Femoral Blood Research Articles

Very bad deep … When the scuba tank smells like smoke

Presentation and discussion of positive tests results for cannabis in a person who died following a diving accident. Scuba diving is a sport that is becoming more and more popular. However, if this sport isn’t practiced properly it can quickly become dangerous. Scuba diving is practiced with a scuba suit composed of the following elements: a stabilizing jacket, a pressure regulator, a mask, palms and a scuba tank. Strict rules must be respected. Given that there is an increase of cannabis users in the general population, it is therefore not uncommon to find some cannabis users practicing scuba diving. This drug has important psychoactive effects that can cause fatal consequences when practicing this activity. The authors report a case of death while scuba diving under the influence of cannabis. A 64-year-old man collapsed after scuba diving, when he was return to the surface. People on the spot took charge of him directly to perform first aid resuscitation, but he rapidly died. Toxicological analyses were requested to document the cause of the death. The matrices collected at the autopsy included femoral blood, cardiac blood, urine, bile, gastric contents, and hair (10 cm, blond, no orientation). After comprehensive toxicological investigations, cannabis exposure was identified. GC-MS/MS (Agilent 7010B) was used for the determination of Δ9-tetrahydrocannabinol (THC), and its metabolites (11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC) and 11-nor Δ9-tetrahydrocannabinol-carboxylic acid (THC-COOH)) in blood. The analysis of THC-COOH in urine and THC and cannabinol (CBN) and cannabidiol (CBD) in hair was performed on a GC/MS (Perkin Elmer). Blood was analyzed after extraction with n-hexane/ethyl acetate (90:10, v/v) and then derivatizated with BSTFA in the presence of THC-D3. In hair, THC, CBN and CBD were extracted in a mixture of n-hexane and ethyl acetate (90:10, v/v) after hydrolysis in alkaline conditions. Δ9-THC was detected with a concentration of 70.5 ng/mL in the femoral blood, 11-OH-THC and THC-COOH were found at 1.6 ng/mL and 27.8 ng/mL respectively. THC-COOH tested positive at 858 ng/mL in the urine. In hair, 2432 pg/mg of THC, 85 pg/mg of CBN and 398 pg/mg of CBD were measured. The concentration of Δ9-THC in blood indicated that the man was under the influence of cannabis when the accident occurred. In addition, the concentrations found in the hair indicate regular cannabis use. In addition to cannabis exposure, amlodipine was found at therapeutic concentration (69 ng/mL) in blood. A very small concentration of ethanol was found in the blood (0.09 g/L) and urine (0.20 g/L). The concentration of strontium was 23 ng/mL, which is inconclusive with respect to vital drowning. The concentrations of cannabis found in blood revealed recent cannabis use before diving, demonstrating that the subject was under the influence during the dive. In addition, regular use of this recreative drug was revealed by the hair test. The recent use of cannabis, ethanol and his hypertension (demonstrated by the amlodipine treatment) could be the cause of this man's discomfort and death.

Bariatric surgery and changes in metformin distribution: About a fatal case

Metformin is an hypoglycemic drug, represented as the « gold standard » for the treatment of maturity-onset diabetes. Lactic acidosis is a well known and feared adverse effect of this substance.The authors present the case of a 65 year-old woman, suffering from mellitus type 2 diabetes and obesity, who died at home, only three days after bariatric surgery (Roux-en-Y gastric by-pass, RYGB). Her treatment, including metformin and dapagliflozin, was stopped before surgery and not postoperatively resumed. The woman was visited by a friend 15 hours before the body discovery, and no information was available about the symptoms presented by the patient before the death, excepted asthenia. A forensic autopsy was performed. Cardiac and femoral blood, urine, gastric content and hair were collected for toxicological investigations. Metformin was specifically tested on a GC-MS/MS system. Liquid matrices (0.2 mL) were subject to protein precipitation with 3 mL of acetonitrile in the presence of merformin-d6 used as internal standard. After agitation and centrifugation, the surpernatant was evaporate to dryness and the dry extract was derivatized with MBTFA. Metformine was extracted from 20 mg of hair with methanol. Detection of metformin was achieved using two transitions: m/z 303 > 288 and m/z 303 > 125. No evidence of digestive perforation or peritonitis was identified during autopsy, excluding an early surgery complication as the cause of death. These findings were confirmed by the histological analysis which showed a physiological intestinal inflammatory reaction related to the recent surgery. Toxicological analysis revealed the presence of acetone in the blood (0.25 g/L) and in the urine (0.58 g/L), confirming diabetes. Metformin tested positive in blood (148 mg/L), urine (49 mg/L), intestinal content (> 100 mg/L) and hair (1.9 ng/mg). Blood concentration was considered potentially fatal. The low concentration in urine indicated rapid death after exposure. Hair concentration was in the therapeutic range. The pathologist concluded that the death was due to lactic acidosis caused by an overdose of metformin. With no evidence for suicide by ingestion of metformin, the hypothesis raised by the authors was that the bariatric surgery might have caused changes in the absorption of metformin, leading to a rapid overdose and death. In the literature, it has been demonstrated in gastric bypass patients that there is a significant increase in the bioavailability of metformin following oral administration. RYGB would increases intestinal transit time and thus the duration of exposure to small intestinal mucosa. Changes induced by the surgical procedure are probably not sufficient to explain the massive concentrations of metformin found in the postmortem samples but the context of the corpse discovery was not suggestive of suicide. As this represents the first case of metformin overdose following bariatric surgery, further cases will be needed to confirm our initial observations. It can be anticipated that a therapeutic dose can become toxic when administrated to a subject who recently modified her digestive equipment. This case highlights the doubts that can be encountered by pathologists and toxicologists in forensic medicine and that evidences must constantly be reconsidered.

Deaths related to the combination of pregabalin and other drugs of abuse: About two cases

The number of deaths involving pregabalin has dramatically increased in recent years. In the majority of the cases described in France, pregabalin is associated with opioids, mainly methadone, as well as benzodiazepines and other psychoactive drugs, notably antidepressants and antipsychotics. Association with alcohol, cannabis and psychostimulants is also mentioned in the literature (Evoy KE et al. Drugs 2021;81:125–56). In this context, pregabalin concentrations are usually high, above therapeutic concentrations, in order to achieve euphoric effects, to enhance the effects of other substances or to improve sociability (Hofmann M & Besson M. Psychiatry Res 2021;35:114193). The authors present two cases of unusual deaths, a few days apart, for which toxicological analyses, showing low concentrations of pregabalin, buprenorphine, clonazepam and psychostimulants, are in favor of new patterns of use. Case 1: a 30-year-old man with a known addiction to pregabalin and cocaine, was seen by a witness snorting buprenorphine. He was found dead two hours later. Case 2: A young homeless man is found dead in the street in the early hours of the morning. No information is available on his identity and medical history. In both cases, the autopsy performed within 48 hours only show diffuse polyvisceral congestion with major cerebral and pulmonary oedema and the pathological examination did not allow to determine the cause of death. The usual toxicological samples, including femoral blood, with sodium fluoride as preservative, were collected and complete toxicological analyses were requested. Analyses were performed in femoral blood. Alcohols were measured by HS-GC-FID. Non-targeted screening and simultaneous quantification was performed by LC-HRMS. Specific detection of drugs of abuse (DOA) was done by LC-MS/MS. In case 1, analyses showed the presence of pregabalin (4900 ng/mL), buprenorphine (2.5 ng/mL), norbuprenorphine (2.5 ng/mL), 7-aminoclonazepam (14 ng/mL), diazepam (51 ng/mL), nordiazepam (137 ng/mL), oxazepam (7 ng/mL), cocaine (4 ng/mL), benzoylecgonine (177 ng/mL), ecgonine methylester (33 ng/mL) and THC-COOH (4 ng/mL). In case 2, results showed the presence of pregabalin (5700 ng/mL), buprenorphine (4.5 ng/mL), norbuprenorphine (0.5 ng/mL), 7-aminoclonazepam (23 ng/mL), MDMA (1140 ng/mL) and MDA (54 ng/mL). Alcohol testing was negative in both cases. In both cases, the concentrations of pregabalin, benzodiazepines and buprenorphine are far below the toxic concentrations. While the risk of death when buprenorphine is combined with benzodiazepines at therapeutic doses, particularly when buprenorphine is injected or absorbed nasally (Sansone RA & Sansone LA. Innov clin NeuroSci 2015;12:32–6), is well described in the literature, the potential role of pregabalin remains to be determined. Indeed, if the combination of gabapentinoids/opioids/other depressants is known to favor the occurrence of respiratory depression, it seems that the favoring factors are a high dose, an advanced age and co-morbidities (chronic pulmonary disease, renal insufficiency in particular) (Gomes T. et al. PLoS Med 2017;14:e1002396). Finally, if the epidemiological profile of the two victims is compatible with the data found in the literature concerning pregabalin misuse (young man, history of addiction for case n o 1), these results are in favor of new patterns of use.

The systematic investigation of the role of adulterants into the postmortem redistribution of heroin

We aimed to investigate systematically the impact of adulterants on postmortem in vivo pharmacokinetic of heroin and its metabolites by determining the postmortem redistribution (PMR) of heroin in this study. Heroin (1 mg/kg) was administered intravenously to the rats. The rats were euthanized 1 hour after drug administrations, followed by placed in the supine position at room temperature. Femoral (right and left), subclavian, cardiac blood samples and liver, lung, kidney, myocardium and brain samples were collected at the time of death, postmortem 1st, 4th and 24th hour. Heroin (1 mg/kg) containing adulterants (1 mg/kg paracetamol, dextromethorphan and caffeine) was administered intravenously to the same number of rat groups, and the same procedures were repeated. All blood and organ tissue samples were analysed with validated liquid-liquid extraction methods using a liquid chromatography tandem-mass spectrometer. The changes in the levels of heroin metabolites during the postmortem interval were determined in different blood and tissue samples included in our study. Accordingly, apart from the minimal statistical differences in morphine concentrations in the left femoral blood, significant differences in each matrix throughout the postmortem interval, especially in heart blood, lung and kidney tissue were observed. In contrast to morphine, there are minimal differences in morphine-3-glucurinide concentrations in heart tissue and other matrices. Graphical differences were observed between morphine concentrations in heart blood, liver, myocardium and lung of animal groups in the presence of adulterants and animal groups in which heroin was administered alone. The findings obtained in this study may play an important role in elucidating heroin-related death cases by forensic toxicologists or pathologists.

Difficulties in interpreting a therapeutic blood concentration of insulin lispro in postmortem blood: A case report

Illustrate the complexity of interpreting a postmortem therapeutic blood concentration of insulin in a probable case of suicide using insulin lispro. The identification and quantification of insulin and its analogues continues to be a challenge in the field of forensic toxicology. Its use is not always limited to the treatment of diabetes but is often used illicitly. Numerous cases of suicides and suicide attempts have been described in the literature and nevertheless, this number still seems to be underestimated due to both the inability of laboratories to identify and discriminate between synthetic analogues of human insulin and due to the chemical instability of insulin especially in postmortem samples. Our case concerns a 63-year-old man who was found dead by his brother at home. Three empty Humalog® insulin pens and a letter mentioning his suicide intention were found next to his bed. At the request of the prosecutor, an external examination of the corpse was performed and revealed only an asphyxial syndrome not specific to a cause of death. The presence of a blood glucose monitoring device on the right arm was also observed, suggesting that the subject was diabetic. External examination was unable to determine the exact cause of death. For this reason, the pathologist was oriented on a toxic death. In order to proceed to further toxicological investigations, peripheral blood, urine and hair samples were collected. It is important to underline that, due to the long period elapsed between the external examination and the pathologist's conclusion (enabling the initiation of the toxicological analysis) and due to administrative delays, insulin testing was performed only 9 months after the samples collection. Blood and urine samples were stored at +4 °C until analysis. The identification and quantification of insulin lispro was performed in the postmortem blood after precipitation in the presence of bovine insulin (internal standard), by a mixture of acetonitrile/methanol (ratio 1:1) followed by a step of ultrafiltration using Amicon® centrifugal filters (3 kDa) and a step of immunopurification using Iso-Insulin ELISA coated wells. The analysis was performed with a UHPLC- Q-TOF (XEVO G2-XS). Detection of insulin lispro was performed using the precursor molecular ion m/z 1162.543 (charge state 5 +). Since human insulin and insulin lispro share the exact same molecular mass (5807 Da), discrimination cannot be made using the multi-charged molecular ion. The distinction is based on fragment m/z 217 which is present only in insulin lispro spectrum. Toxicological tests performed on the blood sample did not reveal any xenobiotic of toxic nature. Insulin lispro was detected in the femoral blood sample at 1,1 ng/mL, whereas human insulin was not detected. Following the administration of a therapeutic dose of 0.2 U/kg of insulin Humalog ®, a maximum concentration of 1.66 ± 0.42 ng/mL is achieved. Consequently, the concentration found in our case was therapeutic. Nevertheless, given the unstable nature of insulin, especially in postmortem blood, it is not possible to exclude that death was caused by an insulin overdose. The long period between collection and analysis (9 months), the hemolyzed state of the blood, the storage of the sample in unsuitable tubes and the fact that we were able to detect even small concentrations of insulin lispro suggest that the concentration at the time of death was much higher. The pathologist concluded that the cause of death was probably an overdose of insulin lispro. This case highlights that the interpretation of a postmortem insulin concentration requires a complex approach involving the assessment of many factors such as the subject's medical history and the time, mode and state of preservation of the sample.

Hydrogen sulfide mitigates skeletal muscle mitophagy‐led tissue remodeling via epigenetic regulation of the gene writer and eraser function

Ketone bodies (KB) serve as the food for mitochondrial biogenetics. Interestingly, probiotics are known to promote KB formation in the gut (especially those that belong to the Lactobacillus genus). Furthermore, Lactobacillus helps produce folate that lowers the levels of homocysteine (Hcy); a hallmark non‐proteinogenic amino acid that defines the importance of epigenetics, and its landscape. In this study, we decided to test whether hydrogen sulfide (H2S), another Hcy lowering agent regulates the epigenetic gene writer DNA methyltransferase (DNMT), eraser FTO and TET2, and thus mitigates the skeletal muscle remodeling. We treated hyperhomocysteinemic (HHcy, cystathionine beta‐synthase heterozygote knockout; CBS+/−) mice with NaHS (the H2S donor). The results suggested multi‐organ damage by HHcy in the CBS+/−mouse strain compared with WT control mice (CBS+/+). H2S treatment abrogated most of the HHcy‐induced damage. The levels of gene writer (DNMT2) and H3K9 (methylation) were higher in the CBS+/− mice, and the H2S treatment normalized their levels. More importantly, the levels of eraser FTO, TET, and associated GADD45, and MMP‐13 were decreased in the CBS+/− mice; however, H2S treatment mitigated their respective decrease. These events were associated with mitochondrial fission, i.e., an increase in DRP1, and mitophagy. Although the MMP‐2 level was lower in CBS+/− compared to WT but H2S could further lower it in the CBS+/− mice. The MMPs levels were associated with an increase in interstitial fibrosis in the CBS+/− skeletal muscle. Due to fibrosis, the femoral artery blood flow was reduced in the CBS+/− mice, and that was normalized by H2S. The bone and muscle strengths were found to be decreased in the CBS+/− mice but the H2S treatment normalized skeletal muscle strength in the CBS+/− mice. Our findings suggest that H2S mitigates the mitophagy‐led skeletal muscle remodeling via epigenetic regulation of the gene writer and eraser function.

Isolated knee extensor exercise training improves skeletal muscle vasodilation, blood flow, and functional capacity in patients with HFpEF.

Patients with HFpEF experience severe exercise intolerance due in part to peripheral vascular and skeletal muscle impairments. Interventions targeting peripheral adaptations to exercise training may reverse vascular dysfunction, increase peripheral oxidative capacity, and improve functional capacity in HFpEF. Determine if 8 weeks of isolated knee extension exercise (KE) training will reverse vascular dysfunction, peripheral oxygen utilization, and exercise capacity in patients with HFpEF. Nine HFpEF patients (66 ± 5 years, 6 females) performed graded IKE exercise (5, 10, and 15 W) and maximal exercise testing (cycle ergometer) before and after IKE training (3x/week, 30 min/leg). Femoral blood flow (ultrasound) and leg vascular conductance (LVC; index of vasodilation) were measured during graded IKE exercise. Peak pulmonary oxygen uptake (V̇O2; Douglas bags) and cardiac output (QC; acetylene rebreathe) were measured during graded maximal cycle exercise. IKE training improved LVC (pre: 810 ± 417, post: 1234 ± 347 ml/min/100 mmHg; p = 0.01) during 15 W IKE exercise and increased functional capacity by 13% (peak V̇O2 during cycle ergometry; pre:12.4 ± 5.2, post: 14.0 ± 6.0 ml/min/kg; p = 0.01). The improvement in peak V̇O2 was independent of changes in Q̇c (pre:12.7 ± 3.5, post: 13.2 ± 3.9 L/min; p = 0.26) and due primarily to increased a‐vO2 difference (pre: 10.3 ± 1.6, post: 11.0 ± 1.7; p = 0.02). IKE training improved vasodilation and functional capacity in patients with HFpEF. Exercise interventions aimed at increasing peripheral oxidative capacity may be effective therapeutic options for HFpEF patients.

Arterial Stiffness and Endothelial Function are Comparable in Young Healthy Vegetarians and Omnivores

Vegetarians (VEG) are reported to have lower body weight, blood pressure (BP), and cardiovascular disease (CVD) risk compared with omnivores (OMN), yet the mechanisms remain unclear. A vegetarian diet may protect the vascular endothelium, reducing the risk of atherosclerosis and CVD. This cross-sectional study compared vascular function between OMN and VEG. We hypothesized that VEG would have greater vascular function compared with OMN. Fifty-eight normotensive young healthy adults participated (40 women [W]/18 men [M]; 28 OMN [15W/13M] and 30 VEG [25W/5M]; 26 ± 7 years; BP: 112 ± 11/67 ± 8 mm Hg). Arterial stiffness, assessed by carotid-to-femoral pulse wave velocity (OMN: 5.6 ± 0.8 m/s, VEG: 5.3 ± 0.8 m/s; P = .17) and wave reflection assessed by aortic augmentation index (OMN: 6.9 ± 12.3%, VEG: 8.8 ± 13.5%; P = .57) were not different between groups. However, central pulse pressure (OMN: 32 ± 5; VEG: 29 ± 5 mm Hg; P = .048) and forward wave reflection were greater in omnivores (OMN: 26 ± 3; VEG: 24 ± 3 mm Hg; P = .048). Endothelial-dependent dilation measured by brachial artery flow-mediated dilation was not different between groups (OMN: 6.0 ± 2.9%, VEG: 6.9 ± 3.3%; P = .29). Percent change in femoral blood flow from baseline during passive leg movement, another assessment of nitric oxide-mediated endothelial dilation, was similar between groups (OMN: 203 ± 88 mL/min, VEG: 253 ± 192 mL/min; P = .50). These data suggest that in healthy young adults, normotensive VEG do not have significantly improved vascular function compared with OMN; however, they have a lower central pulse pressure and forward wave amplitude which may lower the risk of future CVD.

Single Leg Cycling Offsets Reduced Muscle Oxygenation in Hypoxic Environments.

The intensity of large muscle mass exercise declines at altitude due to reduced oxygen delivery to active muscles. The purpose of this investigation was to determine if the greater limb blood flow during single-leg cycling prevents the reduction in tissue oxygenation observed during traditional double-leg cycling in hypoxic conditions. Ten healthy individuals performed bouts of double and single-leg cycling (4, four-minute stages at 50–80% of their peak oxygen consumption) in hypoxic (15% inspired O2) and normoxic conditions. Heart rate, mean arterial pressure, femoral blood flow, lactate, oxygenated hemoglobin, total hemoglobin, and tissue saturation index in the vastus lateralis were recorded during cycling tests. Femoral blood flow (2846 ± 912 mL/min) and oxygenated hemoglobin (−2.98 ± 3.56 au) during single-leg cycling in hypoxia were greater than double-leg cycling in hypoxia (2429 ± 835 mL/min and −6.78 ± 3.22 au respectively, p ≤ 0.01). In addition, tissue saturation index was also reduced in the double-leg hypoxic condition (60.2 ± 3.1%) compared to double-leg normoxic (66.0 ± 2.4%, p = 0.008) and single-leg hypoxic (63.3 ± 3.2, p < 0.001) conditions. These data indicate that while at altitude, use of reduced muscle mass exercise can help offset the reduction in tissue oxygenation observed during larger muscle mass activities allowing athletes to exercise at greater limb/muscle specific intensities.

Femoral head reduction osteotomy to improve femoroacetabular containment in Legg-Calve-Perthes disease

OperationszielWiederherstellung der Hüftkongruenz und des Containments durch zentrale Hüftkopfresektion/Reduktion über eine erweiterte chirurgische Hüftluxation unter Erhaltung/Respektierung der Hüftkopfdurchblutung. Eine gleichzeitige oder spätere Reorientierung des Acetabulums durch Triple-Osteotomie oder PAO bei instabiler Hüfte kann in speziellen Fällen notwendig werden.IndikationenDie Indikation zur chirurgischen Hüftkopfreduktion ist praktisch unabhängig vom Alter bei jeglicher inkongruenten, asphärischen Hüftkopfsituation mit „hinged abduction“ (zu erwartende Endsituation wie Stulberg IV und V) gegeben. Dies kann noch bei aktivem wie auch bei bereits ausgeheiltem Morbus Perthes der Fall sein. Zudem kann bei einer Stulberg-V-Situation selbst im adulten Alter noch eine Verbesserung erziel werden. Nach Resektion muss noch ein tragfähiger Resthüftkopf vorhanden bleiben, d. h. mindestens noch 50 % des ausgeweiteten Hüftkopfes. Die dafür beste Planung erfolgt in der „vergleichenden“ 3‑D-Rekonstruktion.KontraindikationenVöllig zerstörter Knorpel oder Hüftkopf.OperationstechnikIdentisches Operationsvorgehen wie für die klassische chirurgische Hüftluxation. Präparation der retinakulären Flaps. Unter Respektierung und in Kenntnis der Gefäßversorgung Spaltung des Hüftkopfes gemäß dem zu entfernenden, nekrotischen Kopfanteil. Bildung eines möglichst sphärischen Hüftkopfes und Verschraubung der beiden Kopfanteile auf Schenkelhalsniveau. Distalisation und Fixierung des großen Trochanters. Je nach Kongruenz und Stabilität des Hüftkopfs in der Hüftpfanne kann eine primäre oder sekundäre Triple-OT oder PAO notwendig werden.WeiterbehandlungDie intraoperative Stabilität des Femurkopfes im Acetabulum muss erzielt worden sein, um eine beckengipsfreie, funktionelle Nachbehandlung zu gewährleisten: Stockentlastung mit Bodenkontakt ist erlaubt; keine aktive Rotation; Flexion aktiv und passiv bis 90 Grad ist erlaubt; vorerst keine spezifische Physiotherapie; je nach Heilungsverlauf sind diese Maßnahmen 8 bis 10 Wochen einzuhalten.ErgebnisseGemäß unseren publizierten Nachuntersuchungen (aktuell 21 Jahre) sehen wir bei technisch korrekt durchgeführter Operation und korrekter Indikation sowie adäquater Nachbehandlung durchwegs gute Ergebnisse. Nekrosen des reduzierten Hüftkopfs haben wir nie beobachtet. Alle gespaltenen Hüftköpfe, respektive Schenkelhälse sind primär geheilt.

Blood cobalt ion level in patients with different sizes of cobalt/chrome femoral head with the Accolade TMZF stem.

Corrosion of stem-cobalt/chromium (Co/Cr) head interface and subsequent systemic Co ion complication have been a clinical concern after total hip arthroplasty (THA). The aim of this study is to investigate correlation between type of femoral head and blood Co ion level. One hundred four patients with THA using accolade TMZF and Co/Cr femoral head (32-mm; 55 cases, 36-mm; 20 cases, dual mobility system (DM) with 22.2 or 28-mm inner head; 29 cases) participated in this study, and blood Co ion concentration test was performed in patients a minimum of fouryears after THA. DM group was significantly older than 32-mm and 36-mm group (DM: 73.9 ± 4.6, 32-mm: 63.3 ± 8.5, 36-mm: 66.8 ± 10.5). The median (interquartile range) blood Co concentration of each group was 32-mm group 0.26µg/L (0.69), 36-mm group; 0.21µg/L (0.30); and DM group 0.21µg/L (0.13). There was a significant difference of Co concentration between DM and 32-mm group (p = 0.023). Abnormal values of the blood Co concentration (> 1µg/L) were observed in the 32-mm group; 12 cases (21.8%), 36-mm group; 2 cases (10.0%); and DM group; 0 case (0%) (p = 0.018). Co blood concentration differed among the different sizes of Co/Cr femoral head. THA using DM is a safe option with low risk of complication from cobalt ion if it is used for elderly patients.

Fast regulation of the NF-κB signalling pathway in human skeletal muscle revealed by high-intensity exercise and ischaemia at exhaustion: Role of oxygenation and metabolite accumulation

The NF-κB signalling pathway plays a critical role in inflammation, immunity, cell proliferation, apoptosis, and muscle metabolism. NF-κB is activated by extracellular signals and intracellular changes in Ca2+, Pi, H+, metabolites and reactive oxygen and nitrogen species (RONS). However, it remains unknown how NF-κB signalling is activated during exercise and how metabolite accumulation and PO2 influence this process. Eleven active men performed incremental exercise to exhaustion (IE) in normoxia and hypoxia (PIO2:73 mmHg). Immediately after IE, the circulation of one leg was instantaneously occluded (300 mmHg). Muscle biopsies from m. vastus lateralis were taken before (Pre), and 10s (Post, occluded leg) and 60s after exercise from the occluded (Oc1m) and free circulation (FC1m) legs simultaneously together with femoral vein blood samples. NF-κB signalling was activated by exercise to exhaustion, with similar responses in normoxia and acute hypoxia, as reflected by the increase of p105, p50, IKKα, IκBβ and glutathione reductase (GR) protein levels, and the activation of the main kinases implicated, particularly IKKα and CaMKII δD, while IKKβ remained unchanged. Postexercise ischaemia maintained and stimulated further NF-κB signalling by impeding muscle reoxygenation. These changes were quickly reverted at the end of exercise when the muscles recovered with open circulation. Finally, we have shown that Thioredoxin 1 (Trx1) protein expression was reduced immediately after IE and after 1 min of occlusion while the protein expression levels of glutathione peroxidase 1 (Gpx1) and thioredoxin reductase 1 (TrxR1) remained unchanged. These novel data demonstrate that exercising to exhaustion activates NF-κB signalling in human skeletal muscle and regulates the expression levels of antioxidant enzymes in human skeletal muscle. The fast regulation of NF-κB at exercise cessation has implications for the interpretation of published studies and the design of new experiments.

The Comparative Effect of Different Timings of Whole Body Cryotherapy Treatment With Cold Water Immersion for Post-Exercise Recovery.

Despite several established benefits of Whole Body Cryotherapy (WBC) for post-exercise recovery, there is a scarcity of research which has identified the optimum WBC protocol for this purpose. This study investigated the influence of WBC treatment timing on physiological and functional responses following a downhill running bout. An additional purpose was to compare such responses with those following cold water immersion (CWI), since there is no clear consensus as to which cold modality is more effective for supporting athletic recovery. Thirty-three male participants (mean ± SD age 37.0 ± 13.3 years, height 1.76 ± 0.07 m, body mass 79.5 ± 13.7 kg) completed a 30 min downhill run (15% gradient) at 60% VO2 max and were then allocated into one of four recovery groups: WBC1 (n = 9) and WBC4 (n = 8) underwent cryotherapy (3 min, −120°C) 1 and 4 h post-run, respectively; CWI (n = 8) participants were immersed in cold water (10 min, 15°C) up to the waist 1 h post-run and control (CON, n = 8) participants passively recovered in a controlled environment (20°C). Maximal isometric leg muscle torque was assessed pre and 24 h post-run. Blood creatine kinase (CK), muscle soreness, femoral artery blood flow, plasma IL-6 and sleep were also assessed pre and post-treatment. There were significant decreases in muscle torque for WBC4 (10.9%, p = 0.04) and CON (11.3% p = 0.00) and no significant decreases for WBC1 (5.6%, p = 0.06) and CWI (5.1%, p = 0.15). There were no significant differences between groups in muscle soreness, CK, IL-6 or sleep. Femoral artery blood flow significantly decreased in CWI (p = 0.02), but did not differ in other groups. WBC treatments within an hour may be preferable for muscle strength recovery compared to delayed treatments; however WBC appears to be no more effective than CWI. Neither cold intervention had an impact on inflammation or sleep.

Effect of drilling and bone substitute on revascularization of an avascular femoral head

Introduction: Avascular Necrosis (AVN) is pathology in which femoral head undergoes degenerative changes due to reduced blood supply. Osteotomy is surgical procedure that involves cutting bone to reshape or realign your bones. It acts by multiple mechanisms. Newer mechanisms include increased blood flow, Osteoinduction due to new fracture (Osteotomy), Change of shear force into compression force. Novel concept for AVN treatment is blocking of femoral canal near lesser trochanter area by Osteotomy/Drilling and insertion of synthetic absorbable Bone substitute (hydroxyapatite cement). Thereby blocking the trabecular canal system at junction so Blood supply get diverted towards femoral head. Methods and Materials: Study was carried out in New Civil Hospital, Surat in September 2021. The prospective study comprised of 62 year old male having C/o Pain, limp while walking and restriction of Adduction and Internal rotation of Left Hip. Xray, MRI of both hip showed Left Avascular Necrosis Femoral Head (Ficat Arlet Grade 2B). Via Lateral incision of Hip, Drilling and Scooping done at level of Lesser Trochanter. Drilled site was inserted with 10-12 gm of synthetic absorbable Bone substitute (hydroxyapatite cement). Full weight bearing started after 1 week. All patients were followed up immediately, 3rd month, 6th month, 1 year. Follow up X-ray were taken to assess radiological outcome and Harris Hip score for functional outcome. Results: Xray at 3rd, 6th month, 1 year showed No further progression of Grade 2B AVN Femoral head. Improvement in Harris Hip Score with values 38, 67, 81, 89 at Immediate, 3rd, 6th month, 1 year postop respectively. Improvement in Left Hip range of motion. Conclusion: Drilling creates disturbance in trabecular canal system (Creating haematoma callus) so that proximal shaft femoral blood supply is cut off (partially) and that blood supply which was destined for the upper shaft is now directed more proximally Towards femoral head.

Femoral Skeletal Perfusion is Reduced in Male Mice with Type 1 Diabetes

The bone vasculature and blood flow are critical for bone modeling, remodeling, and regeneration. Vascular complications are one of the major health concerns of people with type 1 diabetes (T1D). Moreover, people with T1D have lower bone mineral density and increased bone fragility. The goal of this study was to understand whether bone perfusion was altered in a mouse model of T1D and how this related to changes in bone mass. T1D was induced via the administration of streptozotocin in 12-week-old C57BL/6NHsd male mice. The assessment of bone perfusion utilized the injection of fluorescent microspheres with assessment of levels in the bone. Femoral blood flow and VEGF-A expression in the cortical bone shafts were lower in the T1D mice, compared to healthy controls, in this pattern followed that of changes in bone mass. These data demonstrate a possible association between reduced skeletal perfusion and reduced bone mass in the setting of T1D.

Increased contribution of KCa channels to muscle contraction induced vascular and blood flow responses in sedentary and exercise trained ZFDM rats

In resistance arteries, endothelium-dependent hyperpolarization (EDH)-mediated vasodilatation is depressed in diabetes. We hypothesized that downregulation of KCa channel derived EDH reduces exercise-induced vasodilatation and blood flow redistribution in diabetes. To test this hypothesis, we evaluated vascular function in response to hindlimb muscle contraction, and the contribution of KCa channels in anaesthetised ZFDM, metabolic disease rats with type 2 diabetes. We also tested whether exercise training ameliorated the vascular response. Using in vivo microangiography, the hindlimb vasculature was visualized before and after rhythmic muscle contraction (0.5s tetanus every 3s, 20 times) evoked by sciatic nerve stimulation (40Hz). Femoral blood flow of the contracting hindlimb was simultaneously measured by an ultrasonic flowmeter. The contribution of KCa channels was investigated in the presence and absence of apamin and charybdotoxin. We found that vascular and blood flow responses to muscle contraction were significantly impaired at the level of small artery segments in ZFDM fa/fa rats compared to its lean control fa/+ rats. The contribution of KCa channels was also smaller in fa/fa than in fa/+ rats. Low-intensity exercise training for 12weeks in fa/fa rats demonstrated minor changes in the vascular and blood flow response to muscle contraction. However, the KCa-derived component in the response to muscle contraction was much greater in exercise trained than in sedentary fa/fa rats. These data suggest that exercise training increases the contribution of KCa channels among endothelium-dependent vasodilatory mechanisms to maintain vascular and blood flow responses to muscle contraction in this metabolic disease rat model. KEY POINTS: Microvascular dysfunction in type 2 diabetes impairs blood flow redistribution during exercise and limits the performance of skeletal muscle and may cause early fatigability. Endothelium-dependent hyperpolarization (EDH), which mediates vasodilatation in resistance arteries, is known to be depressed in animals with diabetes. Here, we report that low-intensity exercise training in ZFDM rats increased the KCa channel-derived component in the vasodilator responses to muscle contraction compared to that in sedentary rats, partly as a result of the increase in KCNN3 expression. These results suggest that low-intensity exercise training improves blood flow redistribution in contracting skeletal muscle in metabolic disease with diabetes via upregulation of EDH.

Distributive characteristics of HBV DNA CpG islands in HBsAg positive mothers and its relationship with intrauterine transmission

Objective: To investigate the type, length, and CG loci of HBV DNA CpG islands in HBsAg positive maternal C genotype and its relationship with intrauterine HBV transmission, so as to provide a new perspective for the study of intrauterine transmission of HBV. Methods: From June 2011 to July 2013, HBsAg-positive mothers and their newborns who delivered in the obstetrics and gynecology department of the Third People's Hospital of Taiyuan were collected. Epidemiological data were collected through face-to-face questionnaires and electronic medical records. Serum HBV markers and serum HBV DNA were detected by electrochemiluminescence and quantitative fluorescence PCR, respectively. Intrauterine transmission of HBV was determined by positive HBsAg and/or HBV DNA in femoral venous blood before injection of HBV vaccine/Hepatitis B immunoglobulin within 24 h of birth. A total of 22 mothers and their newborns with HBV DNA load ≥106 IU/ml in intrauterine transmission were selected as the intrauterine transmission group, and 22 mothers with HBV DNA load ≥106 IU/ml without intrauterine transmission were chosen as the control group by random seed method. The distribution prediction of CpG islands of HBV DNA in 39 mothers with genotype C by HBV DNA sequencing was analyzed. Results: Among 39 mothers with HBV C genotype, 19 were in the intrauterine transmission group, and 20 were in the control group. The HBV DNA of 39 patients with genotype C traditional CpG island Ⅱ and Ⅲ, while the control group had traditional CpG island Ⅰ and novel CpG island Ⅳ and Ⅴ. The length of CpG island Ⅱ and Ⅲ and the number of CG loci of CpG island Ⅱ in the intrauterine transmission group differed from those in the control group (P<0.05). The CpG island Ⅱ length ≥518 bp and the number of CG loci ≥40 in the intrauterine transmission group (11/19) were significantly higher than those in the control group (2/20) (P<0.05). The length of CpG island Ⅱ and the number of CG loci in the X gene promoter region (Xp region) were higher than those in the control group (P<0.05). In the HBV intrauterine transmission group, most of maternal (12/19) HBV DNA CpG island Ⅱ completely covered the Xp region, which was significantly higher than that in the control group (5/20), and the number of HBV DNA Xp region CG loci was higher than that in the control group (P<0.05). Conclusions: The distribution of maternal C genotype HBV DNA CpG islands is related to intrauterine transmission. The length of CpG island Ⅱ and the number of CG sites may increase the risk of intrauterine transmission of HBV.

A Rare Finding of Thrombosed Bilateral Giant External Iliac Vein Aneurysms

A 70 year old man presented with severe left lower limb oedema and pain. He had a history of bilateral deep vein thrombosis 20 years ago. (A) Computed tomography phlebography revealed previously unknown bilateral thrombosed external iliac vein aneurysms (arrows), 45 mm and 63 mm on the right and left, respectively. The thrombus extended to both femoral veins being occlusive on the left side. (B) Transpopliteal catheter directed thrombolysis was performed on the symptomatic left side. Although venous aneurysm recanalisation was not achieved, the patient improved due to femoral vein and venous collateral blood clot lysis (arrowheads).Image 1

The study of postmortem blood gamma-hydroxybutyric acid (GHB) concentrations in Thai dead bodies unrelated to GHB use

Background: Gamma-hydroxybutyric acid (GHB) is classified as a category I psychotropic substance in Thai legislation. It can be exogenously consumed or endogenously produced and cut-off concentrations are required. Baseline GHB concentration ranges in Thai postmortem cases are still not available. Objectives: To determine baseline GHB concentrations and factors influencing GHB concentrations in Thai postmortem cases. Materials and methods: Postmortem femoral and cardiac blood samples were collected from Thai cadavers aged 18 years old or older sent for medico-legal autopsies at the Department of Forensic Medicine, Siriraj Hospital, Mahidol University between 18th August 2021 and 25th January 2022 with postmortem interval (PMI) not greater than 24 hours. Case data including sex, age, PMI, sites of collection and causes of death were recorded. Blood GHB concentrations were extracted using protein precipitation and analyzed by gas chromatography-mass spectrometry (GC-MS). Descriptive statistics, paired samples Wilcoxon signed-rank test, Spearman's correlation, Mann-Whitney U test and Kruskal-Wallis H test were analyzed where appropriate. Results: A total of 150 subjects were recruited; 63 (42%) were female with mean age 44.59 years old (range 18-75 years old). Femoral and cardiac GHB concentration ranges were &lt;0.5-20.81 µg/mL and 1.12-39.04 µg/mL, respectively while median GHB concentrations in femoral and cardiac blood were 6.38 and 8.41 µg/mL. Femoral GHB concentrations were significantly lower than cardiac GHB concentrations (p&lt;0.001). Both femoral and cardiac blood showed a positive correlation between GHB concentrations and PMI (Spearman’s correlation =0.52 and 0.32, p&lt;0.001). Most GHB concentrations in femoral blood were less than 10 µg/mL and almost all samples from both sources had GHB concentrations less than 30 µg/mL. Conclusion: GHB concentrations in Thai postmortem cases were &lt;0.5-20.81 µg/mL in femoral blood and 1.12-39.04 µg/mL in cardiac blood, respectively. GHB concentrations in femoral blood were significantly lower than in cardiac blood. GHB concentrations significantly increased with longer PMI

Suppression of long intergenic non-protein coding RNA 1123 constrains lower extremity deep vein thrombosis via microRNA-125a-3p to target interleukin 1 receptor type 1

ABSTRACT Lower extremity deep vein thrombosis (LEDVT) is a disorder of venous return caused by abnormal blood clotting. LEDVT can obstruct the lumen and is the third most common vascular disease after cerebrovascular disease and coronary artery disease. LncRNAs are associated with thrombosis and potentially affect the pathogenesis of DVT. However, no studies have reported the effect of LINC01123 on LEDVT. The aim of this study was to investigate the effect of LINC01123 on LEDVT in rats via the miR-125a-3p/interleukin 1 receptor type 1 (IL1R1) axis. Lentiviral vectors that altering LINC01123, miR-125a-3p and IL1R1 expression were pre-injected into the tail vein of rats, and an LEDVT model was established 1 day later. Detection of LINC01123, miR-125a-3p and IL1R1 expression was performed. Inflammatory factors in femoral venous blood, the length and weight of the thrombus, the histomorphological changes were determined in the rat model. The targeting relation of miR-125a-3p with LINC01123 or IL1R1 was verified. The results presented that LEDVT rats expressed high LINC01123 and IL1R1 and low miR-125a-3p expression levels. After silencing LINC01123 or elevating miR-125a-3p, the rate of thrombosis, length and weight of thrombus, and levels of inflammatory factors were reduced. The targeting relation was presented between miR-125a-3p with LINC01123 or IL1R1. Elevating IL1R1 was available to turn around the action of silence of LINC01123 on LEDVT rats. All in all, suppression of LINC01123 restrains LEDVT via miR-125a-3p to target IL1R1.