HPTLC assay of nicotine and cotinine in biological samples

This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges. Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction. The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis

Persistent left superior vena cava. Prenatal ultrasound diagnosis. Obstetrical approach and considerations in childhood and adulthood.

Centrul Medical Dr. Gh. Iliev, Iaşi, România, Departamentul de Neonatologie, UMF ”Gr.T. Popa” Iasi, România, Laboratorul de Anatomie Patologică, SCOG ”Cuza Vodă”, Iaşi, România, Departmentul de Genetică Umană, of Neonatology, Iaşi, România, Al VI-lea Congres Național de Obstetrică și Ginecologie cu participare internațională, 13-15 septembrie 2018, Chișinău, Republica MoldovaObiective. Vena cavă superioară stânga persistentă (VCSSP) este cea mai frecventă anomalie venoasă toracică. Identificarea postnatală a VCSSP în ecocardiografia transtoracică este dificilă. În general, la copii şi adulţi, malformaţia este descoperită accidental in timpul cateterizării cardiace prin vena subclaviculară stânga (VSCS). Prezentăm importanţa ecocardiografi ei fetale în diagnosticul prenatal al VCSSP şi implicaţiile clinice prenatale şi postnatale ale acesteia. Material şi metode. Am efectuat studiul în perioada 01.01.2008-31.07.2018. Am practicat de rutină ecocardiografia fetală aprofundată în cadrul morfologiei fetale din trimestrul doi şi trei de sarcină conform recomandărilor The International Society of Ultrasound in Obstetrics & Gynecology (ISUOG) Practice Guidelines (2006, 2013). Am identificat VCSSP în incidenţa trei vase ca un vas supranumerar situat în stânga trunchiului pulmonar. Am identificat în incidenţa patru camere sinusul coronar (SC) dilatat. Rezultate. Am diagnosticat 33 de cazuri cu VCSSP: A) 6 cazuri asociate cu malformaţii cardiace severe complexe, dintre care 2 cazuri cu agenezie de venă cavă superioară dreapta (VCSD); B) 4 cazuri asociate cu defect septal ventricular, dintre care 1 caz cu agenezie de VCSD; C) 10 cazuri asociate cu malformaţii extra-cardiace; D) 13 cazuri cu VCSSP izolată, dintre care 2 cazuri cu agenezie de VCSD. Conduita obstetricală a fost stabilită în funcţie de complexitatea cazurilor. Concluzii. Diagnosticul ecografic prenatal al VCSSP este facil. Conduita obstetricală este stabilită în funcţie de complexitatea cazurilor. În acelaşi timp, la copii sau adulţi, identificarea prenatală a VCSSP permite evitarea dificultăţilor sau a complicaţiilor la cateterizarea cordului drept prin VSCS.Summary. Objectives. The persistent left superior vena cava (PLSVC) is the most common thoracic vein abnormality. Postnatal identification of PLSVC in transthoracic echocardiography is difficult. Generally, in children and adults, the malformation is accidentally detected during cardiac catheterization via the left subclavian vein (LSCV). We present the importance of fetal echocardiography in prenatal diagnosis of PLSVC and its prenatal and postnatal clinical implications. Material and methods. We conducted the study between 1st January 2008 – 31st July 2018. We routinely performed fetal echocardiography in the second and third trimester of pregnancy as recommended by The International Society of Ultrasound in Obstetrics and Gynecology Practice Guidelines (2006, 2013). We identified PLSVC in the three vessels view as a supernumerary vessel to the left of the pulmonary trunk. We have identified dilated coronary sinus in the four-chamber view. Results. We have diagnosed 33 cases with PLSVC: A) 6 cases associated with severe cardiac malformations, of which 2 cases with right superior vena cava agenesis (RSVC); B) 4 cases associated with the ventricular septal defect, of which 1 case with RSVC agenesis; C) 10 cases associated with extra-cardiac pathology; D) 13 cases with isolated PLSVC, of which 2 cases with RSVC agenesis. The obstetrical approach was determined according to the complexity of the cases. Conclusions. The prenatal ultrasound diagnosis of PLSVC is easy. The obstetrical approach is determined according to the complexity of the cases. At the same time, in children or adults, prenatal identification of PLSVC allows avoiding difficulties or complications in case of right heart catheterization by LSCV

HPTLC assay of nicotine and cotinine in biological samples

This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges. Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction. The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis

Colorectal Cancer and Inflammatory Bowel Disease

Inflammatory bowel disease (IBD) with its two entities, ulcerative colitis and Crohn’s disease, is at increased risk of developing colorectal cancer (CRC). Risk factors for CRC are represented by the duration of the disease, extent of disease, the association of primary sclerosing cholangitis, family history, and early age at onset. In inflammatory bowel disease, colonic carcinogenesis appears on an inflamed colon, being determined by different genetic alterations. The main element of the process of carcinogenesis is the dysplasia, which is a neoplastic intraepithelial transformation, limited to the basal membrane surrounding the glands around which it appears. The stages of carcinogenesis process start with dysplasia of varying degrees as follows: indefinite dysplasia, low-grade dysplasia, high-grade dysplasia, and finally invasive adenocarcinoma

THE PHYSIOLOGICAL EFFECTS PLAYING BEACH HANDBALL HAS ON JUNIOR HANDBALL INDOOR PLAYERS

Beach handball is a branch of sports that was born on the beaches of Italy in the early 1990s and it was designed to help players under go their physical training in a more dynamic training during the summer. Objectives. The main goal of this study is to describe the physiological state of the athletes that underwent a series of beach handball training sessions and to determine the effects of such training sessions on junior level beach handball athletes. Materials and methods. We have suggested that the evaluation of the athletes’ condition be determined via invasive methods, such as the analysis of their urinary value. Results. The numerical data obtained from our analysis were computed using Student’s t-test for unpaired samples associated with Welch’s unequal variance t-test. The statistical significance was determined for an associated P 95%). The comparison was made between the mean of the initial values and the mean of the final value for all the dosed biochemical markers. The data are expressed as mean ± SD (standard deviation). The statistical processing was done using the GraphPad Prism software. Conclusions. Including beach handball practice sessions in the training program of indoor handball players has no significant effect on their physiological state. Article history: Received 2022 September 20; Revised 2022 September 27; Accepted 2022 September 28; Available online 2022 November 20; Available print 2022 December 20. REZUMAT. Efectele practicării jocului de Beach Handball la nivel fiziologic asupra jucătorilor de handbal junior. Jocul de Beach Handball este o ramură sportivă nouă care a luat naștere pe plajele din Italia la începutul anilor 1990 cu intenția de a face pregătirea fizică a jucătorilor de handbal mult mai dinamică pe perioada verii. Obiective. Obiectivul principal al studiului a fost conturarea statusului fiziologic al sportivilor supuși unor serii de antrenamente organizate de Beach Handball și determinarea efectelor fiziologice în urma aplicării jocului de Beach Handball în pregătirea jucătorilor de handbal la nivel de juniori. Materiale și metode. Am propus evaluare stării sportivilor prin metode invazive, cum ar fi determinarea valorii unitare a acestora. Rezultate. Datele numerice obținute în urma determinărilor efectuate au fost prelucrate cu testul t Student pentru valori neperechi asociat cu corecția lui Welch. Semnificația statistică a fost stabilită la un P asociat 95%). Comparația s-a efectuat între media valorilor inițiale și media valorilor finale, pentru toți markerii biochimici dozați. Datele sunt exprimate ca medie ± ES (eroarea standard a mediei). Prelucrarea statistică a datelor s-a efectuat folosind softul GraphPad Prism 5. Concluzii. Introducerea antrenamentelor de Beach Handball în programul de pregătire al jucătoarelor de handbal (indoor) nu influențează statusul fiziologic a acestora. Cuvinte cheie: handbal pe plajă, juniori, stare fiziologică

Mechanical Characterization and In Vitro Assay of Biocompatible Titanium Alloys

Metals that come into contact with the body can cause reactions in the body, so biomaterials must be tested to avoid side effects. Mo, Zr, and Ta are non-toxic elements; alloyed with titanium, they have very good biocompatibility properties and mechanical properties. The paper aims to study an original Ti20Mo7ZrxTa system (5, 10, 15 wt %) from a mechanical and in vitro biocompatibility point of view. Alloys were examined by optical microstructure, tensile strength, fractographic analysis, and in vitro assay. The obtained results indicate very good mechanical and biological properties, recommending them for future orthopedic medical applications

HPTLC assay of nicotine and cotinine in biological samples

This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges. Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction. The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis

CELIAC DISEASE CASE FINDING STRATEGY IN ROMANIAN SYMPTOMATIC CHILDREN

Objectives. Pediatricians face various challenges in different stages of celiac disease (CD) diagnosis. Whom to test is intensely debated because of CD’s heterogenous clinical spectrum. The main purpose of the study was to identify symptoms or symptom associations that should initiate an active strategy of CD early diagnosis in Romanian children. Material and method. We conducted a prospective study in “Grigore Alexandrescu” Emergency Children’s Hospital. From March 2013 until February 2014, 249 children with symptoms/signs at risk of CD were included. Results. CD was diagnosed in 11 (1/21 patients evaluated). One in 12.6; 16; 18; 18.5 and 18.5 children respectively with chronic diarrhea, low stature, growth failure, recurrent abdominal pain and constipation had CD. Certain symptom associations increased the risk: classical symptom associations (chronic diarrhea and weight loss), as well as other associations: recurrent abdominal pain and weight loss, constipation and weight loss, constipation and refractory iron deficiency anemia. Conclusion. Active screening among patients with symptoms and especially symptom associations at risk of CD would improve diagnosis rates in pediatric CD

Novel Mg-0.5Ca-xMn biodegradable alloys intended for orthopedic application: an in vitro and in vivo study

Mg-based biodegradable materials, used for medical applications, have been extensively studied in the past decades. The in vitro cytocompatibility study showed that the proliferation and viability (as assessed by quantitative MTT-assay—3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyl tetrazolium bromide) were not negatively affected with time by the addition of Mn as an alloying element. In this sense, it should be put forward that the studied alloys don’t have a cytotoxic effect according to the standard ISO 10993-5, i.e., the level of the cells’ viability (cultured with the studied experimental alloys) attained both after 1 day and 5 days was over 82% (i.e., 82, 43–89, 65%). Furthermore, the fibroblastic cells showed variable morphology (evidenced by fluorescence microscopy) related to the alloy sample’s proximity (i.e., related to the variation on the Ca, Mg, and Mn ionic concentration as a result of alloy degradation). It should be mentioned that the cells presented a polygonal morphology with large cytoplasmic processes in the vicinity of the alloy’s samples, and a bipolar morphology in the remote region of the wells. Moreover, the in vitro results seem to indicate that only 0.5% Mn is sufficient to improve the chemical stability, and thus the cytocompatibility; from this point of view, it could provide some flexibility in choosing the right alloy for a specific medical application, depending on the specific parameters of each alloy, such as its mechanical properties and corrosion resistance. In order to assess the in vivo compatibility of each concentration of alloy, the pieces were implanted in four rats, in two distinct body regions, i.e., the lumbar and thigh. The body’s reaction was followed over time, 60 days, both by general clinical examinations considering macroscopic changes, and by laboratory examinations, which revealed macroscopic and microscopic changes using X-rays, CT(Computed Tomography), histology exams and SEM (Scanning Electron Microscopy). In both anatomical regions, for each of the tested alloys, deformations were observed, i.e., a local reaction of different intensities, starting the day after surgery. The release of hydrogen gas that forms during Mg alloy degradation occurred immediately after implantation in all five of the groups examined, which did not affect the normal functionality of the tissues surrounding the implants. Imaging examinations (radiological and CT) revealed the presence of the alloy and the volume of hydrogen gas in the lumbar and femoral region in varying amounts. The biodegradable alloys in the Mg-Ca-Mn system have great potential to be used in orthopedic applications