Intraplaque Neovascularization, CD68+ and iNOS2+ Macrophage Infiltrate Intensity Are Associated with Atherothrombosis and Intraplaque Hemorrhage in Severe Carotid Atherosclerosis.

BACKGROUND: Atherosclerosis is a progressive disease that results from endothelial dysfunction, inflammatory arterial wall disorder and the formation of the atheromatous plaque. This results in carotid artery stenosis and is responsible for atherothrombotic stroke and ischemic injury. Low-grade plaque inflammation determines biological stability and lesion progression. METHODS: Sixty-seven cases with active perilesional inflammatory cell infiltrate were selected from a larger cohort of patients undergoing carotid endarterectomy. CD68+, iNOS2+ and Arg1+ macrophages and CD31+ endothelial cells were quantified around the atheroma lipid core using digital morphometry, and expression levels were correlated with determinants of instability: ulceration, thrombosis, plaque hemorrhage, calcification patterns and neovessel formation. RESULTS: Patients with intraplaque hemorrhage had greater CD68+ macrophage infiltration (p = 0.003). In 12 cases where iNOS2 predominated over Arg1 positivity, the occurrence of atherothrombotic events was significantly more frequent (p = 0.046). CD31 expression, representing neovessel formation, correlated positively with atherothrombosis (p = 0.020). CONCLUSIONS: Intraplaque hemorrhage is often described against the background of an intense inflammatory cell infiltrate. Atherothrombosis is associated with the presence of neovessels and pro-inflammatory macrophages expressing iNOS2. Modulating macrophage polarization may be a successful therapeutic approach to prevent plaque destabilization.

Cervical Pleuropulmonary Blastoma-like Tumor Associated With DICER1 and TP53 Mutations.

We describe a very unusual cervical tumor in a 12-yr-old patient with a clinical history indicative of DICER1 syndrome. Morphologic, immunohistochemical, and molecular genetic analysis together helped to diagnose this lesion as a cervical pleuropulmonary blastoma-like tumor, associated with TP53 and DICER1 mutations. The tumor displayed usual histologic features including mixtures of embryonal rhabdomyosarcoma, sarcomatous cartilage, compact blastema, primitive spindle cells and anaplasia, akin to type III pleuropulmonary blastoma, and trabecular and retiform patterns. In addition to expanding the phenotypic spectrum of DICER1 -associated conditions, we draw attention to genotype-phenotype correlations in DICER1 -associated tumors, particularly as they relate to the discovery of a heritable tumor predisposition syndrome.

When a Chorangioma Becomes a Burden in Fetal Survival: A Reported Case with an Updated Literature Review.

Chorangioma is a rare non-trophoblastic benign vascular neoplasm originating from the primitive chorionic mesenchyme. Usually asymptomatic, it affects approximately 1% of female fetuses. We present the case of a giant placental chorangioma (GPC) in a preterm male pregnancy coexisting with a maternal neuroendocrine carcinoma. A 30-week primigravida was admitted to the Obstetrics and Gynecology Clinic of the Targu-Mures Emergency Clinical Hospital, with abdominal discomfort, and an emergency C-section was performed for fetal congestive heart failure. Medical history revealed an advanced-stage rectal neuroendocrine carcinoma. At 20th gestational week, a well-vascularized placental mass was diagnosed. A 1500g premature male fetus was delivered. Histopathologically, the placental mass revealed an unencapsulated but well-circumscribed tumor with lobular architecture composed of congested vascular capillaries and thin-walled vessels. Diagnosis of giant placental chorangioma (GPC) was rendered. GPC is a challenging condition typically occurring in hypertensive or diabetic primigravidas with female fetuses. Antenatal management is suggested at an early stage for a desirable perinatal outcome.

Perinatal Prognosis of Pregnancies with Single Umbilical Artery in a Romanian Third-Level Unit.

Objectives: Single umbilical artery (SUA) is considered the most common abnormality of the umbilical artery. The objective of the study was to evaluate the perinatal prognosis of fetuses with SUA and to describe the associated malformations. The significance of the study is represented by examining whether our findings are in correlation with data already described.Methods: We performed a prospective cohort study on singleton pregnancies complicated with SUA. The study population was composed of women with singleton pregnancies who were examined at the Department of Obstetrics and Gynecology of the Târgu Mures County Emergency Clinical Hospital between 2012 and 2021.Results: The incidence of SUA in the study population was 0.48%. C-section was performed in 40 cases with SUA and in 5258 cases with no SUA (RR:1.56, P<0.05.) From the total number of 2249 premature deliveries, 23 newborns were diagnosed with SUA (RR:2.12, P<0.05.) From the total number of 869 deliveries with low birth weight (LBW) newborns, 13 were associated with SUA (RR: 3.12, P<0.05.) There were 206 pregnancies noted with antenatal fetal demise after 24 weeks of gestation, and only 2 of them were with SUA (RR:2.01, P>0.05.) Fetal and neonatal malformations were described in 290 cases, and 28 were associated with SUA (R:21.96, P<0.05.) In 57 of 85 cases (67.05%), we found iSUA, and 28 newborns (32.95%) had minor, major, or other associated pathologies. We found two cases of trisomy 18 and one case with trisomy 13 associated with SUA. Investigating the malformations associated with SUA, the most common were cardiac and great vessels malformations (12), followed by limb malformations (8), urogenital malformations (7), digestive tract malformations (7), central nervous system malformations (4), and in one case we found cleft palate.Conclusions: Perinatal prognosis regarding SUA is significantly poorer than in cases without this pathology. One-third of fetuses with SUA were associated with fetal anomalies. The most common pathologies associated with SUA were cardiovascular, limb, urogenital, and digestive system malformations. Our data are similar to those described in other studies; therefore, we conclude, we can implement the general recommendations in our region regarding counselling patients.

Management of recurrent vulvovaginal candidosis: Narrative review of the literature and European expert panel opinion.

Recurrent vulvovaginal candidosis (RVVC) is a chronic, difficult to treat vaginal infection, caused by Candida species, which affects women of all ages and ethnic and social background. A long-term prophylactic maintenance regimen with antifungals is often necessary. In most clinical practice guidelines, oral fluconazole is recommended as the first-line treatment. Although clinical resistance to antifungal agents remains rare, overexposure to azoles may increase the development of fluconazole-resistant C. albicans strains. In addition, non-albicans Candida species are frequently dose-dependent susceptible or resistant to fluconazole and other azoles, and their prevalence is rising. Available therapeutic options to treat such fluconazole-resistant C. albicans and low susceptibility non-albicans strains are limited. Ten experts from different European countries discussed problematic issues of current RVVC diagnosis and treatment in two audiotaped online sessions and two electronic follow-up rounds. A total of 340 statements were transcribed, summarized, and compared with published evidence. The profile of patients with RVVC, their care pathways, current therapeutic needs, and potential value of novel drugs were addressed. Correct diagnosis, right treatment choice, and patient education to obtain adherence to therapy regimens are crucial for successful RVVC treatment. As therapeutic options are limited, innovative strategies are required. Well- tolerated and effective new drugs with an optimized mechanism of action are desirable and are discussed. Research into the impact of RVVC and treatments on health-related quality of life and sex life is also needed.

Primary angiosarcoma of the vulva: report of a case misdiagnosed as a benign tumor.

Primary vulvar angiosarcomas have a propensity for varied macroscopic and histologic appearance that initially may not suggest a vascular malignant neoplasm. Therefore, the rarity of the lesion and it's morphologic diversity may contribute to the high rate of misdiagnosis. We present the case of a 43-year old patient with a primary vulvar lesion, initially misdiagnosed as an angiofibroma. Microscopic examination of the recurrence together with immunohistochemical profile were in favor on a poorly differentiated angiosarcoma. Early diagnoses can improve prognoses in angiosarcomas and, in the case of recurrences, as in the present case, may lead to changes in therapy.

Oxidative stress and peripartum outcomes (Review).

Pregnancy, labor and childbirth are accompanied by excessive oxidative aggression. The excessive formation of free radicals [reactive oxygen species (ROS), reactive nitrogen species (RNS), chlorine reactive species (CRS)] causes cellular oxidative damage, which can be scavenged by enzymatic or non-enzymatic antioxidants in normal healthy pregnancy, physiological labor and delivery without any complications. An imbalance between the pro-oxidant and antioxidant factors may lead to oxidative stress, which contributes to the development of many diseases. This oxidative aggression can be a precursor for pathologies in the pregnant woman including eclampsia, miscarriage, preterm labor, and intrauterine growth retardation; in the offspring it may lead to bronchopulmonary dysplasia/chronic lung disease, necrotizing enterocolitis, retinopathy of prematurity, and periventricular leukomalacia. This review summarizes the studies conducted to identify the mechanisms of oxidative stress and the effect of cell membrane oxidation, the mechanisms that are behind oxidative stress-related diseases, and also those studies which have demonstrated the effect of antioxidants in preventing diseases or diminishing the effects of oxidative stress in the body, in obstetrics and neonatology.

Adolescent smoking and the social capital of local communities in three counties in Romania / El consumo de tabaco en adolescentes y el capital social de las comunidades sociales en tres condados de Rumania

Objective: To identify the effect of social capital on adolescent smoking. Method: A stratified random sample of 1313 7th and 8th grade students from three counties in Transylvania, Romania, completed a self-administered questionnaire on smoking-related knowledge, attitudes and behaviours. The impact of social capital was measured (personal and community activities, school achievements and smoking-related knowledge). Multivariate multinomial logistic regression models were used to measure the association between social participation and smoking. Results: Experimenting with smoking was mostly related to knowledge about smoking, academic performance and second-hand tobacco smoke exposure at home. The strongest risk factor of adolescent smoking was the smoking behaviour of classmates: those who reported a significant proportion of smokers among their classmates were nine times more likely to smoke themselves than in other cases (adjusted odds ratio [aOR]: 9.05). Those who considered smoking to be harmless were 4 times more likely to be smokers than those who considered this behaviour to be dangerous (aOR: 4.28). Poor academic results increased adolescents' smoking (aOR: 3.22 and 2.66). The odds were significantly higher for smoking, if they had an active social life (aOR: 2.54). Regular church attendance proved to be a protective factor (aOR: 0.45). Conclusions: Several social capital factors can play a role in adolescent smoking. The organization and the development of community activities aimed at prevention must strengthen the factors related to the community's social capital to reduce the likelihood of teenage smoking

Objetivo: Evaluar el efecto del capital social sobre el consumo de tabaco en adolescentes. Método: El estudio se realizó en un grupo aleatorizado y estratificado compuesto por 1313 estudiantes de séptimo y octavo grado de tres municipios en Transilvania (Rumanía). Los participantes contestaron un cuestionario autocumplimentado en relación con el consumo de tabaco y sobre actitudes y comportamientos respecto a este. El impacto del capital social, entendido como actividades personales y comunitarias, así como el desarrollo académico y la información sobre el consumo de tabaco, fueron algunas de las medidas. Resultados: Experimentar con el hábito de fumar se relaciona principalmente con el conocimiento sobre dicha adicción, el rendimiento académico y la exposición al consumo de tabaco en el hogar. El mayor riesgo para el consumo se deriva de la interacción con compañeros de clase que incurren en el consumo de tabaco. En tal caso, la probabilidad de consumo aumenta nueve veces (odds ratio ajustada [ORa]: 9,05). Quienes consideraron que fumar es inofensivo tuvieron cuatro veces más probabilidades de ser fumadores que quienes consideraron este comportamiento como peligroso (ORa: 4,28). En cuanto a los/las estudiantes con bajo rendimiento académico se observa un mayor incremento del consumo (ORa: 3,22 y 2,66). Al mismo tiempo, dicho patrón también se observa entre aquellos/as con un entorno social activo (ORa: 2,54). La asistencia a la iglesia de manera regular es un factor protector (ORa: 0,45). Conclusiones: Diferentes aspectos relacionados con el capital social se asocian al consumo de tabaco en adolescentes. La organización de actividades y el desarrollo comunitario deberán tener en cuenta estos aspectos para prevenir el consumo de tabaco. Hay que prestar especial atención a la clase social con el fin de reducir las probabilidades de consumo en adolescentes

Adolescent smoking and the social capital of local communities in three counties in Romania.

OBJECTIVE: To identify the effect of social capital on adolescent smoking. METHOD: A stratified random sample of 1313 7th and 8th grade students from three counties in Transylvania, Romania, completed a self-administered questionnaire on smoking-related knowledge, attitudes and behaviours. The impact of social capital was measured (personal and community activities, school achievements and smoking-related knowledge). Multivariate multinomial logistic regression models were used to measure the association between social participation and smoking. RESULTS: Experimenting with smoking was mostly related to knowledge about smoking, academic performance and second-hand tobacco smoke exposure at home. The strongest risk factor of adolescent smoking was the smoking behaviour of classmates: those who reported a significant proportion of smokers among their classmates were nine times more likely to smoke themselves than in other cases (adjusted odds ratio [aOR]: 9.05). Those who considered smoking to be harmless were 4 times more likely to be smokers than those who considered this behaviour to be dangerous (aOR: 4.28). Poor academic results increased adolescents' smoking (aOR: 3.22 and 2.66). The odds were significantly higher for smoking, if they had an active social life (aOR: 2.54). Regular church attendance proved to be a protective factor (aOR: 0.45). CONCLUSIONS: Several social capital factors can play a role in adolescent smoking. The organization and the development of community activities aimed at prevention must strengthen the factors related to the community's social capital to reduce the likelihood of teenage smoking.

Trends in outcomes of very preterm infants in Romania: A tale of three cities.

BACKGROUND: Although survival of preterm infants has improved, prematurity remains the second most frequent cause of death before 5 years of age in Romania. Data on the changing mortality of Romanian preterm infants born before 29 weeks of gestation have not been available. METHODS: Outcomes of infants of gestational age 25-28 weeks born in 2007-2010 (n = 247) were compared with those born in 2011-2014 (n = 235). Data were analyzed from three tertiary neonatal intensive care centers. Mortality rates and major morbidities were compared between these two epochs. RESULTS: Infants in the later epoch were more likely to have been born by cesarean section and had higher 1 and 5 min Apgar scores. Mortality rate decreased significantly with increasing gestational age at birth. Between the two epochs, the in-hospital mortality rate decreased from 65.6% to 29.4% (P < 0.001); death in the first 48 h decreased from 30.0% to 8.5% (P < 0.001); and prevalence of severe intraventricular hemorrhage decreased from 52.2% to 11.9% (P < 0.001). There were significant increases in the rates of necrotizing enterocolitis and bronchopulmonary dysplasia among survivors but no change in the rate of retinopathy of prematurity. The rate of antenatal corticosteroid use did not change and was only 47% in the more recent epoch (2011-2014). CONCLUSIONS: Overall mortality is decreasing, and infants admitted in the later epoch had substantially different rates of mortality and several serious morbidities. The low rate of antenatal corticosteroid use provides an opportunity for further reductions in mortality and morbidity among very preterm infants born in Romania.

Pregnenolone does not interfere with the effects of cannabinoids on synaptic transmission in the cerebellum and the nucleus accumbens.

The steroid hormone pregnenolone attenuates several in vivo behavioural and somatic effects of the phytocannabinoid Δ9-tetrahydrocannabinol, and it was suggested that pregnenolone can protect the brain from cannabis intoxication. The primary neuronal cannabinoid action behind most of the behavioural and somatic effects of cannabinoids is presynaptic inhibition of synaptic transmission. Therefore, the hypothesis of the present study was that pregnenolone attenuates the inhibition of synaptic transmission elicited by cannabinoids. Brain slices containing the cerebellum or the nucleus accumbens were prepared from brains of mice and rats. Spontaneous and electrically evoked GABAergic inhibitory postsynaptic currents (sIPSCs and eIPSCs) and evoked glutamatergic excitatory postsynaptic currents (eEPSCs) were recorded in superfused brain slices with patch-clamp electrophysiological techniques. Pregnenolone (10-7M) did not affect the spontaneous GABAergic synaptic input (sIPSCs) to Purkinje cells in mouse cerebellar slices. The synthetic mixed CB1/CB2 receptor agonists JWH-210 (5×10-6M) and JWH-018 (5×10-6M) inhibited the spontaneous GABAergic synaptic input (sIPSCs) to Purkinje cells. This inhibition was not affected by pregnenolone (10-7M). Tetrahydrodeoxycorticosterone (THDOC; 10-7M), an in vivo metabolite of pregnenolone, also did not affect the inhibition of the GABAergic synaptic transmission by JWH-018. The depolarization of the Purkinje cells induced suppression of the GABAergic input to Purkinje cells; pregnenolone (10-7M) did not affect this endocannabinoid-mediated form of synaptic suppression. In rat nucleus accumbens slices, glutamatergic and GABAergic synaptic input to medium spiny neurons was activated by electrical stimulation of axons. Δ9-Tetrahydrocannabinol (2×10-5M), which is a partial agonist of both CB1 and CB2 receptors, suppressed the glutamatergic and GABAergic synaptic transmission in the rat nucleus accumbens. These suppressive effects of Δ9-tetrahydrocannabinol were not changed by pregnenolone (10-7M). The suppression of the GABAergic synaptic transmission by Δ9-tetrahydrocannabinol in the rat nucleus accumbens was also not affected by THDOC (10-7M). The results indicate that pregnenolone, a neurosteroid, does not affect GABAergic synaptic transmission. The inhibition of GABAergic and glutamatergic synaptic transmission elicited by synthetic, endogenous and phyto-cannabinoids is also not changed by pregnenolone. Therefore, it is unlikely that interference with cannabinoid-induced inhibition of synaptic transmission is the mechanism by which pregnenolone attenuates behavioural and somatic effects of Δ9-tetrahydrocannabinol in vivo.

The role of TGF-ß1 869 T > C and PPAR γ2 34 C > G polymorphisms, fat mass, and anthropometric characteristics in predicting childhood obesity at birth: A cross-sectional study according the parental characteristics and newborn's risk for child obesity (the newborns obesity's risk) NOR study.

This study proposed to establish a correlation between the risk score for child obesity and anthropometric, genetic, and bioimpedance characteristics in mothers and newborns, and to assess the discriminant ability for anthropometric parameters to classify over-fatness (defined by bioimpedance body fatness %) in pregnant women.We performed a cross-sectional study on 388 couples (mother and father) and their newborns admitted in a Tertiary Hospital from Romania. The measured parameters for mothers and their newborns were risk percentage for child obesity, anthropometric characteristics (mid-upper arm circumference [MUAC], tricipital skinfold thickness [TST] of mother and newborn), genetic polymorphisms (human peroxisome proliferator-activated receptor γ [PPARγ2] 34 C > G and transforming growth factor-beta 1 [TGF-ß1] 869 T > C gene polymorphisms in both mothers and newborns), and mother's bioimpedance characteristics (fat mass [FM] %).The obesity risk score according to standard predictable Northern Finland Birth Cohort equation was in our study 4.07%. We found a monotone positive significant correlation between the newborn's risk of childhood obesity and the mother's TST (P = 0.01), as well as a tendency toward statistical significance concerning correlation with mother's MUAC (P = 0.053), without any correlations with the mothers' bioimpedance parameters and also a positive correlation between the newborn's risk of childhood obesity and the newborn's anthropometrical characteristics like body mass index (BMI), MUAC, and TST (P < 0.001). We observed that the calculated newborn's risk percentage for child obesity was greater for the variant allele of the TGF-ß1 869 T > C polymorphism and also for the wild-type C allele of the PPARγ2 34 C > G gene polymorphism. Our study indicated that the best predictors for over-fatness are BMI and MUAC (P = 0.01 < 0.02 and P = 0.019 < 0.02, respectively).

Radical Trachelectomy Performed During Pregnancy: A Review of the Literature.

OBJECTIVE: Cervical cancer is one of the most frequent malignant diseases diagnosed during pregnancy. Abdominal or vaginal radical trachelectomies are fertility-preserving alternatives to radical hysterectomy for young women with early-stage cervical cancer that can be performed during ongoing pregnancy. METHODS: A literature review of articles on this subject was conducted through a Medline search for articles published in English or French. RESULTS: At this moment, 21 cervical cancer patients, including ours (4 stage IA2, 16 IB1, and 1 IB2) who underwent radical trachelectomy during pregnancy have been reported. Of these, 10 were performed by vaginal route and 11 were abdominal radical trachelectomies. CONCLUSIONS: Radical trachelectomy could be offered as an option for pregnant patients with early invasive cervical cancer. It may help women avoid the triple losses of a desired pregnancy, fertility, and motherhood.

Studies on the possible aetiological role of different Candida species in pathogenesis of dentine caries by monitoring the calcium release from tooth particles.

An in vitro model has been developed for study of cariogenic potential of different Candida species. Slices were prepared from the root of extracted healthy teeth. These disks were covered with inert material, only the central hole, i.e. the root canal dentin surface remained uncovered. These preparates with free root dentin surfaces were incubated in Sabouraud medium in the presence of six-six Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei, Candida inconspicua and Candida norvegensis strains. The calcium release was detected for 15 days. Two types of release could be distinguished. C. albicans deliberated calcium more aggressively (type "A" curve), while other Candidas were characterized by less expressed calcium releasing capacity (type "B" curve). Curves type "A" and "B" were divided into four steps in order to characterize more precisely the different dynamics of calcium release. Analyses of the different steps also suggested the more aggressive behaviour of C. albicans. Our results indicate that in addition to cariogenic role of different bacteria, fungi may also actively take part in the dentinal caries progress.

Abdominal radical trachelectomy: a Romanian series.

OBJECTIVE: Abdominal radical trachelectomy (ART) is one of the fertility-sparing procedures in women with early-stage cervical cancer. The published results of ART, in comparison with vaginal radical trachelectomy, so far are limited. MATERIALS AND METHODS: This retrospective study comprises all cases of female patients referred to ART with early-stage cervical cancer from 2 gynecologic oncology centers in Romania. RESULTS: A total of 29 women were referred for ART, but subsequently, fertility could not be preserved in 3 of them. Eleven women had stage IA2 disease (42.3%), 14 (53.8%) women had stage IB1 disease, and 1 (3.8%) woman had stage IB2 disease. Histologic subtypes were 15 (57.6%) squamous, 8 (30.7%) adenocarcinoma, and 3 (11.5%) adenosquamous. There were no major intraoperative complications in both hospitals. Early postoperative complications were mainly related to the type C parametrectomy-bladder dysfunction for more than 7 days (8 [30.7%] women) and prolonged constipation (6 [23.0%] women). Other complications consisted in symptomatic lymphocele in 2 (7.6%) patients, which were drained. Median follow-up time was 20 months (range, 4-43 months). Up to the present time, there has been 1 (3.8%) recurrence in our series. Most patients did not experience late postoperative complications. Three (11.5%) women are amenorrheic, and 1 (3.8%) woman developed a cervical stenosis. Of the 23 women who have normal menstruation and maintained their fertility, a total of 7 (30.4%) women have attempted pregnancy, and 3 (42.8%) of them achieved pregnancy spontaneously. These pregnancies ended in 2 first trimester miscarriages and 1 live birth at term by cesarean delivery. CONCLUSIONS: Our results demonstrate that ART preserves fertility and maintains excellent oncological outcomes with low complication rates.

Mortality and morbidity of very preterm infants in Romania: how are we doing?

BACKGROUND: Birth before 28 weeks of gestation is associated with high mortality and morbidity. The purpose of this study was to examine characteristics associated with in-hospital mortality and morbidity among extremely low-birthweight neonates admitted to three tertiary care centers in Romania. METHODS: The study was conducted in three Romanian hospitals with level-III neonatal intensive care units. We studied singleton live births at the established Romanian limit of viability (i.e., 25-28 weeks' gestational age) born between January 2007 and December 2010 (n = 227). Infants born in non-level-III facilities transferred to these three centers were included in our study (n = 39). Descriptive and multivariate statistical analyses were used to describe the population and examine outcomes and risk factors. RESULTS: During the study period, 62 neonates (27.3%) were delivered at 25 weeks, 56 (24.7%) were delivered at 26 weeks, 56 (24.7%) at 27 weeks, and 53 (23.3%) at 28 weeks. Overall in-hospital mortality was 65% (from 85% at 25 weeks to 35% at 28 weeks). The rates for major morbidities were necrotizing enterocolitis 8.8%, bronchopulmonary dysplasia 12.5%, and retinopathy of prematurity (stage higher than 2) 26.2%. CONCLUSIONS: During 2007-2010, in-hospital survival of infants admitted to three neonatal intensive care units in Romania was 35% and ranged from 14% at 25 weeks to 64% at 28 weeks.

Activation-dependent plasticity of polarized GPCR distribution on the neuronal surface.

Directionality of information flow through neuronal networks is sustained at cellular level by polarized neurons. However, specific targeting or anchoring motifs responsible for polarized distribution on the neuronal surface have only been identified for a few neuronal G-protein-coupled receptors (GPCRs). Here, through mutational and pharmacological modifications of the conformational state of two model GPCRs, the axonal CB1R cannabinoid and the somatodendritic SSTR2 somatostatin receptors, we show important conformation-dependent variations in polarized distribution. The underlying mechanisms include lower efficiency of conformation-dependent GPCR endocytosis in axons, compared with dendrites, particularly at moderate activation levels, as well as endocytosis-dependent transcytotic delivery of GPCRs from the somatodendritic domain to distal axonal portions, shown by using compartmentalized microfluidic devices. Kinetic modeling predicted that GPCR distribution polarity is highly regulated by steady-state endocytosis, which is conformation dependent and is able to regulate the relative amount of GPCRs targeted to axons and that axonally polarized distribution is an intermediary phenotype that appears at moderate basal activation levels. Indeed, we experimentally show that gradual changes in basal activation-dependent endocytosis lead to highly correlated shifts of polarized GPCR distribution on the neuronal surface, which can even result in a fully reversed polarized distribution of naturally somatodendritic or axonal GPCRs. In conclusion, polarized distribution of neuronal GPCRs may have a pharmacologically controllable component, which, in the absence of dominant targeting motifs, could even represent the principal regulator of sub-neuronal distribution. Consequently, chronic modifications of basal GPCR activation by therapeutic or abused drugs may lead to previously unanticipated changes in brain function through perturbation of polarized GPCR distribution on the neuronal surface.

Acute toxicity due to the confirmed consumption of synthetic cannabinoids: clinical and laboratory findings.

AIMS: Recently, several synthetic cannabinoids were identified in herbal mixtures consumed as recreational drugs alternative to cannabis products. The aim was to characterize the acute toxicity of synthetic cannabinoids as experienced by emergency patients. DESIGN: This was a retrospective study targeting patients seeking emergency treatment after recreational use of synthetic cannabinoids. SETTING AND PARTICIPANTS: Patients were selected from the database of the Poisons Information Center Freiburg between September 2008 and February 2011. The inclusion criteria were: hospitalization, available clinical reports and analytical verification of synthetic cannabinoid uptake. In total, 29 patients were included (age 14-30 years, median 19; 25 males, four females). MEASUREMENTS: Clinical reports were evaluated and synthetic cannabinoids and other drugs were determined analytically. FINDINGS: CP-47,497-C8 (one), JWH-015 (one), JWH-018 (eight), JWH-073 (one), JWH-081 (seven), JWH-122 (11), JWH-210 (11), JWH-250 (four) and AM 694 (one) were quantified in blood samples. JWH-018 was most common in 2008-9, JWH-122 in 2010, and JWH-210 in 2011. Tachycardia, agitation, hallucination, hypertension, minor elevation of blood glucose, hypokalaemia and vomiting were reported most frequently. Chest pain, seizures, myoclonia and acute psychosis were also noted. CONCLUSIONS: There appears to have been an increase in use of the extremely potent synthetic cannabinoids JWH-122 and JWH-210. Acute toxic symptoms associated with their use are also reported after intake of high doses of cannabis, but agitation, seizures, hypertension, emesis and hypokalaemia seem to be characteristic to the synthetic cannabinoids, which are high-affinity and high-efficacy agonists of the CB(1) receptor. Thus, these effects are due probably to a strong CB(1) receptor stimulation.