Recent advances in neoadjuvant immunotherapy for urothelial bladder cancer: What to expect in the near future.

Urothelial bladder cancer (UC) is the most common malignancy involving the urinary system and represents a significant health problem. Immunotherapy has been used for decades for UC with intravesical bacillus Calmette-Guérin (BCG) set as the standard of care for non-muscle-invasive bladder cancer (NMIBC). The advent of immune checkpoint inhibitors (ICIs) has completely transformed the treatment landscape of bladder cancer enabling to expand the treatment strategies. Novel ICIs have successfully shown improved outcomes on metastatic disease to such an extent that the standard of care paradigm has changed leading to the development of different trials with the aim of determining whether ICIs may have a role in early disease. The localized muscle-invasive bladder cancer (MIBC) scenario remains challenging since the recurrence rate continues to be high despite all therapeutic efforts. This article will review the current experience of ICIs in the neoadjuvant setting of UC, the clinical trials landscape and finally, an insight of what to expect in the immediate and mid-term future.

Three-way multivariate analysis of metal fractionation results from sediment samples obtained by different sequential extraction procedures and ICP-OES.

Three typical schemes for metal fractionation were applied to analyse coastal surface sediment samples from Bahía Blanca estuary, where an important industrial emplacement is located. Also, three certified reference materials for total metal concentrations were analysed. The studied metals were cadmium, chromium, copper, lead and zinc because of their hazardous potential and related abundance in the estuary. The concentration of metals was determined by inductively coupled plasma-optical emission spectrometry (ICP-OES). A three-way multivariate analysis was performed in order to obtain a better visualization of the experimental data. The extracted information was used to evaluate the equivalence among the results obtained by the three sequential extraction schemes. The data were analysed by parallel factor analysis (PARAFAC). PARAFAC models with two factors describe appropriately the data sets (explained variance about 54% and core consistency of 100%). The multivariate decomposition showed that the three applied schemes are able to describe equally well the behaviour of the metals in the different sediment fractions.

Synchronous fluorescence for simultaneous determination of hydroquinone and resorcinol in air samples.

Several phenolic compounds are present in tobacco smoke. They are formed from the pyrolysis of tobacco during the smoking process and all of them are toxic. Therefore, the determination of these compounds in air samples is important. A rapid, simple, and sensitive method using a synchronous spectrofluorimetry technique was developed to quantify hydroquinone and resorcinol simultaneously. One of the advantages of this method is the simple and rapid sampling technique, which uses water as the absorption solution of the analytes in the air sample. The precision of the method (%RSD) was 1.8% and the detection limits were 0.125 mg m(-3) and 0.292 mg m(-3) for hydroquinone and resorcinol, respectively.

Evaluation of the mutagenic, antimutagenic and antiproliferative potential of Croton lechleri (Muell. Arg.) latex.

Sangre de Drago is a red viscous latex extracted from Croton lechleri (Euphorbiaceae) cortex, renowned in South American popular medicine for its wound-healing properties. The in vitro antiproliferative effects were determined on the human myelogenous leukemia K562 cells line (IC50 = 2.5 +/- 0.3 microg ml(-1)). The mutagenic and antimutagenic activity of C. lechleri sap was examined by means of the Ames/Salmonella test. No mutagenic activity was found on the Salmonella typhimurium strains T98 and T100, either with or without S9 activation. On the other hand, the sap showed an inhibitory effect against the mutagenic activity of the indirectly acting mutagen 2-Aminoanthracene in presence of S9 and a moderate protective activity against directly acting mutagens Sodium Azide and 2-Nitrofluorene. Therefore we suggest that C. lechleri sap interacts with the enzymes of the S9 mix, thereby inhibiting the transformation of 2-Aminoantracene into its active forms.

Fluorimetric determination of fluoride in a flow assembly integrated on-line to an open/closed FIA system to remove interference by solid phase extraction.

A simple flow injection fluorimetric method for fluoride determination is proposed. The method is based on the enhanced fluorescence of quercitin-Zr(IV) complex when fluoride ion is present in the sample. An open/closed FIA manifold with a mini-column of Dowex 50W X8 resin was used to remove the most important interference (aluminum). The two FIA assemblies were integrated on-line to automate the pretreatment of the water sample and fluoride determination. The calibration graph was linear over the range 0.1-3.0 mug ml(-1) of fluoride with a correlation coefficient of 0.999 and LOD 0.06 mug ml(-1). The relative standard deviation was 2.5% and the sample throughput was 52 h(-1) without pretreatment and 10 h(-1) with pretreatment of the sample. The method was applied to the determination of fluoride in water samples.

Use of eosin as a fluorophore in capillary electrophoresis with laser detection.

Eosin has been used to generate the background signal for indirect fluorimetric detection of inorganic and organic ions, simultaneously separated by capillary zone electrophoresis (CZE). This reagent provides constant fluorescence over the pH range of 5-10 and is compatible with the excitation by an argon ion laser at 488 nm with emission at 520 nm. The use of esosine as fluorophore, H3BO3, and Na2B4O7 as electrolyte and diethylentriamine as modifier of the electroosmotic flow in CZE were optimised. The analytical potential of the studied buffer was tested on a group of 12 anions, used as model compounds. Both, hydrodynamic and electrokinetic injection mode were optimised. The detection limits determined by the last injection mode, were in the range 0.008-0.037 mg l(-1). By using this method, the quantitation of the common anions in tap and mineral water has been carried out successfully.

Flow-injection fluorimetric determination of mercury(II) with calcein.

A flow-injection spectrofluorimetric method for the determination of Hg(II) is described. The method is based on the complex that is formed between Hg(II) and calcein at pH 11-12. This reagent was not used before for determining Hg(II). The excitation wavelength is 324 nm and the emission wavelength is 522 nm. The calibration graph shows a linear range between 7.7 and 128 microg l(-1) (relative standard deviation 2.0%), the sampling rate was 90 h(-1). The effect of several interferents has been examined. Trace mercury in natural waters were determined satisfactorily using a modified manifold, which has an EDTA stream in order to minimize the calcium and magnesium interference.

Differential properties of pre- and postsynaptic 5-hydroxytryptamine1A receptors in the dorsal raphe and hippocampus: II. Effect of pertussis and cholera toxins.

The injection of 1 microgram of pertussis toxin, which inactivates Gi/o proteins, in the rat dorsal raphe nearly abolished the responsiveness of serotonin (5-HT) neurons to microiontophoretic applications of 5-HT and selective 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) without altering their responsiveness to of gamma-aminobutyric acid (GABA). In contrast, the injection of 1 microgram of cholera toxin, which causes an activation of Gs proteins, did not alter the responsiveness of 5-HT neurons to 5-HT, 8-OH-DPAT or GABA. Such in situ injection of either toxin in the dorsal hippocampus decreased by about 90% the responsiveness of CA3 pyramidal neurons to microiontophoretic applications onto their cell body of 5-HT and 8-OH-DPAT, but not of GABA. The effectiveness of the stimulation of the ascending 5-HT pathway in suppressing the firing activity of the same neurons, which results from the release of 5-HT at the level of their dendritic tree, was also markedly decreased in the cholera toxin-treated rats, but intriguingly not in the pertussis toxin-treated rats. These results indicate that, on 5-HT neurons, the somato-dendritic 5-HT1A autoreceptor is coupled to Gi/o, but insensitive to the persistent activation of Gs proteins. In the CA3 region of the hippocampus, there would be two subsets of postsynaptic 5-HT1A receptors on the pyramidal neurons: those apposed to 5-HT terminals on their dendritic tree (denoted intrasynaptic) and those located on their cell body (denoted extrasynaptic). The former are cholera toxin sensitive, whereas the latter are sensitive to both pertussis and cholera toxins.

Differential properties of pre- and postsynaptic 5-hydroxytryptamine1A receptors in the dorsal raphe and hippocampus: I. Effect of spiperone.

The i.v. administration of the 5-hydroxytryptamine (5-HT)1A antagonist spiperone (1 mg/kg) antagonized the suppressant effect of microiontophoretic applications of 5-HT and of the selective 5-HT1A agonist 8-OH-DPAT on the firing activity of the rat dorsal raphe 5-HT neurons. In contrast, the same dose of spiperone did not alter the responsiveness to microiontophoretic applications of 5-HT and 8-OH-DPAT of pyramidal neurons in the CA3 region of the hippocampus, therefore indicating that spiperone does not act as a classical antagonist at these postsynaptic 5-HT1A receptors. After two daily injections of spiperone (5 mg/kg, i.p.) and a third one immediately before the experiment, the responsiveness of CA3 pyramidal neurons to 5-HT applied by microiontophoresis and to that released by the electrical stimulation of the ascending 5-HT pathway was markedly reduced. After 24, but not 12 or 48 h after a single injection of spiperone (5 mg/kg, i.p.), the responsiveness to exogenous and endogenous 5-HT was decreased. Smaller doses (1 and 2.5 mg/kg, i.p.) were ineffective. Haloperidol and ketanserin (5 mg/kg, i.p., 24 h before the experiment) were ineffective in blocking the responsiveness of CA3 pyramidal neurons to 5-HT, thereby demonstrating that the dopamine D2 and 5-HT2 properties of spiperone could not account for the attenuated responsiveness of the hippocampus neurons to 5-HT. Methiothepin (5 mg/kg, i.p., 24 h before recording), as for spiperone, antagonized the suppressant effect of 5-HT applied by microiontophoresis.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of the 5-hydroxytryptamine receptor modulating the release of 5-[3H]hydroxytryptamine in slices of the human neocortex.

In the rat brain, the presynaptic 5-hydroxytryptamine (5-HT) autoreceptors located on 5-HT terminals correspond to the 5-HT1B subtype. The presence of a 5-HT receptor probably located on 5-HT nerve endings and modulating transmitter release in the human neocortex has been reported, but its detailed pharmacological characterization is not yet available. On the other hand, receptor binding and autoradiographic results indicate that the 5-HT1B receptor subtype is not present in the human brain. We, therefore, studied the modulation of the electrically evoked release of [3H]5-HT by various 5-HT receptor agonists and antagonists in preloaded slices of human neocortex obtained from 18 patients undergoing neurosurgery. The nonselective 5-HT1A/1B/1D receptor agonist 5-carboxamidotryptamine produced a potent inhibition (70% at 0.03 microM) of the electrically evoked release of [3H]5-HT which was blocked by 5-HT receptor antagonists with the following relative order of potency: methiothepin greater than metergoline = methysergide greater than propranolol. The selective 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin at 0.1 microM did not modify the electrically evoked release of [3H]5-HT. The 5-HT1A/1B receptor agonist RU 24969 was 10 times more potent at inhibiting [3H]5-HT overflow in the rat frontal cortex than in the human neocortex. The potent 5-HT1B receptor antagonist cyanopinodolol did not modify the 5-carboxamidotryptamine-induced inhibition of the electrically evoked release of [3H]5-HT in slices of the human neocortex, but produced by itself a small inhibition of [3H]5-HT overflow.(ABSTRACT TRUNCATED AT 250 WORDS)

Benzodiazepine receptors modulating serotonergic neurotransmission in rat hippocampus do not desensitize after long-term diazepam treatment.

Previous studies have shown in the rat that long-term diazepam treatment induces a desensitization of gamma-aminobutyric acid (GABA) benzodiazepine receptors which modulate serotonergic (5-HT) neuron firing activity in the dorsal raphe. We have examined the effect of a 21-day treatment with diazepam (5 mg/kg/day, i.p.) on the benzodiazepine receptors (BZR) which modulate the efficacy of the electrical stimulation of the ascending 5-HT pathway in suppressing the firing activity of rat dorsal hippocampus pyramidal neurons. The efficacy of the stimulation was not modified 24 h after the last dose of diazepam. Moreover, the potentiating effect of acute diazepam (1 mg/kg, i.v.) on the efficacy of the stimulation was similar in diazepam- and vehicle-treated rats. These results indicate that GABA-benzodiazepine receptors that modulate the release of 5-HT in the rat dorsal hippocampus are not desensitized by long-term diazepam treatment.

Benzodiazepine receptors modulate 5-hydroxytryptamine neurotransmission in the rat hippocampus: in vivo electrophysiological evidence.

In the present electrophysiological studies the effects of the activation of putative presynaptic benzodiazepine receptors (BZR) in modulating serotonergic (5-HT) transmission in dorsal hippocampus were investigated in chloral hydrate-anesthetized rats. The effectiveness of the electrical stimulation of the ascending 5-HT pathway on the firing activity of CA3 hippocampus pyramidal neurons was modulated by the intravenous administration of BZR ligands. The BZR agonists, diazepam, lorazepam and CL 218,872, dose-dependently enhanced the effectiveness of the stimulation. In contrast, the BZR inverse agonist, FG 7142, dose-dependently decreased it. The effects of agonist and inverse agonist BZR ligands were blocked by the specific BZR antagonist flumazenil. Diazepam and FG 7142 did not modify the efficacy of microiontophoretic applications of gamma-aminobutyric acid and 5-HT onto hippocampal CA3 neurons. BMY 7378, a 5-HT1A receptor antagonist which blocks the effect of the endogenous 5-HT released by the electrical stimulation of 5-HT fibers in this paradigm, also blocked the enhancing action of diazepam on the effectiveness of the stimulation. These results indicate that the activation of BZR of type I modulates 5-HT neurotransmission, presumably through a presynaptic regulatory mechanism.

The benzodiazepine receptor inverse agonist DMCM decreases serotonergic transmission in rat hippocampus: an in vivo electrophysiological study.

The intravenous administration of the benzodiazepine inverse agonist DMCM (6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylic acid methyl ester) dose-dependently reduced the efficacy of electrical stimulation of the 5-HT pathway in depressing the firing activity of rat hippocampus pyramidal neurons. This effect was prevented by the benzodiazepine receptor antagonist flumazenil and reversed by the benzodiazepine receptor agonist diazepam. This indicates that DMCM alters 5-HT transmission in the rat dorsal hippocampus via benzodiazepine receptors and that these receptors have both agonist and inverse agonist sites.

Differential rates of urinary noradrenalin excretion in affective disorders: utility of a short time sampling procedure.

Levels of morning urinary noradrenalin (NA) excretion were compared in 61 patients with unipolar major depression (MDE), 25 patients with minor depression (mDE), 39 patients with anxiety disorder (AD), and 21 healthy control subjects (C). The following differences in the level of urinary NA excretion rate were found: MDE greater than mDE = AD greater than C. In addition, a significant positive correlation was found in the MDE group between severity of illness, expressed as total Hamilton depression score, in diagnostic subclasses from the Research Diagnostic Criteria, and urinary NA excretion rates (first episode, MDE total group, recurrent, with psychotic features). These data support the utility of a short time urine sampling procedure to measure NA excretion as a peripheral marker monitoring sympathetic activity in inpatient as well as outpatient conditions.

In vivo presynaptic modulation of serotonergic neurotransmission in the rat hippocampus by diazepam.

The effect of the benzodiazepine agonist, diazepam, on serotonergic (5-HT) neurotransmission was assessed in in vivo electrophysiological experiments. Diazepam enhanced, in a dose-dependent manner (0.05-2 mg/kg i.v.), the effect of electrical stimulation of the ascending 5-HT pathway on the firing activity of dorsal hippocampus pyramidal neurons. This effect was blocked by the benzodiazepine antagonist, flumazenil. Diazepam did not modify the efficacy of microiontophoretic application of 5-HT and GABA. The results indicate that the activation of benzodiazepine receptors facilitates of 5-HT neurotransmission, presumably through a presynaptic modulatory mechanism.

Modulation of the electrically evoked release of 5-[3H]hydroxytryptamine from rat cerebral cortex: effects of alpidem, CL 218872, and diazepam.

The effect of omega (benzodiazepine)-receptor agonists, antagonists, and inverse agonists on the electrically evoked release of 5-[3H]hydroxytryptamine ([3H]5-HT) was studied in superfused slices of the rat frontal cerebral cortex. The electrically evoked release of [3H]5-HT was enhanced by nanomolar concentrations of diazepam and the selective omega 1-receptor agonists alpidem and CL 218872. The omega 1/omega 2- and omega 1-receptor antagonists flumazenil and CGS 8216, respectively, did not modify the electrically evoked release of [3H]5-HT. The omega 3-receptor agonist Ro 5-4864 and the omega 1-receptor inverse agonist ethyl-beta-carboline-3-carboxylate on their own did not affect the electrically evoked release of [3H]5-HT. On the other hand, the inverse agonist 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylic acid methyl ester (DMCM), at micromolar concentrations, inhibited both the spontaneous and the evoked release of [3H]5-HT. The facilitation of the electrically evoked release of [3H]5-HT by diazepam, alpidem, or CL 218872 was potentiated by gamma-aminobutyric acid (GABA). Exposure to flumazenil and CGS 8216 antagonized the facilitation by diazepam, alpidem, or CL 218872 of [3H]5-HT release. The inhibition of the release of [3H]5-HT by DMCM was not modified by exposure to either flumazenil, CGS 8216, or GABA. The inhibitory effect of DMCM was not observed when monoamine oxidase activity was inhibited by pargyline.(ABSTRACT TRUNCATED AT 250 WORDS)

Rat striatal dopamine and tetrahydrobiopterin content following an intrastriatal injection of manganese chloride.

Injection of manganese into the rat corpus striatum causes a rapid fall in the biopterin and dopamine (DA) content ipsilateral to the lesion. Two weeks after the lesion both biopterin and DA are partially recovered. Controls, injected with saline or magnesium, do not show alterations in their DA or cofactor levels. It is proposed that the fall in DA levels results from a rapid displacement of the amine from its storage sites by manganese followed by a decrease in the rate of DA synthesis causes by the drop in cofactor levels.

High urinary norepinephrine excretion in major depressive disorders: effects of a new type of MAO inhibitor (Moclobemide, RO 11-1163).

The urinary excretion rates of norepinephrine were assayed in 26 patients diagnosed as major depressive disorders (primary, unipolar), before and after 14 days of treatment with the monoamine oxidase inhibitor Moclobemide (Ro 11-1163). A standardized 1-h urine collection procedure was used and norepinephrine was assayed by liquid chromatography with electrochemical detection. Norepinephrine was found significantly increased in depressed patients when compared with a control population. The psychotic patients showed the highest excretion rates although they were not significantly different from the endogenous (non-psychotic) group. Urinary norepinephrine output significantly decreased after 14 days of treatment with Moclobemide. This decrease was also marked in those patients that did not show any therapeutic effect. A clear antidepressant effect, shown by a significant decrease of the Hamilton Scale scores for depression, was apparent as early as the 7th day. Increased norepinephrine in melancholic patients was taken as a presumptive indication of altered sympathetic activity.

Resting urinary catecholamine excretion in schizophrenics: methodology and interpretation of results.

Plasma and urinary catecholamines were measured in a group of schizophrenics and in normal subjects by high performance liquid chromatography with electrochemical detection. A brief procedure for urine sampling performed under standardized environmental and physical conditions was used. Plasma levels and urinary excretion rates of noradrenaline were significantly higher in schizophrenics than in a control population. When plotted for linear regression analysis, both plasma and urinary noradrenaline values were positively correlated (r = 0.75, p less than 0.05). Results reported suggest that urinary catecholamine measurement could be used for a reliable assessment of sympatho-adrenal function in schizophrenia.