Étude de faisabilité du ganglion sentinelle dans le cancer de la prostate par cœlioscopie : premiers résultats

Objectives

The standard lymphadenectomy is currently a challenge in the management of prostate cancer. The aim of this prospective study was to evaluate the performance of the sentinel lymph node (SLN) by laparoscopy in patients with localized prostate cancer, candidates for local treatment.

Patients and methods

Patients were injected transrectal ultrasound-guided with 0.3 mL/100MBq 99^mTc-Sulfur rhenium colloid in each prostatic lobe, the day before surgery. Lymphoscintigraphy was performed after 2 hours. The detection was realized intraoperatively with a laparoscopic probe (Gamma Sup Clerad^®) followed by extensive dissection. Counts of SLN were performed in vivo and confirmed ex vivo. The histological analysis was performed by HPS staining and followed by immunochemistry if SLN was free.

Results

Seventy patients with carcinoma of the prostate at intermediate or high risk of lymph node metastases (D’Amico), PSA median 9.5 ng/mL [6–130], were included in the study. The lymphoscintigraphic detection rate was 94.2% (66/70) and intraoperative of 97.0% (68/70). Fourteen patients had lymph node metastases, six only in SLN. The false negative rate was 2/14 (14.0%). The internal iliac region is the first metastatic site (40.9%). Limited or standard lymph node dissection would have ignored respectively 72.7% and 59.0% of lymph node metastases.

Conclusion

The laparoscopy is adapted to a broad identification of SLN and targeted dissection of these lymph nodes significantly limits the risk of surgical extended dissection while maintaining the accuracy of the information.     

Valeur pronostique du curage ganglionnaire prophylactique dans le cancer différencié de la thyroïde

Objective

Although the efficacy of total thyroidectomy as the first step in differentiated thyroid cancer (DTC) management is well proven, it is always a matter of controversy whether prophylactic lymph node dissection improves prognosis and survival in DTC patients without suspicion of lymph node metastases either clinically or radiologically. The aim of this work is to study relapse rates in patients according to the initial presentation and type of surgery performed, in order to analyze the prognostic impact of lymph node dissection.

Patients and methods

One hundred and thirty patients with DTC were classified according to clinical presentation and type of surgery performed. They were monitored for six years to watch for recurrence.

Results

In patients with no clinical evident lymph nodes, the relapse rate was lower among patients who underwent total thyroidectomy in one step with prophylactic lymph node dissection.

Conclusion

Based on the results of other studies, and on our department experience of thyroid cancer management and monitoring, we strongly advocate careful and systematic exploration of the central lymph node compartments with exploration of the lateral side of the lesion if in doubt.     

Fixations extra-osseuses du Tc MDP dans le cas d’un cancer du sein métastatique à l’os

Aim

To report and discuss an unusual visceral uptake on bone scan in a case of breast cancer with bone metastases.

Patient and methods

A 40-year-old woman, with untreated bilateral breast cancer was referred to our department for a bone scan.

Results

The bone scan evidenced multiple metastases over the axial skeleton. Uncommonly, visceral uptake was noted associating diffuse bilateral lung uptake and intense myocardium, stomach and kidneys uptakes. Serum calcium level was high: 4.08 mmol/L (normal: 2.38–2.70 mmol/L).

Conclusion

The incidental observation of metastasic calcifications on bone scan is often related to severe hypercalcemia. Such pattern should alert the physician on the existence and the severity of calcium metabolism disturbances that had not been suggested before.     

Clinical and Laboratory Toxicity after Intra-Arterial Radioembolization with 90Y-Microspheres for Unresectable Liver Metastases

Objective

To investigate clinical and laboratory toxicity in patients with unresectable liver metastases, treated with yttrium-90 radioembolization (⁹⁰Y-RE).

Methods

Patients with liver metastases treated with ⁹⁰Y-RE, between February 1^st 2009 and March 31^st 2012, were included in this study. Clinical toxicity assessment was based on the reporting in patient’s charts. Laboratory investigations at baseline and during a four-month follow-up were used to assess laboratory toxicity according to the Common Terminology Criteria for Adverse Events version 4.02. The occurrence of grade 3–4 laboratory toxicity was stratified according to treatment strategy (whole liver treatment in one session versus sequential sessions). Response assessment was performed at the level of target lesions, whole liver and overall response in accordance with RECIST 1.1 at 3- and 6 months post-treatment. Median time to progression (TTP) and overall survival were calculated by Kaplan-Meier analysis.

Results

A total of 59 patients, with liver metastases from colorectal cancer (n = 30), neuroendocrine tumors (NET) (n = 6) and other primary tumors (n = 23) were included. Clinical toxicity after ⁹⁰Y-RE treatment was confined to grade 1–2 events, predominantly post-embolization symptoms. No grade 3–4 clinical toxicity was observed, whereas laboratory toxicity grade 3–4 was observed in 38% of patients. Whole liver treatment in one session was not associated with increased laboratory toxicity. Three-months disease control rates for target lesions, whole liver and overall response were 35%, 21% and 19% respectively. Median TTP was 6.2 months for target lesions, 3.3 months for the whole liver and 3.0 months for overall response. Median overall survival was 8.9 months.

Conclusion

The risk of severe complications or grade 3–4 clinical toxicity in patients with liver metastases of various primary tumors undergoing ⁹⁰Y-RE is low. In contrast, laboratory toxicity grade 3–4 can be expected to occur in more than one-third of patients without any clinical signs of radiation induced liver disease.     

A phase 1, open-label study of LCAR-B38M,a chimeric antigen receptor T cell therapy directed against B cell maturation antigen,in patients with relapsed or refractory multiple myeloma

Background

Chimeric antigen receptor (CAR) T cell therapy has demonstrated proven efficacy in some hematologic cancers. We evaluated the safety and efficacy of LCAR-B38M, a dual epitope-binding CAR T cell therapy directed against 2 distinct B cell maturation antigen epitopes, in patients with relapsed/refractory (R/R) multiple myeloma (MM).

Methods

This ongoing phase 1, single-arm, open-label, multicenter study enrolled patients (18 to 80?years) with R/R MM. Lymphodepletion was performed using cyclophosphamide 300?mg/m². LCAR-B38M CAR T cells (median CAR+ T cells, 0.5?×?10⁶ cells/kg [range, 0.07 to 2.1?×?10⁶]) were infused in 3 separate infusions. The primary objective is to evaluate the safety of LCAR-B38M CAR T cells; the secondary objective is to evaluate the antimyeloma response of the treatment based on the general guidelines of the International Myeloma Working Group.

Results

At data cutoff, 57 patients had received LCAR-B38M CAR T cells. All patients experienced ≥?1 adverse events (AEs). Grade ≥?3 AEs were reported in 37/57 patients (65%); most common were leukopenia (17/57; 30%), thrombocytopenia (13/57; 23%), and aspartate aminotransferase increased (12/57; 21%). Cytokine release syndrome occurred in 51/57 patients (90%); 4/57 (7%) had grade ≥?3 cases. One patient reported neurotoxicity of grade 1 aphasia, agitation, and seizure-like activity. The overall response rate was 88% (95% confidence interval [CI], 76 to 95); 39/57 patients (68%) achieved a complete response, 3/57 (5%) achieved a very good partial response, and 8/57 (14%) achieved a partial response. Minimal residual disease was negative for 36/57 (63%) patients. The median time to response was 1?month (range, 0.4 to 3.5). At a median follow-up of 8?months, median progression-free survival was 15?months (95% CI, 11 to not estimable). Median overall survival for all patients was not reached.

Conclusions

LCAR-B38M CAR T cell therapy displayed a manageable safety profile and demonstrated deep and durable responses in patients with R/R MM.

Trial registration

ClinicalTrials.gov, NCT03090659; Registered on March 27, 2017, retrospectively registered

     

Activation and route of administration both determine the ability of bone marrow-derived dendritic cells to accumulate in secondary lymphoid organs and prime CD8<Superscript>+</Superscript> T cells against tumors

Aims

To examine the effects of route of administration and activation status on the ability of dendritic cells (DC) to accumulate in secondary lymphoid organs, and induce expansion of CD8⁺ T cells and anti-tumor activity.

Methods

DC from bone marrow (BM) cultures were labeled with fluorochromes and injected s.c. or i.v. into naïve mice to monitor their survival and accumulation in vivo. Percentages of specific CD8⁺ T cells in blood and delayed tumor growth were used as readouts of the immune response induced by DC immunization.

Results

The route of DC administration was critical in determining the site of DC accumulation and time of DC persistence in vivo. DC injected s.c. accumulated in the draining lymph node, and DC injected i.v. in the spleen. DC appeared in the lymph node by 24 h after s.c. injection, their numbers peaked at 48 h and declined at 96 h. DC that had spontaneously matured in vitro were better able to migrate compared to immature DC. DC were found in the spleen at 3 h and 24 h after i.v. injection, but their numbers were low and declined by 48 h. Depending on the tumor cell line used, DC injected s.c. were as effective or more effective than DC injected i.v. at inducing anti-tumor responses. Pre-treatment with LPS increased DC accumulation in lymph nodes, but had no detectable effect on accumulation in the spleen. Pre-treatment with LPS also improved the ability of DC to induce CD8⁺ T cell expansion and anti-tumor responses, regardless of the route of DC administration.

Conclusions

Injection route and activation by LPS independently determine the ability of DC to activate tumor-specific CD8⁺ T cells in vivo.

     

Association hyperthyroïdie et cancer thyroïdien différencié (médullaire exclu). À propos de sept observations

The aim of this study was to draw attention to the fact that hyperthyroidism does not exclude the possibility of concurrent thyroid carcinoma. We analyzed the frequency and consequences of this association through seven cases of thyrotoxic patients with differentiated thyroid carcinoma followed by a review of literature.

Patients and methods

The nuclear medicine department of the CHU Ibn Sina has more than 3000 cases of differentiated thyroid carcinoma where 10 patients with association of hyperthyroidism could be retrieved. Our study focused on seven cases well documented and regularly followed. The main parameters collected were the following: age, sex, personal and family history of thyroid disease, clinical presentation, etiology of hyperthyroidism, preoperative paraclinical explorations, surgery performed, histopahological findings, complementary therapy and follow-up.

Results

Group included six females and one male, average age was 41.4 years. There were four cases of Graves’ disease, two cases of single toxic nodule and one case of functional multinodular goiter. All patients underwent total thyroidectomy. Final pathologic examination disclosed five papillary carcinomas and two follicular carcinomas. During 2 to 8 years follow-up, there was neither local recurrence nor metastasis.

Conclusion

The diagnosis of hyperthyroidism does not preclude concurrent thyroid carcinoma. Careful assessment of such patients is mandatory to exclude an associated thyroid malignancy particularly in the presence of a nodule in a hyperfunctioning thyroid.     

Ecophysiological response of Crambe maritima to airborne and soil-borne salinity

Background and Aims

There is a need to evaluate the salt tolerance of plant species that can be cultivated as crops under saline conditions. Crambe maritima is a coastal plant, usually occurring on the driftline, with potential use as a vegetable crop. The aim of this experiment was to determine the growth response of Crambe maritima to various levels of airborne and soil-borne salinity and the ecophysiological mechanisms underlying these responses.

Methods

In the greenhouse, plants were exposed to salt spray (400 mm NaCl) as well as to various levels of root-zone salinity (RZS) of 0, 50, 100, 200 and 300 mm NaCl during 40 d. The salt tolerance of Crambe maritima was assessed by the relative growth rate (RGR) and its components. To study possible salinity effects on the tissue and cellular level, the leaf succulence, tissue Na⁺ concentrations, Na⁺ : K⁺ ratio, net K⁺/Na⁺ selectivity, N, P, K⁺, Ca²⁺, Mg²⁺, proline, soluble sugar concentrations, osmotic potential, total phenolics and antioxidant capacity were measured.

Key Results

Salt spray did not affect the RGR of Crambe maritima. However, leaf thickness and leaf succulence increased with salt spray. Root zone salinities up to 100 mm NaCl did not affect growth. However, at 200 mm NaCl RZS the RGR was reduced by 41 % compared with the control and by 56 % at 300 mm NaCl RZS. The reduced RGR with increasing RZS was largely due to the reduced specific leaf area, which was caused by increased leaf succulence as well as by increased leaf dry matter content. No changes in unit leaf rate were observed but increased RZS resulted in increased Na⁺ and proline concentrations, reduced K⁺, Ca²⁺ and Mg²⁺ concentrations, lower osmotic potential and increased antioxidant capacity. Proline concentrations of the leaves correlated strongly (r = 0·95) with RZS concentrations and not with plant growth.

Conclusions

Based on its growth response, Crambe maritima can be classified as a salt spray tolerant plant that is sensitive to root zone salinities exceeding 100 mm NaCl.     

Molecular species of membrane phospholipids containing arachidonic acid and linoleic acid contribute to the interindividual variability of red blood cell Na+-Li+ countertransport: In vivo and in vitro evidence

Summary Previous studies indicate a particular sensitivity of red blood cell Na⁺-Li⁺ countertransport activity to small variations in the fatty acid composition of membrane phospholipids. To assess whether the interindividual variability of Na⁺-Li⁺ countertransport is related to differences in the species pattern of erythrocyte phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in vivo, the molecular species composition of PC and PE as well as the kinetics of Na⁺-Li⁺ countertransport were analyzed in parallel in normo- and hyperlipidemic donors. Both in diacyl PC and in diacyl-PE the species 160/204 and 160/182 were, respectively, positively and negatively related to the apparent maximal velocity of Na⁺-Li⁺ countertransport. The sum of all species with 204 at sn₂ of diacyl-PE exhibited a strong positive (r = 0.82, 2p < 0.001), and those containing 182 a negative correlation (r = –0.63, 2p < 0.01) to the transport activity. Essentially similar connections were observed between these species and the apparent affinity of the transport system for intracellular Na⁺. To evaluate whether the associations between molecular species of membrane phospholipids and Na⁺-Li⁺ countertransport activity were indicative of a causal relationship, the species 160/204-PC and 160/182-PC were selectively introduced into the erythrocyte membrane by means of the PC-specific transfer protein. Replacement of 11% of native PC by 160/182-PC inhibited the transport rate by about 25%. Exchange of 6 and 9% of PC with 160/204-PC, in contrast, accelerated the transport rate by 30 and 60%, respectively. The accordance between the in vivo relations and the results of the in vitro modification strongly suggests that elevations and reductions in the arachidonic acid and linoleic acid content of membrane PC and PE contribute to the interindividual variability of red blood cell Na⁺-Li⁺ counter-transport activity and its acceleration in hyperlipidemias.The authors wish to thank Dr. W.O. Richter (II. Medizinische Klinik, Klinikum Großhadern, Universität München) for selection of the patients and Dr. T. Brosche (Universität ErlangenNürnberg) for gaschromatographic analyses. This study was supported in part by a grant of the Wilhelm-Sander-Stiftung to B.E.     

Caméras hybrides TEMP-TDM et semi-quantification du I-FPCIT : évaluation de l’apport de la TDM

Objectives

Tomoscintigraphy of dopamine transporters with ¹²³I-FP-CIT is nowadays essential to visualise impairment of nigro-striatal system for the diagnosis of parkinsonism and for the differential diagnosis of dementia. With the development of hybrid cameras (SPECT-CT), the CT contribution in nuclear neurology needs to be assessed in diagnostic and semi-quantification performances. The main purpose of our study is to compare attenuation correction using CT to attenuation correction using the linear algorithm of Chang. SPECT-CT with parallel collimation results were also weighed against fan beam collimation and the contribution of partial volume effect correction was studied in secondary objective.

Materials and methods

We used a trilinear phantom to define spatial resolution and an anthropomorphic striatal phantom to quantify the activity in striatal cavities. We compared the impact of attenuation and scatter correction on spatial resolution and semi-quantification in striatum. We performed the partial volume effect correction on reconstructed images according to the method of Rousset.

Results

Attenuation correction by CT did not improve significantly spatial resolution compared to the algorithm of Chang. The semi-quantification of ¹²³I-FPCIT in striata was not significantly different according to the various CA, but was significantly improved with CT attenuation and scatter correction. Partial volume effect correction improved the quantification from 40 to 60% in the striatal structures, when the activity was superior in at least twice the background noise.

Conclusion

SPECT-CT hybrid cameras increase spatial resolution and improve semi-quantification of ¹²³I-FPCIT because of CT attenuation and scatter correction. Another use of CT is the possibility of calibrating anatomic segmentation of striata for partial volume effect correction. Partial volume effect correction improves quantification and is essential for early diagnosis of nigro-striatal disease.     

Effects of Krill-derived phospholipid-enriched n − 3 fatty acids on Ca regulation system in cerebral arteries from ovariectomized rats

Aims

To investigate the effects of n − 3 polyunsaturated fatty acids on cerebral circulation, ovariectomized (OVX) rats were administered with phospholipids in krill oil (KPL) or triglycerides in fish oil (FTG); effects on the Ca^2 + regulating system in their basilar artery (BA) were then analyzed.

Main methods

The rats were divided into 4 groups: control, OVX, OVX given KPL (OVXP), and OVX given FTG (OVXT) orally, daily for 2 weeks. Time dependent relaxation (TDR) of contractile response to 5HT in BA was determined myographically, Na⁺/Ca^2 + exchanger (NCX) 1 mRNA expression was determined by real time PCR, and nucleotides were analyzed by HPLC.

Key findings

The level of TDR in OVX that was significantly lower in the control was inhibited by l-NAME and indomethacin; TEA inhibited TDR totally in the control but only partly in OVXP and OVXT. Relaxation induced by the addition of 5 mM KCl to the BA pre-contracted with 5-HT was inhibited by TEA in the controls, OVXP and OVXT, but not in OVX. Overexpression of NCX1 mRNA in the BA from OVX was significantly inhibited by FTG. The ratio of ADP/ATP in cerebral arteries from OVX was significantly inhibited by KPL and FTG. Levels of triglyceride and arachidonic acid in the plasma of OVX increased, but were significantly inhibited by KPL and FTG.

Significance

Ovarian dysfunction affects Ca^2 + activated-, ATP-sensitive-K⁺ channels and NCX1, which play crucial roles in the autoregulation of cerebral blood flow. Also, KPL may become as good a supplement as FTG for postmenopausal women.     

Coordination compounds of Cd(II) with several bidentate-NN′ and tridentate-NN′N nitrogen donor ligands. Cd NMR studies of monomeric compounds containing nitrogen donor atoms

Reaction of CdCl₂ with N-alkylaminopyrazole ligands 1-[(2-ethylamino)ethyl]-3,5-dimethylpyrazole (deae), 1-[(2-(tert-butylamino)ethyl)]-3,5-dimethylpyrazole (deat), bis-[(3,5-dimethylpyrazolyl)methyl]ethylamine (bdmae), and bis-[(3,5-dimethylpyrazolyl)ethyl]ethylamine (ddae) in absolute ethanol yields [CdCl₂(NN′)] (NN′ = deae (1), deat (2)), [CdCl₂(bdmae)] (3), and [CdCl(ddae)]₂[CdCl₄] (4). The Cd(II) complexes have been characterised by elemental analyses, conductivity measurements, IR, ¹H, ¹³C{¹H} and ¹¹³Cd NMR spectroscopies, and X-ray diffraction methods. ¹H and ¹¹³Cd NMR experiments at variable temperature for 3 and 4 show that dynamic processes are taking place in solution. We report the measurements of ¹¹³Cd NMR chemical shift data for complexes 1-4 in solution. X-ray crystal structures for complexes 2 and 3 have been determined. The Cd(II) is coordinated to the deat ligand, in 2, by one nitrogen atom of the pyrazolyl group and one nitrogen atom of the amine. It finishes a tetrahedral geometry with two chlorine atoms. The bdmae ligand is linked to Cd(II), in 3, by two nitrogens atoms of the pyrazolyl groups and one amine nitrogen, along with two chlorine atoms, in a distorted trigonal bipyramidal geometry.     

Biophysical characterization of rat cardiac Ca2+/Mg2+ ecto-ATPase (Myoglein)

Sarcolemmal Ca²⁺/Mg²⁺ ecto-ATPase (Myoglein; MW 180 kD) is a membrane bound enzyme which requires a millimolar concentration of either Ca²⁺ or Mg²⁺ for maximal hydrolysis of ATP. The isoelectric point (pI) of the cardiac ecto-ATPase was 5.7. The purified Ca²⁺/Mg²⁺ ecto-ATPase from the rat heart sarcolemmal appeared as a single band with MW 90 kD in the SDS-PAGE. In order to understand the nature of this enzyme, the 90 kD band in the SDS-PAGE was electroeluted; the analysis of the eluate showed 2 prominent bands with MW 90 and 85 kD. The presence of 2 bands was further confirmed by gradient gel (10-20%) electrophoresis in 0.375 M Tris-HCl buffer, pH 8.8. Analysis of the purified Ca²⁺/Mg²⁺ ecto-ATPase as well as the electroeluted protein in a non-equilibrium linear two dimensional electrophoresis (Ampholyte pI 3.0-10.0) also showed two distinct bands. Mass spectroscopic analysis of the enzyme using different matrix combinations revealed the presence of multi-components indicating microheterogeneity in the protein structure. Treatment of the ecto-ATPase with DL-dithiothreitol did not alter the pattern of mass spectroscopic analysis and this indicated that the microheterogeneity may be due to some posttranslational modifications. It is concluded that rat cardiac Ca²⁺/Mg²⁺ ecto-ATPase is an acidic protein having two subunits. Furthermore, the enzyme shows microheterogeneity in its molecular structure.     

Synthesis and spectroscopic and X-ray structural characterisation of some para-substituted triaryltin(pentacarbonyl)manganese(I) complexes

A series of para-substituted triaryltin(pentacarbonyl)manganese(I) compounds [(p-XC₆H₄)₃SnMn(CO)₅: II, X=CH₃; III, X=CH₃O; IV, X=CH₃S; V, X=F; VI, X=Cl; VII, X=CH₃S(O₂)] is reported for comparison with the known phenyl analogue I. IR data [ν(CO)] as well as complete ¹¹⁹Sn/⁵⁵Mn/¹³C solution NMR results are given for I-VII. Chemical shifts, ¹¹⁹Sn versus ⁵⁵Mn, except I, correlate well, but have differing single parameter (SP) correlations, ¹¹⁹Sn versus σ_I and ⁵⁵Mn versus σ°_p. These results are compared with previous SP studies of the ¹¹⁹Sn solution NMR spectra of the series, (p-XC₆H₄)₄Sn and (p-XC₆H₄)₃SnY (Y=Cl, Br, I). Full crystal structures are reported for compounds II-VI. All are similar to that of I, with the Mn(CO)₅ moiety being a distorted tetragonal pyramid, and having a quasi-mirror plane through the central C₄MnSnC₃ skeleton. The Ar₃Sn are distorted trigonal propellers with ring torsion angles in the range 30-80°, the exception being IV with one torsion angle of 22°.     

Evidence for two modes of Ca2+ entry following muscarinic stimulation of a human salivary epithelial cell line

Summary We have investigated muscarinic receptor-operated Ca²⁺ mobilization in a salivary epithelial cell line, HSG-PA, using an experimental approach which allows independent evaluation of intracellular Ca²⁺ release and extracellular Ca²⁺ entry. The carbachol (Cch) dose response of intracellular Ca²⁺ release indicates the involvement of a single, relatively low-affinity, muscarinic receptor site (K _0.510 or 30 m, depending on the method for [Ca²⁺] _i determination). However, similar data for Ca²⁺ entry indicate the involvement of two Cch sites, one consistent with that associated with Ca²⁺ release and a second higher affinity site withK _0.52.5 m. In addition, the Ca²⁺ entry response observed at lower concentrations of Cch (2.5 m) was completely inhibited by membrane depolarization induced with high K⁺ (>55mm) or gramicidin D (1 m), while membrane depolarization had little or no effect on Ca²⁺ entry induced by 100 m Cch. Another muscarinic agonist, oxotremorine-M (100 m; Oxo-M), like Cch, also induced an increase in the [Ca²⁺] _i of HSG-PA cells (from 72±2 to 104±5nm). This response was profoundly blocked (75%) by the inorganic Ca²⁺ channel blocker La³⁺ (25–50 m) suggesting that Oxo-M primarily mobilizes Ca²⁺ in these cells by increasing Ca²⁺ entry. Organic Ca²⁺ channel blockers (verapamil or diltiazem at 10 m, nifedipine at 1 m), had no effect on this response. The Oxo-M induced Ca²⁺ mobilization response, like that observed at lower doses of Cch, was markedly inhibited (70–90%) by membrane depolarization (high K⁺ or gramicidin D). At 100 m Cch the formation of inositol trisphosphate (IP₃) was increased 55% above basal levels. A low concentration of carbachol (1 m) elicited a smaller change in IP₃ formation (25%), similar to that seen with 100 m Oxo-M (20%). Taken together, these results suggest that there are two modes of muscarinic receptor-induced Ca²⁺ entry in HSG-PA cells. One is associated with IP₃ formation and intracellular Ca²⁺ release and is independent of membrane potential; the other is less dependent on IP₃ formation and intracellular Ca²⁺ release and is modulated by membrane potential. This latter pathway may exhibit voltage-dependent gating.     

Synthesis, characterisation and solution behaviour of fluoroalkyl phosphoryl complexes of tin tetrachloride

The synthesis, characterisation and solution behaviour of a series of octahedral complexes SnCl₄·2L (L = R₂NP(O)(OCH₂CF₃)₂; R = Me (1); Et (2) or L = P(O)(OCH₂R_f)₃; R_f = CF₃ (3); C₂F₅ (4)) are described. Complexes 1-4 were prepared from SnCl₄ and 2 equiv. of the ligand, L, in anhydrous CH₂Cl₂ solution. The adducts have been characterised by multinuclear (¹H, ³¹P and ¹¹⁹Sn) NMR, IR spectroscopy and elemental analysis. In dichloromethane solution, the NMR data showed the presence of a mixture of cis and trans isomers for 1 and 2 and only the cis isomer for 3 and 4. The difference could be interpreted in terms of the electronic effects of the substituents on the phosphorus atom of the ligand. In addition, the solution structure of the complexes studied by variable temperature ³¹P-{¹H} and ¹H NMR in the presence of excess ligand indicated that the ligand exchange on the cis isomer dominates the chemistry. The metal-ligand exchange barriers were estimated to be 13.38 and 11.39 kcal/mol for 1 and 3, respectively. The results are discussed and compared with those previously reported for the related hexamethylphosphoramide adduct, SnCl₄·2HMPA.     

Microsampling of Fish Otoliths: A Comparison Between DM 2800 and Dremel in Stable Isotope Analysis

This paper describes and compares two microsampling methods, DM 2800 and Dremel, which were used in taking aragonite samples from otolith zones of cod, Gadus morhua, for stable isotope analysis. There were no significant differences between the two sampling methods both in carbon and oxygen isotope ratios (¹³C/¹²C, ¹⁸O/¹⁶O) and in annual and seasonal otolith zones. The possible finite effect with Dremel could be minimized or negligible by selecting otoliths of younger fishes. Thus, Dremel may be accepted as a convenient microsampling method in stable isotope analysis of otoliths.     

<Superscript>1</Superscript>H NMR based metabolomic approach to monitoring of the head and neck cancer treatment toxicity

Introduction

Anticancer treatment results in temporary or permanent toxicity considered as changes in normal tissues and/or involved regions. The net effect is mirrored in morphological, functional and molecular disturbances—thus in a systemic response of the human body. To date, specific NMR biomarkers of radiation therapy toxicity in head and neck squamous cell carcinoma (HNSCC) patients are scarce or even missing.

Objectives

We aimed to investigate molecular processes reflecting acute radiation sequelae (ARS) in HNSCC patients using NMR-based metabolomics of blood serum.

Methods

45 patients with HNSCC were treated with radiotherapy (RT) or chemoradiotherapy (CHRT). Blood samples were collected within a week after RT/CHRT completion. Patients were divided into two classes (of high and low ARS) on the basis of the highest individual ARS value observed during the treatment. ¹H NMR spectra of serum samples were acquired on a Bruker 400.13 MHz spectrometer at 310 K and analyzed using principal component analysis and orthogonal partial least squares discriminant analysis. Additional statistical analyses were performed on quantified metabolites.

Results

1D projections of the J-resolved NMR spectra seem to be of the great potential in the quest for the HNSCC treatment toxicity biomarker. The metabolic features characteristic for high ARS are the increased signals of N-acetyl-glycoprotein and acetate, as well as decrease of choline and the metabolites involved in energy metabolism: branched chain amino acids (BCAAs), alanine, creatinine and carnitine. Furthermore, we observed significant correlations between N-acetyl-glycoprotein and clinical markers of inflammation as well as acetate and a percentage-weight-loss during the treatment. CRP was also negatively correlated with alanine and BCAAs.

Conclusion

NMR-based metabolomics provides relevant biomarkers of RT/CHRT toxicity (ARS) in HNSCC patients. The results indicate at least three concomitant processes related to high ARS: inflammation, altered energy metabolism and disturbed membrane metabolism, and indicate an exciting potential of J-resolved NMR spectroscopy combined with multivariate projection techniques.