Transfection of hypoxia-inducible factor-1α mRNA upregulates the expression of genes encoding angiogenic growth factors.

Hypoxia-Inducible Factor-1α (HIF-1α) has presented a new direction for ischemic preconditioning of surgical flaps to promote their survival. In a previous study, we demonstrated the effectiveness of HIF-1a DNA plasmids in this application. In this study, to avoid complications associated with plasmid use, we sought to express HIF-1α through mRNA transfection and determine its biological activity by measuring the upregulation of downstream angiogenic genes. We transfected six different HIF-1a mRNAs-one predominant, three variant, and two novel mutant isoforms-into primary human dermal fibroblasts using Lipofectamine, and assessed mRNA levels using RT-qPCR. At all time points examined after transfection (3, 6, and 10 h), the levels of HIF-1α transcript were significantly higher in all HIF-1α transfected cells relative to the control (all p < 0.05, unpaired Student's T-test). Importantly, the expression of HIF-1α transcription response genes (VEGF, ANG-1, PGF, FLT1, and EDN1) was significantly higher in the cells transfected with all isoforms than with the control at six and/or ten hours post-transfection. All isoforms were transfected successfully into human fibroblast cells, resulting in the rapid upregulation of all five downstream angiogenic targets tested. These findings support the potential use of HIF-1α mRNA for protecting ischemic dermal flaps.

Induced Pluripotent Stem Cell-Derived Extracellular Vesicles Promote Wound Repair in a Diabetic Mouse Model via an Anti-Inflammatory Immunomodulatory Mechanism.

Extracellular vesicles (EVs) derived from mesenchymal stem/stromal cells (MSCs) have recently been explored in clinical trials for treatment of diseases with complex pathophysiologies. However, production of MSC EVs is currently hampered by donor-specific characteristics and limited ex vivo expansion capabilities before decreased potency, thus restricting their potential as a scalable and reproducible therapeutic. Induced pluripotent stem cells (iPSCs) represent a self-renewing source for obtaining differentiated iPSC-derived MSCs (iMSCs), circumventing both scalability and donor variability concerns for therapeutic EV production. Thus, it is initially sought to evaluate the therapeutic potential of iMSC EVs. Interestingly, while utilizing undifferentiated iPSC EVs as a control, it is found that their vascularization bioactivity is similar and their anti-inflammatory bioactivity is superior to donor-matched iMSC EVs in cell-based assays. To supplement this initial in vitro bioactivity screen, a diabetic wound healing mouse model where both the pro-vascularization and anti-inflammatory activity of these EVs would be beneficial is employed. In this in vivo model, iPSC EVs more effectively mediate inflammation resolution within the wound bed. Combined with the lack of additional differentiation steps required for iMSC generation, these results support the use of undifferentiated iPSCs as a source for therapeutic EV production with respect to both scalability and efficacy.

Induced pluripotent stem cell-derived extracellular vesicles promote wound repair in a diabetic mouse model via an anti-inflammatory immunomodulatory mechanism.

Extracellular vesicles (EVs) derived from mesenchymal stem/stromal cells (MSCs) have recently been widely explored in clinical trials for treatment of diseases with complex pathophysiology. However, production of MSC EVs is currently hampered by donor-specific characteristics and limited ex vivo expansion capabilities before decreased potency, thus restricting their potential as a scalable and reproducible therapeutic. Induced pluripotent stem cells (iPSCs) represent a self-renewing source for obtaining differentiated iPSC-derived MSCs (iMSCs), circumventing both scalability and donor variability concerns for therapeutic EV production. Thus, we initially sought to evaluate the therapeutic potential of iMSC EVs. Interestingly, while utilizing undifferentiated iPSC EVs as a control, we found that their vascularization bioactivity was similar and their anti-inflammatory bioactivity was superior to donor-matched iMSC EVs in cell-based assays. To supplement this initial in vitro bioactivity screen, we employed a diabetic wound healing mouse model where both the pro-vascularization and anti-inflammatory activity of these EVs would be beneficial. In this in vivo model, iPSC EVs more effectively mediated inflammation resolution within the wound bed. Combined with the lack of additional differentiation steps required for iMSC generation, these results support the use of undifferentiated iPSCs as a source for therapeutic EV production with respect to both scalability and efficacy.

Comparison of four formulas for nasotracheal tube length estimation in pediatric patients: an observational study.

BACKGROUND: Correct endotracheal intubation results in better ventilation, prevents hypoxia and its possible damages, such as brain injury, and minimizes attempts for re-intubation. Up to now, several formulas have been published to estimate nasotracheal intubation tube length. This study aims to compare the accuracy of different suggested formulas to find the one that better estimates the tube insertion distance. METHODS: This cross-sectional retrospective study was carried out in 102 (51 female, 51 male) children who underwent cardiac surgery under general anesthesia. Inclusion criteria were correct nasotracheal intubation according to the postintubation chest X-ray (CXR). The estimated tracheal tube length was calculated by four different formulas. Pearson...s correlation coefficient was used to find the correlations between the estimated length of each formula and the correct nasotracheal tube length. Also, linear regression was used to obtain a formula to estimate nasotracheal tube length by weight, height, and age. RESULTS: The formula L=3*tube size+2 had the best correlation with tube length (r ...=...0.81, Confidence Interval: 0.732...0.878, p-value < 0.001). Among demographic variables, height had the highest correlation coefficient with the tube length (r...=...0.83, Confidence Interval: 0.788...0.802, p-value < 0.001). Therefore, considering the height as an independent variable and tube length as a dependent variable, using linear regression, the following formula was achieved for determining tube length: nasotracheal tube length...=...0.1*Height+7. CONCLUSIONS: The formula L=3*tube size+2 and the new suggested formula in this study can be used to estimate nasotracheal tube length in children under 4 years old. However, these formulas are only guides and require confirmation by auscultation and CXR.

Comparison of four formulas for nasotracheal tube length estimation in pediatric patients: an observational study

Abstract Background: Correct endotracheal intubation results in better ventilation, prevents hypoxia and its possible damages, such as brain injury, and minimizes attempts for re-intubation. Up to now, several formulas have been published to estimate nasotracheal intubation tube length. This study aims to compare the accuracy of different suggested formulas to find the one that better estimates the tube insertion distance. Methods: This cross-sectional retrospective study was carried out in 102 (51 female, 51 male) children who underwent cardiac surgery under general anesthesia. Inclusion criteria were correct nasotracheal intubation according to the postintubation chest X-ray (CXR). The estimated tracheal tube length was calculated by four different formulas. Pearson's correlation coefficient was used to find the correlations between the estimated length of each formula and the correct nasotracheal tube length. Also, linear regression was used to obtain a formula to estimate nasotracheal tube length by weight, height, and age. Results: The formula L=3*tube size+2 had the best correlation with tube length (r =0.81, Confidence Interval: 0.732-0.878, p-value < 0.001). Among demographic variables, height had the highest correlation coefficient with the tube length (r = 0.83, Confidence Interval: 0.788-0.802, p-value < 0.001). Therefore, considering the height as an independent variable and tube length as a dependent variable, using linear regression, the following formula was achieved for determining tube length: nasotracheal tube length =0.1*Height+7. Conclusions: The formula L=3*tube size+2 and the new suggested formula in this study can be used to estimate nasotracheal tube length in children under 4 years old. However, these formulas are only guides and require confirmation by auscultation and CXR.

Revolutionary transformation lowering the mortality of pancreaticoduodenectomy: a historical review.

The History Maker paper focuses on the extraordinary revolution that dramatically improved the surgical results for the Whipple procedure (pancreaticoduodenectomy) in the 1980s and identifies Dr. Cameron as the leader of this revolution, who reported a mortality rate of approximately 1%. The revolutionary reduction of postoperative mortality for the Whipple procedure was achieved by adherence to gentle and precise Halstedian surgical techniques with adequate drainage of pancreatico-jejunal anastomosis with closed-suction silastic drains, along with the development of high-volume surgeons and hospitals. Excellent teamwork in patient care, including but not limited to preoperative evaluation by multidisciplinary teams, intraoperative communication between surgeons and anaesthesiologists, and postoperative management, contributed to a successful Whipple procedure.

Efficacy of laparoscopic transversus abdominis plane block on postoperative pain management and surgery side effects in laparoscopic bariatric surgeries.

PURPOSE: Transversus abdominis plane (TAP) block is a new option for reducing postoperative pain. This study investigated the effects of laparoscopic TAP block on postoperative adverse events and analgesia and antiemetics requirements after bariatric surgery. METHODS: In this randomized clinical trial study, patients were randomly divided into control (N = 20) or TAP block (N = 20) groups. In the TAP block group, the block was performed under direct laparoscopic guidance after surgery and before the removal of trocars. RESULTS: Fifteen patients (75%) versus four patients (20%) received opioids within the first 6 h in the control and TAP groups, respectively (p-value < 0.001). The cumulative amount of opioids consumed in the 24 and 48 h after surgery was lower in the TAP group (p-value < 0.001). The visual analog scale (VAS) regarding general and wound-specific pain was significantly lower in the TAP group compared to the control group at 6 and 24 h both at rest and in movement. However, there was no significant difference at 48 h postoperatively. The percentages of patients having postoperative nausea and vomiting (PONV), pruritus, and resumption of bowel movement were not significantly different between the two groups at any time (6, 24, and 48 h) postoperatively. CONCLUSION: Laparoscopic-guided TAP block is a pragmatic, applicable, and minimally invasive regional technique and can be part of effective postoperative pain management in morbidly obese patients undergoing bariatric surgery. Applying it laparoscopically without the need for ultrasound is also useful and effective.

Association of Pro-inflammatory Cytokine Gene Polymorphism with Meniere's Disease in an Iranian Sample.

Meniere's disease (MD) is known as a rare chronic disorder of the inner ear with elevated serum levels of pro-inflammatory cytokines like tumor necrosis factor (TNF)-α, Interleukin (IL)-1, and IL-6. This study aims to evaluate genes polymorphism in some pro-inflammatory cytokines in a group of Iranian MD patients compared to the healthy controls. In this case-control study, 25 MD patients and 139 healthy controls were enrolled. DNA was extracted from blood samples, and single nucleotide polymorphisms were detected using polymerase chain reaction with sequence-specific primers assay. MD patients and controls were examined in terms of allele, genotype, and haplotype frequency of pro-inflammatory cytokine genes. Only the frequencies of alleles A/G at position -238 in the promoter of the TNF-α gene differed significantly between MD patients and healthy controls. G to A allele ratio was 23 and 3.6 in MD and controls, respectively. In individuals with MD, genotype GG was found to be significantly more prevalent at position -238 of the TNF-α gene promoter sequence. In addition, the heterozygote AG variant of -238 A/G TNF-α gene polymorphism was lower in MD patients than controls. Compared to the control group, the haplotype TNF- (-308, -238) AG was higher in MD patients, although not statistically significant. This is the first study that we know of that evaluates the frequencies of pro-inflammatory cytokine genes in an Iranian MD sample. This study shows the association between TNF-α and susceptibility to MD.

Role of hypothalamic-pituitary adrenal-axis, toll-like receptors, and macrophage polarization in pre-atherosclerotic changes induced by social isolation stress in mice.

It has been well documented that chronic stress can induce atherosclerotic changes, however, the underlying mechanisms is yet to be established. In this regard, this study aimed to elucidate the relation between hypothalamic-pituitary adrenal-axis (HPA-axis), toll-like receptors (TLRs), as well as M1/M2 macrophage ratio and pre-atherosclerotic changes in social isolation stress (SIS) in mice. We used small interfering RNA against the glucocorticoid receptor (GR) to evaluate the relation between HPA-axis and TLRs. C57BL/6J mice were subjected to SIS and RT-PCR, ELISA, flow cytometry, and immunohistochemistry were used to assess the relations between pre-atherosclerotic changes and TLRs, macrophage polarization, pro-inflammatory cytokines, and cell adhesion molecules in aortic tissue. We used TAK-242 (0.3 mg/kg, intraperitoneally), a selective antagonist of TLR4, as a possible prophylactic treatment for atherosclerotic changes induced by SIS. We observed that isolated animals had higher serum concentration of corticosterone and higher body weight in comparison to normal animals. In isolated animals, results of in vitro study showed that knocking-down of the GR in bone marrow-derived monocytes significantly decreased the expression of TLR4. In vivo study suggested higher expression of TLR4 on circulating monocytes and higher M1/M2 ratio in aortic samples. Pathological study showed a mild pre-atherosclerotic change in isolated animals. Finally, we observed that treating animals with TAK-242 could significantly inhibit the pre-atherosclerotic changes. SIS can possibly increase the risk of atherosclerosis through inducing abnormal HPA-axis activity and subsequently lead to TLR4 up-regulation, vascular inflammation, high M1/M2 ratio in intima. Thus, TLR4 inhibitors might be a novel treatment to decrease the risk of atherosclerosis induced by chronic stress.

The possible role of nitric oxide in anti-convulsant effects of Naltrindole in seizure-induced by social isolation stress in male mice.

Social isolation stress (SIS) as a chronic model of early-life stress could induce proconvulsant effects in mice. In the current study, we evaluated the role of opioid receptors (OPRs) agonists and antagonists in pro-conversant effects of SIS and the common pathway between delta-opioid receptors (DORs) and nitric oxide (NO) in stress-induced seizure. For reaching to this goal, we used pentylenetetrazol (PTZ) model of clonic-seizure to measure seizure threshold and administrated selective and non-selective OPRs agonists and antagonists in both social condition (SC) and isolated condition (IC) animals. In the next step, we administrated sub effective dose of naltrindole (NLT, 0.3 mg/kg) with sub-effective doses of nitric oxide synthesis (NOS) inhibitors including L-NAME (10 mg/kg), aminoguanidine (50 mg/kg) and 7-NI (15 mg/kg). Also, we co-administrated sub-effective dose of SNC80 (0.5 mg/kg) with sub-effective dose of l-arg (25 mg/kg) to assess the seizure threshold. In addition, we measured nitrite levels of hippocampus following administration of mentioned drugs in both SC and IC mice. Our findings showed that L-NAME and 7-NI (but not AG) increased anti-convulsant activity of NLT and l-arg increased proconvulsant effects of SNC80 in IC animals. Nitrite assay showed that co-administration of NLT plus sub-effective doses of L-NAME and 7-NI (but not AG) decreased and co-administration of SNC80 with sub-effective dose of l-arg increased nitrite levels of hippocampus in IC mice. This study suggests the role of n-NOS in anti-convulsant effects of NLT and pro-convulsant effects of SNC80 in stress-induced seizure.