Chloroprocaine 3% Gel as a Novel Ocular Topical Anesthetic: Results from a Multicenter, Randomized Clinical Trial in Patients Undergoing Cataract Surgery.

Purpose: To compare the efficacy and safety of a novel ophthalmic anesthetic, chloroprocaine 3% gel to tetracaine 0.5% eye drops in patients undergoing cataract surgery with phacoemulsification. Methods: This was a prospective, randomized, multicenter, active-controlled, masked-observer, parallel group competitive equivalence study. The study comprised 338 patients having routine cataract extraction by clear corneal phacoemulsification, randomized to receive 3 drops of chloroprocaine gel (n = 166) or tetracaine eye drops (n = 172) before surgery. The primary objective of the study was to assess the equivalence of chloroprocaine gel to tetracaine eye drops as proportion of patients with successful ocular surface anesthesia, without any supplementation just before intraocular lens implantation. Safety measurements were pain, irritation, burning, stinging, photophobia, and foreign body sensation, graded by the patient and objective ocular signs. Results: Equivalence was demonstrated, with a somewhat higher success rate of chloroprocaine gel: 152/166 (92.0%) chloroprocaine versus 153/172 (90.5%) tetracaine patients achieved ocular surface anesthesia with no supplementation. Difference in proportions was 1.5% confidence interval [95% CI: (-3.6 to 6.6)] and 90% CI fell within (-10 to 10). Mean onset of anesthesia was 1.35 ± 0.87 min for chloroprocaine and 1.57 ± 1.85 for tetracaine (P = 0.083). Mean duration of anesthesia was 21.57 ± 12.26 min for chloroprocaine and 22.04 ± 12.58 for tetracaine (P = 0.574). No treatment emergent adverse events related to chloroprocaine were reported and no relevant findings related to local tolerance or vital signs were observed in both arms. Conclusions: Results obtained from the present cataract study demonstrated that chloroprocaine 3% ophthalmic gel is safe and effective, representing a valid alternative in ocular topical anesthesia. Clinical Trial Registration number: NCT04685538.

In situ sono-rheometric assessment of procaine-loaded calcium pectinate hydrogel for enhanced drug releasing under ultrasound stimulation.

Ultrasound (US) triggered alterations in the viscoelastic behavior of the procaine-loaded ionically gelatinized pectin hydrogel matrix, and drug release was observed using a sono-device rheometer. The gel softened immediately upon activation of the ultrasound operated at 43 kHz and remained in a softened state throughout the irradiation. Upon cessation of ultrasound, the gel promptly reverted to its original hardness. This cycle of softening was consistently observed in ionically crosslinked pectin hydrogels, resulting in the promotion of procaine release, particularly with higher US power and lower calcium concentration. As the amount of loaded procaine increased, the gel weakened due to ion exchange with the calcium crosslinker and procaine. The most substantial release efficiency, reaching 82 % with a concentration of 32 µg/ml, was achieved when the hydrogels contained 0.03 % procaine within the gelatinized hydrogel medicine at a calcium concentration of 0.9 M, representing a six-fold increase compared to that without US. Notably, US exposure affected the 3D porous structure and degradation rate, leading to hydrogel collapse and facilitating medicine release. Additionally, the procaine-loaded pectin hydrogels with 0.9 M calcium exhibited improved fibroblast cell viability, indicating non-toxicity compared to those hydrogels prepared at a higher Ca2+ concentration of 2.4 M.

[The effectiveness of the use of non-drug treatment complexes and their impact on quality of life indicators, the psychosomatic status of patients with residual brucellosis with lesions of the musculoskeletal system]. / Effektivnost' primeneniya kompleksov nemedikamentoznogo lecheniya i ikh vliyanie na pokazateli kachestva zhizni, psikhosomaticheskii status patsientov s rezidual'nym brutsellezom s porazheniem oporno-dvigatel'nogo apparata.

Secondary focal lesions of the musculoskeletal system that occur with residual brucellosis are characterized by a variety of localizations and simultaneous damage to several groups of joints, a deterioration in the quality of life and a high percentage of disability in people of working age. At present, there are many different schemes for the treatment and rehabilitation of developed residual brucellosis, in which, in addition to the "basic" course, including systemic anti-inflammatory therapy, much attention is paid to physiotherapeutic procedures, but the choice of the most effective treatment tactics remains an unresolved problem. PURPOSE OF THE STUDY: To determine the effectiveness of the use of options for non-drug methods of treatment in the complex rehabilitation of patients with residual brucellosis with lesions of musculoskeletal system. MATERIAL AND METHODS: Study included 140 patients treated for osteoarthritis of brucellosis etiology, who were divided into three groups matched by age, gender, average duration and stage of the disease, place of residence. Patients of all groups received standard medical treatment and different sets of physiotherapeutic procedures: in the 1st group (45 patients) - electrophoresis of novocaine on the knee joints, therapeutic massage of the cervical-collar zone; in the 2nd group (45 patients) - magnetotherapy on the area of the knee joints, sinusoidal modulated currents (SMC) on the shoulder joints, decimeter wave therapy of the lumbosacral zone (DMW-therapy); in the 3rd group (50 patients) - magnetic laser therapy on the shoulder, elbow, knee joints, therapeutic massage of the lumbosacral zone. Complaints, clinical symptoms, goniometry results, and blood parameters were assessed: ESR, C-reactive protein, fibrinogen, before the start of treatment, immediately after the course of rehabilitation, and after 6 and 12 months. At the same time, testing was carried out according to the SF-36 Health Status Survey questionnaire to monitor the quality of life. RESULTS: The applied scheme of drug treatment in combination with magnetic laser therapy and therapeutic massage in the 3rd group made it possible to achieve a significant reduction in arthralgic syndrome, a statistically significant increase in the range of motion in the joints, positive dynamics of laboratory data and an improvement in the psychological state and quality of life of patients compared to other observation groups. CONCLUSION: The results of the study indicate the high efficiency of magnetic laser therapy in the complex treatment of patients with residual brucellosis with lesions of the musculoskeletal system.

Improvement of Left Ventricular Graft Function Using an Iron-Chelator-Supplemented Bretschneider Solution in a Canine Model of Orthotopic Heart Transplantation.

Demand for organs is increasing while the number of donors remains constant. Nevertheless, not all organs are utilized due to the limited time window for heart transplantation (HTX). Therefore, we aimed to evaluate whether an iron-chelator-supplemented Bretschneider solution could protect the graft in a clinically relevant canine model of HTX with prolonged ischemic storage. HTX was performed in foxhounds. The ischemic time was standardized to 4 h, 8 h, 12 h or 16 h, depending on the experimental group. Left ventricular (LV) and vascular function were measured. Additionally, the myocardial high energy phosphate and iron content and the in-vitro myocyte force were evaluated. Iron chelator supplementation proved superior at a routine preservation time of 4 h, as well as for prolonged times of 8 h and longer. The supplementation groups recovered quickly compared to their controls. The LV function was preserved and coronary blood flow increased. This was also confirmed by in vitro myocyte force and vasorelaxation experiments. Additionally, the biochemical results showed significantly higher adenosine triphosphate content in the supplementation groups. The iron chelator LK614 played an important role in this mechanism by reducing the chelatable iron content. This study shows that an iron-chelator-supplemented Bretschneider solution effectively prevents myocardial/endothelial damage during short- as well as long-term conservation.

Custodiol® Supplemented with Synthetic Human Relaxin Decreases Ischemia-Reperfusion Injury after Porcine Kidney Transplantation.

Objective. Ischemia-reperfusion injury (IRI) is inevitable after kidney transplantation (KT), impairing outcomes. Relaxin-2 (RLX) is a promising insulin-related peptide hormone that protects against renal IRI in rodents, although large animal models are needed before RLX can be tested in a human setting. Methods. In this blinded, randomized, and placebo-controlled experimental study kidneys from 19 donor pigs were retrieved after perfusion with Custodiol® ± RLX (5 or 20 nmol/L) and underwent static cold storage (SCS) for 24 and 48 h, respectively. Subsequently, KT was performed after unilateral right nephrectomy. Study outcomes included markers for kidney function, oxidative stress, lipid peroxidation, and endothelial cell damage. PCR analysis for oxidative stress and apoptosis-related gene panels as well as immunohistochemistry were performed. Results. RLX upregulated SOD2 and NFKB expression to 135% (p = 0.042) and 125% (p = 0.019), respectively, while RIPK1 expression was downregulated to 82% (p = 0.016) of corresponding controls. Further RLX significantly downregulated RIPK1 and MLKL expression and decreased the number of Caspase 3- and MPO-positive cells in grafts after SCS. Conclusions. RLX supplemented Custodiol® significantly decreased IRI via both antioxidant and anti-apoptotic mechanisms. Clinical trials are warranted to implement synthetic human RLX as a novel additive to preservation solutions against IRI.

Dairy calves show behavioural responses to hot iron disbudding after local anaesthesia with procaine.

BACKGROUND: The efficacy of procaine as anaesthetic for disbudding of calves is largely unknown. Likewise, information about effect of interval from local anaesthesia to disbudding and the size of the hot iron used is missing. METHODS: A total of 167 dairy calves were disbudded after cornual nerve block with procaine. Time from administration of anaesthetic to disbudding varied, and calves were disbudded using two different sizes of hot iron. The calves' behavioural responses to disbudding were recorded based on (1) getting up, (2) kicking, and (3) lifting head. Using logistic regression, the associations between calves showing at least one of these three behaviours and interval from administration of anaesthetic to disbudding and size of the hot iron were evaluated. RESULTS: Overall, 42% of calves showed at least one of the three behaviours, even though they had received a cornual nerve block prior to disbudding. Using the large hot iron was associated with a 2.3 times higher odds of the calf reacting behaviourally compared to the small iron. Interval from administration of anaesthetic to disbudding was not significantly associated with the odds of behavioural responses. CONCLUSION: Approximately 40% of calves disbudded after cornual nerve block with procaine showed signs of inadequate local anaesthesia. This was especially problematic when hot irons with larger tips were used.

Evaluation of corneal anaesthesia after the application of topical 0.5% bupivacaine, 2% lidocaine and 0.4% oxybuprocaine in normal horses.

OBJECTIVE: To compare the corneal anaesthetic effect of 0.5% bupivacaine, 2% lidocaine and 0.4% oxybuprocaine on normal equine eyes. STUDY DESIGN: Prospective, blinded crossover study. ANIMALS: A group of 10 clinically healthy horses. METHODS: Corneal sensitivity was determined in each eye by measuring corneal touch threshold (CTT). The study had three phases. Each subject was randomly given one of the three treatments followed by a 72 hour washout period. Every horse received all treatments. Baseline CTT was recorded prior to anaesthetic instillation (T0) then CTT was measured 5 and 10 minutes after (T1 and T2, respectively), then 20 to 90 minutes (T3 to T10) at 10 minute intervals. CTT data were compared among treatments at each time point using the Friedman test p < 0.05. RESULTS: Median (range) baseline CTT was 51.3 (25.0-60.0) mm for bupivacaine, 50.0 (40.0-55.0) mm for oxybuprocaine and 55.0 (30.0-60.0) mm for lidocaine. All treatments caused a significant decrease in CTT at T1. The lowest CTT was observed at T3 with bupivacaine and oxybuprocaine treatments. Median CTTs at this time point were 18.7 (5.0-25.0) mm and 28.7 (25.0-40.0) mm, respectively. The lowest CTT with lidocaine treatment was 28.7 (20.0-50.0) mm at T6 (50 minutes). At T3, CTT was significantly lower with the bupivacaine treatment compared with oxybuprocaine and lidocaine treatments (p < 0.0074). There was no significant difference in CTT values between T1 and T6 for bupivacaine, between T1 and T7 for lidocaine, and between T1 and T8 for oxybuprocaine. Duration of the maximum effect was 45 minutes for the bupivacaine, 55 minutes for the lidocaine and 65 minutes for the oxybuprocaine treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of a 0.5% injectable solution of bupivacaine or a 2% lidocaine had similar anaesthetic effect to the commonly used oxybuprocaine. Therefore, they might be used as alternatives for corneal anaesthesia.

Treatment of Localized Vulvar Pain with Neural Therapy: A Case Series and Literature Review.

BACKGROUND: Localized vulvar pain (LVP) is a common condition among fertile women, with physical and psychosexual implications. Treatment is complex with limited benefits. Neural therapy is a regulatory therapy that uses injections of local anesthetics in low concentrations in specific points to treat different conditions. CASE PRESENTATION: We present the cases of 5 women, ages 33-44 years, with LVP treated with procaine 0.5% injections in painful points. Complete relief from pain occurred in 2 patients, and significant improvement in 3. Only 1 or 2 sessions were required. Initial VAS score was ≥70 and decreased to ≤30 after the intervention. The improvement was maintained over time, with a minimum follow-up period of 6 months. None of the patients were able to have sex or use tampons due to pain, but they were able to resume after the intervention. CONCLUSIONS: In this case series, local injections of procaine showed a favorable outcome. Future randomized clinical trials could help elucidate the role of this intervention in LVP.

Use of the new preservation solution Custodiol-MP for ex vivo reconditioning of kidney grafts.

Organ shortage and the increasing use of extended criteria donor grafts for transplantation drives efforts for more efficient organ preservation strategies from simple cold storage toward dynamic organ reconditioning. The choice of a suitable preservation solution is of high relevance in different organ preservation or reconditioning situations. Custodiol-MP is a new machine perfusion solution giving the opportunity to add colloids according to organ requirements. The present study aimed to compare new Custodiol-MP with clinically established Belzer MPS solution. Porcine kidneys were ischemically predamaged and cold stored for 20 hours. Ex vivo machine reconditioning was performed either with Custodiol-MP (n = 6) or with Belzer MPS solution (n = 6) for 90 minutes with controlled oxygenated rewarming up to 20°C. Kidney function was evaluated using an established ex vivo reperfusion model. In this experimental setting, differences between both types of perfusion solutions could not be observed. Machine perfusion with Custodiol-MP resulted in higher creatinine clearance (7.4 ± 8.6 mL/min vs. 2.8 ± 2.5 mL/min) and less TNC perfusate levels (0.22 ± 0.25 ng/mL vs. 0.09 ± 0.08 ng/mL), although differences did not reach significance. For short-term kidney perfusion Custodiol-MP is safe and applicable. Particularly, the unique feature of flexible colloid supplementation makes the solution attractive in specific experimental and clinical settings.

HTK-N: Modified Histidine-Tryptophan-Ketoglutarate Solution-A Promising New Tool in Solid Organ Preservation.

To ameliorate ischemia-induced graft injury, optimal organ preservation remains a critical hallmark event in solid organ transplantation. Although numerous preservation solutions are in use, they still have functional limitations. Here, we present a concise review of a modified Histidine-Tryptophan-Ketoglutarate (HTK) solution, named HTK-N. Its composition differs from standard HTK solution, carrying larger antioxidative capacity and providing inherent toxicity as well as improved tolerance to cold aiming to attenuate cold storage injury in organ transplantation. The amino acids glycine, alanine and arginine were supplemented, N-acetyl-histidine partially replaced histidine, and aspartate and lactobionate substituted chloride. Several in vitro studies confirmed the superiority of HTK-N in comparison to HTK, being tested in vivo in animal models for liver, kidney, pancreas, small bowel, heart and lung transplantation to adjust ingredients for required conditions, as well as to determine its innocuousness, applicability and potential advantages. HTK-N solution has proven to be advantageous especially in the preservation of liver and heart grafts in vivo and in vitro. Thus, ongoing clinical trials and further studies in large animal models and consequently in humans are inevitable to show its ability minimizing ischemia-induced graft injury in the sequel of organ transplantation.

Reduction of Cold Ischemic Injury with the Addition of Compound Glycyrrhizin in HTK Solution in a Mouse Heart Transplantation Model.

Cold ischemic injury in heart storage is an important issue pertaining to heart transplantation. This study aims to evaluate the addition of compound glycyrrhizin (CG) in histidine-tryptophan-ketoglutarate (HTK) solution on chronic isograft injury in comparison to traditional HTK solution.Hearts of mouse were stored for 8 h in 4°C cold preservation solution and then transplanted heterotopically into mouse. Five groups were evaluated: HTK, low dose of CG solution (LCG), medium dose of CG solution (MCG), high dose of CG solution (HCG), and hearts without cold ischemia (sham). Survival was assessed. Time to restoration of heartbeat and strength of the heartbeat was measured. Lactate dehydrogenase (LDH) and creatine kinase (CK) levels in the preservation solution were determined. The myocardial damage and interstitial fibrosis of transplanted hearts were evaluated. TGF-ß1 expression in the transplanted hearts was assessed.Addition of CG to HTK solution significantly attenuated cold ischemic injury during cold storage, as evidenced by the lower time to restoration of heartbeat, higher strength of the heartbeat, lower LDH, and CK leakage. After transplantation, hearts stored in HTK solution containing CG had decreased the myocardial damage and interstitial fibrosis, compared with those stored without CG. The percentage of TGF-ß1-positive cells and TGF-ß1 level in the transplanted hearts were also decreased when stored in CG-containing HTK solution.The addition of CG to HTK solution attenuates cold ischemic injury during cold storage.

Pentadecapeptide BPC 157 shortens duration of tetracaine- and oxybuprocaine-induced corneal anesthesia in rats.

We focused on the relationship of 0.5% tetracaine- and 0.4% oxybuprocaine-induced corneal anesthesia in rats, and pentadecapeptide BPC 157 (0.4 µg/eye), along with nitric oxide synthase (NOS) inhibitor N(gamma)-nitro-L-arginine methyl ester (L-NAME) (0.1 mg/eye) and/or NOS substrate L-arginine (2 mg/eye), applied in the form of eye drops. We assessed corneal sensitivity recovery (Cochet-Bonnet esthesiometer), corneal lesion elimination (staining with 10% fluorescein) and decrease in tear volume (Schirmer test). BPC 157 administration had a full counteracting effect. Recovery also occurred in the presence of NOS blockade and NOS substrate application. L-arginine eventually shortened duration of corneal insensitivity and exerted corneal lesion counteraction (and counteraction of tetracaine-induced decrease of tear volume) only in earlier but not in later period. L-NAME application led to longer duration of corneal insensitivity, increase in corneal lesions and decrease in tear volume. When L-NAME and L-arginine were applied together, they antagonized each other's effect. These distinctions may indicate particular NOS involvement (corneal insensitivity vs. corneal lesion along with tear production), distinctively affected by the administration of NO agents. However, additional BPC 157 co-administration would re-establish counteraction over topical ophthalmic anesthetic-induced effect, be it in its early or late course. We suggest BPC 157 as an antidote to topical ophthalmic anesthetics.

pH-dependent effects of procaine on equine gamete activation†.

Procaine directly triggers pH-dependent cytokinesis in equine oocytes and induces hypermotility in stallion spermatozoa, an important event during capacitation. However, procaine-induced hyperactivated motility is abolished when sperm is washed to remove the procaine prior to sperm-oocyte co-incubation. To understand how procaine exerts its effects, the external Ca2+ and Na+ and weak base activity dependency of procaine-induced hyperactivation in stallion spermatozoa was assessed using computer-assisted sperm analysis. Percoll-washed stallion spermatozoa exposed to Ca2+-depleted (+2 mM EGTA) procaine-supplemented capacitating medium (CM) still demonstrated hyperactivated motility, whereas CM without NaCl or Na+ did not. Both procaine and NH4Cl, another weak base, were shown to trigger a cytoplasmic pH increase (BCECF-acetoxymethyl (AM)), which is primarily induced by a pH rise in acidic cell organelles (Lysosensor green dnd-189), accompanied by hypermotility in stallion sperm. As for procaine, 25 mM NH4Cl also induced oocyte cytokinesis. Interestingly, hyperactivated motility was reliably induced by 2.5-10 mM procaine, whereas a significant cytoplasmic cAMP increase and tail-associated protein tyrosine phosphorylation were only observed at 10 mM. Moreover, 25 mM NH4Cl did not support the latter capacitation characteristics. Additionally, cAMP levels were more than 10× higher in boar than stallion sperm incubated under similar capacitating conditions. Finally, stallion sperm preincubated with 10 mM procaine did not fertilize equine oocytes. In conclusion, 10 mM procaine causes a cytoplasmic and acidic sperm cell organelle pH rise that simultaneously induces hyperactivated motility, increased levels of cAMP and tail-associated protein tyrosine phosphorylation in stallion spermatozoa. However, procaine-induced hypermotility is independent of the cAMP/protein tyrosine phosphorylation pathway.

[Pain and distress responses of suckling piglets to injection and castration under local anaesthesia with procaine and lidocaine - Part 1: Cortisol, chromogranin A, wound healing, weights, losses]. / Schmerz- und Stressbestimmung bei der Injektion und Kastration von Saugferkeln unter Lokalanästhesie mit Procain und Lidocain.

OBJECTIVE: Comparison of the effectiveness of local anaesthesia (LA) in piglet castration by procaine 2 % and lidocaine 5 % both through combined inguinal and scrotal application as well as by intratesticular application of lidocaine 1 %. The parameters used were serum cortisol and chromogranin A (CgA) concentrations as well as wound healing, body weight and animal losses. MATERIAL AND METHODS: In 2 substudies, a total of 232 male piglets aged 3-6 days were included. Substudy 1 (112 piglets): Group H: fixation of piglets as for an injection; group L5: inguinal and scrotal injection of lidocaine 5 %; group P2: inguinal and scrotal injection of procaine 2 %; group L1: intratesticular injection of lidocaine 1 %. In all the groups, blood samples were taken 45 minutes before and 30 minutes post-injection (p. i.). Substudy 2 (120 piglets): Group H: handling only as for an injection and castration; group K: handling as for an injection and castration without LA after 30 minutes. Groups L5, P2 and L1: management as in substudy 1 and castration after 30 minutes. Blood samples were taken 75 minutes before as well as 30, 60 and 240 minutes post-castration (p. c.). The evaluated parameters were serum cortisol and CgA concentrations, wound healing, body weight and piglet losses. RESULTS: Substudy 1: The elevation of the cortisol and CgA concentrations in group P2 p. i. were significantly higher than in the other groups. The mean total cortisol concentration of group P2 p. i. was significantly higher than those of the other groups. Substudy 2: At 30 minutes p. c., all the groups displayed a significant increase in the cortisol concentration compared to group H. In group P2, the highest total cortisol concentration was measured 60 minutes p. c. and the elevation of the cortisol level was significantly higher than in the other groups. In group L1, a significantly greater increase in the CgA level was observed at 60 minutes p. c. when compared to the other groups. Regarding wound healing, body weight and losses, there were no significant differences between the groups. CONCLUSION: The combined inguinal and scrotal injection of procaine 2 % induced a greater neuroendocrine stress response than the inguinal and scrotal injection of lidocaine 5 % and the intratesticular injection of lidocaine 1 %. LA using procaine 2 %, lidocaine 5 % or lidocaine 1 % did not completely eliminate pain during castration. Castration under LA with procaine 2 % induced a greater pain reaction than castration without LA. Both groups castrated with LA using lidocaine (L1, L5) tended to display lower pain responses after castration than group K. On the basis of the findings of this study, other local anaesthetics that have a stronger effect could be further investigated according to their pain-killing effects in an appropriate application route.

Cold storage of porcine hepatocyte spheroids for spheroid bioartificial liver.

BACKGROUND AND AIMS: Cell-based therapies for liver disease such as bioartificial liver rely on a large quantity and high quality of hepatocytes. Cold storage was previously shown to be a better way to preserve the viability and functionality of hepatocytes during transportation rather than freezing, but this was only proved at a lower density of rat hepatocytes spheroids. The purpose of this study was to optimize conditions for cold storage of high density of primary porcine hepatocyte spheroids. METHODS: Porcine hepatocytes were isolated by a three-step perfusion method; hepatocyte spheroids were formed by a 24 hours rocked culture technique. Hepatocyte cell density was 5 × 106 /mL in 1000 mL spheroid forming medium. Spheroids were then maintained in rocked culture at 37°C (control condition) or cold stored at 4°C for 24, 48 or 72 hours in four different cold storage solutions: histidine-tryptophan-ketoglutarate (HTK) alone; HTK + 1 mM deferoxamine (DEF); HTK + 5 mM N-acetyl-L-cysteine (NAC); and HTK + 1 mM DEF + 5 mM NAC. The viability, ammonia clearance, albumin production, gene expression, and functional activity of cytochrome P450 enzymes were measured after recovery from the cold storage. RESULTS: In this study, we observed that cold-induced injury was reduced by the addition of the iron chelator. Viability of HTK + DEF group hepatocyte spheroids was increased compared with other cold storage groups (P < 0.05). Performance metrics of porcine hepatocyte spheroids cold stored for 24 hours were similar to those in control conditions. The hepatocyte spheroids in control conditions started to lose their ability to clear ammonia while production of albumin was still active at 48 and 72 hours (P < 0.05). In contrast, the viability and functionality of hepatocyte spheroids including ammonia clearance and albumin secretion were preserved in HTK + DEF group at both 48- and 72-hour time points (P < 0.05). CONCLUSIONS: The beneficial effects of HTK supplemented with DEF were more obvious after cold storage of high density of porcine hepatocyte spheroids for 72 hours. The porcine hepatocyte spheroids were above the cutoff criteria for use in a spheroid-based bioartificial liver.

Serotonin enhances oxybuprocaine- and proxymetacaine-induced cutaneous analgesia in rats.

The aim of the study was to investigate the analgesic effects of adding serotonin to oxybuprocaine or proxymetacaine preparations. We employed a rat model of the cutaneous trunci muscle reflex (CTMR) to conduct the dose-response curves and duration of drugs (oxybuprocaine, proxymetacaine, or serotonin) as an infiltrative anesthetic. The use of isobolographic methods to analyze the drug-drug interactions. We showed that oxybuprocaine and proxymetacaine, as well as serotonin produced dose-dependent skin antinociception. On the basis of 50% effective dose (ED50), the rank order of drug potency was serotonin [7.22 (6.45-8.09) µmol/kg] < oxybuprocaine [1.03 (0.93-1.15) µmol/kg] < proxymetacaine [0.59 (0.53-0.66) µmol/kg] (P < 0.01 for each comparison). The sensory block duration of serotonin was longer (P < 0.01) than that of oxybuprocaine or proxymetacaine at the equipotent doses (ED25, ED50, and ED75). The mixture of serotonin with oxybuprocaine or proxymetacaine produced a better analgesic effect than the drug itself. We have concluded that oxybuprocaine, proxymetacaine, or serotonin displays dose-related cutaneous analgesia. Oxybuprocaine or proxymetacaine is more potent and has a shorter duration of cutaneous analgesia than serotonin. Serotonin produces a synergistic antinociceptive interaction with oxybuprocaine or proxymetacaine.

Antiarrhythmic effect of sevoflurane as an additive to HTK solution on reperfusion arrhythmias induced by hypothermia and ischaemia is associated with the phosphorylation of connexin 43 at serine 368.

BACKGROUND: Reperfusion ventricular arrhythmia (RA) associated with hypothermic ischaemic storage is increasingly recognized as a substantial contributor to adverse consequences after heart transplantation. Ischemia- or hypothermia-induced gap junction (GJ) remodelling is closely linked to RA. Reducing GJ remodelling contributes to RA attenuation and is important in heart transplantation. However, sevoflurane has an antiarrhythmic effect associated with the connexin 43 (Cx43) protein that has not yet been fully established. METHODS: Hearts were divided into two groups according to a random number table: all hearts were arrested by an infusion of histidine-tryptophan-ketoglutarate (HTK) solution (4 °C) followed by (1) storage in HTK solution (4 °C) alone for 6 h (n = 8, Control group) or (2) storage in HTK solution supplemented with sevoflurane (2.5%) (4 °C) for 6 h (n = 8, Sevo-HTK group). First, the total Cx43 level and the phosphorylation of Cx43 at Ser368 (Cx43-pS368) were assessed by Western blotting, and the distribution of Cx43 was assessed by immunohistochemistry. Second, programmed electrical stimulation (PES) and monophasic action potential (MAP) recording were used to analyse the MAP duration (MAPD), conduction velocity (CV) and transmural repolarization dispersion (TDR). In addition, haematoxylin and eosin (HE) and terminal deoxynucleotidyl transferase-dUTP nick end labelling (TUNEL) staining were individually used to investigate the degree of myocardial pathological damage and cell apoptosis. Finally, bipolar electrograms were used to record the graft re-beating time and monitor RA during reperfusion for 15 to 30 min. RESULTS: Sevo-HTK solution relatively increased the total Cx43 (P < 0.01) and Cx43-pS368 (P < 0.01) levels and prevented Cx43 redistribution (P < 0.05) and CV slowing (P < 0.001) but did not change TDR (P > 0.05). Additionally, the Cx43-pS368/total Cx43 ratio (P>0.05) was similar in the two groups. However, with Sevo-HTK solution, the graft re-beating times were shortened, myocardial pathological damage was ameliorated, and the number of apoptotic cells was markedly decreased. CONCLUSION: The reduction in hypothermia and ischaemia-induced reperfusion arrhythmias by the addition of sevoflurane to HTK solution may be related to the phosphorylation of Cx43 at serine 368.

Using chloroprocaine for spinal anaesthesia in outpatient knee-arthroscopy results in earlier discharge and improved operating room efficiency compared to mepivacaine and prilocaine.

PURPOSE: Knee arthroscopies are regularly carried out in an outpatient setting. The purpose of this retrospective analysis was to investigate the impact of different local anaesthetics for spinal anaesthesia on operating room efficiency (perioperative process times) and postoperative recovery. This study aims to determine the optimal LA for SPA in patients undergoing knee arthroscopy at a day-surgery centre. METHODS: Anaesthesia records of all patients undergoing knee arthroscopy under spinal anaesthesia from 2010 until 2017 were analysed. Patients were categorised as having received spinal anaesthesia with prilocaine, mepivacaine or chloroprocaine. RESULTS: Three-hundred and nine patients were included. Postoperative recovery was significantly faster for chloroprocaine 1% compared with both other local anaesthetics regarding all stages of recovery until discharge. Perioperative processes and surgery time were significantly shorter when chloroprocaine was used. Early postoperative pain occurred more frequently and earlier after spinal anaesthesia with chloroprocaine. Nevertheless, pain intensity did not differ between groups. CONCLUSION: Spinal anaesthesia provides reliable blocks for outpatient knee arthroscopy. Considerations on the choice of local anaesthetic for spinal anaesthesia must include not only the recovery profile, but also the impact on operating room efficiency. Due to a superior recovery profile, low incidences of adverse side effects and raised operating room efficiency, chloroprocaine is the recommendable local anaesthetic for spinal anaesthesia in patients undergoing knee arthroscopy in an ambulatory setting. Since the frequency of SPA in patients undergoing outpatient knee arthroscopy is rising yearly, the results of this study are of high clinical relevance. The use of chloroprocaine leads to improved recovery, optimized perioperative processes and consecutively to a raised OR efficiency. LEVEL OF EVIDENCE: III.

Resistance to local anesthesia in people with the Ehlers-Danlos Syndromes presenting for dental surgery

BACKGROUND: People with the Ehlers-Danlos Syndromes (EDS), a group of heritable disorders of connective tissue, often report experiencing dental procedure pain despite local anesthetic (LA) use. Clinicians have been uncertain how to interpret this apparent LA resistance, as comparison of EDS and non-EDS patient experience is limited to anecdotal evidence and small case series. The primary goal of this hypothesis-generating study was to investigate the recalled adequacy of pain prevention with LA administered during dental procedures in a large cohort of people with and without EDS. A secondary exploratory aim asked people with EDS to recall comparative LA experiences. METHODS: We administered an online survey through various social media platforms to people with EDS and their friends without EDS, asking about past dental procedures, LA exposures, and the adequacy of procedure pain prevention. Among EDS respondents who both received LA and recalled the specific LA used, we compared agent-specific pain prevention for lidocaine, procaine, bupivacaine, mepivacaine, and articaine. RESULTS: Among the 980 EDS respondents who had undergone a dental procedure LA, 88% (n = 860) recalled inadequate pain prevention. Among 249 non EDS respondents only 33% (n = 83) recalled inadequate pain prevention (P < 0.001 compared to EDS respondents). The agent with the highest EDS-respondent reported success rate was articaine (30%), followed by bupivacaine (25%), and mepivacaine (22%). CONCLUSIONS: EDS survey respondents reported nearly three times the rate of LA non-response compared to non-EDS respondents, suggesting that LAs were less effective in preventing their pain associated with routine office dental procedures.

Emotional release and physical symptom improvement: a qualitative analysis of self-reported outcomes and mechanisms in patients treated with neural therapy.

BACKGROUND: Neural Therapy (NT) is a common complementary treatment approach using injections with short-acting local anesthetics to treat pain and chronic diseases. However, little is known about the underlying mechanisms and the domains of treatment response. This study therefore analyzed patient experiences following NT injections with procaine. METHODS: Maximum variation sampling was used to collect data from semi-structured interviews conducted with 22 hospital inpatients aged 59.6 ± 14.9 years (81.8% female). Each had multiple (9.4 ± 6.9) diagnoses. They were undergoing two weeks of integrative treatment, which included individualized NT. The interview data were analyzed in MAXQDA using qualitative content analysis. RESULTS: With injection, patients first described local anesthetic effects including temporary blocking of pain and increased local warmth. Second, patients reported on vegetative reactions frequently leading to turmoil within the body like initial aggravation of existing symptoms or the appearance of new, concealed or phantom symptoms. This often required the need for rest to deal with the treatment stimulus. As a third step, many patients could gain physical and emotional release and relief in symptoms, mood and functioning. Emotional release was often accompanied by weeping and initially overwhelmed affected patients with dissociated memories. However, in cases where patients were able to experience those memories with a new distance, a fourth step of integration was achievable. It included reframing processes as well as a gain in pain perception and body-awareness. As a possible fifth step, patients experienced improved mood, increased pain acceptance and empowerment. Adverse events of NT included pain from the injections, vegetative complaints and emotional turmoil that lasted for minutes or hours, with a maximum of two days. CONCLUSIONS: Patients treated with procaine injections reported different psychophysiological outcomes contributing to the understanding of the mechanisms underlying NT. Further efficacy studies should separate specific NT from non-specific/placebo effects. TRIAL REGISTRATION: DRKS00004567 .