Circadian rhythmicity in substance use disorder male patients with and without comorbid depression under ambulatory and therapeutic community treatment

Although there have been described alterations of circadian rhythmicity both in patients with substance use disorder (SUD) and patients with major depressive disorder (MDD), the circadian characteristics of SUD patients with comorbid MDD (SUD-MDD) are unknown. Likewise, the possible influence of the different modalities of treatments (ambulatory or therapeutic community) upon the circadian rhythmicity of SUD patients has not been characterized. Therefore, this study analyzes the circadian rhythmic profiles of SUD and SUD-MDD patients under ambulatory and therapeutic community treatment. The sample was composed of 40 SUD and 40 SUD-MDD men, aged 22–55 yrs, under treatment and with abstinence for at least three months (including each group 20 ambulatory and 20 from therapeutic community). Patients completed a sociodemographic, clinical and sleep-wake schedules interview, the Composite Scale of Morningness, and wore on the wrist an ambulatory device known as iButton® Thermochron DS1921H, which registered their distal skin temperature every two minutes for 48 hours. All the groups showed a tendency to morningness without differences among them in concordance with their sleep-wake schedules. With regard to distal skin temperature circadian rhythm, SUD patients showed higher values than SUD-MDD in amplitude, relative amplitude, percentage rhythm, and first harmonic power, and lower minimum temperature in 10 consecutive hours (p < .043, in all cases). Therapeutic community group values were lower in minimum temperature and higher in amplitude, relative amplitude, and 12 harmonic accumulated power (p < .028, in all cases) as compared to ambulatory ones. Moreover, all groups showed higher Rayleigh vector and rhythm stability as compared to normative population (p < .043, in both cases). The circadian rhythmic differences observed for diagnosis and type of treatment are indicative of a higher circadian rhythmicity robustness in SUD and therapeutic community patients as compared to SUD-MDD and ambulatory ones, respectively. Although drug consumption exerts a negative effect on the circadian rhythmicity, our results (high amplitude and rhythm stability) are indicative of an adequate circadian functioning as well as of an adjustment to the light-dark cycle in both diagnosis and type of treatment which may constitute a marker of the adherence to treatment and recovery status.     

The historical biogeography of the southern group of the sucker genus Moxostoma (Teleostei: Catostomidae) and the colonization of central Mexico

The historical biogeography of the southern group of Moxostoma Rafinesque, 1820, a genus of Nearctic freshwater fishes belonging to the Catostomidae, along its entire distribution in North America was inferred to: (1) determine the biogeographical events responsible for its current pattern of diversity and distribution; (2) correlate the climatic and geologic history of the region with the biogeographical pattern observed; and (3) trace the colonization route into central Mexico and the western Pacific slope drainages. The sequences of mitochondrial cytochrome b and the third intron of the growth hormone were obtained for the members of the southern group and related species of the Catostomidae. Phylogenetic analyses and relaxed molecular clock analyses were performed to determine the relatedness of the species and to estimate divergence times. To uncover biogeographical patterns, a dispersal–extinction–cladogenesis (DEC) analysis was conducted. The phylogenetic analyses were consistent with the historical hydrographic scenario in the region. The divergence times show that the southern group evolved during the Pliocene–Pleistocene. The DEC analyses showed that vicariance and dispersal played an important role in the current distribution patterns of the lineages in central Mexico, and allow us to trace an independent route of colonization from the northern areas of North America into central Mexico.     

Bone marrow mesenchymal stem cell donors with a high body mass index display elevated endoplasmic reticulum stress and are functionally impaired

Bone marrow mesenchymal stem cells (BM‐MSCs) are promising candidates for regenerative medicine purposes. The effect of obesity on the function of BM‐MSCs is currently unknown. Here, we assessed how obesity affects the function of BM‐MSCs and the role of endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) therein. BM‐MSCs were obtained from healthy donors with a normal (<25) or high (>30) body mass index (BMI). High‐BMI BM‐MSCs displayed severely impaired osteogenic and diminished adipogenic differentiation, decreased proliferation rates, increased senescence, and elevated expression of ER stress–related genes ATF4 and CHOP. Suppression of ER stress using tauroursodeoxycholic acid (TUDCA) and 4‐phenylbutyrate (4‐PBA) resulted in partial recovery of osteogenic differentiation capacity, with a significant increase in the expression of ALPL and improvement in the UPR. These data indicate that BMI is important during the selection of BM‐MSC donors for regenerative medicine purposes and that application of high‐BMI BM‐MSCs with TUDCA or 4‐PBA may improve stem cell function. However, whether this improvement can be translated into an in vivo clinical advantage remains to be assessed.     

Laboratory Effects of Ingestion of Azadirachtin by Two Pests ( Ceratitis capitata and Spodoptera exigua ) and Three Natural Enemies ( Chrysoperla carnea , Opius concolor and Podisus maculiventris )

The effects of azadirachtin on two pests: neonate larvae and newly emerged adults of Ceratitis capitata (Wiedemann) and last instar larvae of Spodoptera exigua (Hubner); and three natural enemies: newly emerged adults of Opius concolor Szepligeti, second instar larvae of Chrysoperla carnea (Stephens), and fifth instar nymphs of Podisus maculiventris (Say) were studied in laboratory. Adult insects were exposed to a non-oil formulation of azadirachtin (Align, emulsifiable concentrate, 3.2% azadirachtin, Sipcam Inagra, Spain) via their drinking water and immature instars were reared in the presence of the insecticide-treated diet. The natural enemies were exposed to at least the maximum field recommended concentration of the insecticide (0.15% v/v). Azadirachtin was highly toxic to neonate larvae of C. capitata and prevented adult emergence at a concentration of 1 mg a.i. l -1 . When adults were fed the insecticide at the maximum recommended concentration, their survival was not affected but egg laying was totally inhibited. Last instar S. exigua larvae were also very susceptible (LC 50 = 7.7 mg a.i. l -1 ) and at a concentration of 10 mg a.i. l -1 fecundity of surviving adults, and egg fertility, were reduced by 72 and 85%, respectively. Effects on O. concolor were large, and significant reductions in longevity, percentage of attacked hosts, and progeny size per female, were recorded. The predator P. maculiventris was much less sensitive to azadirachtin, but slight reductions in survival of emerged adults and of reproductive parameters occurred. The insecticide had no significant effect on C. carnea larvae fed with treated Sitotroga cerealella (Oliver) eggs, probably because of its inability to penetrate inside the egg.     

Tuning sperm chemotaxis by calcium burst timing

Marine invertebrate oocytes establish chemoattractant gradients that guide spermatozoa towards their source. In sea urchin spermatozoa, this relocation requires coordinated motility changes initiated by Ca²⁺-driven alterations in sperm flagellar curvature. We discovered that Lytechinus pictus spermatozoa undergo chemotaxis in response to speract, an egg-derived decapeptide previously noted to stimulate non-chemotactic motility alterations in Strongylocentrotus purpuratus spermatozoa. Sperm of both species responded to speract gradients with a sequence of turning episodes that correlate with transient flagellar Ca²⁺ increases, yet only L. pictus spermatozoa accumulated at the gradient source. Detailed analysis of sperm behavior revealed that L. pictus spermatozoa selectively undergo Ca²⁺ fluctuations while swimming along negative speract gradients while S. purpuratus sperm generate Ca²⁺ fluctuations in a spatially non-selective manner. This difference is attributed to the selective suppression of Ca²⁺ fluctuations of L. pictus spermatozoa as they swim towards the source of the chemoattractant gradient. This is the first study to compare and characterize the motility components that differ in chemotactic and non-chemotactic spermatozoa. Tuning of Ca²⁺ fluctuations and associated turning episodes to the chemoattractant gradient polarity is a central feature of sea urchin sperm chemotaxis and may be a feature of sperm chemotaxis in general.     

Synthesis of cyclic α-MSH peptides

Summary Cyclic lactam analogs of α-melanocyte stimulating hormone (α-MSH) have been shown to be potent agonists in the frog skin bioassay [Al-Obeidi, F. et al., J. Med. Chem., 32 (1989) 2555], demonstrating melanocortin-1 (MC1) receptor activity. We synthesized cyclic α-MSH(1–13) and α-MSH(4–10) lactam analogs. The peptides were synthesized using Fmoc chemistry. We improved the cyclization procedure: side chains of Asp⁵ and Lys¹⁰ from the deprotected peptide were coupled in DMF to form a cyclic lactam, using an excess of PyBOP reagent and DIEA as a base. The cyclization reaction was completed within 1 h and was almost quantitative. We also synthesized an α-MSH analog cyclized via a disulphide bridge. The peptides were tested for their selectivity for the rat MC4 receptor. Cyclization and substitutions at position 7 dramatically influenced the selectivity for the rMC4 receptor.     

Flavin containing monooxygenase 3 genetic polymorphisms Glu158Lys and Glu308Gly and their relation to ischemic stroke

Ischemic stroke is a multifactorial disease leading to severe long-term disability and it is the third leading cause of death in developed countries. Although many studies have been reported to elucidate etiological and pathological mechanisms of stroke, the genetic and molecular basis of disease remains poorly understood. Recent studies have shown that reactive oxygen species causing oxidative stress play a pivotal role in the pathogenesis of atherosclerosis that is the main cause of a group of cardiovascular diseases including ischemic stroke. In this study, we aimed to investigate the relationship between FMO3 Glu158Lys and Glu308Gly variants, and the risk of incidence of ischemic stroke in Turkish population. Two single nucleotide polymorphisms (SNPs) within the FMO3 gene were genotyped by using PCR-RFLP technique in a sample set of 245 cases and 145 controls. In the case-control analysis, no significant difference was observed between stroke patients and controls with respect to FMO3 Glu158Lys and Glu308Gly polymorphisms' genotype and allele frequency distribution. However, heterozygote 158Glu/Lys (OR = 6.110, P < 0.001) and 308Glu/Gly (OR = 6.000, P = 0.006) genotypes increase the risk of stroke 6 times in hypertensive subjects. On the other hand, the wild type genotypes 158Glu/Glu and 308Glu/Glu had 6.2-fold and 4.8-fold higher risk of ischemic stroke in obese subgroup, respectively. Our results clearly showed that the risk of hypertension-related ischemic stroke was higher in the heterozygote genotype carriers. This is the first study conducted regarding the association of FMO3 Glu158Lys and Glu308Gly genetic polymorphisms and ischemic stroke risk in Turkish population.     

Combined Use of the Canine Adenovirus-2 and DREADD-Technology to Activate Specific Neural Pathways In Vivo

We here describe a technique to transiently activate specific neural pathways in vivo. It comprises the combined use of a CRE-recombinase expressing canine adenovirus-2 (CAV-2) and an adeno-associated virus (AAV-hSyn-DIO-hM₃D(G_q)-mCherry) that contains the floxed inverted sequence of the designer receptor exclusively activated by designer drugs (DREADD) hM₃D(G_q)-mCherry. CAV-2 retrogradely infects projection neurons, which allowed us to specifically express hM₃D(G_q)-mCherry in neurons that project from the ventral tegmental area (VTA) to the nucleus accumbens (Acb), the majority of which were dopaminergic. Activation of hM₃D(G_q)-mCherry by intraperitoneal (i.p.) injections of clozapine-N-oxide (CNO) leads to increases in neuronal activity, which enabled us to specifically activate VTA to Acb projection neurons. The VTA to Acb pathway is part of the mesolimbic dopamine system and has been implicated in behavioral activation and the exertion of effort. Injections of all doses of CNO led to increases in progressive ratio (PR) performance. The effect of the lowest dose of CNO was suppressed by administration of a DRD1-antagonist, suggesting that CNO-induced increases in PR-performance are at least in part mediated by DRD1-signaling. We hereby validate the combined use of CAV-2 and DREADD-technology to activate specific neural pathways and determine consequent changes in behaviorally relevant paradigms.     

Polysialic Acid on Neuropilin-2 Is Exclusively Synthesized by the Polysialyltransferase ST8SiaIV and Attached to Mucin-type O-Glycans Located between the b2 and c Domain

Neuropilin-2 (NRP2) is well known as a co-receptor for class 3 semaphorins and vascular endothelial growth factors, involved in axon guidance and angiogenesis. Moreover, NRP2 was shown to promote chemotactic migration of human monocyte-derived dendritic cells (DCs) toward the chemokine CCL21, a function that relies on the presence of polysialic acid (polySia). In vertebrates, this posttranslational modification is predominantly found on the neural cell adhesion molecule (NCAM), where it is synthesized on N-glycans by either of the two polysialyltransferases, ST8SiaII or ST8SiaIV. In contrast to NCAM, little is known on the biosynthesis of polySia on NRP2. Here we identified the polySia attachment sites and demonstrate that NRP2 is recognized only by ST8SiaIV. Although polySia-NRP2 was found on bone marrow-derived DCs from wild-type and St8sia2^−/− mice, polySia was completely lost in DCs from St8sia4^−/− mice despite normal NRP2 expression. In COS-7 cells, co-expression of NRP2 with ST8SiaIV but not ST8SiaII resulted in the formation of polySia-NRP2, highlighting distinct acceptor specificities of the two polysialyltransferases. Notably, ST8SiaIV synthesized polySia selectively on a NRP2 glycoform that was characterized by the presence of sialylated core 1 and core 2 O-glycans. Based on a comprehensive site-directed mutagenesis study, we localized the polySia attachment sites to an O-glycan cluster located in the linker region between b2 and c domain. Combined alanine exchange of Thr-607, -613, -614, -615, -619, and -624 efficiently blocked polysialylation. Restoration of single sites only partially rescued polysialylation, suggesting that within this cluster, polySia is attached to more than one site.