Background: Achilles-tendon rupture prevails as a common tendon pathology. Adipose-derived mesenchymal stem cells (ADMSCs) are multipotent stem cells derived from adipose tissue with attractive regeneration properties; thus, their application in tendinopathies could be beneficial. Methods: Male rabbit ADMSCs were obtained from the falciform ligament according to previously established methods. After tenotomy and suture of the Achilles tendon, 1 × 106 flow-cytometry-characterized male ADMSCs were injected in four female New Zealand white rabbits in the experimental group (ADMSC group), whereas four rabbits were left untreated (lesion group). Confirmation of ADMSC presence in the injured site after 12 weeks was performed with quantitative sex-determining region Y (SRY)-gene RT-PCR. At Week 12, histochemical analysis was performed to evaluate tissue regeneration along with quantitative RT-PCR of collagen I and collagen III mRNA. Results: Presence of male ADMSCs was confirmed at Week 12. No statistically significant differences were found in the histochemical analysis; however, statistically significant differences between ADMSC and lesion group expression of collagen I and collagen III were evidenced, with 36.6% and 24.1% GAPDH-normalized mean expression, respectively, for collagen I (p < 0.05) and 26.3% and 11.9% GAPDH-normalized mean expression, respectively, for collagen III (p < 0.05). The expression ratio between the ADMSC and lesion group was 1.5 and 2.2 for collagen I and collagen III, respectively. Conclusion: Our results make an important contribution to the understanding and effect of ADMSCs in Achilles-tendon rupture.
<abstract> <p>The pH Low Insertion Peptide (pHLIP) has versatile applications in several diseases due to its differential behavior at slightly different pH values. pHLIP is an unstructured and peripheral membrane-associated peptide at neutral pH and an α-helical transmembrane peptide at acidic values. Similar to what happened to insulin and growth hormone, pHLIP´s expanding applications require high-yield production to further scale-up its usefulness. To date, synthesis of the pHLIP has not been reported in a prokaryotic platform, mainly relying on solid-phase synthesis. Bacterial production arises as an option for high-amount peptide generation and larger pHLIP fusion protein-synthesis; however, cell-based pH-responsive peptide production could be challenging due to intracellular peptide interactions or degradation due to unstructured conformations. An <italic>Escherichia coli</italic> (E. coli)-BL21 cell culture was induced with Isopropyl ß-D-1-thiogalactopyranoside (IPTG) in order to produce a Glutathione S-transferase-pHLIP (GST-pHLIP) fusion construct. Purification was done with Glutathione (GSH)-decorated magnetic beads using 4 ml of the induced cell culture. The production was quantified with Bradford reagent and characterized with SDS-PAGE and Western blot, contrasting Bradford results with densitometry analysis to obtain production approximate absolute values. A purified approximate total yield of ~26 µg with an apparent GSH-bead saturation and a total production of ~82 µg was obtained. Our Western Blot assay confirmed the presence of the GST-pHLIP construct in all the IPTG-induced fractions. Conclusion: A high-yield pHLIP production irrespective of its membrane affinity in acidic environments or its unstructured nature was achieved. Our study could be useful to scale up pHLIP synthesis for future applications.</p> </abstract>
Background: Currently, completed suicide, suicide ideation, suicide behavior, and suicide attempts are major public health problems worldwide. Major Depressive Disorder (MDD) is one of the most common mental disorders associated with an increased risk of suicide. Since the relationship between suicide and cholesterol levels is still controversial, in this study, we explore the association between SNPs rs754203 and rs4900442 of CYP46A1 with suicide risk in Mexican patients with major depressive disorder. Methods: We evaluated 188 unrelated suicide completers and compared them to 144 non-suicidal individuals (controls) and 126 MDD patients. Genotypes were analyzed using the Real Time-polymerase chain reaction method and two allele-specific probes to detect specific SNP targets. A chi-square test was used to identify a possible risk genotype or allele type for suicide. Results: Statistical analysis showed significant differences between completed suicide and controls in their allelic and genotypic frequencies in rs754203 SNP. The genotype G/G of CYP46A1 rs754203 was significantly associated with suicide. Also, the G allele was associated with an increased risk of suicide (OR= 1.370, 95% CI= 1.002-1.873). No differences in either genotype distribution or allele frequencies of CYP46A1 rs4900442 were observed. Conclusions: The results of the current study report the first association between G allele carriers (A/G + G/G) of rs754203 and increased risk for suicide, especially in males.
Chikungunya is a public health problem in Africa, Asia, and Latin America. Innate immune response (IIR) possesses enzymatic cleavage of nucleic acids of the agent as an important mechanism of viral counteraction. Oligoadenylate synthetase (OAS) enzymes are key regulators of RNase L to develop IIR against RNA viruses. OAS´ Single Nucleotide Polymorphisms (SNPs) have been related to susceptibility toward RNA-virus diseases. A cross-sectional study was done of 187 patients. OAS1 SNPs (rs1131454), OAS2 SNPs (rs1293762, rs15895, and rs1732778) and OAS3 SNPs (rs2285932 and rs2072136) genes were studied by qPCR in check and symptomatic patients to associate SNPs with susceptibility to disease. Relative risk (RR), Chi2, and Hardy-Weinberg equilibrium indicated p < 0.05 statistical significance. SNPs rs1131454 and rs1293762 showed statistically significant differences between cases and check. Homozygous genotypes GG of rs1131454 and CC of rs1293762 were considered risk factors (RR 1.59; 95% CI 1.19-2.14 and RR 1.91; 95% CI 1.31-2.78, respectively). Heterozygous genotypes GA of rs1131454 and AC of rs1293762 were protective factors (RR 0.67; 95% CI 0.48-0.94 and RR 0.56; 95% CI 0.37-0.83, respectively). Moreover, rs1293762 was statistically significant in allelic frequencies between cases and checks, with A allele a protective factor (RR = 0.55; 95% CI 0.39-.078) and C allele a susceptibility factor (RR = 1.80; 95% CI difference (p < 0.05) in the allelic frequencies between cases and checks, with A allele as protection factor (RR = 1.28-2.54). Our results add to previously reported evidence of susceptibility of certain populations with specific OAS-family SNPs against CHIKV.
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