An Explorative Study on Calcium Electroporation for Low-risk Basal Cell Carcinoma.

In electrochemotherapy, permeabilization of the cell membrane by electric pulses increases the anti-tumour effect of chemotherapeutics. In calcium electroporation, chemotherapy is replaced by calcium chloride with obvious benefits. This study explores the effect and underlying mechanisms of calcium electroporation on basal cell carcinomas using either high- or low-frequency electroporation. Low-risk primary basal cell carcinomas were treated in local anaesthesia with intratumoral calcium chloride followed by electroporation with high (167 kHz) or low (5 kHz) frequencies. Non-complete responders were retreated after 3 months. The primary endpoint was tumour response 3 months after last calcium electroporation. Plasma membrane calcium ATPase was examined in various cell lines as plasma membrane calcium ATPase levels have been associated with calcium electroporation efficacy. Twenty-two out of 25 included patients complete the study and 7 of these (32%) achieved complete response at 3 months with no difference in efficacy between high- and low-frequency pulses. High-frequency calcium electroporation was significantly less painful (p=0.03). Plasma membrane calcium ATPase was increased 16-32-fold in basal cell carcinoma cell lines compared with 4 other cancer cell lines. Calcium electroporation for low-risk basal cell carcinomas does not fulfil the requirements of a new dermatological basal cell carcinoma treatment but may be useful as adjuvant treatment to surgery in more advanced basal cell carcinomas. The elevated PMCA levels in basal cell carcinomas may contribute to low efficacy.

Excessive EP4 Signaling in Smooth Muscle Cells Induces Abdominal Aortic Aneurysm by Amplifying Inflammation.

OBJECTIVE: Excessive prostaglandin E2 production is a hallmark of abdominal aortic aneurysm (AAA). Enhanced expression of prostaglandin E2 receptor EP4 (prostaglandin E receptor 4) in vascular smooth muscle cells (VSMCs) has been demonstrated in human AAAs. Although moderate expression of EP4 contributes to vascular homeostasis, the roles of excessive EP4 in vascular pathology remain uncertain. We aimed to investigate whether EP4 overexpression in VSMCs exacerbates AAAs. Approach and Results: We constructed mice with EP4 overexpressed selectively in VSMCs under an SM22α promoter (EP4-Tg). Most EP4-Tg mice died within 2 weeks of Ang II (angiotensin II) infusion due to AAA, while nontransgenic mice given Ang II displayed no overt phenotype. EP4-Tg developed much larger AAAs than nontransgenic mice after periaortic CaCl2 application. In contrast, EP4fl/+;SM22-Cre;ApoE-/- and EP4fl/+;SM22-Cre mice, which are EP4 heterozygous knockout in VSMCs, rarely exhibited AAA after Ang II or CaCl2 treatment, respectively. In Ang II-infused EP4-Tg aorta, Ly6Chi inflammatory monocyte/macrophage infiltration and MMP-9 (matrix metalloprotease-9) activation were enhanced. An unbiased analysis revealed that EP4 stimulation positively regulated the genes binding cytokine receptors in VSMCs, in which IL (interleukin)-6 was the most strongly upregulated. In VSMCs of EP4-Tg and human AAAs, EP4 stimulation caused marked IL-6 production via TAK1 (transforming growth factor-ß-activated kinase 1), NF-κB (nuclear factor-kappa B), JNK (c-Jun N-terminal kinase), and p38. Inhibition of IL-6 prevented Ang II-induced AAA formation in EP4-Tg. In addition, EP4 stimulation decreased elastin/collagen cross-linking protein LOX (lysyl oxidase) in both human and mouse VSMCs. CONCLUSIONS: Dysregulated EP4 overexpression in VSMCs promotes inflammatory monocyte/macrophage infiltration and attenuates elastin/collagen fiber formation, leading to AAA exacerbation.

Effect of Intravenous or Intraosseous Calcium vs Saline on Return of Spontaneous Circulation in Adults With Out-of-Hospital Cardiac Arrest: A Randomized Clinical Trial.

Importance: It is unclear whether administration of calcium has a beneficial effect in patients with cardiac arrest. Objective: To determine whether administration of calcium during out-of-hospital cardiac arrest improves return of spontaneous circulation in adults. Design, Setting, and Participants: This double-blind, placebo-controlled randomized clinical trial included 397 adult patients with out-of-hospital cardiac arrest and was conducted in the Central Denmark Region between January 20, 2020, and April 15, 2021. The last 90-day follow-up was on July 15, 2021. Interventions: The intervention consisted of up to 2 intravenous or intraosseous doses with 5 mmol of calcium chloride (n = 197) or saline (n = 200). The first dose was administered immediately after the first dose of epinephrine. Main Outcomes and Measures: The primary outcome was sustained return of spontaneous circulation. The secondary outcomes included survival and a favorable neurological outcome (modified Rankin Scale score of 0-3) at 30 days and 90 days. Results: Based on a planned interim analysis of 383 patients, the steering committee stopped the trial early due to concerns about harm in the calcium group. Of 397 adult patients randomized, 391 were included in the analyses (193 in the calcium group and 198 in the saline group; mean age, 68 [SD, 14] years; 114 [29%] were female). There was no loss to follow-up. There were 37 patients (19%) in the calcium group who had sustained return of spontaneous circulation compared with 53 patients (27%) in the saline group (risk ratio, 0.72 [95% CI, 0.49 to 1.03]; risk difference, -7.6% [95% CI, -16% to 0.8%]; P = .09). At 30 days, 10 patients (5.2%) in the calcium group and 18 patients (9.1%) in the saline group were alive (risk ratio, 0.57 [95% CI, 0.27 to 1.18]; risk difference, -3.9% [95% CI, -9.4% to 1.3%]; P = .17). A favorable neurological outcome at 30 days was observed in 7 patients (3.6%) in the calcium group and in 15 patients (7.6%) in the saline group (risk ratio, 0.48 [95% CI, 0.20 to 1.12]; risk difference, -4.0% [95% CI, -8.9% to 0.7%]; P = .12). Among the patients with calcium values measured who had return of spontaneous circulation, 26 (74%) in the calcium group and 1 (2%) in the saline group had hypercalcemia. Conclusions and Relevance: Among adults with out-of-hospital cardiac arrest, treatment with intravenous or intraosseous calcium compared with saline did not significantly improve sustained return of spontaneous circulation. These results do not support the administration of calcium during out-of-hospital cardiac arrest in adults. Trial Registration: ClinicalTrials.gov Identifier: NCT04153435.

Surgical castration versus chemical castration in donkeys: response of stress, lipid profile and redox potential biomarkers.

BACKGROUND: Castration is a husbandry practice raising important questions on the welfare and physiological status of farm animals. Searching for effective castration methods that minimally compromise the body physiology is worthy of attention. Therefore, this study aimed to evaluate the differential response of biological systems in donkeys to surgical castration versus the chemical one by CaCl2 with special emphasis on stress, lipid profile, and oxidative stress biomarkers. Donkeys were divided randomly and equally into two groups; the chemical (Ch) and surgical (S) groups (n = 6). The Ch group was chemically castrated by intratesticular injection of 20% CaCl2 dissolved in absolute ethanol. Blood samples were collected prior to castration and at 15, 30, 45, and 60 days after the beginning of experiment. RESULTS: Surprisingly, the Ch group at the end of the experiment was characterized by significantly higher cortisol level compared to the S group. TC and LDL-C levels in the S group significantly decreased at day 45, while TG levels significantly increased at days 45 and 60 in comparison with day 0. HDL-C levels at days 30 and 60 in the Ch group significantly increased in comparison with day 0. At day 30 post-castration, HDL-C was significantly higher and LDL-C was significantly lower in the Ch group than the S group. A significant elevation in TC and LDL-C was observed at day 45 and in HDL-C at the end of experimental duration in the Ch group when compared with the S group. TPX level was significantly lower and TAC was significantly higher in the Ch group at day 45 than the S group. CONCLUSION: Surgical castration evoked less stress and minor changes in lipid profile and oxidant/antioxidant balance relative to chemical castration by intratesticular 20% CaCl2 dissolved in absolute ethanol.

Solving the calcium gluconate shortage in real-time: Mistakes made and lessons learned.

PURPOSE: During a national shortage of calcium gluconate, we switched to calcium chloride for routine supplementation for peripheral blood stem cell (PBSC) collections. Subsequently, we analyzed the postprocedure ionized calcium level, as we aimed for an equivalent result compared to before the shortage. METHODS: Pharmacy representatives helped us to find an "equivalent" substitute for calcium gluconate at 46.5 mEq in 500 mL normal saline, infused at 100 mL/hour. After instituting a presumably comparable protocol using calcium chloride (40.8 mEq in 250 mL normal saline at a rate of 100 mL/hour), we reviewed ionized calcium results post-PBSC procedures to compare with those obtained with calcium gluconate. Having noticed a difference in the mean values, we adjusted the rate of calcium chloride to reach our desired outcome. RESULTS: Twenty-seven procedures were analyzed on 15 unique patients. We used the Spectra OPTIA with a whole blood: anticoagulant ratio of 13:1. Ionized calcium levels post-PBSC collection with the first calcium chloride protocol were significantly higher (P = 0.003) in nine patients treated. Subsequently, we decreased the calcium chloride infusion rate to 75 mL/hour and achieved similar mean levels to calcium gluconate (P = 0.382). CONCLUSION: Changes in replacement fluids for apheresis procedures can be complex, particularly when dealing with electrolytes that could be clinically significant at critically high or low levels. Once we recognized the need to take into account the amount of elemental calcium infused, we achieved the desired postprocedure ionized calcium results. This study can serve as a lesson for future shortages of infusions used during apheresis procedures.

Effect of treatment with oral Ca boluses following calving on concentrations of Ca in serum in pasture-based dairy cows.

AIMS To assess the effect of the administration of two oral Ca boluses on concentrations of total Ca, ß-hydoxybutyrate (BHB) and non-esterified fatty acids (NEFA) in serum, and urine pH, in recently calved pasture-fed dairy cows. METHODS Friesian or Friesian cross Jersey cows from one dairy farm were blocked by age and randomly assigned to no treatment (control; n=14), or treatment (n=13) with two oral Ca boluses administered approximately 12 hours apart, with the first bolus being given within 14 hours of calving. Each bolus weighed 198 g and contained 43 g of Ca; 31 g of Ca from calcium chloride and 12 g of Ca from calcium sulfate. Cows were enrolled over three calendar days, and all cows were managed in one group during the 24-hour study period. Blood samples were collected at 0, 1, 2, 4, 8, 12, 13, 14, 16, 20 and 24 hours after the initial treatment. Serum from each time point was analysed for concentrations of total Ca, and from 0, 12, and 24 hours for NEFA and BHB. Urine was collected at 0, 12 and 24 hours for pH measurement and pH was categorised as <7 or ≥7. The effect of treatment on percentage change in concentrations of Ca in serum relative to 0 hours, and concentrations of NEFA, BHB and urine pH, was examined using multivariable repeated measures mixed models with cow as a random effect. RESULTS In the final multivariable model for percentage change in concentrations of Ca, there was an interaction between time and treatment (p=0.004), with the percentage increase being higher in treatment than control cows at 1, 2, 4, 8 and 13 hours. At 12 hours, 5/13 (41%) treated cows had a urine pH <7compared to 0/12 (0%) control cows (p<0.001), and at 24 hours 13/13 (100%) treated cows had urine pH <7 compared to 0/12 (0%) control cows (p<0.001). Over the 24-hour period, mean concentrations of NEFA or BHB in serum were similar in treated and control cows (p>0.3). CONCLUSIONS AND CLINICAL RELEVANCE Oral treatment with two Ca boluses increased concentrations of total Ca in serum and decreased urine pH in pasture-fed cows. This bolus has the potential to reduce the prevalence and duration of subclinical hypocalcaemia in recently calved cows.

Regional citrate anticoagulation in membrane based plasma exchange: Safety, efficacy and effect on calcium balance.

AIM: To assess the efficacy, safety and calcium balance of a membrane based regional citrate anticoagulation plasma exchange protocol. METHODS: This was an observational, prospective, single centre study of membrane separation plasma exchange using regional citrate anticoagulation. It was performed using a fixed dose pre-filter citrate infusion that was based on the plasma flow rate. Patients received a post filter calcium infusion that was modified during treatment based on systemic ionized calcium monitoring. Post filter ionized calcium was not assessed. Safety and efficacy were assessed by extraction of clinical events and laboratory data contemporaneously recorded in electronic health records. RESULTS: Thirty-six sessions in five patients were performed. No patients developed symptomatic hypocalcaemia, and no patient had a recorded ionized calcium below 0.81 mmol/L. Filter clotting occurred in two sessions. The mean net calcium gained was 9.6 ± 1.8 mmol per session. CONCLUSION: Regional citrate anticoagulated membrane separation plasma exchange can be performed safely and effectively without the need for post filter ionized calcium monitoring. The algorithm employed resulted in a net calcium gain.

Effects of oral administration of acidogenic boluses at dry-off on performance and behavior of dairy cattle.

The aim of this study was to evaluate the potential of oral acidogenic mineral boluses (196 g) containing anionic salts to facilitate the transition from lactation to the dry stage by inducing a mild and temporary metabolic acidosis at dry-off. In experiment 1, 84 lactating cows were randomly allocated to 1 of 3 treatment groups consisting of an oral administration of 0, 1, or 2 boluses 5 d before dry-off to evaluate the effects on milk production. In experiment 2, 16 lactating cows were involved in a crossover study to evaluate the effects of the administration of 2 boluses on milk production, feed intake, and urine pH. In experiment 3, 152 lactating cows were allocated to 1 of 2 treatments (control: no treatment; bolus: 2 oral boluses the day before last milking) to evaluate udder pressure, incidence of milk leakage, and lying behavior during the first days following dry-off. Also, milk yield in the subsequent lactation for all enrolled cows was recorded during the first 60 DIM. In experiment 1, cows receiving 2 boluses had the greatest reduction in milk production (-2.56 kg/d of milk) compared with those receiving 1 bolus or no treatment (-1.15 and -0.23 kg/d, respectively) the second day after bolus application. In experiment 2, the application of oral boluses decreased feed intake of cows during the first 3 d following treatment, and milk production was reduced on d 2 and 3 after bolus application. Reduced urine pH at 8 and 24 h after treatment was observed in bolus cows compared with control cows. In experiment 3, bolus cows had lower udder pressure after drying off, but incidence of milk leakage did not differ between treatments. Bolus cows had an additional 85 min of lying time in the 24 h following dry-off. Serum P and ß-OH-butyrate concentrations were lower in bolus cows than in control cows after dry-off, but no other differences in blood parameters between treatments were observed. Also, no differences in milk yield in the subsequent lactation were observed between treatments. It is concluded that oral bolus application diminishes feed intake and milk production, and, if applied at dry-off, it decreases udder pressure and increases lying time during the first 24 h after dry-off.

Calcium chloride combined with dimethyl sulphoxide for the chemical sterilization of dogs.

The aim of this study was to evaluate the effect of intratesticular injection of calcium chloride (CaCl2 ) combined with dimethyl sulphoxide (DMSO) as a chemical sterilization in dogs. Twelve dogs were divided into two groups: the treated group (n = 6), in which 15 mg/kg of a 7.5% CaCl2 solution combined with 0.5% DMSO was injected into each testicle (volume range 1.0-4.76 ml); and a control group (n = 6) that received the same volume/kg of 0.9% sodium chloride solution (NaCl). Semen characteristics pre- and post-treatment were evaluated. Serum testosterone concentration was determined before the injection (D-1) and at 15 (D15), 30 (D30) and 60 (D60) days after intratesticular injection. Testicle sizes and local pain were evaluated for 7 consecutive days (D1 to D7) and at D15, D30 and D60 after injection. At D60, testicle histological evaluation was performed after orchiectomy. No pain was observed by testicular palpation, with the exception of one dog in the treated group; this dog then received analgesic therapy. An increase in testicular volume was evident within 24 hr after treatment, followed by gradual reduction for 3 weeks. Five of 6 dogs from the treated group presented azoospermia at D15; the remaining dog presented at D30. There was no significant difference in testosterone concentrations in the treated group during the experimental period. Histological evaluation showed testicular degenerative lesions, especially at the proximal and middle portions. The results indicated that one injection of 7.5% CaCl2 combined with 0.5% DMSO into each testis is a viable alternative for canine sterilization.

Implications of CaCl2 application to plants in LID facilities.

Low impact development (LID) technologies mimic the natural water cycle through the physico-chemical and biological interactions of plants, filter media and soil, and microorganisms, thereby reducing the release of pollutants. In LID facilities, plants carry out photosynthesis, facilitate microbial growth, and uptake pollutants contained in stormwater runoff. However, de-icers (CaCl2) used to melt snow during winter slow the growth of plants and even increase plant mortality. In addition, de-icers change the soil structure, causing changes in soil content and affecting the growth of plants and microorganisms. Therefore, this study examined the effects of CaCl2 on the resistance of plants, the removal efficiency of non-point source pollutants, and water circulation. The mortality rate of the tree and shrubs caused by CaCl2 was found to be in the order of Rhododendron indicum > Spiraea prunifolia var. simpliciflora > Metasequoia glyptostroboides. For herbaceous plants, mortality rate was in the order of Pratia pedunculata > Aquilegia japonica > Tagetes erecta > Sedum makinoi aurea > Hosta longipes > Dianthus chinensis > Acorus gramineus > Liriope platyphylla. In addition, it was found that the amount of chlorophyll decreases with high concentrations of CaCl2. The findings of this research will be useful for plant selection considering CaCl2 concentrations applied to paved areas during the winter.

Heparin-Free Prolonged Intermittent Hemodialysis Using Calcium-Free Citrate Dialysate in Critically Ill Patients.

OBJECTIVES: Critically ill patients who have a high risk of bleeding but require prolonged intermittent dialysis need a heparin-free easy-to-use alternative type of anticoagulation within the dialysis circuit. We assessed the safety and efficiency of heparin-free regional citrate anticoagulation of the dialysis circuit using a calcium-free citrate-containing dialysate, with calcium reinjected according to ionic dialysance. DESIGN: Prospective cohort study. SETTING: Critical care units. PATIENTS: Critically ill patients who required renal replacement therapy. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 101 dialysis sessions were performed in 35 patients (mechanical ventilation n = 78; norepinephrine n = 13). Median duration of dialysis was 294 minutes (interquartile range, 240-300), and median ultrafiltration volume was 2.3 L (1-2.8). Urea and ß2-microglobulin reduction rates were 64.5% ± 0.4% and 48% ± 0.13%, respectively. Postfilter ionized calcium was 0.35 ± 0.17 and 0.38 ± 0.14 mmol/L at 1 and 3 hours, respectively, within the extracorporeal circuit. A major clotting event that led to premature termination of the session occurred in only three of 101 sessions. In these three cases, major catheter dysfunction occurred before clotting within the circuit. Prefilter ionized calcium remained within narrow ranges (before/after change +0.07 ± 0.006 mmol/L), and total-to-ionized calcium ratio, a surrogate marker for citratemia, was unchanged. CONCLUSIONS: Dialysis anticoagulation with calcium-free citrate-containing dialysate and calcium reinjection according to ionic dialysance is an easy-to-use, efficient, and inexpensive form of heparin-free regional anticoagulation. It allows prolonged hemodialysis sessions in critically ill patients without the need to systemically monitor ionized calcium. Furthermore, sessions can be safely extended according to the hemodynamic tolerance to ensure an adequate dose of dialysis and a negative water balance, a major point in patients with severe acute kidney disease.

Effects of a Lidocaine-Loaded Poloxamer/Alginate/CaCl2 Mixture on Postoperative Pain and Adhesion in a Rat Model of Incisional Pain.

BACKGROUND: Pain and adhesion are problematic issues after surgery. Lidocaine has analgesics and anti-inflammatory properties, and poloxamer/alginate/CaCl2 (PACM) is a known antiadhesive agent. We hypothesized that the novel combination of lidocaine as chemical barrier and PACM as physical barrier would be beneficial for both postoperative pain and adhesion. The purpose of this study was to investigate the effects of lidocaine-loaded PACM in a rat model of incisional pain. Primary outcome was to evaluate between-group differences for the mechanical withdrawal threshold (MWT) measured by von Frey filament in various concentrations of lidocaine-loaded PACM applied, PACM applied, and sham-operated groups. METHODS: Male Sprague-Dawley rats were used for the postoperative pain model. After plantar incision and adhesion formation, 0.5%, 1%, 2%, and 4% lidocaine-loaded PACM, PACM only, nothing, and 4% lidocaine only were applied at the incision site in groups PL0.5, PL1, PL2, PL4, P, S, and L4, respectively. MWT using a von Frey filament and serum levels of tumor necrosis factor-α, interleukin (IL)-1ß, IL-6, and high-sensitivity C-reactive protein were measured. Rats were euthanized 2 weeks after surgery, and inflammation and fibrosis were assessed with microscopy. Data were analyzed using the Kruskal-Wallis test, multivariate analysis of variance, and linear mixed-effect model. To compare MWT at each time point, analysis of variance with Bonferroni correction was used. RESULTS: Multivariate analysis of variance showed that 4% lidocaine-loaded PACM significantly raised the MWT up to 6 and 8 hours after surgery compared with lidocaine-unloaded groups S and P, respectively; 2% lidocaine-loaded PACM significantly increased the MWT at 4 hours after surgery compared with groups S and C. Linear mixed-effect model showed that the MWT (estimated difference in means [95% confidence interval]) was significantly increased in groups PL2 and PL4 (6.58 [2.52-10.63], P = .002; 11.46 [7.40-15.51], P < .001, respectively) compared with group P. Inflammation and fibrosis seen on microscopic evaluation were significantly decreased in groups PL2 and PL4 compared with group S. Four percent of lidocaine only showed a significant reduction in inflammation. Serum levels of tumor necrosis factor-α, IL-1ß, IL-6, and high-sensitivity C-reactive protein were decreased in lidocaine-loaded groups compared with group S or P at 1, 2, and 48 hours, and 2 weeks after surgery, respectively. CONCLUSIONS: Lidocaine-loaded PACM reduced postoperative pain, and lidocaine strengthened the antiadhesive effect of PACM.

Platelet-Rich Plasma for the Treatment of Clean Diabetic Foot Ulcers.

BACKGROUND: Diabetic foot ulcer is considered as a major health problem that predisposes to limb amputation. Among the different methods to achieve ulcer healing, platelet-rich plasma (PRP) gel is gaining popularity. It is thought to stimulate wound closure by providing essential growth factors for healing. This study aims to evaluate the value of autologous PRP gel in the treatment of diabetic ulcers. METHODS: The study included 56 patients of both sex from 18 to 80 years, with clean chronic diabetic foot ulcers divided into 2 equal groups. The first group was treated by antiseptic ointment dressing, and the second group was treated by autologous platelet gel. PRP together with thrombin were prepared by centrifugation at each dressing session. Thrombin and calcium chloride were used to activate the PRP. The formed platelet gel was applied to the wound twice weekly. RESULTS: Statically significant increase in healing rate was found in the PRP-treated group, and complete healing was achieved in 86% of them in comparison to 68% of the control group. In the study group, rate of healing per week was greater during the first 8 weeks and starts to decline afterward. The use of platelet gel showed a lower rate of wound infection. CONCLUSIONS: Autologous platelet gel is more effective than the local antiseptic dressing in terms of healing rate and prevention of infection in clean diabetic ulcers.

Effects of calcium chloride coadministered with neostigmine on neuromuscular blockade recovery: A double-blind randomised study.

BACKGROUND: Ionised calcium plays an important role in neuromuscular transmission, but its effects on the reversal of nondepolarising neuromuscular blockade have not been fully evaluated. OBJECTIVE: We examined whether calcium chloride coadministered with neostigmine could enhance the rate of neuromuscular recovery. DESIGN: Randomised double-blind trial. SETTING: A tertiary teaching hospital. PATIENTS: In total, 53 patients undergoing elective surgery under general anaesthesia with neuromuscular monitoring by acceleromyography using a TOF-Watch SX monitor. INTERVENTIONS: Patients were randomly allocated to receive either 5 mg kg of calcium chloride (calcium group, n = 26) or the same volume of normal saline (control group, n = 27) coadministered with 25 µg kg of neostigmine and 15 µg kg of atropine at the end of surgery. MAIN OUTCOME MEASURES: The primary end point was the neuromuscular recovery time [time from neostigmine administration to recovery of the TOF ratio (TOFr) to 0.9]. Secondary end points included the TOFr at 5, 10 and 20 min after neostigmine administration and the incidence of postoperative residual curarisation (PORC), defined as a TOFr less than 0.9 at each time point. RESULTS: The neuromuscular recovery time was significantly faster in the calcium group than in the control group (median [Q1 to Q3]; 5.0 [3.0 to 7.0] vs. 6.7 [5.7 to 10.0] min, respectively; P = 0.007). At 5 min after neostigmine administration, the TOFr was higher [87 (74 to 100) vs. 68 (51 to 81)%, respectively; P = 0.002] and the incidence of PORC was lower (50.0 vs. 81.5%, respectively; P = 0.016) in the calcium group than in the control group. There were no differences between the two groups with respect to the TOFr or incidence of PORC at 10 and 20 min after neostigmine administration. CONCLUSION: Calcium chloride coadministered with neostigmine enhanced neuromuscular recovery in the early period of nondepolarising neuromuscular blockade reversal.

Hemostatic Wound Dressing for Postinterventional Hemostasis in Large Femoral Artery Access Sites: An Initial Efficacy and Safety Study.

PURPOSE: To present the results of a prospective single-center study that evaluated the safety and efficacy of a hemostatic dressing following femoral artery access. METHODS: Within a 9-month period, 80 patients (mean age 68±14 years; 55 men) were treated with a hemostatic dressing patch (Hematrix Active Patch) containing aminocaproic acid, calcium chloride, and thrombin after endovascular procedures via a 6- to 8-F femoral artery access. After removing the sheath, the wound dressing was placed on the puncture site followed by constant manual compression adapted to the sheath size (specified pressure times: 8 minutes for 6-F, 9 minutes for 7-F, and 10 minutes for 8-F). Patients were treated with an additional pressure bandage for 24 hours. Hemostasis was checked clinically and with duplex ultrasound after patch removal and at 24 hours. Patient characteristics [platelets, systolic blood pressure, international normalized ratio (INR), and partial thromboplastin time (PTT)], sheath sizes, and approach direction were compared among patients with successful hemostasis (within specified pressure times) vs those with prolonged compression. RESULTS: A total of 39 6-F, 19 7-F, and 22 8-F sheaths were employed. In 73 (91.2%) of 80 patients, hemostasis was reached within the prespecified pressure times (mean 8.8±0.8 minutes). In 7 patients (4 6-F, 1 7-F, 2 8-F) a longer compression time was necessary (mean 34±30 minutes). No serious major complication occurred. Twelve (15.0%) minor and 5 (6.3%) moderate subcutaneous hematomas were observed. Two (2.5%) false aneurysms were treated successfully. Ambulation and discharge was possible within 24 hours in 79 (98.7%) cases. Patients with initial hemostasis and those with prolonged compression did not differ substantially (p>0.05) according to sheath size, approach direction, INR (1.09±0.3 vs 1.11±0.3), platelets (234±47×10(3)/µL vs 249±93×10(3)/µL), systolic blood pressure (150±26 vs 152±17 mm Hg), or PTT (31±7.9 vs 34.8±10.0 seconds). CONCLUSION: The evaluated wound dressing seems to be safe and effective in reducing time to hemostasis in large arterial access sites. However, a randomized trial with a larger population and an active control group is necessary to confirm these preliminary data. Moreover, additional focus on shortening the time to ambulation is required in future studies.

Evaluation of chemical castration with calcium chloride versus surgical castration in donkeys: testosterone as an endpoint marker.

BACKGROUND: For the last few years, researchers have been interested in developing a method for chemical sterilization which may be a better alternative to surgical castration. An ideal chemical sterilant would be one that effectively arrests spermatogenesis and androgenesis as well as libido with absence of toxic or other side effects. Calcium chloride in various solutions and concentrations has been tested in many animal species, but few studies have been evaluated it in equines as a chemical sterilant. So, the objective of this study was to evaluate the clinical efficacy of chemical castration with 20% calcium chloride dissolved in absolute ethanol in comparison with surgical castration in donkeys based on the changes in the serum testosterone level and the histopathological changes in treated testes. METHODS: Twelve clinically healthy adult male donkeys were used in this study. Donkeys were divided randomly and equally into two groups: a surgical (S) group (n = 6) and a chemical (C) group (n = 6). Animals in the (S) group were subjected to surgical castration while those in the (C) group received a single bilateral intratesticular injection of 20% calcium chloride dissolved in absolute ethanol (20 ml/testis). Animals were kept under clinical observation for 60 days. Changes in animals' behavior and gross changes in external genitalia were monitored daily. Serum concentrations of testosterone were measured prior to treatment and at 15, 30, 45 and 60 days post-treatment. Testicles in the (C) group were examined histopathologically at the end of the experiment. RESULTS: Chemical castration with intratesticular calcium chloride vs. surgical castration failed to reduce serum concentrations of testosterone throughout the whole duration of the study; however it induced orchitis that was evident by focal necrotic areas in seminiferous tubules, cellular infiltration of neutrophils, proliferative intertubular fibrosis with a compensatory proliferation of Leydig cells. Donkeys tolerated the intratesticular injection of calcium chloride. There were no detectable changes in the general health status of the animals with the exception of swelling in external genitalia, scrotal ulcerations and fistulas. Food and water consumption and the gait of animals remained unaffected. CONCLUSION: Intratesticular calcium chloride can't be considered an effective method for chemical castration in donkeys.

Effects of oral calcium supplementation on mineral and acid-base status, energy metabolites, and health of postpartum dairy cows.

Two experiments were conducted to characterize blood concentrations of minerals and acid-base status after oral dosing of Ca salts and to determine the effects of oral Ca on mineral and metabolic status and incidence diseases. The hypotheses were that administration of oral Ca as CaCl2 and CaSO4 maintains blood total Ca (tCa) concentrations ≥2.125 mM and reduces the incidence of diseases in early lactation. In experiment 1, 18 Holstein cows on the day of calving were assigned to receive a single dose of 0, 43, or 86g of Ca as an oral bolus. Blood was sampled before and after treatments to characterize acid-base status and concentrations of minerals. In experiment 2, 450 Holstein cows considered of low (LRM; normal calving) or high risk (HRM; dystocia, twins, stillbirth, retained placenta, vulvo-vaginal laceration, or a combination of these) of metritis (primiparous-LRM=84; primiparous-HRM=84; multiparous-LRM=138; multiparous-HRM=138) on the day of calving were blocked by parity and then randomly assigned to control, no Ca supplementation; 86g of Ca on d 0 and 1 postpartum (CaS1); or 86g of Ca on d 0 and 1 postpartum followed by 43g/d on d 2 to 4 postpartum (CaS4). Blood was sampled before and 30 min after treatment on d 0, and 30 min after treatments on d 1 to 4, and d 7 and 10 for determination of concentrations of minerals and metabolites and blood acid-base responses. Disease incidence was evaluated for the first 30 DIM. Concentrations of ionized Ca (iCa) increased for 2h in cows supplemented with 43g of Ca and fewer than 8h in cows supplemented with 86g of Ca. The changes in iCa concentrations from pretreatment to 30 min after 86g of Ca supplemented on d 0 were 0.11±0.03 mM in multiparous cows and 0.25±0.03 mM in primiparous cows. Oral Ca reduced the incidence of subclinical hypocalcemia (SCH; tCa <2.125mM) in the first 4 d in the experiment (control=69.3%; CaS1=57.5%; CaS4=34.2%). Calcium supplementation decreased the prevalence of SCH on d 0 and 1 postpartum in all cows. Stopping oral Ca in CaS1 on d 1 postpartum, however, caused a rebound in SCH on d 2 to 4 postpartum in primiparous cows. Oral Ca increased the incidence of metritis (control=22.7%; CaS1=34.8%; CaS4=32.8%), primarily because of an increase in LRM primiparous cows (control=17.9%; CaS1=35.7%; CaS4=42.9%). Oral Ca increased morbidity in primiparous cows (control=38.1%; CaS1=61.8%; CaS4=60.3%) but had no effect on multiparous cows (control=38.2%; CaS1=35.1%; CaS4=30.1%). Large doses of oral Ca as salts of chloride and sulfate in the first days postpartum should be avoided in primiparous cows and used only in cows at risk of clinical hypocalcemia.

Initial Observations of the Effects of Calcium Chloride Infusions in Pediatric Patients with Low Cardiac Output.

Myocardial contractility and relaxation are highly dependent on calcium homeostasis. Immature myocardium, as in pediatric patients, is thought to be more dependent on extracellular calcium for optimal function. For this reason, intravenous calcium chloride infusions may improve myocardial function in the pediatric patient. The objectives of this study were to report the hemodynamic changes seen after administration of continuous calcium chloride to critically ill children. We retrospectively identified pediatric patients (newborn to 17 years old) with hemodynamic instability admitted to the cardiac ICU between May 2011 and May 2012 who received a continuous infusion of calcium chloride. The primary outcome was improvement in cardiac output, assessed by arterial-mixed venous oxygen saturation (A-V) difference. Sixty-eight patients, mean age 0.87 ± 2.67 years, received a total of 116 calcium infusions. Calcium chloride infusions resulted in significant improvements in primary and secondary measures of cardiac output at 2 and 6 h. Six hours after calcium initiation, A-V oxygen saturation difference decreased by 7.4 % (32.6 ± 2.1 to 25.2 ± 2.0 %, p < 0.001), rSO2 increased by 5.5 % (63.1 vs 68.6 %, p < 0.001), and serum lactate decreased by 0.9 mmol/l (3.3 vs 2.4 mmol/l, p < 0.001) with no change in HR (149.1 vs 145.6 bpm p = 0.07). Urine output increased 0.66 ml/kg/h in the 8-h period after calcium initiation when compared to pre-initiation (p = 0.003). Neonates had the strongest evidence of effectiveness with other age groups trending toward significance. Calcium chloride infusions improve markers of cardiac output in a heterogenous group of pediatric patients in a cardiac ICU. Neonates appear to derive the most benefit from utilization of these infusions.

Micromolar changes in lysophosphatidylcholine concentration cause minor effects on mitochondrial permeability but major alterations in function.

Mice deficient in group 1b phospholipase A2 have decreased plasma lysophosphatidylcholine and increased hepatic oxidation that is inhibited by intraperitoneal lysophosphatidylcholine injection. This study sought to identify a mechanism for lysophosphatidylcholine-mediated inhibition of hepatic oxidative function. Results showed that in vitro incubation of isolated mitochondria with 40-200µM lysophosphatidylcholine caused cyclosporine A-resistant swelling in a concentration-dependent manner. However, when mitochondria were challenged with 220µM CaCl2, cyclosporine A protected against permeability transition induced by 40µM, but not 80µM lysophosphatidylcholine. Incubation with 40-120µM lysophosphatidylcholine also increased mitochondrial permeability to 75µM CaCl2 in a concentration-dependent manner. Interestingly, despite incubation with 80µM lysophosphatidylcholine, the mitochondrial membrane potential was steady in the presence of succinate, and oxidation rates and respiratory control indices were similar to controls in the presence of succinate, glutamate/malate, and palmitoyl-carnitine. However, mitochondrial oxidation rates were inhibited by 30-50% at 100µM lysophosphatidylcholine. Finally, while 40µM lysophosphatidylcholine has no effect on fatty acid oxidation and mitochondria remained impermeable in intact hepatocytes, 100µM lysophosphatidylcholine inhibited fatty acid stimulated oxidation and caused intracellular mitochondrial permeability. Taken together, these present data demonstrated that LPC concentration dependently modulates mitochondrial microenvironment, with low micromolar concentrations of lysophosphatidylcholine sufficient to change hepatic oxidation rate whereas higher concentrations are required to disrupt mitochondrial integrity.

Comparatively Evaluating Medication Preparation Sequences for Treatment of Hyperkalemia in Pediatric Cardiac Arrest: A Prospective, Randomized, Simulation-Based Study.

OBJECTIVES: To determine whether time to prepare IV medications for hyperkalemia varied by 1) drug, 2) patient weight, 3) calcium salt, and 4) whether these data support the Advanced Cardiac Life Support recommended sequence. DESIGN: Prospective randomized simulation-based study. SETTING: Single pediatric tertiary medical referral center. SUBJECTS: Pediatric nurses and adult or pediatric pharmacists. INTERVENTIONS: Subjects were randomized to prepare medication doses for one of four medication sequences and stratified by one of three weight categories representative of a neonate/infant, child, or adult-sized adolescent: 4, 20, and 50 kg. Using provided supplies and dosing references, subjects prepared doses of calcium chloride, calcium gluconate, sodium bicarbonate, and regular insulin with dextrose. Because insulin and dextrose are traditionally prepared and delivered together, they were analyzed as one drug. Subjects preparing medications were video-recorded for the purpose of extracting timing data. MEASUREMENTS AND MAIN RESULTS: A total of 12 nurses and 12 pharmacists were enrolled. The median (interquartile range) total preparation time for the three drugs was 9.5 minutes (6.4-13.7 min). Drugs were prepared significantly faster for larger children (50 kg, 6.8 min [5.6-9.1 min] vs 20 kg, 9.5 min [8.6-13.0 min] vs 4 kg, 16.3 min [12.7-18.9 min]; p = 0.001). Insulin with dextrose took significantly longer to prepare than the other medications, and there was no difference between the calcium salts: (sodium bicarbonate, 1.9 [0.8-2.6] vs calcium chloride, 2.1 [1.2-3.1] vs calcium gluconate, 2.4 [2.1-3.0] vs insulin with dextrose, 5.1 min [3.7-7.7 min], respectively; p < 0.001). Forty-two percent of subjects (10/24) made at least one dosing error. CONCLUSIONS: Medication preparation for hyperkalemia takes significantly longer for smaller children and preparation of insulin with dextrose takes the longest. This study supports Pediatric Advanced Life Support guidelines to treat hyperkalemia during pediatric cardiac arrest similar to those recommended per Advanced Cardiac Life Support (i.e., first, calcium; second, sodium bicarbonate; and third, insulin with dextrose).