An LC-MS/MS method for the determination of drugs of abuse included THC-COOH, EtG, and NPS, using a single hair extraction sample.

Hair analysis plays an important role in the determination of drugs of abuse in both forensic and clinical toxicology investigations. The analysis of different substances often requires the use of different sample preparation methods, thereby increasing the amount of hair sample and time required. In the present study, a fast method involving a combination of a single 25 mg hair extraction procedure and four liquid chromatography-tandem mass spectrometry methods using the same chromatographic phases and column was developed and validated. The target was the identification and quantification of various commonly abused drugs and their metabolites, including amphetamines, cocaine, opioids, cannabinoids, THC-COOH and EtG, and more than 140 new psychoactive substances, including synthetic cannabinoids, phenethylamines, synthetic opioids, methylphenidate, cathinone, piperidine, and tryptamines.

Comparison of two methods for the extraction of ethylglucuronide from hair.

The aim was the comparison between the Society of Hair Testing (SoHT) consensus for the use of alcohol markers which powdering hair for the extraction of ethylglucuronide (EtG) in water and extraction using the patented M3 Reagent Test kit on cut hair. Hair samples were cut into small segments and washed twice with methanol and diethyl ether. The SoHT-Consensus entails the extraction of pulverised hair in water. This is obtained by incubation of 25 mg of hair at room temperature overnight and 2 h sonication, even if the overnight incubation is not mandatory. The M3 method entails incubation of 25 mg of cut hair with the M3-Reagent at 100°C for 60 min. After centrifugation, the supernatant is injected into a liquid chromatography-tandem mass spectrometry (LC-MS/MS). Samples (191) were collected in the APSS laboratory in Trento, Italy, between 2021 and 2022. The limit of quantification (LOQ) was set at 5 pg/mg for the pulverised and M3-Reagent methods. Assays showed good linearity above the range of LOQ-300 pg/mg. Precision (within 20%) values were also obtained using both methods. In the Passing-Bablock linear regression, the final regression curve between M3 (y) and the pulverising method (x) showed good agreement; the Bland-Altman analysis did not show any significant bias between the two methods. The M3-Reagent method, due to cut hair use, is easy to perform, saves time and allows for a smaller sample quantity loss with use of nondisposable grinding jars for the ball mill to obtain the extraction of EtG.

Autogenous side-to-side brachial-basilic fistulas without vein transposition: a valuable option?

An autogenous brachial-basilic arteriovenous fistula (BBAVF) in the upper arm must be considered before placing prosthetic grafts in hemodialysis patients with multiple failures of forearm AVFs. The aim of this observational study was to compare technical and clinical outcomes of a new construction technique for BBAVF (n-BBAVF) with that of the standard one-stage side-artery to end-vein transposed BBAVF (t-BBAVF). A n-BBAVF is constructed in the following way: basilic vein and brachial artery are isolated. Patency of the proximal and distal vein is verified by injecting warmed (37 degrees C) saline solution. A venotomy and an arterotomy of 4-5 mm are performed. The two vessels are prepared for a side-to-side anastomosis without transposition of the vein. The latter allows both an antegrade and retrograde flow along the basilic vein, both proximally and distally to the anastomosis with more sites available for the venipunctures of the dialysis. Thirty BBAVFs were constructed as the secondary or tertiary vascular access in 30 patients over a 4-year period: 17 patients with adequate forearm basilic vein underwent the construction of a n-BBAVF; 13 underwent the construction of a t-BBAVF. The construction of a n-BBAVF requires a significantly lesser surgical time (55.0 +/- 9.0 minutes vs. 115.0 +/- 18.0, p < 0.0001), has fewer surgical complications (5.9% vs. 46.2%, p < 0.0001), and a reduced time to first use (24.5 +/- 6.3 vs. 37.7 +/- 9.1 days, p < 0.0001) than that of a t-BBAVF. n-BBAVFs showed a relatively low rate of thrombosis per patient-year at risk (0.067 at 1 year and 0.099 at 2 years). The latter was significantly lower at 1 year when compared with t-BBAVFs (0.067 vs. 0.285; p < 0.004). Our policy of "all AVFs should be autogenous" led us to the construction of a vascular access which is based on a side-to-side anastomosis between the brachial artery and the basilic vein without transposition of the vein allowing both antegrade and retrograde flow into the basilic vein. The results of this surgical technique appear satisfactory.

Cinacalcet is effective in relapses of secondary hyperparathyroidism after parathyroidectomy.

BACKGROUND: Relapses of secondary hyperparathyroidism (SHPTH) after parathyroidectomy (PTx) in haemodialysis patients are relatively frequent. Calcimimetics (cinacalcet HCl) offer a new therapeutic opportunity for their treatment. However, no data about the treatment with cinacalcet of relapses of SHPTH after PTx are available in literature. The aim of this single-centre prospective study was to evaluate the therapeutic efficacy of cinacalcet in this high-risk category of patients. METHODS: Twelve haemodialysis patients of our Dialysis Unit had a relapse of SHPTH after PTx, defined as serum levels of immunoreactive intact parathyroid hormone (iPTH)>300 pg/ml. They were stratified into a treatment group (the six patients having the highest serum levels of iPTH) and a control group (the remaining six patients): the former were treated for 6 months with a dose of cinacalcet ranging from 30 mg every other day to 60 mg a day; the latter continued to be administered the conventional treatment. Serum levels of albumin, iPTH, calcium (Ca), phosphate (P) and alkaline phosphatase were determined monthly. The treatment group included four cases of nodular hyperplasia and two cases of carcinoma of parathyroid glands, whereas the control group included four cases of nodular hyperplasia and two cases of diffuse hyperplasia. RESULTS: At the start of the study, the six patients treated with cinacalcet showed a more severe picture of biochemical abnormalities when compared with the control patients. After 6 months of treatment, a statistically significant reduction in the serum levels of iPTH, Ca, P and CaxP product was obtained only in the patients treated with cinacalcet. Symptomatic episodes of hypocalcaemia (serum Ca<7.0 mg/dl) were observed in three patients of this group. The six patients undergoing the conventional treatment showed at 6 months a not significant decrease in the mean serum levels of iPTH and a not significant increase in the mean serum levels of Ca, P and CaxP product, when compared with the baseline values. CONCLUSIONS: Our single-centre prospective study, even though small and of short duration, shows that cinacalcet is effective also in controlling relapses of SHPTH after PTx, thus representing a solid, and sometimes unique, therapeutic opportunity for this high-risk category of patients.

Is there a place for duplex screening of the brachial artery in the maturation of arteriovenous fistulas?

Clinically there are some autogenous arteriovenous fistulas (AVFs) that are obviously mature. The real problem in clinical evaluation is in predicting the ultimate outcome of AVFs that are not clearly mature. Thus it would be advantageous to develop objective quantitative criteria to be applied early after vascular access placement in order to evaluate the suitability of AVFs for dialysis. The goal of this study was to document the blood flow rate modifications that the construction and maturation of a radiocephalic wrist AVF produce in the brachial artery by means of duplex Doppler ultrasonography. All incident uremic patients who needed the construction of a radiocephalic wrist AVF in the last 9 months of 2003 were enrolled in the study: 18 patients underwent such an operation. A linear color Doppler ultrasound scan was performed with a 7.0 MHz imaging/5.0 MHz Doppler probe by sampling the brachial artery 2 cm above the elbow: the internal diameter of the artery was measured and its blood flow rate calculated just before AVF construction and 1, 7, 28 days, and at least 6 months after AVF construction. The internal diameter and blood flow rate of the brachial artery were, respectively, 4.3 +/- 0.7 mm and 56.1 +/- 19.2 ml/min at baseline. A new AVF was constructed in one patient whose brachial artery blood flow rate was 80.0 ml/min at 28 days. When excluding this AVF, the mean brachial artery blood flow rate of the 17 AVFs was 720.4 +/- 132.8 ml/min (median 750 ml/min, range 480-890 ml/min) at 28 days and 997.6 +/- 259.7 ml/min 258.0 +/- 63.0 days after AVF construction. When analyzing the percent increase in brachial artery blood flow rate of the 17 AVFs at the different time points, the most dramatic one occurred at day 1 compared to the baseline (549.0%; mean blood flow rate at day 1, 365.0 +/- 129.3 ml/min). Thus the blood flow rate at day 1 represents more than half (50.7%) of the blood flow rate that will be measured at day 28. Then the increase was less steep, with a 20.1% increase between day 7 and day 1 (mean blood flow rate at day 7, 438.4 +/- 86.0 ml/min), a 64.3% increase between day 28 and day 7, and a 38.5% increase at 258.0 +/- 63.0 days compared to 28 days. The present study was able to document the changes in brachial blood flow rate consequent to a radiocephalic wrist AVF maturation by means of duplex Doppler ultrasonography of the brachial artery. This measure may be helpful in monitoring which AVFs will probably fail. This screening should integrate clinical assessment, thus allowing sound judgment of the level of maturation of an AVF and of its outcome.

End-stage renal disease in leprosy.

BACKGROUND: Leprosy or Hansen's disease (HAD) undoubtedly remains an emergency in certain countries. It is an ancient deforming disease caused by Mycobacterium leprae. The countries with the highest endemic leprosy rate in 2000 were Brazil, India and Madagascar. In Italy, the old epidemic has been defeated and there are approximately 400 patients under constant monitoring with three to four new cases per year involving Italian residents. The kidney is one of the target organs during the splanchnic localization of leprosy. The histopathological renal lesion spectrum includes glomerulonephritis (GN), renal amyloidosis (RA) and interstitial nephritis (IN). Both proteinuria and chronic renal failure are the main clinical expressions of renal damage in leprosy. To the best of our knowledge, very little is reported concerning end-stage renal disease (ESRD) in leprosy patients both in the most important national and international renal registries and in the available literature. This study aimed to report the long-term experience of our department in this field. METHODS: To achieve this, we analyzed retrospectively the HAD Center (Gioia del Colle) database at ourhospital. RESULTS: Eight leprosy patients were dialyzed from 1980 to June 2003 (six males and two females), with a mean age of 61.0+/-8.9 SD yrs (range: 51-76) and a mean HAD duration of 36.1+/-5.1 yrs. The first clinical nephropathymanifestations were non-nephrotic proteinuria associated with chronic renal failure in four patients, and nephrotic proteinuria in four patients. Kidney biopsies performed in three patients showed two had RA, and one had IN. Two patients were treated initially by peritoneal dialysis; they were then switched to hemodialysis (HD) after 3 and 10 months because of recurrent peritonitis. HD treatment lasted 40.6+/-31.4 months (range: 9-101). Six patients died, one due to hyperkalemia, one because of a technical dialysis accident, and the remainder due to causes unrelated to the dialysis treatment. Two patients are still alive, treated with HD for 17 and 44 months. CONCLUSIONS: Uremia represents a late complication of leprosy and has a multifactorial genesis, although RA is among the most frequent causes, conventional bicarbonate HD appears to offer good results in the treatment of uremia in leprosy patients.