The traceability of meat origin has become a necessity to safeguard consumer's confidence in commercial meat products. Our study recommends a meat species detection using qualitative approach based ...on PCR–RFLP and species-specific primers PCR. Here, we targeted a 359 bp fragment of the mitochondrial cytochrome b gene amplified by PCR using universal primers followed by three enzymatic digestions. Seven animal species including dromedary, rabbit, goat, turkey, rat, donkey and pork, have been efficiently detected in pure and mixed samples. The combination of PCR–RFLP and triplex PCR assays offers, in addition, the identification of chicken, dog and cat species in meat. In conclusion, by the mean of PCR-based techniques using universal primers followed by enzymatic digestion and multiplex primer-specific approach, we developed an extensible protocol by which we identified 10 animal species that could be integrated in meat analysis daily routine.
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CEKLJ, EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
The 14-kilodalton human growth hormone (14 kDa hGH) N-terminal fragment derived from the proteolytic cleavage of its full-length counterpart has been shown to sustain antiangiogenic potentials. This ...study investigated the antitumoral and antimetastatic effects of 14 kDa hGH on B16-F10 murine melanoma cells. B16-F10 murine melanoma cells transfected with 14 kDa hGH expression vectors showed a significant reduction in cellular proliferation and migration associated with an increase in cell apoptosis in vitro. In vivo, 14 kDa hGH mitigated tumor growth and metastasis of B16-F10 cells and was associated with a significant reduction in tumor angiogenesis. Similarly, 14 kDa hGH expression reduced human brain microvascular endothelial (HBME) cell proliferation, migration, and tube formation abilities and triggered apoptosis in vitro. The antiangiogenic effects of 14 kDa hGH on HBME cells were abolished when we stably downregulated plasminogen activator inhibitor-1 (PAI-1) expression in vitro. In this study, we showed the potential anticancer role of 14 kDa hGH, its ability to inhibit primary tumor growth and metastasis establishment, and the possible involvement of PAI-1 in promoting its antiangiogenic effects. Therefore, these results suggest that the 14 kDa hGH fragment can be used as a therapeutic molecule to inhibit angiogenesis and cancer progression.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Purpose
Congenital hypogonadotropic hypogonadism (CHH) is a rare genetic disorder mostly characterized by gonadotropins release and/or action deficiencies. Both isolated (idiopathic hypogonadotropic ...hypogonadism) and syndromic (Kallmann) forms are identified depending on the olfactory ability. Clinical and genetic heterogeneities of CHH have been widely explored, thus improving our understanding of the disease’s pathophysiology. This work aims to (1) provide a detailed clinical and hormonal description of normosmic CHH patients and (2) identify the mutation linked to the studied phenotype.
Participants and methods
We investigated three affected patients with normosmic CHH, belonging to a consanguineous Tunisian family. Patients underwent an insulin-induced hypoglycemia test. We performed whole exome sequencing to identify the causal mutation.
Results
At first diagnosis, a total gonadotropic deficiency was identified in all patients. The insulin-induced hypoglycemia test has also revealed a reduced cortisol secretion and complete growth hormone deficiency. At 20.8 years, one female exhibited a spontaneous recovery of the hypothalamic–pituitary–adrenal axis function, unlike her affected siblings who still depend on corticosteroid replacement therapy. Herein, we identified a novel homozygous nonstop mutation (c.1195T>C) in
KISS1R
gene in all affected subjects. This mutation led to the substitution of the physiologic stop codon by an arginine (p.X399R).
Conclusions
Our study highlights the importance of the KISS1R signaling, in gonadotropin-releasing hormone neurons, in the control of reproductive function. Additionally, our data suggests a complex central and peripheral metabolic control of puberty, through the hypothalamic KISS1R signaling. We suggest a mutual link between the hypothalamic–pituitary–gonadal, –adrenal, and –somatotropic axes.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Pemphigus foliaceus (PF) is a bullous autoimmune skin disease diagnosed through sera and skin analyses. PF severity is associated with maintained anti‐Dsg1 sera levels and its prognosis is ...unpredictable. MicroRNA (miRNA), dynamic regulators of immune function, have been identified as potential biomarkers for some autoimmune diseases. This study aimed to assess the miRNA expression of miR‐17‐5p, miR‐21‐5p, miR‐146a‐5p, miR‐155‐5p and miR‐338‐3p using quantitative real‐time PCR in peripheral blood mononuclear cells (PBMC) and lesional skin samples from untreated and treated PF patients (both remittent and chronic) over 3 months. Overall, miRNA expression was significantly higher in PBMC than in biopsy samples. Blood miR‐21 expression was increased in untreated patients compared to controls and had a diagnostic value with an AUC of 0.78. After 6 weeks, it decreased significantly, similar to anti‐Dsg1 antibodies and the PDAI score. In addition, a positive correlation was observed between cutaneous miR‐21 expression and the disease activity score. Conversely, cutaneous expressions of miR‐17, miR‐146a and miR‐155 were significantly higher in treated chronic patients compared to remittent ones. The cutaneous level of miR‐155 positively correlated with pemphigus activity, making it a potential predictive marker for patients' clinical stratification with an AUC of 0.86.These findings suggest that blood miR‐21 and cutaneous miR‐155 can be used as supplemental markers for PF diagnosis and activity, respectively in addition to classical parameters.
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BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Fighting food industry standard violations has become an important stake especially in developing countries. In this work, different common DNA extraction protocols, namely NaOH, Phenol Chloroform ...Isoamyl alcohol, CTAB, Chloroform and the commercialized DNeasy blood and tissue kit were tested and compared for turkey salami genomic authentication in mitochondrial genes species detection by Polymerase Chain Reaction (PCR). The obtained results proved that a protocol based on NaOH after an advanced grinding step was the most adequate alternative in terms of simplicity, rapidity and detection threshold even in swabs. The selected protocol encourages wide range cost-effective routine authentications.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Background: Non-syndromic combined pituitary hormone deficiency (CPHD) occurs due to defects in transcription factors that govern early pituitary development and the specification of ...hormone-producing cells. The most common mutations are in the Prophet of Pit-1 (ProP1) gene. This work aims to (1) report findings of genetic analyses of Tunisian patients with non-syndromic CPHD and (2) describe their phenotype patterns and their evolution through life. Methods: Fifteen patients from twelve unrelated families with variable clinical phenotypes were included after excluding autoimmune and acquired forms of non-syndromic CPHD. Detailed pedigree charts and auxological, hormonal, radiological, and therapeutic details were recorded. Sanger sequencing was performed, and sequences were analyzed with a specific focus on coding and splice site regions of the ProP1 gene. Retained variants were classified using several in silico pathogenicity prediction tools and the VarSome platform. Results: We identified the common p.Arg73Cys mutation in seven patients from four unrelated pedigrees. We found a novel homozygous mutation (c.340C>T) in one sporadic case. This mutation generates a truncated ProP1 protein, predicted to be non-functional, lacking the last 112 codons (p.(Gln114Ter)). We confirmed by polymerase chain reaction (PCR) the absence of large exon deletions or insertions in the remaining sporadic patients (7/8). Conclusions: We report two mutations {one newly identified p.(Gln114Ter) and one previously reported (p.Arg73Cys)} in five unrelated Tunisian families with non-syndromic CPHD. This work is of clinical importance as it reports the high frequency of the p.Arg73Cys mutation in Tunisian CPHD families. Our study also illuminated the involvement of novel gene(s) in the emergence of non-syndromic CPHD.
Determination of pork DNA is essential to avoid the serious allergens imposed by it as well as for halal authentication for certain community. The needed sensor should be able to identify the pork ...DNA in rapid way and sensitive to ultra-low molarity detection. In this work we developed a surface enhanced Raman spectroscopy sensor (SERS) using silver nanoparticles (AgNPs) on Si matrix. A deep investigation of the effect of the laser wavelength on the sensing efficiency has been performed. Wavelength dependent sensing mechanisms are deliberately studied in the current research. Further the selectivity of sensor towards Pork DNA discrimination is exhibited with Chicken DNA analysis. The developed SERS sensor identifies the Pork DNA down to 13.6 fg/ μL proving its excellent sensitivity. The statistical outfit studies with regard to the developed sensor shows the low values of RSD (∼3%) adding advantage of the developed SERS sensor biosensor. The SERS sensor will definitely serve to be an excellent fast, selective and sensitive tool for onsite screening of meat products to cease the adulteration.
•SERS Biosensor to target pork adulterant in food products was developed.•Si/AgNPS is employed to amplify the EM enhancement mechanism for sensing.•Ultra-low detectability is achieved with sensing of pork DNA (13.6 fg/ μL).•Selectivity is illustrated with chicken DNA analysis.•Mixture of analysis with Pork and chicken DNA is exhibited.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
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•Highly-sensitive SERS sensor based on AgNPr/SiNP hybrid arrays was fabricated.•Simple low-cost production of spherical Ag nanoprisms and Ag nanoparticles.•AgNPr/SiNP hybrid arrays ...exhibit substantial amplification of fingerprint bands of R6G compared to flat bulk silicon.•Detection of human genomic DNA with a lower detection limit of 1.5 pg/µL.
In the present work, we have fabricated silver nanoprism (AgNPrs)/silicon nanoparticle (SiNPs) hybrid arrays for highly sensitive detection of biomolecules via surface-enhanced Raman spectroscopy (SERS) technique. SiNPs having 7 to 37 nm in size and with phosphorous doping varying from 1 × 1019 to 1 × 1020 cm−3 were synthesized in nonthermal plasma synthesis. SiNPs were further immobilized on glass substrates using spin-coating, followed by deposition of AgNPrs using the drop-casting method. SERS studies showed that AgNPrs/SiNPs hybrid arrays exhibit substantial amplification of fingerprint bands of rhodamine 6G (R6G) compared to bare silicon as the reference. Raman signal intensity was found to be dependent on the size of SiNPs, with the largest nanoparticles exhibiting the highest SERS enhancement. In addition, an increase in phosphorous doping concentration was found to reduce R6G peak intensities. AgNPrs/SiNPs hybrid arrays showed excellent stability over time and high spot-to-spot reproducibility as well. Moreover, hybrid arrays enabled DNA detection through intense vibrational modes of human genomic DNA, with a lower detection limit of 1.5 pg/µL; indicating that AgNPrs/SiNPs sensors can serve as a reliable and cost-effective biosensing platform for rapid and label-free analysis of biomolecules.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Retraction: ‘Genotyping of Tunisian azoospermic men with Sertoli cell‐only and maturation arrest‘ by Hadjkacem‐Loukil, L., Hadj‐kacem, H., Hadj Salem, I., Bahloul, A., Fakhfakh, F. and Ayadi, H.
The ...above article, published online on 6 July 2011 in Wiley Online Library (www.onlinelibrary.wiley.com), has been retracted by agreement between the journal Editor‐in‐Chief, Ralf Henkel, and Blackwell Verlag GmbH. The retraction has been agreed due to unverifiable data and authorship issues.
Reference
Hadjkacem‐Loukil, L., Hadj‐kacem, H., Hadj Salem, I., Bahloul, A., Fakhfakh, F. and Ayadi, H. (2011), Genotyping of Tunisian azoospermic men with Sertoli cell‐only and maturation arrest. Andrologia. doi:10.1111/j.1439‐0272.2010.01088.x
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DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Mesenchymal stem cells (MSCs) isolated from bone marrow have different developmental origins, including neural crest. MSCs can differentiate into neural progenitor-like cells (NPCs) under the ...influence of bFGF and EGF. NPCs can terminally differentiate into neurons that express beta-III-tubulin and elicit action potential. The main aim of the study was to identify key genetic markers involved in differentiation of MSCs into NPCs through transcriptomic analysis.
Total RNA was isolated from MSCs and MSCs-derived NPCs followed by cDNA library construction for transcriptomic analysis. Sample libraries that passed the quality and quantity assessments were subjected to high throughput mRNA sequencing using NextSeq®500. Differential gene expression analysis was performed using the DESeq2 R package with MSC samples being a reference group. The expression of eight differentially regulated genes was counter validated using real-time PCR.
In total, of the 3,252 differentially regulated genes between MSCs and NPCs with two or more folds, 1,771 were upregulated genes, whereas 1,481 were downregulated in NPCs. Amongst these differential genes, 104 transcription factors were upregulated, and 45 were downregulated in NPCs. Neurogenesis related genes were upregulated in NPCs and the main non-redundant gene ontology (GO) terms enriched in NPCs were the autonomic nervous system, cell surface receptor signalling pathways), extracellular structure organisation, and programmed cell death. The main non-redundant GO terms enriched in MSCs included cytoskeleton organisation cytoskeleton structural constituent, mitotic cell cycle), and the mitotic cell cycle process Gene set enrichment analysis also confirmed cell cycle regulated pathways as well as Biocarta integrin pathway were upregulated in MSCs. Transcription factors enrichment analysis by ChEA3 revealed Foxs1 and HEYL, amongst the top five transcription factors, inhibits and enhances, respectively, the NPCs differentiation of MSCs.
The vast differences in the transcriptomic profiles between NPCs and MSCs revealed a set of markers that can identify the differentiation stage of NPCs as well as provide new targets to enhance MSCs differentiation into NPCs.
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