In situ Reprogramming as a Pro-Angiogenic Inducer to Rescue Ischemic Tissues

Background: Enhanced regenerative therapeutic strategies are required to treat intractable ischemic heart disease. Summary: Since the discovery of putative endothelial progenitor cells (EPCs) in 1997, many studies have focused on their extraction, ex vivo processing, and autotransplantation under ischemic conditions. Nonetheless, numerous randomized clinical trials involving thousands of patients have yielded only marginal treatment effects, highlighting the need for advances regarding insufficient dosage and complex ex vivo processing. The prevailing paradigm of cellular differentiation highlights the potential of direct cellular reprogramming, which paves the way for in situ reprogramming. In situ reprogramming holds the promise of significantly enhancing current therapeutic strategies, yet its success hinges on the precise targeting of candidate cells for reprogramming. In this context, the spleen emerges as a pivotal “in situ reprogramming hub,” owing to its dual function as both a principal site for nanoparticle distribution and a significant reservoir of putative EPCs. The in situ reprogramming of splenic EPCs offers a potential solution to overcome critical challenges, including the aforementioned insufficient dosage and complex ex vivo processing. Key Messages: This review explores the latest advancements in EPC therapy and in situ reprogramming, spotlighting a pioneering study that integrates those two strategies with a specific focus on the spleen. Such an innovative approach will potentially herald a new era of regenerative therapy for ischemic heart disease

Influence of welding passes on grain orientation - The example of a multi-pass V-weld

International audienceThe accurate modelling of grain orientations in a weld is important, when accurate ultrasonic test predictions of a welded assembly are needed. To achieve this objective, Electricité de France (EDF) and the Laboratoire de Caractérisation Non Destructive (LCND) have developed a dedicated code, which makes use of information recorded in the welding procedure. Among the welding parameters recorded, although the order in which the welding passes are made is of primary importance in the welding process, this information is not always well known or accurately described. In the present paper we analyse in greater detail the influence of the order of welding passes, using data obtained from the Centre for Advanced Non Destructive Evaluation (CANDE), derived from a dissimilar metal weld (DMW) with buttering. Comparisons are made using grain orientation measurements on a macrograph

Arterial Injury Associated with Tension-Free Vaginal Tapes-SECUR Procedure Successfully Treated by Radiological Embolization

Various postoperative complications have been reported after the use of tension-free vaginal tapes (TVT). The transobturator approach was introduced to minimize the potential complications. The next generation of recently introduced TVT-SECUR is intended to minimize the incidence of complications. Herein we report a case of internal pudendal artery injury sustained during this procedure that was successfully treated by radiological embolization. Angiography with vessel embolization, when available, should be considered when the arterial injury is suspected

Fluid-filled Giant Bulla Treated with Percutaneous Drainage and Talc Sclerotherapy: A Modified Brompton Technique

A 75-year-old man who was diagnosed as having a fluid-filled giant bulla was treated with a modified Brompton technique due to his poor performance status. Percutaneous drainage, suction, and talc sclerotherapy through a Foley catheter can be good treatment options for patients with conditions that are too poor to allow surgical intervention, especially if there is adhesion between a giant bulla and parietal pleura. Talc can also be used safely when mixed with normal saline as a sclerosant

A Surgically Treated Case of Chronic Necrotizing Aspergillosis with Pleural Invasion

Aspergillus is a ubiquitous fungus and can cause many levels of disease severity. Chronic necrotizing aspergillosis is a rare disease and few cases have been reported in Korea. We experienced a case of pleural aspergillosis that was treated successfully with medical and surgical interventions. The 52-year-old man who was diagnosed with chronic necrotizing pulmonary aspergillosis underwent surgical treatment including a lobectomy, decortication, and myoplasty. The patient was also medically treated with amphotericin B followed by voriconazole. Pleural irrigation with amphotericin B was also performed. A multi-dimensional approach should be considered for treating chronic necrotizing pulmonary aspergillosis

Directing lineage specification of human mesenchymal stem cells by decoupling electrical stimulation and physical patterning on unmodified graphene

The organization and composition of the extracellular matrix (ECM) have been shown to impact the propagation of electrical signals in multiple tissue types. To date, many studies with electroactive biomaterial substrates have relied upon passive electrical stimulation of the ionic media to affect cell behavior. However, development of cell culture systems in which stimulation can be directly applied to the material – thereby isolating the signal to the cell-material interface and cell–cell contracts – would provide a more physiologically-relevant paradigm for investigating how electrical cues modulate lineage-specific stem cell differentiation. In the present study, we have employed unmodified, directly-stimulated, (un)patterned graphene as a cell culture substrate to investigate how extrinsic electrical cycling influences the differentiation of naïve human mesenchymal stem cells (hMSCs) without the bias of exogenous biochemicals. We first demonstrated that cyclic stimulation does not deteriorate the cell culture media or result in cytotoxic pH, which are critical experiments for correct interpretation of changes in cell behavior. We then measured how the expression of osteogenic and neurogenic lineage-specific markers were altered simply by exposure to electrical stimulation and/or physical patterns. Expression of the early osteogenic transcription factor RUNX2 was increased by electrical stimulation on all graphene substrates, but the mature marker osteopontin was only modulated when stimulation was combined with physical patterns. In contrast, the expression of the neurogenic markers MAP2 and β3-tubulin were enhanced in all electrical stimulation conditions, and were less responsive to the presence of patterns. These data indicate that specific combinations of non- biological inputs – material type, electrical stimulation, physical patterns – can regulate hMSC lineage specification. This study represents a substantial step in understanding how the interplay of electrophysical stimuli regulate stem cell behavior and helps to clarify the potential for graphene substrates in tissue engineering applications

The Association of Maximum Body Weight on the Development of Type 2 Diabetes and Microvascular Complications: MAXWEL Study

Background: Obesity precedes the development of type 2 diabetes (T2D). However, the relationship between the magnitude and rate of weight gain to T2D development and complications, especially in non-White populations, has received less attention. Methods and Findings: We determined the association of rate and magnitude of weight gain to age at T2D diagnosis (AgeT2D), HbA1c at T2D diagnosis (HbA1cT2D), microalbuminuria, and diabetic retinopathy after adjusting for sex, BMI at age 20 years, lifestyles, family history of T2D and/or blood pressure and lipids in 2164 Korean subjects aged ≥30 years and newly diagnosed with diabetes. Body weight at age 20 years (Wt20y) was obtained by recall or from participants’ medical, school, or military records. Participants recalled their maximum weight (Wtmax) prior to T2D diagnosis and age at maximum weight (Agemax_wt). The rate of weight gain (Ratemax_wt) was calculated from magnitude of weight gain (ΔWt = Wtmax–Wt20y) divided by ΔTime (Agemax_wt –20 years). The mean Agemax_wt and AgeT2D were 41.5±10.9 years and 50.1±10.5 years, respectively. The Wt20y and Wtmax were 59.9±10.5 kg and 72.9±11.4 kg, respectively. The Ratemax_wt was 0.56±0.50 kg/year. After adjusting for risk factors, greater ΔWt and higher Ratemax_wt were significantly associated with earlier AgeT2D, higher HbA1cT2D after additional adjusting for AgeT2D, and microalbuminuria after further adjusting for HbA1cT2D and lipid profiles. Greater ΔWt and higher Ratemax_wt were also significantly associated with diabetic retinopathy. Conclusions: This finding supports public health recommendations to reduce the risk of T2D and its complications by preventing weight gain from early adulthood

MG-63 Cell Proliferation with Static or Dynamic Compressive Stimulation on an Auxetic PLGA Scaffold

The effect of dynamic compressive stimulation on MG-63 cell proliferation on an auxetic PLGA scaffold was investigated. The estimated Poisson ratio of the prepared auxetic scaffold specimens was approximately (−)0.07, while the Poisson ratio estimated for conventional scaffold specimens was (+)0.12 under 10% strain compression on average. Three stimulus groups were examined: control (no stimulation), static compression, and dynamic compression. In preparation for proliferation testing, cells were seeded at 2.2 × 105 cells/80 μL on each scaffold specimen. The average proliferation rates of the static and dynamic groups were higher than those of the control group: 13.4% and 25.5% higher at culture day 1, 34.7% and 56.2% at culture day 3, and 17.5% and 43.0% at culture day 5, respectively. The static and dynamic group results at culture day 5 were significantly different (p<0.01). Moreover, proliferation rate of the dynamic stimulation group was 1.22 times higher than that of the static group (p<0.01). Conclusively, proliferation of osteoblast-like cells was enhanced through compressive stimulation, but the enhancement was maximal with dynamic compressive stimulation of auxetic scaffolds

Nitric oxide induces MUC5AC mucin in respiratory epithelial cells through PKC and ERK dependent pathways

BACKGROUND: Nitric oxide (NO) is generally increased during inflammatory airway diseases. This increased NO stimulates the secretion of mucin from the goblet cell and submucosal glands but the mechanism is still unknown precisely. In this study, we investigated potential signaling pathways involving protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) in the NO-induced MUC5AC mucin gene and protein expression in A549 cells. METHODS: Nitric oxide was donated to the A549 cells by NOR-1. MUC5AC mucin levels were assayed by enzyme-linked immunosorbent assay (ELISA). MUC5AC promoter activity was determined by measuring luciferase activity after the lysing the transfected cells. Activation of PKC isoforms were measured by assessing the distribution of the enzyme between cytosolic and membrane fractions using immunoblotting. Immunoblotting experiments using a monoclonal antibody specific to PKC isoforms were performed in the cytosol and membrane fractions from A549 cells. Western blot analysis for pERK and p38 were performed using the corresponding antibodies from the cell lysates after donating NO to the A549 cells by NOR-1. RESULTS: The transcriptional activity of MUC5AC promoter was maximal at the concentration of 0.1 mM NOR-1 for 1 hour incubation in transfected A549 cells. (±)-(E)-methyl-2-((E)-hydroxyimino)-5-nitro-6-methoxy-3-hexenamide (NOR-1) markedly displaced the protein kinase C (PKC)α and PKCδ from the cytosol to the membrane. Furthermore, the PKC-α,βinhibitors, GÖ6976 (10 nM) and PKCδ inhibitors, rottlerin (4 μM) inhibited the NOR-1 induced migration of PKCα and PKCδ respectively. NOR-1 also markedly increased the MUC5AC promoter activity and mRNA expression, mucin synthesis and ERK1/2 phosphorylation. The PKC inhibitors also inhibited the NOR-1 induced MUC5AC mRNA and MUC5AC protein synthesis by inhibiting the activation of PKCα and PKCδ with ERK1/2 pathways. CONCLUSION: Exogenous NO induced the MUC5AC mucin gene and protein through the PKCα and PKCδ – ERK pathways in A549 cells. Inhibition of PKC attenuated NO-mediated MUC5AC mucin synthesis. In view of this findings, PKC inhibitors might be useful in the treatment of bronchial asthma and chronic bronchitis patients where NO and mucus are increased in the bronchial airways

Anti-atherosclerotic vaccination against Porphyromonas gingivalis as a potential comparator of statin in mice

Background Porphyromonas gingivalis (Pg) is an oral anaerobe which damages teeth and periodontal tissues. Its body infection is known to cause chronic inflammation, thereby inducing an early stage of atherosclerosis through humoral immune actions. Hence, vaccination by immunizing the proteins of P. gingivalis (Pg) post sonication with heating may prevent atherosclerosis. This study aimed to compare the effect of its vaccination with statin, which effectively prevents atherosclerosis by lowering lipids. Methods The vaccine was produced by sonicating P. gingivalis through heating, and a total of 32 male APOE-/-mice (8-week old) were subjected Western diet for 8 weeks, in order to induce atherosclerosis in a physiological manner. Then, the mice were grouped to undergo four treatment conditions (i.e., no treatment, pitavastatin, vaccine, or pitavastatin with vaccine). Vaccination was conducted through nasal immunization and confirmed by a Pg-specific humoral immune reaction. Then, half of the mice in each group were orally injected with P. gingivalis for the next 5 weeks while the other half remained uninfected, generating a total of eight groups (n = 4/group). The mice were sacrificed at 3 weeks after the last injection. After harvesting the aorta, Oil Red O staining of en face was conducted with imaging and image analysis, and plaque formation was quantitatively determined. Results Compared to no treatment, the vaccination through nasal immunization significantly reduced the atherosclerotic plaque sizes in APOE -/- mice under Western diet to the comparable level of statin group. When both vaccine and statin were used, no clear synergistic effect was observed as opposed to expectation. Conclusions This study revealed that nasal immunization of heat shock P. gingivalis has a significant impact on the prevention of arteriosclerosis and acts as a potential comparator of statin