These findings identify loss of miR-34b as a robust biomarker for prostate cancer progression in androgen-sensitive tumors, and anticipate a potential role of progenitor/stem cell signaling in this stage of disease

Biomimetic graphene for enhanced interaction with the external membrane of astrocytes

Graphene and graphene substrates display huge potential as material interfaces for devices and biomedical tools targeting the modulation or recovery of brain functionality. However, to be considered reliable neural interfaces, graphene-derived substrates should properly interact with astrocytes, favoring their growth and avoiding adverse gliotic reactions. Indeed, astrocytes are the most abundant cells in the human brain and they have a crucial physiological role to maintain its homeostasis and modulate synaptic transmission. In this work, we describe a new strategy based on the chemical modification of graphene oxide (GO) with a synthetic phospholipid (PL) to improve interaction of GO with brain astroglial cells. The PL moieties were grafted on GO sheets through polymeric brushes obtained by atom-transfer radical-polymerization (ATRP) between acryloyl-modified PL and GO nanosheets modified with a bromide initiator. The adhesion of primary rat cortical astrocytes on GO–PL substrates increased by about three times with respect to that on glass substrates coated with standard adhesion agents (i.e. poly-D-lysine, PDL) as well as with respect to that on non-functionalized GO. Moreover, we show that astrocytes seeded on GO–PL did not display significant gliotic reactivity, indicating that the material interface did not cause a detrimental inflammatory reaction when interacting with astroglial cells. Our results indicate that the reported biomimetic approach could be applied to neural prosthesis to improve cell colonization and avoid glial scar formation in brain implants. Additionally, improved adhesion could be extremely relevant in devices targeting neural cell sensing/modulation of physiological activity

Valutazione di marcatori urinari nel cane: KIM-1 e GGT

Parole chiave: cane, danno renale, biomarker urinari, KIM-1, GGT. Obiettivo: valutazione del KIM-1 urinario e delle GGT urinario nel cane come marcatori precoci di danno renale, valutandone principalmente la loro sensibilità e specificità nel discriminare soggetti sani da pazienti nefropatici (AKI, CKD) o con patologie delle basse vie urinarie (LUTD). Materiali e metodi:Inclusione di soggetti sani con esame clinico obiettivo generale negativo e in buono stato di salute; pazienti AKI con insufficienza renale acuta divisi in base alla classificazione IRIS, pazienti CKD secondo classificazione IRIS, pazienti LUTD che presentano esame urine indice di patologie delle basse vie, sintomatologia e/o rilevanza ecografica; Di tutti i soggetti sono stati presi campioni di urina per minzione spontanea, cateterismo o cistocentesi sui quali è stato effettuato l'esame delle urine completo con lettura del sedimento e valutazione della uGGT e uKIM-1 urinari. Risultati: in questo studio sono stati inclusi 95 soggetti, di cui 37 sani, 21 AKI.1, 11 AKI 2-5, 11 CKD, 15 LUTD. È risultata una differenza statisticamente significativa (p=0.0007) tra i valori mediani di KIM-1/Cu urinario tra pazienti sani, AKI-1, AKI-2-5, CKD e LUTD. Tale differenza è risultata particolarmente significativa tra soggetti sani ed AKI-1 (p=0.0013); Tra i valori mediani di GGT/Cu urinario tra pazienti sani, è risultata una differenza statisticamente significativa (p<0,0001) tra AKI-1, AKI-2-5, CKD e LUTD. Tale differenza è risultata particolarmente significativa tra soggetti sani ed AKI-1 (p<0,0001); sani e AKI-2-5 (p=0,0385); sani e CKD (p=0,0450); AKI-1 e LUTD (p=0,0042); l'elaborazione della ROC curve per il valore di KIM-1/Cu dei soggetti sani e AKI-1 ha evidenziato una differenza significativa (p<0,0001). La ROC curve elaborata per il valore delle GGT/Cu dei soggetti inclusi come sani (n=37) e AKI 1 (n=21) ha dimostrato una differenza significativa (p<0,0001). Conclusioni: la valutazione dei due marcatori urinari (GGT e KIM-1) nei pazienti affetti da patologie renale/urinarie, ha evidenziato dei risultati promettenti in quanto entrambi i marcatori sembrano avere un forte potere diagnostico nel discriminare il paziente sano da quello affetto da AKI ad uno stadio iniziale (non azotemico) ma non per discriminare le forme azotemiche acute da quelle croniche. Sebbene siano necessari ulteriori studi e una casistica maggiore per rafforzare tale conclusione, c’è da dire che la metodica per ottenere KIM-1 risulta assai più indaginosa e costosa rispetto a quella utilizzata per la determinazione delle GGT urinarie. Non necessariamente però, deve essere scelto un marcatore piuttosto che un altro, infatti, è già risaputa l’importanza di associare più marcatori per migliorare la diagnosi, la classificazione e la prognosi di AKI. Inoltre, sebbene questo sia il primo studio che cerca di determinare un cut off del KIM-1 nel soggetto sano, in quanto ancora non riportato in medicina veterinaria, ulteriori studi sono comunque necessari, soprattutto per approfondire la valutazione di questo nuovo marker emergente e per incrementare la casistica in medicina veterinaria, ad oggi ancora povera di questi dati. Key words: dog, kidney injury, urinary biomarkers, KIM-1, GGT. Objective: evaluation of Urinary KIM-1 urinary and urinary GGT in dogs as early markers of renal damage, primarily evaluating their sensitivity and specificity in discriminating healthy subjects from nephropathic patients (AKI, CKD) or low urinary tract pathologies (LUTD). Materials and methods: inclusion of healthy subjects with clinical examination overall negative and good health; AKI patients with acute renal impairment divided by IRIS classification, CKD patients according to IRIS classification, LUTD patients with urine screening index of low pathology, symptomatology and / or ultrasound relevance; Spontaneous, catheterized or cystocentric urine samples were taken from all subjects on which full urine examination was performed with sediment reading and UGGT and uKIM-1 urinary assessment. Results: In this study, 95 subjects were included, including 37 healthy, 21 AKI.1, 11 AKI 2-5, 11 CKD, 15 LUTD. There was a statistically significant difference (p = 0.0007) between urinary KIM-1 / Cu median values between healthy patients, AKI-1, AKI-2-5, CKD and LUTD. This difference was particularly significant among healthy subjects and AKI-1 (p = 0.0013); Among median GGT / Cu urinary values among healthy patients, there was a statistically significant difference (p <0.0001) between AKI-1, AKI-2-5, CKD and LUTD. This difference was particularly significant between healthy subjects and AKI-1 (p <0.0001); healthy and AKI-2-5 (p = 0.0385); healthy and CKD (p = 0.0450); AKI-1 and LUTD (p = 0.0042); the development of curve ROC for the KIM-1 / Cu value of healthy subjects and AKI-1 showed a significant difference (p <0.0001). The ROC curves processed for the GGT / Cu value of subjects included as healthy (n = 37) and AKI 1 (n = 21) showed significant differences (p <0.0001). Conclusion: The evaluation of two urinary markers (GGT and KIM-1) in patients with renal / urinary tract disorders has shown promising results as both markers seem to have a strong diagnostic power in discriminating healthy patients with those affected by AKI at an initial stage (non-azotemic) but not to discriminate acute and chronic forms of azotemic forms. Though further studies and a more casual case are needed to reinforce this conclusion, it is to be said that the method for obtaining KIM-1 is much more prudent and expensive than that used to determine urinary GGT. Not necessarily, however, a marker should be chosen rather than another, in fact, it is already recognized the importance of associating multiple markers to improve AKI's diagnosis, classification and prognosis. In addition, although this is the first study attempting to determine a cut-off of KIM-1 in the healthy subject, as it is not yet reported in veterinary medicine, further studies are still needed, especially in order to deepen the evaluation of this emerging marker and to increase the casework in veterinary medicine, which is still poor in these data

Determination of Prolactin in Canine Saliva: Is it Possible to Use a Commercial ELISA kit?

Prolactin has been reported to be a remarkable index of stress response, both acute and chronic, in several species. The use of biological matrixes other than blood is receiving increasing interest in the study of hormones, due to the lower invasiveness in collection. This research aimed to investigate the possibility of using a commercial ELISA (enzyme-linked immunosorbent assay) kit for measuring canine prolactin in blood for the quantification of canine prolactin in saliva. Study 1 consisted of a validation protocol, using saliva samples collected from lactating and non-lactating dogs. Study 2 was conducted to investigate a possible correlation between prolactin concentration in saliva and plasma in sheltered dogs by using the same kit. Prolactin values were reliably read only when they came from blood samples, not from saliva, but tended to be low in most of the cases. Study 1 showed that saliva had a matrix effect. In study 2, saliva prolactin levels were low and in 42.9% of cases, not readable. No correlation between prolactin values in plasma and saliva was found (ρ=0.482; p=0.274). These findings suggested that the determination of prolactin in dog saliva through an ELISA kit created for measuring prolactin in dog blood was unreliable

Poly(Alkylene 2,5-Thiophenedicarboxylate) Polyesters: A New Class of Bio-Based High-Performance Polymers for Sustainable Packaging

In the present study, 100% bio-based polyesters of 2,5-thiophenedicarboxylic acid were synthesized via two-stage melt polycondensation using glycols containing 3 to 6 methylene groups. The so-prepared samples were characterised from the molecular point of view and processed into free-standing thin films. Afterward, both the purified powders and the films were subjected to structural and thermal characterisation. In the case of thin films, mechanical response and barrier properties to O2 and CO2 were also evaluated. From the results obtained, it emerged that the length of glycolic sub-units is an effective tool to modulate the chain mobility and, in turn, the kind and amount of ordered phases developed in the samples. In addition to the usual amorphous and 3D crystalline phases, in all the samples investigated it was possible to evidence a further phase characterised by a lower degree of order (mesophase) than the crystalline one, whose amount is strictly related to the glycol sub-unit length. The relative fraction of all these phases is responsible for the different mechanical and barrier performances. Last, but not least, a comparison between thiophene-based homopolymers and their furan-based homologues was carried out

Oxytocin Receptor Gene Polymorphism in Lactating Dogs

Genetic variations in the oxytocinergic system, known to regulate social behavior throughout the evolution of mammals, are believed to account for differences in mammalian social behavior. Particularly, polymorphic variants of the oxytocin receptor (OXTR) gene have been associated with behavioral variations in both humans and dogs. In this study, we offered evidence of the correlation between levels of salivary oxytocin (sOXT), maternal behavior and a single-nucleotide gene variant in OXTR (rs8679684) in nineteen lactating Labrador Retriever dogs. Carriers of at least one copy of the minor A allele showed higher levels of sOXT and maternal care in comparison with the homozygous T allele carriers. Considering the relevance of mother care in newborn development, these findings could help us to better understand the possible impact of variants in the OXTR gene in selecting dams

Poly(propylene 2,5-thiophenedicarboxylate) vs. Poly(propylene 2,5-furandicarboxylate): Two Examples of High Gas Barrier Bio-Based Polyesters

Both academia and industry are currently devoting many efforts to develop high gas barrier bioplastics as substitutes of traditional fossil-based polymers. In this view, this contribution presents a new biobased aromatic polyester, i.e., poly(propylene 2,5-thiophenedicarboxylate) (PPTF), which has been compared with the furan-based counterpart (PPF). Both biopolyesters have been characterized from the molecular, thermo-mechanical and structural points of view. Gas permeability behavior has been evaluated with respect to 100% oxygen, carbon dioxide and nitrogen at 23 &deg;C. In case of CO2 gas test, gas transmission rate has been also measured at different temperatures. The permeability behavior at different relative humidity has been investigated for both biopolyesters, the thiophen-containing sample demonstrating to be better than the furan-containing counterpart. PPF&rsquo;s permeability behavior became worse than PPTF&rsquo;s with increasing RH, due to the more polar nature of the furan ring. Both biopolyesters under study are characterized by superior gas barrier performances with respect to PEF and PET. With the simple synthetic strategy adopted, the exceptional barrier properties render these new biobased polyesters interesting alternatives in the world of green and sustainable packaging materials. The different polarity and stability of heterocyclic rings was revealed to be an efficient tool to tailor the ability of crystallization, which in turn affects mechanical and barrier performances