Sex and age differences in AMPK phosphorylation, mitochondrial homeostasis, and inflammation in hearts from inflammatory cardiomyopathy patients.

Linked to exacerbated inflammation, myocarditis is a cardiovascular disease, which may lead to dilated cardiomyopathy. Although sex and age differences in the development of chronic myocarditis have been postulated, underlying cellular mechanisms remain poorly understood. In the current study, we aimed to investigate sex and age differences in mitochondrial homeostasis, inflammation, and cellular senescence. Cardiac tissue samples from younger and older patients with inflammatory dilated cardiomyopathy (DCMI) were used. The expression of Sirt1, phosphorylated AMPK, PGC-1α, Sirt3, acetylated SOD2, catalase, and several mitochondrial genes was analyzed to assess mitochondrial homeostasis. The expression of NF-κB, TLR4, and interleukins was used to examine the inflammatory state in the heart. Finally, several senescence markers and telomere length were investigated. Cardiac AMPK expression and phosphorylation were significantly elevated in male DCMI patients, whereas Sirt1 expression remained unchanged in all groups investigated. AMPK upregulation was accompanied by a preserved expression of all mitochondrial proteins/genes investigated in older male DCMI patients, whereas the expression of TOM40, TIM23, and the mitochondrial oxidative phosphorylation genes was significantly reduced in older female patients. Mitochondrial homeostasis in older male patients was further supported by the reduced acetylation of mitochondrial proteins as indicated by acetylated SOD2. The inflammatory markers NF-κB and TLR4 were downregulated in older male DCMI patients, whereas the expression of IL-18 was increased in older female patients. This was accompanied by progressed senescence in older DCMI hearts. In conclusion, older women experience more dramatic immunometabolic disorders on the cellular level than older men.

The impact of the PCSK-9/VLDL-Receptor axis on inflammatory cell polarization.

BACKGROUND: Studies have shown that lipoproteins, such as LDL and VLDL, as well as its major protein component ApoE2 impact on macrophage polarization important in atherosclerosis. Proprotein convertase subtilisin/kexin 9 (PCSK9) is a key regulator of lipoprotein receptor expression. The present study investigated the effect of the VLDL/VLDL-receptor (VLDL-R) axis on mononuclear cell polarization, as well as the role of PCSK9 and PCSK9 inhibitors (PCSK9i) within this network. METHODS: Human monocytic THP-1 cells and human monocyte-derived macrophages isolated from peripheral blood mononuclear cells (PBMC) were treated with either LPS/IFN-γ to induce a pro-inflammatory phenotype, or with IL-4/IL-13 to induce an anti-inflammatory phenotype. Cells were then subjected to further treatments by lipoproteins, PCSK9, PCSK9i and lipoprotein receptor blockers. RESULTS: LPS/IFN-γ treatment promoted a pro-inflammatory state with an increased expression of pro-inflammatory mediators such as TNF-α, CD80 and IL-1ß. VLDL co-treatment induced a switch of this pro-inflammatory phenotype to an anti-inflammatory phenotype. In pro-inflammatory cells, VLDL significantly decreased the expression of pro-inflammatory markers e.g., TNF-α, CD80, and IL-1ß. These effects were eliminated by PCSK9 and restored by co-incubation with a specific anti-PCSK9 monoclonal antibody (PCSK9i). Migration assays demonstrated that pro-inflammatory cells displayed a significantly higher invasive capacity when compared to untreated cells or anti-inflammatory cells. Moreover, pro-inflammatory cell chemotaxis was significantly decreased by VLDL-mediated acquisition of the anti-inflammatory phenotype. PCSK9 significantly lessened this VLDL-mediated migration inhibition, which was reversed by the PCSK9i. CONCLUSION: VLDL promotes mononuclear cell differentiation towards an anti-inflammatory phenotype. PCSK9, via its capacity to inhibit VLDL-R expression, reverses the VLDL-mediated anti-inflammatory action, thereby promoting a pro-inflammatory phenotype. Thus, PCSK9 targeting therapies may exert anti-inflammatory properties within the vessel wall.

Cardiovascular Inflammaging: Mechanisms and Translational Aspects.

Aging is one of the major non-reversible risk factors for several chronic diseases, including cancer, type 2 diabetes, dementia, and cardiovascular diseases (CVD), and it is a key cause of multimorbidity, disability, and frailty (decreased physical activity, fatigue, and weight loss). The underlying cellular mechanisms are complex and consist of multifactorial processes, such as telomere shortening, chronic low-grade inflammation, oxidative stress, mitochondrial dysfunction, accumulation of senescent cells, and reduced autophagy. In this review, we focused on the molecular mechanisms and translational aspects of cardiovascular aging-related inflammation, i.e., inflammaging.

Male Macrophages and Fibroblasts from C57/BL6J Mice Are More Susceptible to Inflammatory Stimuli.

Mounting evidence argues for the significant impact of sex in numerous cardiac pathologies, including myocarditis. Macrophage polarization and activation of cardiac fibroblasts play a key role in myocardial inflammation and remodeling. However, the role of sex in these processes is still poorly understood. In this study, we investigated sex-specific alterations in the polarization of murine bone marrow-derived macrophages (BMMs) and the polarization-related changes in fibroblast activation. Cultured male and female murine BMMs from C57/BL6J mice were polarized into M1 (LPS) and M2 (IL-4/IL-13) macrophages. Furthermore, male and female cardiac fibroblasts from C57/BL6J mice were activated with TNF-α, TGF-ß, or conditioned medium from M1 BMMs. We found a significant overexpression of M1 markers (c-fos, NFκB, TNF-α, and IL-1ß) and M2 markers (MCP-1 and YM1) in male but not female activated macrophages. In addition, the ROS levels were higher in M1 male BMMs, indicating a stronger polarization. Similarly, the pro-fibrotic markers TGF-ß and IL-1ß were expressed in activated cardiac male fibroblasts at a significantly higher level than in female fibroblasts. In conclusion, the present study provides strong evidence for the male-specific polarization of BMMs and activation of cardiac fibroblasts in an inflammatory environment. The data show an increased inflammatory response and tissue remodeling in male mice.

Sex-Specific Differences of the Inflammatory State in Experimental Autoimmune Myocarditis.

Increasing evidence suggests male sex as a potential risk factor for a higher incidence of cardiac fibrosis, stronger cardiac inflammation, and dilated cardiomyopathy (DCM) in human myocarditis. Chronic activation of the immune response in myocarditis may trigger autoimmunity. The experimental autoimmune myocarditis (EAM) model has been well established for the study of autoimmune myocarditis, however the role of sex in this pathology has not been fully explored. In this study, we investigated sex differences in the inflammatory response in the EAM model. We analyzed the cardiac function, as well as the inflammatory stage and fibrosis formation in the heart of EAM male and female rats. 21 days after induction of EAM, male EAM rats showed a decreased ejection fraction, stroke volume and cardiac output, while females did not. A significantly elevated number of infiltrates was detected in myocardium in both sexes, indicating the activation of macrophages following EAM induction. The level of anti-inflammatory macrophages (CD68+ ArgI+) was only significantly increased in female hearts. The expression of Col3A1 and fibrosis formation were more prominent in males. Furthermore, prominent pro-inflammatory factors were increased only in male rats. These findings indicate sex-specific alterations in the inflammatory stage of EAM, with a pro-inflammatory phenotype appearing in males and an anti-inflammatory phenotype in females, which both significantly affect cardiac function in autoimmune myocarditis.

Dose Escalation and Safety of Capsaicin for Cerebral Perfusion Augmentation: A Pilot Study.

Background and Purpose: Sphenopalatine ganglion (SPG) electrical stimulation has been studied in the setting of acute ischemic stroke to enhance collateral flow. Capsaicin poses an alternative to chemically stimulate the sphenopalatine ganglion. Therefore, the objective of this study was to determine the safety and effect of increasing doses of capsaicin upon serial transcranial Doppler markers of cerebral blood flow. Methods: We performed serial transcranial Doppler testing in 30 healthy volunteers divided into 5 equal groups. Capsaicin doses ranged from 33 to 165 µMol. We recorded peak systolic and end-diastolic velocities in the middle cerebral artery, arterial pressure, and perceived pungency in 5-minute intervals up to 20 minutes. We then calculated the mean velocity, the pulsatility index, and the cerebral blood flow index. Results: The participants' median age was 21 years (range, 5 years); all reported consumption of capsaicin in their diets. After and during the study, none reported side effects. Perceived pungency peaked at 5 minutes, and by the 20-minute mark, none perceived any pungency. All the tested doses produced the same pattern, consisting of augmentation of the middle cerebral artery mean velocity with the pulsatility index's diminution. The effects peaked between the 5- and the 10-minute measurements and then returned to basal levels except for the 66-µMol doses, which produced a sustained effect. We found no correlation between perceived pungency and dose, but the middle cerebral artery mean velocity was strongly correlated with the dose administered. Conclusions: This study provides evidence supporting the safety and tolerability of oral capsaicin in a population of healthy volunteers. Capsaicin appears to produce effects similar to those of sphenopalatine ganglion electrical stimulation. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT04545892.

Conversion from Duodenal Switch to Single Anastomosis Duodenal Switch to Deal with Postoperative Malnutrition.

BACKGROUND: Duodenal switch (DS) is considered one of the most effective bariatric techniques for long-term weight and comorbidity control. After these operations, some patients may get severe complications related to malnutrition and a few of them may need surgical revision. Lengthening the common channel (CC) is usually the solution: changing the Roux anastomosis or with a side-to-side anastomosis (kissing X). We propose that when simplified construction of the DS is used, conversion to single anastomosis DS (SADI-S/OADS) is an easy and safe choice. OBJECTIVES: To evaluate the safety and effectiveness of conversion from DS to SADI-S in cases of malnutrition. METHODS: We report three patients with severe malnutrition after a DS at 9, 74, and 84 months. One of them had also liver failure related to alcohol abuse and malnutrition. Laparoscopic reoperations included a new ileo-ileal anastomosis and takedown of the Roux-en-Y anastomosis with the aim of lengthening the CC. RESULTS: All three patients were successfully converted by laparoscopy. After a median follow-up of 54.6 months [32-76 months], all of them had moderate weight regain and returned to normal biochemical nutritional parameters. Two patients with type 2 diabetes (T2DM) before DS had complete remission before conversion; one of them had recurrence of T2DM after conversion. The patient with liver failure improved significantly after conversion. CONCLUSIONS: Conversion from DS to SADI-S/OADS is a simple operation with excellent results in resolving malnutrition in those patients. However, weight regain and recurrence of comorbidities may arise.

Dilated cardiomyopathy impairs mitochondrial biogenesis and promotes inflammation in an age- and sex-dependent manner.

Dilated cardiomyopathy (DCM) belongs to the myocardial diseases associated with a severe impairment of cardiac function, but the question of how sex and age affect this pathology has not been fully explored. Impaired energy homeostasis, mitochondrial dysfunction, and systemic inflammation are well-described phenomena associated with aging. In this study, we investigated if DCM affects these phenomena in a sex- and age-related manner. We analyzed the expression of mitochondrial and antioxidant proteins and the inflammatory state in DCM heart tissue from younger and older women and men. A significant downregulation of Sirt1 expression was detected in older DCM patients. Sex-related differences were observed in the phosphorylation of AMPK that only appeared in older males with DCM, possibly due to an alternative Sirt1 regulation mechanism. Furthermore, reduced expression of several mitochondrial proteins (TOM40, TIM23, Sirt3, and SOD2) and genes (cox1, nd4) was only detected in old DCM patients, suggesting that age has a greater effect than DCM on these alterations. Finally, an increased expression of inflammatory markers in older, failing hearts, with a stronger pro-inflammatory response in men, was observed. Together, these findings indicate that age- and sex-related increased inflammation and disturbance of mitochondrial homeostasis occurs in male individuals with DCM.

Quantitative comparison of taxa and taxon concepts in the diatom genus Fragilariopsis: a case study on using slide scanning, multiexpert image annotation, and image analysis in taxonomy1.

Semiautomated methods for microscopic image acquisition, image analysis, and taxonomic identification have repeatedly received attention in diatom analysis. Less well studied is the question whether and how such methods might prove useful for clarifying the delimitation of species that are difficult to separate for human taxonomists. To try to answer this question, three very similar Fragilariopsis species endemic to the Southern Ocean were targeted in this study: F. obliquecostata, F. ritscheri, and F. sublinearis. A set of 501 extended focus depth specimen images were obtained using a standardized, semiautomated microscopic procedure. Twelve diatomists independently identified these specimen images in order to reconcile taxonomic opinions and agree upon a taxonomic gold standard. Using image analyses, we then extracted morphometric features representing taxonomic characters of the target taxa. The discriminating ability of individual morphometric features was tested visually and statistically, and multivariate classification experiments were performed to test the agreement of the quantitatively defined taxa assignments with expert consensus opinion. Beyond an updated differential diagnosis of the studied taxa, our study also shows that automated imaging and image analysis procedures for diatoms are coming close to reaching a broad applicability for routine use.

A shift in the biogenic silica of sediment in the Larsen B continental shelf, off the Eastern Antarctic Peninsula, resulting from climate change.

In 2002, section B of the Larsen ice shelf, off of the Eastern Antarctic Peninsula, collapsed and created the opportunity to study whether the changes at the sea surface left evidence in the sedimentary record. Biogenic silica is major constituent of Antarctic marine sediment, and its presence in the sediment column is associated with diatom production in the euphotic zone. The abundance of diatom valves and the number of sponge spicules in the biogenic silica was analyzed to determine how the origin of the biogenic silica in the upper layers of the sediment column responded to recent environmental changes. Diatom valves were present only in the upper 2 cm of sediment, which roughly corresponds to the period after the collapse of the ice shelf. In contrast, sponge spicules, a more robust form of biogenic silica, were also found below the upper 2 cm layer of the sediment column. Our results indicate that in this region most of the biogenic silica in the sedimentary record originated from sponge spicules rather than diatoms during the time when the sea surface was covered by the Larsen ice shelf. Since the collapse of the ice shelf, the development of phytoplankton blooms and the consequent influx of diatom debris to the seabed have shifted the biogenic silica record to one dominated by diatom debris, as occurs in most of the Antarctic marine sediment. This shift provides further evidence of the anthropogenic changes to the benthic habitats of the Antarctic and will improve the interpretation of the sedimentary record in Polar Regions where these events occur.

Intraneural catheterization of the sciatic nerve in humans: a pilot study.

BACKGROUND AND OBJECTIVES: Imaging studies in humans have shown that intraneural injection of local anesthetic may be relatively frequent. The incidence of intraneural catheterization is unknown. We speculated that early neural blockade after the injection of a small dose of local anesthetic might be a common finding produced by unintended intraneural catheterization. We investigated the clinical effect of a small dose of local anesthetic injected through a stimulating sciatic catheter, and also the placement of the tip of the catheters by computed tomography (CT) scan. METHODS: In this descriptive study, a dose of 3 to 5 mL of 1.5% mepivacaine was injected through a stimulating sciatic catheter in 45 patients undergoing hallux valgus repair. Patients with early neural blockade at the foot, within 5 minutes after injection, were suspected to have an intraneural placement of the catheter. A CT scan of the thighs was performed in 10 additional patients to assess catheter tip placement. RESULTS: Nine of 45 patients had early neural blockade with the small dose of local anesthetic. Seven patients underwent surgery without further supplementation. There was a statistically significant lower intensity of stimulating current through the catheter in the group of patients with early neural blockade. Of 10 CT scans, 3 showed clear intraneural placement of the catheter. No postoperative sequelae were observed. CONCLUSIONS: Intraneural catheterization of the sciatic nerve may be a frequent finding and was not followed by nerve injury under the conditions of our small pilot study.

A comparison of stimulation patterns in axillary block: part 2.

BACKGROUND AND OBJECTIVES: Radial plus musculocutaneous nerve stimulation may have a predominant role in the success of an axillary block, producing more extensive anesthesia of the upper limb than median plus musculocutaneous nerve stimulation. However, no comparison has been made with ulnar plus musculocutaneous nerve stimulation. We compared the extent of both sensory and motor block after ulnar plus musculocutaneous nerve stimulation or radial plus musculocutaneous nerve stimulation. METHODS: Sixty patients were randomly assigned to receive an axillary block using either radial plus musculocutaneous or ulnar plus musculocutaneous nerve stimulation with 40 mL plain 1.5% mepivacaine. Patients were assessed for sensory block by the pinprick method at 5 and 20 minutes. RESULTS: No statistically significant differences were found in the rates of anesthesia at 20 minutes in the cutaneous nerve distributions of the upper limb between radial plus musculocutaneous and ulnar plus musculocutaneous nerve stimulation except for the following nerves: radial (90% and 63.3%, respectively), medial cutaneous of the forearm (83.3% and 100%, respectively), and medial cutaneous of the arm (73.3% and 93.3%, respectively). Global sensory score (minimum: 0; maximum: 12 points) at 20 minutes was significantly higher after radial plus musculocutaneous than after ulnar plus musculocutaneous nerve stimulation: 12 (11-13) and 11 (10-12), respectively. The rates of median nerve blockade were 50% and 53%, respectively. CONCLUSIONS: Radial plus musculocutaneous nerve stimulation produced more extensive anesthesia of the upper limb than did ulnar plus musculocutaneous nerve stimulation. However, there is not an optimal combination of 2 responses in axillary brachial plexus block.

Stimulating popliteal catheters for postoperative analgesia after hallux valgus repair.

Stimulating catheters have been recently introduced in clinical practice. We assessed the efficacy of stimulating and nonstimulating catheter placement for pain control and local anesthetic requirements after hallux valgus repair with continuous sciatic popliteal nerve block in this comparative, randomized, blinded-to-observer study of 48 patients. A stimulating catheter was placed in groups S-125 and S-0625. The same catheter was inserted without stimulation in group NS-125. An infusion of 0.125% levobupivacaine was given in groups S-125 and NS-125, whereas 0.0625% levobupivacaine was used in group S-0625. All patients received an infusion of the test drug at a basal rate of 3 mL/h, with the possibility of an additional bolus of 3 mL every hour. Verbal analog scale (VAS) scores for pain were assessed between 6-8 h and between 19-23 h postoperatively. Multiple attempts were required for catheter insertion in all patients in groups S-125 and S-0625. Lower median (range) VAS scores for pain (0-100 points) were found in group S-125 at 6-8 h postoperatively when compared with groups S-0625 and NS-125: 5 (0-17.5) versus 60 (15-80) and 70 (25-80), respectively (P < 0.05); and lower VAS scores for pain were also found in group S-125 at 19-23 h when compared with group NS-125: 0 (0-0) and 7.5 (0-10), respectively (P < 0.05). Fewer patients required IV opioid analgesia in group S-125 than in groups S-0625 and NS-125: 0, 5, and 7 patients, respectively (P < 0.05). We conclude that efficacy in pain control was increased with stimulating catheter placement.

Horner syndrome after unintended subdural block. A report of 2 cases.

Horner syndrome is considered a common finding after epidural analgesia or anesthesia in obstetric patients. Conversely, Horner syndrome is very uncommon in nonobstetric patients. We report 2 cases of Horner syndrome after attempted epidural lumbar anesthesia in 2 patients undergoing peripheral vascular surgery. Spinal fluoroscopy with contrast medium showed subdural catheterization in both cases. Horner syndrome after lumbar epidural anesthesia can be an indicator of inadvertent subdural catheter placement.

A comparison of four stimulation patterns in axillary block.

BACKGROUND AND OBJECTIVES: Insufficient spread of the local anesthetic toward the retroarterial region of the neurovascular space may be responsible for inconsistent anesthesia of the upper limb after single-injection axillary block. We hypothesized that injection of the local anesthetic on a single radial-nerve stimulation would produce the same extent of anesthesia as either a single median-nerve stimulation, a double-stimulation technique (radial and musculocutaneous nerves), or a triple-stimulation technique (radial, musculocutaneous, and median nerves). METHODS: One hundred twenty patients were randomly assigned to receive an axillary block by either median-nerve, radial-nerve, radial-nerve plus musculocutaneous-nerve, or triple-nerve stimulation with 40 mL of plain 1.5% mepivacaine. Patients were assessed for sensory block by the pinprick method at 5 and 20 minutes. RESULTS: Radial-nerve stimulation produced more extensive anesthesia than did median-nerve stimulation. The rate of anesthesia at 20 minutes in the median-nerve cutaneous distribution was similar after median-nerve stimulation or radial-nerve stimulation. The ulnar nerve was more frequently blocked at 20 minutes after radial-nerve stimulation than after median-nerve stimulation. Extent of anesthesia at 20 minutes after radial-nerve plus musculocutaneous-nerve stimulation was similar to that produced by triple-nerve stimulation, except for lower rates of anesthesia that corresponded to the median nerve. All of the differences were statistically significant. CONCLUSIONS: Musculocutaneous-nerve stimulation and radial-nerve stimulation play predominant roles in the success of axillary brachial plexus block, although a triple-nerve stimulation technique is still required to produce complete anesthesia of the upper limb.

Median versus musculocutaneous nerve response with single-injection infraclavicular coracoid block.

BACKGROUND AND OBJECTIVES: Local anesthetic injection after elicitation of a distal motor response with a nerve stimulator is believed to produce a more clinically efficient infraclavicular coracoid block than after elicitation of a proximal motor response. The aim of this study was to investigate whether elicitation of a median or of a musculocutaneous-type nerve response influenced the quality of anesthesia. METHODS: Randomized, prospective, single-blind study. One hundred thirty patients received a coracoid block with 40 mL plain mepivacaine 1.5% after stimulation of median nerve fibers (group 1) or musculocutaneous nerve fibers (group 2). Patients were assessed for sensory and motor block at 5 and 20 minutes. RESULTS: Significantly higher rates of complete anesthesia at 20 minutes were found in the cutaneous distributions of the radial and ulnar nerves in group 1. Significantly higher rates of complete paralysis were found for elbow extension, wrist flexion, and finger and thumb movements in group 1 at 20 minutes. Differences in the extent of anesthesia and paralysis were more remarkable at 5 minutes than at 20 minutes. CONCLUSIONS: Elicitation of a median nerve response improved the efficacy of infraclavicular coracoid block when compared with a musculocutaneous nerve response. Complete paralysis and complete anesthesia of the upper limb were low in both groups.

Restricted infraclavicular distribution of the local anesthetic solution after infraclavicular brachial plexus block.

BACKGROUND AND OBJECTIVES: The distribution of local anesthetic after different approaches for brachial plexus anesthesia could be responsible for the varying rates of side effects, such as phrenic block, hoarseness, and Horner's syndrome associated with each approach. We compared the distribution of local anesthetic within the neurovascular space in infraclavicular block with that of interscalene and supraclavicular block. METHODS: In a prospective analysis using fluoroscopy, we studied the distribution of a solution of local anesthetic containing radiologic contrast medium in 18 patients. Six patients received an interscalene block, another 6 patients received a perpendicular supraclavicular block, and another 6 patients, a perpendicular coracoid block. RESULTS: Distribution of the anesthetic solution in the interscalene and supraclavicular groups extended to both supraclavicular and infraclavicular spaces in all patients. This distribution was significantly different (P <.05) compared with that of the infraclavicular group. In this group, the solution remained below the clavicle in every patient. CONCLUSIONS: Spread of the local anesthetic from the infraclavicular space after infraclavicular coracoid block appears to be limited to below the level of the clavicle. Conversely, local anesthetic solution passes below the clavicle in all patients given interscalene or supraclavicular blocks.