Several methodologies have been developed over the past several years for super-resolution fluorescence microscopy including saturated structured-illumination microscopy (SSIM), stimulated emission ...depletion microscopy (STED), photoactivated localization microscopy (PALM), fluorescence photoactivation localization microscopy (FPALM), and stochastic optical reconstruction microscopy (STORM). While they have shown great promise for biological research, these techniques all have individual strengths and weaknesses. This review will describe the basic principles for achieving super resolution, demonstrate some applications in biology, and provide an overview of technical considerations for implementing these methods.
The complex formation between silver(I) and cysteine (H2Cys), penicillamine (H2Pen), and glutathione (H3Glu) in alkaline aqueous solution was examined using extended X-ray absorption fine structure ...(EXAFS) and 109Ag NMR spectroscopic techniques. The complexes formed in 0.1 mol dm–3 Ag(I) solutions with cysteine and penicillamine were investigated for ligand/Ag(I) (L/Ag) mole ratios increasing from 2.0 to 10.0. For the series of cysteine solutions (pH 10–11) a mean Ag–S bond distance of 2.45 ± 0.02 Å consistently emerged, while for penicillamine (pH 9) the average Ag–S bond distance gradually increased from 2.40 to 2.44 ± 0.02 Å. EXAFS and 109Ag NMR spectra of a concentrated Ag(I)–cysteine solution (C Ag(I) = 0.8 mol dm–3, L/Ag = 2.2) showed a mean Ag–S bond distance of 2.47 ± 0.02 Å and δ(109Ag) 1103 ppm, consistent with prevailing, partially oligomeric AgS3 coordinated species, while for penicillamine (C Ag(I) = 0.5 mol dm–3, L/Ag = 2.0) the mean Ag–S bond distance of 2.40 ± 0.02 Å and δ(109Ag) 922 ppm indicate that mononuclear AgS2 coordinated complexes dominate. For Ag(I)–glutathione solutions (C Ag(I) = 0.01 mol dm–3, pH ∼11), mononuclear AgS2 coordinated species with a mean Ag–S bond distance of 2.36 ± 0.02 Å dominate for L/Ag mole ratios from 2.0 to 10.0. The crystal structure of the silver(I)–cysteine compound (NH4)Ag2(HCys)(Cys)·H2O (1) precipitating at pH ∼10 was solved and showed a layer structure with both AgS3 and AgS3N coordination to the cysteinate ligands. A redetermination of the crystal structure of Ag(HPen)·H2O (2) confirmed the proposed digonal AgS2 coordination environment to bridging thiolate sulfur atoms in polymeric intertwining chains forming a double helix. A survey of Ag–S bond distances for crystalline Ag(I) complexes with S-donor ligands in different AgS2, AgS2(O/N), and AgS3 coordination environments was used, together with a survey of 109Ag NMR chemical shifts, to assist assignments of the Ag(I) coordination in solution.
The complex formation between cadmium(II) and the ligands cysteine (H2Cys) and penicillamine (H2Pen = 3,3′-dimethylcysteine) in aqueous solutions, having C Cd(II) ∼ 0.1 mol dm−3 and C H2L = 0.2−2 mol ...dm−3, was studied at pH = 7.5 and 11.0 by means of 113Cd NMR and Cd K- and L3-edge X-ray absorption spectroscopy. For all cadmium(II)−cysteine molar ratios, the mean Cd−S and Cd−(N/O) bond distances were found in the ranges 2.52−2.54 and 2.27−2.35 Å, respectively. The corresponding cadmium(II)−penicillamine complexes showed slightly shorter Cd−S bonds, 2.50−2.53 Å, but with the Cd−(N/O) bond distances in a similar wide range, 2.28−2.33 Å. For the molar ratio C H2L/C Cd(II) = 2, the 113Cd chemical shifts, in the range 509−527 ppm at both pH values, indicated complexes with distorted tetrahedral CdS2N(N/O) coordination geometry. With a large excess of cysteine (molar ratios C H2Cys/C Cd(II) ≥ 10), complexes with CdS4 coordination geometry dominate, consistent with the 113Cd NMR chemical shifts, δ ∼ 680 ppm at pH 7.5 and 636−658 ppm at pH 11.0, and their mean Cd−S distances were 2.53 ± 0.02 Å. At pH 7.5, the complexes are almost exclusively sulfur-coordinated as Cd(S-cysteinate)4 n−, while at higher pH, the deprotonation of the amine groups promotes chelate formation. At pH 11.0, a minor amount of the Cd(Cys)34− complex with CdS3N coordination is formed. For the corresponding penicillamine solutions with molar ratios C H2Pen/C Cd(II) ≥ 10, the 113Cd NMR chemical shifts, δ ∼ 600 ppm at pH 7.5 and 578 ppm at pH 11.0, together with the average bond distances, Cd−S 2.53 ± 0.02 Å and Cd−(N/O) 2.30−2.33 Å, indicate that Cd(penicillaminate)3 n− complexes with chelating CdS3(N/O) coordination dominate already at pH 7.5 and become mixed with CdS2N(N/O) complexes at pH 11.0. The present study reveals differences between cysteine and penicillamine as ligands to the cadmium(II) ion that can explain why cysteine-rich metallothionines are capable of capturing cadmium(II) ions, while penicillamine, clinically useful for treating the toxic effects of mercury(II) and lead(II) exposure, is not efficient against cadmium(II) poisoning.
Cutaneous immune-related adverse events (cirAEs) occur in up to 40% of immune checkpoint inhibitor (ICI) recipients. However, the association of cirAEs with survival remains unclear.
To investigate ...the association of cirAEs with survival among ICI recipients.
ICI recipients were identified from the Mass General Brigham healthcare system and Dana-Farber Cancer Institute. Patient charts were reviewed for cirAE development within 2 years after ICI initiation. Multivariate time-varying Cox proportional hazards models, adjusted for age, sex, race/ethnicity, Charlson Comorbidity Index, ICI type, cancer type, and year of ICI initiation were utilized to investigate the impact of cirAE development on overall survival.
Of the 3731 ICI recipients, 18.1% developed a cirAE. Six-month landmark analysis and time-varying Cox proportional hazards models demonstrated that patients who developed cirAEs were associated with decreased mortality (hazardratio HR = 0.87, P = .027), particularly in patients with melanoma (HR = 0.67, P = .003). Among individual morphologies, lichenoid eruption (HR = 0.51, P < .001), psoriasiform eruption (HR = 0.52, P = .005), vitiligo (HR = 0.29, P = .007), isolated pruritus without visible manifestation of rash (HR = 0.71, P = .007), acneiform eruption (HR = 0.34, P = .025), and non-specific rash (HR = 0.68, P < .001) were significantly associated with better survival after multiple comparisons adjustment.
Retrospective design; single geography.
CirAE development is associated with improved survival among ICI recipients, especially patients with melanoma.
Curative intent therapy of stage II NSCLC may include surgical resection or definitive radiotherapy. Primary management with surgery or radiotherapy may be influenced by patient and disease ...characteristics. We sought to perform a comparison of patients receiving surgery or radical radiation therapy as their curative treatment, and explore the impact of known prognostic factors on outcome.
A retrospective review was completed of all patients with stage II NSCLC referred to the BC Cancer Agency from 2005 to 2012. Cases were filtered to identify those receiving curative intent therapy including surgery or radiotherapy. Information was collected on known prognostic and predictive factors. The primary outcome measure was overall survival. We compared survival among patients receiving curative intent radiotherapy versus surgical intervention.
A total of 535 patients were referred. Of these, 245 (46%) received curative intent surgery, 132 (25%) curative intent radiotherapy, and 158 (30%) did not receive curative therapy. There were significant differences between cohorts with respect to median age, histology, ECOG PS, smoking status, and weight loss. Median OS was significantly different between cohorts: 61.4 m surgery, 26.5 m curative RT, and 13.1 m non-curative therapy. In a case-matched analysis, median OS remained superior for surgery at 101.6 m vs 28.1 m for curative RT. In a multivariate analysis, ECOG PS, weight loss, and treatment cohort all influenced survival. Among patients receiving curative intent radiotherapy, the use of concurrent chemotherapy and RT dose > = 60Gy were associated with improved outcomes.
Among patients with stage II NSCLC, many are unable to undergo standard of care surgical resection. Radiotherapy provides an inferior yet still curative option in the management of inoperable patients. Further work is needed to optimize outcomes in this population.
Metastatic non-small cell lung cancer (NSCLC) has a poor prognosis. Most patients present with stage IV, and many patients treated curatively with stage I to III develop recurrent metastatic disease. ...It is unknown whether the natural history differs between patients with recurrent versus de novo metastatic NSCLC. We hypothesized that de novo metastatic status is associated with decreased overall survival compared with recurrent metastatic disease.
A retrospective review was completed of all patients with NSCLC referred to BC Cancer from 2005 to 2012. Two cohorts were created; de novo metastatic disease and patients treated with curative intent (surgery or radiotherapy) that developed recurrent, metastatic disease. Information was collected on known prognostic and predictive factors. Overall survival was calculated from the date of diagnosis of metastatic disease.
A total of 9651 patients were evaluated, 5782 (60%) with de novo stage IV disease, and 3869 (40%) with stage I to III disease. Of the 1658 patients who received curative therapy for stage I to III disease, 757 (46%) developed metastases. Patients in the de novo cohort versus recurrent cohort were more likely male (52% vs. 48%), have poorer performance status (Eastern Cooperative Oncology Group≥2 50% vs. 44%), and receive no palliative systemic therapy (67% vs. 61%). The median overall survival in the de novo cohort was 4.7 versus 6.9 m in the recurrent cohort (P<0.001). De novo status was associated with shorter overall survival and this remained significant in a multivariate model that incorporated known prognostic factors.
In a large population-based study of NSCLC, de novo metastatic status was independently associated with decreased overall survival from the time of metastatic disease diagnosis.
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