High Impact AMPAkines Induce a Gq-Protein Coupled Endoplasmic Calcium Release in Cortical Neurons: A Possible Mechanism for Explaining the Toxicity of High Impact AMPAkines.

α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) positive allosteric modulators (AMPAkines) have a multitude of promising therapeutic properties. The pharmaceutical development of high impact AMPAkines has, however, been limited by the appearance of calcium-dependent neuronal toxicity and convulsions in vivo. Such toxicity is not observed at exceptionally high concentrations of low impact AMPAkines. Because most AMPAR are somewhat impermeable to calcium, the current study sought to examine the extent to which different mechanisms contribute to the rise in intracellular calcium in the presence of high impact ampakines. In the presence of AMPA alone, cytosolic calcium elevation is shown to be sodium-dependent. In the presence of high impact AMPAkines such as cyclothiazide (CTZ) or CX614, however, AMPAR potentiation also activates an additional mechanism that induces calcium release from endoplasmic reticular (ER) stores. The pathway that connects AMPAR to the ER system involves a Gq-protein, phospholipase Cß-mediated inositol triphosphate (InsP3) formation, and ultimately stimulation of InsP3-receptors located on the ER. The same linkage was not observed using high concentrations of the low impact AMPAkines, CX516 (Ampalex), and CX717. We also demonstrate that CX614 produces neuronal hyper-excitability at therapeutic doses, whereas the newer generation low impact AMPAkine CX1739 is safe at exceedingly high doses. Although earlier studies have demonstrated a functional linkage between AMPAR and G-proteins, this report demonstrates that in the presence of high impact AMPAkines, AMPAR also couple to a Gq-protein, which triggers a secondary calcium release from the ER and provides insight into the disparate actions of high and low impact AMPAkines.

Myo-inositol impact on sperm motility in vagina and evaluation of its effects on foetal development.

OBJECTIVE: To counteract the arising problem of couple infertility, having good quality gametes is increasingly important. A molecule that seems to be useful to favor this condition is myo-inositol (MI), the most common stereoisomer of the inositol family, involved as second messenger in several cell pathways (osmoregulation, chromatin remodeling, gene expression, etc.). To evaluate this possibility, a treatment with myo-inositol in idiopathic infertile couples was performed in this randomized, placebo-controlled study. PATIENTS AND METHODS: 86 couples were enrolled and randomly assigned to two groups, treated either with MI (Xyminal®, Lo.Li. Pharma Srl, Rome, Italy) or placebo suppositories, to evaluate the effects on sperm motility, cervical mucus quality and pregnancy rate. Moreover, in pregnancy cases, all routine controls on gestation progress and foetal health were performed to confirm the safety of this treatment. RESULTS: As showed in this study, MI treatment allows an increase of total sperm motility (54.42 ± 8.72) in comparison to placebo group (46.21 ± 5.33). Moreover, MI mildly improves cervical mucus quality and increases the number of pregnancies (18.60%) in comparison to the placebo group (6.97%). CONCLUSIONS: MI improves sperm motility and cervical mucus quality, increasing the probability of conception. The absence of adverse events both for the mother and the foetus confirmed the safety of this molecule in pregnancy, supporting even more its use for couples seeking pregnancy.

Assisted reproduction and Coronavirus in Italy.

OBJECTIVE: A novel type of Coronavirus was identified in China in December 2019. The first cases of a form of pneumonia of unknown etiology were detected at the beginning of that month in Wuhan. The virus is believed to have emerged at the Wuhan Huanan Seafood Market, where transmission of a zoonotic pathogen to humans occurred. PATIENTS AND METHODS: Some studies conducted in China during the epidemic report small numbers of pregnant women infected with SARS-CoV-2 and some pregnancy complications in patients with COVID-19. However, they fail to document the transplacental passage of the virus from mother to fetus. RESULTS: Following the COVID-19 outbreak, guidelines for couples who are undergoing treatments of assisted reproduction have been issued by the International Federation for Fertility Societies (IFFS), the American Society for Reproductive Medicine (ASRM), the European Society of Human Reproduction and Embryology (ESHRE) and the Latin American Network of Assisted Reproduction (REDLARA). They recommend couples to discuss assisted reproduction with their doctors while those at risk or with SARS-CoV-2 should consider freezing oocytes or embryos and retransferring them later. CONCLUSIONS: Other than the US, Italy is the country with the highest number of cases (197675 positives, 26644 deaths) (updated on April 26). The Italian National Transplant Centre and the Higher Institute of Health advised on March 17 to complete the cycles already started and not to start new cycles. Phase 2 will begin on 4 May with an increase in freedom of action and fertilization treatments will start again. The Society that brings together embryologists (SIERR) has issued the guidelines to be followed when this happens.

The cholinergic hypothesis of geriatric memory dysfunction.

Biochemical, electrophysiological, and pharmacological evidence supporting a role for cholinergic dysfunction in age-related memory disturbances is critically reviewed. An attempt has been made to identify pseudoissues, resolve certain controversies, and clarify misconceptions that have occurred in the literature. Significant cholinergic dysfunctions occur in the aged and demented central nervous system, relationships between these changes and loss of memory exist, similar memory deficits can be artificially induced by blocking cholinergic mechanisms in young subjects, and under certain tightly controlled conditions reliable memory improvements in aged subjects can be achieved after cholinergic stimulation. Conventional attempts to reduce memory impairments in clinical trials hav not been therapeutically successful, however. Possible explanations for these disappointments are given and directions for future laboratory and clinical studies are suggested.

Efficacy of the synergic action of myoinositol, tyrosine, selenium and chromium in women with PCOS.

OBJECTIVE: The aim of the study is to investigate the efficacy of a treatment with myoinositol plus L-tyrosine, selenium, and chromium in women with polycystic ovarian syndrome (PCOS). PATIENTS AND METHODS: One hundred and eighty-six women, with diagnosis of PCOS, were divided in four groups according to their clinical features. Phenotype A: androgen excess + ovulatory dysfunction + polycystic ovarian morphology. Phenotype B: androgen excess + ovulatory dysfunction. Phenotype C: androgen excess + polycystic ovarian morphology. Phenotype D: ovulatory dysfunction + polycystic ovarian morphology. All patients were given daily for six months a compound with 2 g myo-inositol, 0.5 mg L-Tyrosine, 0.2 mg folic acid, 55 mcg selenium, 40 mcg chromium. Hormonal assessment, BMI, Ferriman-Gallway Gallway score, HOMA index, and follicular monitoring were reported before starting the therapy, three months and six months after. RESULTS: Phenotype A showed an improvement, consistent with restored ovulation: more regular length of the menstrual cycle, detection of periovulatory follicle at ultrasound, and rising of progesterone in the luteal phase. A total of 45 patients (65.2%) ovulated after six months. In the same period glucose and HOMA index decreased. In the phenotype B, 80% of patients ovulated after six months. An improvement of the clinical and biochemical sign of hyperandrogenism was also reported. In the phenotype C, after BMI had followed the treatment for six months, it decreased in a statistically significant manner. In the phenotype D, 49 out of 82 women (59.7%) restored their regular menstrual period and ovulated. CONCLUSIONS: Our study reported how the synergistic action of myoinositol, L-tyrosine, selenium, and chromium could restore normal menstrual cycle, ovulation, and decrease weight in these patients.

[Diagnosis of thoracic outlet syndrome. Value of angiography in the sitting position]. / Diagnostic des syndromes du défilé cervico-thoraco-brachial: apport de l'artériographie réalisée en position assise.

PURPOSE: Thoracic outlet syndrome includes arterial, venous or neurological symptoms. Frequently difficult to diagnose clinically, confirmatory imaging studies are usually required. The purpose of this study is to review the diagnostic work-up during management of patients with thoracic outlet syndrome and demonstrate the value of angiography in the sitting position. MATERIALS AND METHODS: Retrospective study of 81 surgical procedures for thoracic outlet syndrome, between 1997 and 2005, in 56 patients aged 17-57 years. Surgery was bilateral in 26 cases, with bilateral surgery in a single setting for 1 patient. All patients presented clinical symptoms confirmed on US, angiography, venogram or EMG. Angiography, from a transfemoral approach, was initially performed in the supine position, without and with dynamic maneuver, and in the sitting position with dynamic maneuver when needed. RESULTS: In 48 patients, supine rest angiography showed stenosis in 6% of cases and supine dynamic angiography showed stenosis in 81% of cases, with severe stenosis in only 35% of cases (stenosis>80% or arterial occlusion). Angiography in the sitting position was performed in 33 patients, showing worsening of stenosis in 91% of cases, with severe stenosis in 87%. CONCLUSION: Angiography in the sitting position with dynamic maneuver improves the sensitivity for detection of thoraci coutlet syndrome. This procedure may be considered in addition to other imaging modalities routinely used including Doppler US, CT and MRI.

[Trans-vesical incarceration of a small bowel loop after pelvic trauma]. / Incarcération transvésicale d'une anse grêle lors d'un traumatisme du bassin.

The authors report the case of a patient presenting with small bowel obstruction after a pelvic trauma with associated bladder injury. While urinary bladder and bowel injuries are frequent following this type of trauma, their association is rare. Small bowel obstruction secondary to trans-vesical incarceration of a small bowel loop at the fracture site is exceptional. CT is invaluable for diagnosis.

AL721, a novel membrane fluidizer.

AL721, which is a novel lipid mixture extracted from egg yolks, is believed to be a therapeutic pharmacologic agent. AL721 interacts with membranes of various types of cells with a common mode of action. AL721 modifies cellular membrane composition and fluidity through passive extraction and/or exchange of cholesterol. Physiologically diminished cell function due to rigidification of its membrane is reversible both in vitro and in vivo by AL721. Fluidization of aged membranes with AL721 has been shown to restore brain serotonin receptor function both in vitro and in vivo. AL721 can also successfully restore deficient immune responsiveness of lymphocytes to mitogen stimulation in aged subjects. Drug tolerance to morphine and ethanol develops upon elevation of the viscosity of neuronal cell membranes in order to counteract the fluidization effect of the drug. Treatment of rigidified cellular membranes with AL721 in vivo can markedly reduce withdrawal symptoms. The virucidal effect of AL721 on the human immunodeficiency virus is believed to operate by lowering of viral membrane cholesterol thus interfering with the binding of the viral antigen to the host cell. Non-toxicity of AL721 is clearly demonstrated in animal and human safety studies.

Reduction of motor behavioral deficits in senescent animals via chronic prolactin administration. II. Non-stereotypic behaviors.

The effects of chronic prolactin administration on three non-stereotypic psychomotor behaviors (inclined screen performance, rod walking and wire hanging) were examined in senescent (24 month) Fisher 344 rats. Prolactin (150 ng/hr) was administered for 7 days via Alzet minipumps to rats which had been pretested on the three tasks. The animals were tested on days 4 and 7 following the implants and the pumps were removed after testing on day 7. In order to assess the persistence of any prolactin effects on psychomotor performance the animals were tested again on days 7 and 14 after pump removal. Control animals were implanted with Alzet pumps containing only saline and tested in a similar manner. Results demonstrated that the prolactin treated animals showed improvements on both the inclined screen and rod walking tasks but not the wire hanging test. Moreover, the improved performance seen on the inclined screen test persisted for as long as 7 days after pump removal. Results are discussed in terms of the importance of techniques which increase striatal dopaminergic responsivity on enhancing psychomotor performance in senescence.

Reduction of motor behavioral deficits in senescent animals via chronic prolactin administration. I. Rotational behavior.

The effects of chronic prolactin administration on amphetamine or dopamine (DA) induced rotational behavior was examined in mature (6 month) and senescent (24 month) Wistar rats which were unilaterally lesioned in the left substantia nigra with 6-hydroxydopamine. Prolactin (150 ng/hr) was administered for 7 days via subcutaneously implanted Alzet minipumps. Amphetamine (AMPH) (0, 10 micrograms) or DA (0, 25 micrograms) was administered through cannula which had been implanted into the right (intact) striatum. Both DA-active agents were given prior to pump implantation and on day 4 of prolactin administration. The AMPH was dissolved in saline (1 microliter; pH, 5.5-6.0), while DA was dissolved in N2 bubbled distilled H2O (1 microliter; pH, 5.5-6.0) and the animals were pretreated with nialamide (50 mg/kg) intraperitoneally 1 hr before DA or DA-vehicle injection. The order of drug administration was counterbalanced within the age groups. Results showed that both groups of animals exhibited higher rotational behavior scores following prolactin treatment. In fact, there was a trend toward greater enhancement of rotational behavior in the senescent animals following prolactin treatment than that seen in mature animals. These results parallel those reported previously wherein it was found that striatal DA receptor concentrations (as assessed with [3H]spiperone binding) were higher in prolactin treated mature and senescent animals than in their respective controls. The findings suggest that there is a relationship between increases in the density of striatal DA receptors and improvement in motor performance tasks in senescent animals.

Brain cholinergic dysfunction and memory in aged rats.

Age related alterations in mnemonic ability and in the functional status of muscarinic receptors were evaluated and compared to biochemical measures of pre and post-synaptic cholinergic functioning. Retention of a single trial passive avoidance task was considerably disturbed as a function of aging. The functional status of muscarinic receptors, as measured by the ability of microiontophoretically applied acetylcholine to stimulate the firing of hippocampal pyramidal cells, was similarly disturbed in aged rats. A small, but significant decrease in muscarinic receptors was detected in the dorsal hippocampi of these same aged rats, while choline acetyltransferase activity did not change. When considered with prior psychopharmacological studies, these data suggest that specific muscarinic receptor impairments may play a critical role in the memory disturbances associated with old age.

Conformational changes in muscarinic receptors may produce diminished cholinergic neurotransmission and memory deficits in aged rats.

Both clinical and laboratory studies suggest that age-related memory deficits may be due, at least in part, to disturbances in muscarinic acetylcholine (mAChR) receptors. In order to further evaluate this premise, the present studies examined the electrophysiological responses rates of hippocampal pyramidal cells to iontophoretically applied ACh in young, middle-age and aged animals. The relationship between age and muscarinic agonist and antagonist binding in the hippocampus was also examined. In addition, possible age-related changes in receptor-effector coupling were assessed by determining calmodulin levels and the activities of phospholipid methyl-transferase I and II. Analysis of electrophysiological data showed selective age-related decrements in the ability of ACh to alter burst rate but not simple spike rate. These age-related decreases in the efficacy of ACh to increase burst rate were not paralleled by decreases in mAChR density as assessed by 3H-QNB binding, but they were temporally paralleled by age-related changes in the ability of oxotremorine to inhibit 3H-QNB binding. In the young animals, the resultant Hill coefficients derived from these analyses approached 1, while in the middle and old aged animals, the Hill coefficients deviated significantly from 1, indicating the possible existence of 2 or more receptor states with differential affinity for oxotremorine in the 2 older age groups. When carbamylcholine was used to inhibit 3H-QNB, these complex binding patterns were seen even in the young, since carbamylcholine induces conformational/orientational changes in the mAChR while oxotremorine does not.(ABSTRACT TRUNCATED AT 250 WORDS)

Synthesis and anxiolytic activity of 6-(substituted-phenyl)-1,2,4-triazolo[4,3-b]pyridazines.

The synthesis of a series of 6-(substituted-phenyl)-1,2,4-triazolo[4,3-b]pyridazines (VIII) is reported. Some of these derivatives show activity in tests predictive of anxiolytic activity [(a) protection against pentylenetetrazole-induced convulsions; (b) thirsty rat conflict procedure]. They also represent a new class of compound which inhibits [3H]diazepam binding. Structure--activity correlations, as well as the ability of structures VIII to inhibit [3H]diazepam binding (in vitro), are discussed.

Development of benzodiazepine binding subtypes in three regions of rat brain.

An examination of the development of benzodiazepine binding of the high and low affinity triazolopyridizine (TPZ) type was undertaken in rat cortex, hippocampus and cerebellum. Various concentrations of a typical triazolopyridizine, CL-218-872, were used to displace [3H]flunitrazepam from synaptosomal fractions from rats of postnatal ages day 0, 7, 14, 21, 35 and 70. In contrast to the cortex and cerebellum, hippocampus displays high affinity TPZ binding at birth, prior to the periods of dendritic elaboration and synaptogenesis, suggesting that adult proportions of high and low affinity sites are maintained throughout postnatal development. Delayed postnatal development of high affinity TPZ sites is observed in the cerebellum, similar to the postnatal development previously observed in the cortex.

Desensitization of muscarinic acetylcholine receptors: possible relation to receptor heterogeneity and phosphoinositides.

The increases in firing rates of hippocampal cells were examined following microiontophoretic application of several muscarinic cholinergic receptor agonists. The agonists studied had been pharmacologically characterized previously and divided into two classes: class A agonists (e.g. acetylcholine, carbamylcholine, and oxotremorine-M) which maximally stimulate PI turnover and reveal mAChR heterogeneity, and class B agonists (e.g. bethanecol and oxotremorine-1) which poorly stimulate PI turnover and do not alter mAChR conformation/orientation in the hippocampus. While comparable stimulatory effects on hippocampal pyramidal cell firing rates were seen with both classes of agonists during short (20 s) ejection periods, longer applications (greater than 25 s) produced class-dependent differential firing patterns. Prolonged ejection of class A agonists selectively desensitized cells to further, continued application in the same ejection period, and the firing rates declined. Class B agonists produced stimulatory responses in hippocampal cells during the entire ejection period, and DE was not observed. This desensitization effect (DE) was observed only for bursts and not for simple spikes.

Effect of acute and chronic pentylenetetrazol treatment on benzodiazepine and cholinergic receptor binding in rat brain.

The binding of [3H] diazepam and [3H] flunitrazepam in rat cerebral cortex was not altered by either acute or chronic administration of pentylenetetrazol except in rats made to convulse 30 min before sacrifice. Rats treated for up to 6 months with doses of pentylenetetrazol which are below seizure threshold in naive rats, became increasingly sensitive to the CNS stimulant effect of pentylenetetrazol as demonstrated by the development of myoclonus and convulsions during treatment periods. These effects were not correlated with any changes in benzodiazepine binding in cerebral cortex or cerebellum and [3H] quinuclidinyl benzilate binding in cerebral cortex. Acute convulsant doses of pentylenetetrazol increased benzodiazepine binding in cerebral cortex, but only in those rats which actively convulsed. Benzodiazepine and cholinergic receptors of the cortex, and benzodiazepine receptors of the cerebellum, therefore, do not appear to change with either the acute or chronic subconvulsive administration of pentylenetetrazol.

Lateral olfactory tract lesions reveal neuronal localization of benzodiazepine recognition sites.

The effects of neuronal degeneration on benzodiazepine binding parameters were assessed following bilateral electrolytic lesions of rat brain lateral olfactory tracts. [3H]Flunitrazepam binding was measured in olfactory bulb homogenates 1, 7 and 14 days post-lesion. Specific [3H]flunitrazepam binding was significantly decreased two weeks after lesions. Diminution in binding was associated with a decreased population of sites (Bmax) rather than affinity alterations (KD). The time course for the loss of receptors is compatible with retrograde degeneration and suggests that specific benzodiazepine binding sites are in part located on mitral cell-bodies.

Antagonism of the anxiolytic action of diazepam and chlordiazepoxide by the novel imidazopyridines, EMD 39593 and EMD 41717.

The imidazopyridines EMD 35993 and EMD 41717 antagonized the anticonflict actions of diazepam and chlordiazepoxide in rodent models which are predictive for anxiolytic action in man. In contrast to other described benzodiazepine antagonists, these compounds did not antagonize either the anticonvulsant or muscle relaxant properties of either benzodiazepine. Both EMD 39593 and EMD 41717 competitively inhibit the binding of [3H]diazepam to brain membranes, but do not exhibit regional differences in potency. These observations suggest that both EMD 39593 and EMD 41717 display some selectivity in antagonizing the anxiolytic properties of benzodiazepines, and as such may be useful tools in identifying neuronal substrates of anxiety.

Spermatozoa with chromosomal abnormalities may result in a higher rate of recurrent abortion.

OBJECTIVE: To determine whether sperm aneuploidy can lead to abortion. DESIGN: Clinical study. SETTING: Couples with reproductive problems evaluated in a private diagnostic laboratory. PATIENT(S): Two men whose wives had histories of multiple abortions. INTERVENTION(S): Whole and Percoll-processed semen samples were analyzed. MAIN OUTCOME MEASURE(S): The results of fluorescence in situ hybridization. RESULT(S): Aneuploidy rates in Percoll-processed samples were higher than those found in whole specimens. Aneuploid spermatozoa also displayed greater motility. CONCLUSION(S): Sperm aneuploidy should be studied before and after Percoll capacitation in all couples with unexplained infertility.