Vibrational line shape effects in plasmon-enhanced stimulated Raman spectroscopies.

A density matrix treatment of plasmon-enhanced (PE) stimulated Raman spectroscopies is developed. Specifically, PE stimulated Raman Gain/Loss (PE-SRG/L) and coherent anti-Stokes Raman scattering (PE-CARS) due to monochromatic excitation and PE femtosecond stimulated Raman spectroscopy (PE-FSRS) are considered. A Lorentz oscillator model is used to explicitly describe the time dependence of plasmon-enhanced optical fields. These temporal characteristics are required for a density matrix based description of all plasmon-enhanced nonlinear molecular spectroscopies. Dispersive vibrational line shapes in PE-SRG/L and PE-FSRS spectra are shown to result primarily from terms proportional to the square of the complex optical field enhancement factor. The dependence on the plasmon resonance, picosecond and femtosecond pulse characteristics, and molecular vibrational properties are evident in the density matrix derived PE-FSRS intensity expression. The difference in signal detection mechanisms accounts for the lack of dispersive line shapes in PE spontaneous Raman spectroscopy. This density matrix treatment of PE-FSRS line shapes is compared with prior coupled wave results.

Surface enhanced Raman scattering for robust, sensitive detection and confirmatory identification of dried bloodstains.

An optimized procedure is described for the acquisition of 785 nm excited SERS spectra of dried bloodstains and shown to offer great potential for rapid, portable, highly sensitive and specific, confirmatory identification for forensic applications. Following extraction in 1 µL of 50% acetic acid, a robust, highly reproducible SERS spectrum is observed from dried bloodstains resulting from a hematin-like heme moiety (ferric, high spin). As anticipated, this blood signature can be classified with 100% specificity and sensitivity with respect to the SERS spectra of other body fluids. High quality SERS spectra can be observed from stains of blood diluted by as much as 105. Dried blood spectra acquired on Au and Ag SERS active substrates exhibit very different relative intensities at this electronically, non-resonant excitation wavelength (785 nm) indicating that a strong chemical effect contributes to the SERS enhancement of this body fluid. DFT calculations further confirm the vibrational band assignments of the features seen in these SERS spectra of dried blood.

Anomalous pH-Dependent Enhancement of p-Methyl Benzoic Acid Sum-Frequency Intensities: Cooperative Surface Adsorption Effects.

Vibrational sum-frequency generation (SFG) spectroscopy is used to determine the surface pKa of p-methyl benzoic acid (pMBA) at the air-water interface by monitoring the carbonyl and carboxylate stretching modes over the pH range of 2 to 12. The SFG intensities of pMBA and its conjugate base, p-methyl benzoate (pMBA-), exhibit an anomalously large enhancement over a narrow pH range (∼0.5) centered at pH 6.3 near the SFG-determined surface pKa, 5.9 ± 0.1. The increase in the surface pKa relative to the bulk value of 4.34 is consistent with the trend previously observed for long chain carboxylic acids in which the surface pKa is higher than the bulk solution pKa. SFG polarization studies help distinguish the orientation and number density contributions to this observed anomalous surface phenomenon. The large SFG intensity increase is attributed to an increase in the pMBA and pMBA- surface concentrations in this narrow pH range due to a cooperative adsorption effect between pMBA and pMBA-. This cooperativity is manifested only on the 2D air-water interface, where the interactions between the acid and base are not as dielectrically screened as in the aqueous bulk phase. Surface effects are critical to understanding and controlling the reactivity, solubility, and behavior of organic acids at interfaces and can have an impact on biomedical applications.

Two-dimensional infrared spectroscopy from the gas to liquid phase: density dependent J-scrambling, vibrational relaxation, and the onset of liquid character.

Ultrafast 2DIR spectra and pump-probe responses of the N2O ν3 asymmetric stretch in SF6 as a function of density from the gas to supercritical phase and liquid are reported. 2DIR spectra unequivocally reveal free rotor character at all densities studied in the gas and supercritical region. Analysis of the 2DIR spectra determines that J-scrambling or rotational relaxation in N2O is highly efficient, occurring in ∼1.5 to ∼2 collisions with SF6 at all non-liquid densities. In contrast, N2O ν3 vibrational energy relaxation requires ∼15 collisions, and complete vibrational equilibrium occurs on the ∼ns scale at all densities. An independent binary collision model is sufficient to describe these supercritical state point dynamics. The N2O ν3 in liquid SF6 2DIR spectrum shows no evidence of free rotor character or spectral diffusion. Using these 2DIR results, hindered rotor or liquid-like character is found in gas and all supercritical solutions for SF6 densities ≥ρ* = 0.3, and increases with SF6 density. 2DIR spectral analysis offers direct time domain evidence of critical slowing for SF6 solutions closest to the critical point density. Applications of 2DIR to other high density and supercritical solution dynamics and descriptions are discussed.

Ultrafast Two-Dimensional Infrared Spectroscopy of a Quasifree Rotor: J Scrambling and Perfectly Anticorrelated Cross Peaks.

Ultrafast two-dimensional infrared (2DIR) spectra of the N_{2}O ν_{3} mode in moderately dense SF_{6} gas exhibit complex line shapes with diagonal and antidiagonal features in contrast to condensed phase vibrational 2DIR spectroscopy. Observed spectra for this quasifree rotor system are well captured by a model that includes all 36 possible rovibrational pathways and treats P (ΔJ=-1) and R (ΔJ=+1) branch resonances as distinct Kubo line shape features. Transition frequency correlation decay is due to J scrambling within one to two gas collisions at each density. Studies of supercritical solvation and relaxation at high pressure and temperature are enabled by this methodology.

Rapid urinary tract infection diagnostics by surface-enhanced Raman spectroscopy (SERS): identification and antibiotic susceptibilities.

SERS spectra of 12 bacterial strains of urinary tract infection (UTI) clinical isolates grown and enriched from urine are reported. A partial least squares-discriminant analysis (PLS-DA) classification treatment of these SERS spectra results in strain level identification with >95% sensitivity and >99% specificity. The classification model successfully identified the SERS spectra of a urine-cultured strain not used to build this statistical model. Enrichment was accomplished by a filtration and centrifugation protocol. The predetermined drug susceptibility profiles of these clinical isolates thus allowed the SERS methodology to provide appropriate UTI antibiotic information in less than 1 h. Most of this time was used for sample preparation procedures (enrichment and washing) for this proof of principle study. SERS spectra of the enriched bacterial samples are dominated by nucleotide degradation metabolites: adenine, hypoxanthine, xanthine, guanine, uric acid, AMP, and guanosine. Strain-specific specificity is due to the different relative amounts of these purines contributing to the corresponding SERS spectra of these clinical isolates. All measurements were made at the minimal bacterial concentration in urine for UTI diagnosis (105 cfu/mL). Graphical abstract The relative contribution of each of the seven purines found to contribute to the bacterial SERS spectra are summarized in this bar graph. Although strain specific differences are evident, it can be see how the pattern of contributing purines is more different between the four species than between strains of a given species.

NIR Raman spectra of whole human blood: effects of laser-induced and in vitro hemoglobin denaturation.

Care must be exercised in the use of Raman spectroscopy for the identification of blood in forensic applications. The Raman spectra of dried whole human blood excited at 785 nm are shown to be exclusively due to oxyhemoglobin or related hemoglobin denaturation products. Raman spectra of whole blood are reported as a function of the incident 785-nm-laser power, and features attributable to heme aggregates are observed for fluences on the order of 10(4) W/cm(2) and signal collection times of 20 s. In particular, the formation of this local-heating-induced heme aggregate product is indicated by a redshifting of several heme porphyrin ring vibrational bands, the appearance of a large broad band at 1,248 cm(-1), the disappearance of the Fe-O2 stretching and bending bands, and the observation of a large overlapping fluorescence band. This denaturation product is also observed in the low-power-excitation Raman spectrum of older ambient-air-exposed bloodstains (2 weeks or more). The Raman spectrum of methemoglobin whole blood excited at 785 nm is reported, and increasing amounts of this natural denaturation product can also be identified in Raman spectra of dried whole blood particularly when the blood has been stored prior to drying. These results indicate that to use 785-nm-excitation Raman spectra as an identification method for forensic applications to maximum effect, incident laser powers need to be kept low to eliminate variable amounts of heme aggregate spectral components contributing to the signal and the natural aging process of hemoglobin denaturation needs to be accounted for. This also suggests that there is a potential opportunity for 785-nm-excitation Raman spectra to be a sensitive indicator of the age of dried bloodstains at crime scenes.

Ultra-sensitive, rapid detection of dried bloodstains by surface enhanced Raman scattering on Ag substrates.

A simple water extraction and transfer procedure is found to result in reproducible and highly sensitive 785 nm excited SERS spectra of 24 h dried bloodstains on Ag nanoparticle substrates. This protocol allows confirmatory detection and identification of dried stains of blood that have been diluted by up to 105 in water on Ag substrates. While previous SERS results demonstrated similar performance on Au substrates when a 50% acetic acid extraction and transfer procedure was used, the water/Ag methodology avoids any potential DNA damage when the sample size is extremely small (≤∼1 µL) due to low pH exposure. The water only procedure is not effective on Au SERS substrates. This metal substrate difference results from the efficient red blood cell lysis and hemoglobin denaturation effects of the Ag nanoparticle surfaces as compare to that of Au nanoparticles. Consequently, the 50% acetic acid exposure is required for the acquisition of 785 nm SERS spectra of dried bloodstains on Au substrates.

Electron correlation effects on the femtosecond dephasing dynamics of E22 excitons in (6,5) carbon nanotubes.

Highly nonlinear pump fluence dependence was observed in the ultrafast one-color pump-probe responses excited by 38 fs pulses resonant with the E(22) transition in a room-temperature solution of (6,5) carbon nanotubes. The differential probe transmission (ΔT/T) at the peak of the pump-probe response (τ = 20 fs) was measured for pump fluences from ∼10(13) to 10(17) photons/pulse cm(2). The onset of saturation is observed at ∼2 × 10(15) photons/pulse cm(2) (∼8 × 10(5) excitons/cm). At pump fluences >4 × 10(16) photons/pulse cm(2) (∼1.6 × 10(6) excitons/cm), ΔT/T decreases as the pump fluence increases. Analogous signal saturation behavior was observed for all measured probe delays. Despite the high exciton density at saturation, no change in the E(22) population decay rate was observed at short times (<300 fs). The pump probe signal was modeled by a third-order perturbation theory treatment that includes the effects of inhomogeneous broadening. The observed ΔT/T signal is well-fit by a pump-fluence-dependent dephasing rate linearly dependent on the number of excitons created by the pump pulse. Therefore, the observed nonlinear pump intensity dependence is attributed to the effects of quasi-elastic exciton-exciton interactions on the dephasing rates of single carbon nanotubes. The low fluence total dephasing time is 36 fs, corresponding to a homogeneous width of 36 meV (290 cm(-1)), and the derived E(22) inhomogeneous width is 68 meV (545 cm(-1)). These results are contrasted with photon-echo-derived parameters for the E(11) transition.

Conjugate Acid-Base Interaction Driven Phase Transition at a 2D Air-Water Interface.

A lattice model is described to explain a recent striking Sum Frequency Generation (SFG) observation of a cooperative surface adsorption effect for an organic acid system at an air-water interface. The reported anomalous pH-dependent enhancement in p-methylbenzoic acid (pmBA) arises from an interaction between the acid (HA) and its conjugate base anion (A-), which competes with strong Coulombic repulsion between the conjugate bases (A--A -). Using a statistical mechanical approach, this lattice gas model reveals an analogy to well-studied magnetic systems in which the attraction between the two different molecular species leads to a phase transition to a two-dimensional checkerboard phase consisting of a network of anion-acid complexes formed at the low-dielectric air-water interface. Cooperative acid-anion interactions that control partitioning at solution and aerosol interfaces are of interest to fields ranging from oceanic and atmospheric chemistry, pharmacology, and chemical engineering.

Surface enhanced Raman scattering specificity for detection and identification of dried bloodstains.

Surface enhanced Raman spectroscopy (SERS) provides highly specific vibrational signatures identifying dried blood for a variety of forensic applications. SERS spectra on Au nanoparticle substrates excited at 785 nm are found to identify dried stains of human and nonhuman blood from seven animals, and distinguish stains due to menstrual and peripheral blood. In addition, the unique SERS bloodstain spectrum is distinct from the SERS spectra of thirty red-brown stains of potential household substances that could be visually mistaken for bloodstains and from food stains that have been shown to give positive results with presumptive colorimetric blood tests. Finally, a SERS swab procedure has been developed and demonstrates that the substrates that a blood sample dried on does not offer any Raman or fluorescence interference for the SERS identification of dried blood. Such bloodstains on porous and nonporous materials are all identical and exclusively due to the heme moiety of hemoglobin. Optimized selection of the extraction solvent is found to control the chemical composition of molecular components appearing in the SERS spectrum of complex, multicomponent biological mixtures, such as body fluids.

Ultrafast H2 and D2 rotational Raman responses in near critical CO2: an experimental and theoretical study of anisotropic solvation dynamics.

The optical heterodyne detected anisotropic rotational Raman responses of H(2) and D(2) (22 mol %) in a near critical CO(2) (rho(*) = rho/rho(c) = 0.8, T = 308 K) solution are reported. J-specific rotational Raman correlation functions (RCFs) for the S(J) transitions of H(2) (J = 0,1,2) and D(2) (J = 0,1,2,3) in this CO(2) solution are determined from these measurements. A mixed classical-quantum simulation methodology results in RCFs that are in excellent agreement with the experimentally derived J-specific responses. The observed S(J) coherence decay time scales, J-dependence, rotor mass dependence, and solvent-induced transition frequency shifts are well captured by these simulations. Pure dephasing of these rotational Raman transitions is shown to be close to the homogeneous limit of the standard Kubo line shape analysis and attributable to the rotor center-of-mass translation in an anisotropic solvent cage. Rotor translational motion in the vicinity of a single CO(2) appears to dominate this dephasing mechanism. Mixed classical-quantum simulations, incorporating the effects of solution fluctuation driven nonadiabatic coupling of instantaneous adiabatic states, including full J-mixing, are required for the agreement between theory and experiment obtained here. Simulations of the classically excited angular kinetic energy of D(2) rotors are used as an estimate of T(1) relaxation rates and are found to be negligible compared to the D(2) rotational Raman coherence time scale. These results are discussed in the context of previous mixed classical-quantum and rotational friction calculations of the dephasing and energy relaxation contributions to H(2) rotational Raman coherence decays. Advantages of time domain acquisition of these rotational Raman responses as compared to spontaneous Raman measurements are illustrated here.

Surface enhanced Raman spectroscopy of Chlamydia trachomatis and Neisseria gonorrhoeae for diagnostics, and extra-cellular metabolomics and biochemical monitoring.

SERS spectra excited at 785 nm of the bacteria Chlamydia trahomatis (elementary bodies, EB) and Neisseria gonorrheoae, the causative pathogens for the two most common sexually transmitted diseases (STD), chlamydia and gonorrhea, respectively, are reported. Although both are Gram-negative bacteria, the SERS signatures of C. trachomatis and N. gonorrheoae are completely different. N. gonorrheoae SERS spectra are due to the starvation induced nucleotide metabolites adenine and guanine, and the surface associated co-enzyme nicotinamide adenine dinucleotide and are very similar on Au and Ag although the spectrum appears more rapidly on Ag. The C. trachomatis SERS spectrum is dominated by the vibrational features of cell surface proteins. While features attributable to specific residues and the amide backbone characterize the C. trachomatis spectrum on Ag, the corresponding SERS spectrum on Au substrates displays vibrational characteristics of aggregated proteins. The prospects for the development of a SERS based platform for rapid (

Structure Making and Breaking Effects of Cations in Aqueous Solution: Nitrous Oxide Pump-Probe Measurements.

Ultrafast IR pump-probe responses resonant with the ν3 asymmetric stretch of nitrous oxide (N2O) at ∼2230 cm-1 are reported for 2 M aqueous salt solutions of MgCl2, CaCl2, NaCl, KCl, and CsCl at room temperature. The solvated cations of these chloride solutions span the range from strongly to weakly hydrating ions, and correspondingly are often categorized as structure makers and structure breakers, respectively. The observed salt dependent trends of the N2O ν3 vibrational energy relaxation (VER) and rotational reorientation anisotropy (R(t)) decays are consistent with the categorization of these cations as structure breakers or makers, and show evidence of effects on the water hydrogen bonding network beyond the first solvation shell of these ions. This N2O mode is resonant with the H2O bend-libration band region. The corresponding FTIR is fitted well by a two Gaussian plus sloping continuum baseline model that allows a framework for characterizing the salt perturbations of the solvent spectral density in the ν3 resonant region. Both coupling strengths and density of states effects appear to contribute the systematic cation dependent T1 effects reported here. R(t) decays follow bulk viscosity values. These results are contrasted with previous IR pump-probe studies predominantly based on the relaxation dynamics of the OH/OD vibrational stretch of HOD hydrogen bonded to anions in salt solutions.

Phase I trial of murine monoclonal antibody L6 in combination with subcutaneous interleukin-2 in patients with advanced carcinoma of the breast, colorectum, and lung.

PURPOSE: A phase I trial was undertaken to determine the toxicity and biologic effects of a combination of murine monoclonal antibody L6 (MoAb L6) plus subcutaneous (SC) interleukin-2 (IL-2). PATIENTS AND METHODS: Fifteen patients with refractory adenocarcinoma (five breast, five lung, five colorectal), received L6 at 200 mg/m2 intravenously (IV) daily on days 1 to 7, followed by a 1-week rest period. IL-2 was given at either 2, 3, or 4.5 x 10(6) U/m2 daily doses times 4 days for a total duration of 3 weeks. RESULTS: Side effects of L6 consisted of mild fever and chills along with a rash and serum sickness in one patient. One patient developed dyspnea and urticaria, that resolved with antihistamines. Maximum-tolerated dose (MTD) of SC IL-2 was 3 x 10(6) U/m2, with dose-limiting toxicities that consisted of grade 4 fatigue and dyspnea. Significant decreases in complement levels along with increases in absolute lymphocyte count and eosinophil count were observed. Mean antibody-dependent cellular cytotoxicity from mononuclear cells taken from patients who received IL-2 was elevated significantly compared with baseline in all patients independent of IL-2 dose (P less than .05). Serum IL-2 levels were elevated in 13 of 14 patients (range, 0.9 to 100 U/mL). Human antimouse antibody (HAMA) titers were elevated in nine of 14 (64%) patients who were tested between 3 and 8 weeks after L6 infusion. One patient with breast cancer had a transient mixed response, and one patient with colorectal cancer had a partial response. CONCLUSIONS: L6 and SC IL-2 were well tolerated in the majority of patients when given in the outpatient setting. In view of the clinical efficacy of this combination, more phase II trials are warranted.

Long-term follow-up of patients with complete remission following combination chemotherapy for metastatic breast cancer.

PURPOSE: To determine the long-term clinical course of patients with metastatic breast cancer (MBC) who achieved a complete remission with doxorubicin-alkylating agent-containing combination chemotherapy programs. PATIENTS AND METHODS: To assess the long-term prognosis of MBC, we reviewed our experience with 1,581 patients treated on consecutive doxorubicin and alkylating agent-containing front-line treatment protocols between 1973 and 1982. Treatment was administered for a maximum duration of 2 years. Characteristics of long-term survivors were evaluated, and hazard rates for progression were calculated. RESULTS: From this group, 263 (16.6%) achieved complete responses (CR) and 49 (3.1%) remained in CR for more than 5 years. After a median duration of 191 months, 26 patients remain in first CR, four patients died in CR at times ranging from 118 to 234 months, 18 patients died of breast cancer, and one is alive with metastatic disease. Compared with the overall CR and total patient populations, the long-term CR group had more premenopausal patients, a younger median age, a lower tumor burden, and better performance status. The hazard function shows a substantial drop in risk of progression after approximately 3 years from initiation of therapy. Ten long-term CR patients developed second primary cancers: breast (3), ovary (2), pancreas (1), endometrium (1), colon (1), head and neck (1), and lung (1). CONCLUSION: Most patients with MBC treated with systemic therapies have only temporary responses to treatment, but some patients continue in CR following initial treatment. These data show that a small percentage of patients achieve long-term remissions with standard chemotherapy regimens. Remission consolidation strategies are needed.

Characterization of the surface enhanced raman scattering (SERS) of bacteria.

The surface enhanced Raman scattering (SERS) of a number of species and strains of bacteria obtained on novel gold nanoparticle (approximately 80 nm) covered SiO(2) substrates excited at 785 nm is reported. Raman cross-section enhancements of >10(4) per bacterium are found for both Gram-positive and Gram-negative bacteria on these SERS active substrates. The SERS spectra of bacteria are spectrally less congested and exhibit greater species differentiation than their corresponding non-SERS (bulk) Raman spectra at this excitation wavelength. Fluorescence observed in the bulk Raman emission of Bacillus species is not apparent in the corresponding SERS spectra. Despite the field enhancement effects arising from the nanostructured metal surface, this fluorescence component appears "quenched" due to an energy transfer process which does not diminish the Raman emission. The surface enhancement effect allows the observation of Raman spectra of single bacterial cells excited at low incident powers and short data acquisition times. SERS spectra of B. anthracis Sterne illustrate this single cell level capability. Comparison with previous SERS studies reveals how the SERS vibrational signatures are strongly dependent on the morphology and nature of the SERS active substrates. The potential of SERS for detection and identification of bacterial pathogens with species and strain specificity on these gold particle covered glassy substrates is demonstrated by these results.

Current status of adjuvant therapy of early breast cancer.

Adjuvant therapy in operable breast cancer (stage I and II) can significantly reduce the risk of recurrences and improve survival. In stage I disease, 20-30% of patients will eventually recur. Several prognostic factors may help in identifying poor prognostic subgroups of stage I patients, including ER and PR status, flow cytometry data, nuclear grade, neu oncogene expression, and perhaps haptoglobin-related protein, Cathepsin-D, and Ki-67 expression. Single-agent chemotherapy and oophorectomy have not resulted in prolongation of survival. Combination chemotherapy regimens are superior to single agents, and doxorubicin-containing regimens may be superior to non-doxorubicin-containing regimens. Tamoxifen is effective in improving survival in patients who are ER positive, particularly those women older than 50 years. It appears that six cycles of an effective regimen is as effective as more prolonged administration of the same drugs, and drugs should be given at the optimal dose rate. Preliminary results of alternating non-cross-resistant chemotherapy regimens show promise, but additional data are needed to determine its impact on survival.

Recent advances in the treatment of breast cancer.

Multidisciplinary efforts have defined a number of prognostic factors and newer strategies to improve the outcome of patients with breast cancer. Conservative surgery has led to improved functional and cosmetic results. The development of a number of effective adjuvant regimens has led to improved survival. In patients with stage I disease, several biological characteristics of tumor have been identified that are associated with increased risk of relapse. A multimodality approach to patients with locally advanced disease and inflammatory cancer has resulted in improved survival. A number of hormonal and cytotoxic drug contaminations can palliate metastatic disease, with a small fraction of patients remaining in extended remission. Dose-intensive programs may lead to further improvements in survival of selected patients with this disease.

Dispersed three-pulse infrared photon echoes of nitrous oxide in water and octanol.

Dispersed IR three-pulse photon echoes due to the antisymmetric (ν3) stretch mode of N2O dissolved in H2O and 1-octanol at room temperature are reported and analyzed. The experimentally determined transition frequency-frequency correlation function (FFCF) in these two solvents is explained in terms of inertial solvent contributions, hydrogen bond network fluctuations, and, for octanol, the motions of the alkyl chains. The H2O hydrogen bond fluctuations result in 1.5 ps FFCF decay, in agreement with relaxation rates determined from photon echo based measurements of other aqueous solutions including salt solutions. In octanol, hydrogen bond fluctuations decay on a slower time scale of 3.3 ps and alkyl chain motions result in an inhomogeneous broadening contribution to the ν3 absorption spectrum that decays on a 35 ps time scale. Rotational reorientation of N2O is nearly 3 times faster in octanol as compared to water. Although the vibrational ν3 N2O absorption line shapes in water and octanol are similar, the line widths result from different coherence loss mechanisms. A hot band contribution in the N2O in octanol solution is found to have a significant effect on the echo spectrum due to its correspondingly stronger transition moment than that of the fundamental transition. The dephasing dynamics of the N2O ν3 stretch mode is of interest as a probe in ultrafast studies of complex or nanoconfined systems with both hydrophobic and hydrophilic regions such as phospholipids, nucleic acids, and proteins. These results demonstrate the value of the N2O molecule to act as a reporter of equilibrium fluctuations in such complex systems particularly due to its solubility characteristics and long vibrational lifetime.