Use of acupuncture with acupressure in addition to standard-of-care cryotherapy to decrease chemotherapy-associated neuropathy in patients with gastrointestinal malignancies receiving oxaliplatin-based chemotherapy: Study protocol for a randomized, controlled pilot and feasibility study.

BACKGROUND: Oxaliplatin is a key chemotherapeutic agent in the treatment of local and metastatic gastrointestinal (GI) malignancies. Dose density and treatment adherence can be limited by chemotherapy-induced peripheral neuropathy (CIPN). Early research suggests CIPN incidence and severity may be mitigated by acupuncture, but rigorous data in GI oncology patients is limited. Here, we describe the protocol of a randomized, waitlist-controlled pilot study testing the use of preemptive of acupuncture plus acupressure to decrease CIPN and chemotherapy-related toxicities. METHODS: Patients with a GI malignancy (n = 56) with planned 5-fluorouracil (5-FU) and oxaliplatin IV (FOLFOX, FOLFIRINOX) every 2 weeks are being recruited. Additional concurrent anti-neoplastic agents may be used. Enrolled patients are randomized 1:1 to a 3-month intervention of Arm A: acupuncture with acupressure and standard-of-care treatment, or Arm B: standard-of-care alone. In Arm A, on days 1 and 3 of each chemotherapy cycle a standardized acupuncture protocol is administered and patients are taught self-acupressure to perform daily between chemotherapy treatments. Patients in both arms are given standard-of-care oral and peripheral (hands/feet) ice chip cryotherapy during oxaliplatin administration. CIPN and other symptoms are assessed at baseline, 6 weeks, and 3 months from registration. The primary endpoint is CIPN severity at 3 months (EORTC-CIPN 20). Additional endpoints evaluate CIPN incidence (CTCAE, Neuropen, tuning fork); incidence of pain, fatigue, nausea, oral dysesthesia, and anxiety; and feasibility (recruitment, retention, adherence, acceptability). If warranted, trial results will inform the design of a multi-center trial to expand testing of the intervention to a larger patient cohort.

Label-free imaging of human cells: algorithms for image reconstruction of Raman hyperspectral datasets.

Raman microspectroscopy-based, label-free imaging methods for human cells at sub-micrometre spatial resolution are presented. Since no dyes or labels are used in this imaging modality, the pixel-to-pixel spectral variations are small and multivariate methods of analysis need to be employed to convert the hyperspectral datasets to spectral images. Thus, the main emphasis of this paper is the introduction and comparison of a number of multivariate image reconstruction methods. The resulting Raman spectral imaging methodology directly utilizes the spectral contrast provided by small (bio)chemical compositional changes over the spatial dimension of the sample to construct images that can rival fluorescence images in terms of spatial information, yet without the use of any external dye or label.

Detection of breast micro-metastases in axillary lymph nodes by infrared micro-spectral imaging.

We report the ability of infrared micro-spectral imaging, coupled with completely unsupervised methods of multivariate statistical analysis, to accurately reproduce the histological architecture of axillary lymph nodes and detect metastatic breast cancer cells. The acquisition of spectral data from tissue embedded in paraffin provided spectra free of dispersive artefacts that may be observed for infrared microscopic measurements using a 'reflection/absorption' methodology. As a consequence, superior tissue classification and identification of cellular abnormality unattainable for deparaffinised tissue was achieved.

Infrared micro-spectral imaging: distinction of tissue types in axillary lymph node histology.

BACKGROUND: Histopathologic evaluation of surgical specimens is a well established technique for disease identification, and has remained relatively unchanged since its clinical introduction. Although it is essential for clinical investigation, histopathologic identification of tissues remains a time consuming and subjective technique, with unsatisfactory levels of inter- and intra-observer discrepancy. A novel approach for histological recognition is to use Fourier Transform Infrared (FT-IR) micro-spectroscopy. This non-destructive optical technique can provide a rapid measurement of sample biochemistry and identify variations that occur between healthy and diseased tissues. The advantage of this method is that it is objective and provides reproducible diagnosis, independent of fatigue, experience and inter-observer variability. METHODS: We report a method for analysing excised lymph nodes that is based on spectral pathology. In spectral pathology, an unstained (fixed or snap frozen) tissue section is interrogated by a beam of infrared light that samples pixels of 25 mum x 25 mum in size. This beam is rastered over the sample, and up to 100,000 complete infrared spectra are acquired for a given tissue sample. These spectra are subsequently analysed by a diagnostic computer algorithm that is trained by correlating spectral and histopathological features. RESULTS: We illustrate the ability of infrared micro-spectral imaging, coupled with completely unsupervised methods of multivariate statistical analysis, to accurately reproduce the histological architecture of axillary lymph nodes. By correlating spectral and histopathological features, a diagnostic algorithm was trained that allowed both accurate and rapid classification of benign and malignant tissues composed within different lymph nodes. This approach was successfully applied to both deparaffinised and frozen tissues and indicates that both intra-operative and more conventional surgical specimens can be diagnosed by this technique. CONCLUSION: This paper provides strong evidence that automated diagnosis by means of infrared micro-spectral imaging is possible. Recent investigations within the author's laboratory upon lymph nodes have also revealed that cancers from different primary tumours provide distinctly different spectral signatures. Thus poorly differentiated and hard-to-determine cases of metastatic invasion, such as micrometastases, may additionally be identified by this technique. Finally, we differentiate benign and malignant tissues composed within axillary lymph nodes by completely automated methods of spectral analysis.

Cytology by Infrared Micro-Spectroscopy: Automatic Distinction of Cell Types in Urinary Cytology.

We report microscopically collected infrared spectra of cells found in human urine in an effort to develop automatic methods for bladder cancer screening. Unsupervised multivariate analysis of the observed spectral patterns reveals distinct spectral classes, which correlated very well with visual cytology. Therefore, we believe that spectral analysis of individual cells can aid cytology in rendering reliable diagnoses based on objective measurements and discriminant algorithms.

Infrared micro-spectroscopic studies of epithelial cells.

We report results from a study of human and canine mucosal cells, investigated by infrared micro-spectroscopy, and analyzed by methods of multivariate statistics. We demonstrate that the infrared spectra of individual cells are sensitive to the stage of maturation, and that a distinction between healthy and diseased cells will be possible. Since this report is written for an audience not familiar with infrared micro-spectroscopy, a short introduction into this field is presented along with a summary of principal component analysis.

Cell-cycle-dependent variations in FTIR micro-spectra of single proliferating HeLa cells: principal component and artificial neural network analysis.

We have previously reported spectral differences for cells at different stages of the eukaryotic cell division cycle. These differences are due to the drastic biochemical and morphological changes that occur as a consequence of cell proliferation. We correlate these changes in FTIR absorption and Raman spectra of individual cells with their biochemical age (or phase in the cell cycle), determined by immunohistochemical staining to detect the appearance (and subsequent disappearance) of cell-cycle-specific cyclins, and/or the occurrence of DNA synthesis. Once spectra were correlated with their cells' staining patterns, we used methods of multivariate statistics to analyze the changes in cellular spectra as a function of cell cycle phase.

Raman and infrared microspectral imaging of mitotic cells.

We report the first ever Raman and infrared microspectroscopic images of human cells at different stages of mitosis. These spectroscopic methods monitor the distribution of condensed nuclear chromatin, and other biochemical components, utilizing inherent protein and DNA spectral markers, and, therefore, do not require the use of any stains. In conjunction with previously reported data from the G1, S, and G2 phases of the cell cycle, the complete cell division cycle has now been mapped by spectroscopic methods. Although the results reported here do not offer new insights into the distribution of biochemical components during mitosis, the recognition of cell division without the use of stains, and the possibility of doing so on living cells, may be useful for an automatic, spectroscopic determination of the proliferation rates of cells and tissues. Spectral images were constructed by plotting spectral intensities of DNA or protein versus the coordinates from which spectra were recorded. We found that both Raman and infrared intensities depend on the overall chromatin density variation among the individual subphases of mitosis.

Infrared micro-spectroscopy of human cells: Causes for the spectral variance of oral mucosa (buccal) cells.

We discuss the causes contributing to the variance of the spectra of individual human epithelial cells. This aspect has largely been ignored in previous studies, but needs to be understood for diagnostic applications of infrared micro-spectroscopy. We attribute the spectral variance to Mie scattering, and to variations of nuclear contributions to the overall spectra caused by different nuclear size.

Infrared spectral imaging of lymph nodes: Strategies for analysis and artifact reduction.

In this contribution, we discuss state-of-the-art methodology for the collection and analysis of hyperspectral images of tissue that will become useful in complementing classical histopathology. In particular, we discuss sampling strategies, data collection methods, and computational approaches to produce pseudo-color maps of large tissue sections of lymph nodes, up to about 100 mm(2) in size. The latter efforts include methods to reduce the presence of dispersion artifacts in IR transflection micro-spectra which can greatly impact the statistical analyzes performed on the data, such as hierarchical cluster analysis and principal components analysis.

Acupuncture to Treat the Symptoms of Patients in a Palliative Care Setting.

INTRODUCTION: Patients enrolled in hospice and palliative care programs experience a vast array of symptoms requiring the expertise of a multidisciplinary team to address. Acupuncture can be an effective addition to a hospice team whose goal is maximum comfort and quality of life (QOL). The objective of this project was to examine the effectiveness of acupuncture to relieve symptoms commonly observed in patients in a hospice program. METHODS: All over 26 patients participated in the acupuncture trial, receiving a course of weekly treatments that ranged from 1 to 14 weeks. The average number of treatments was five. The Edmonton Symptom Assessment Scale (ESAS) was used to assess the severity of pain, tiredness, nausea, depression, anxiety, drowsiness, appetite, well-being, and dyspnea. A two-tailed, paired t test was applied to the data to compare symptom scores pre- versus post-acupuncture treatment. SETTINGS/PARTICIPANTS: Patients enrolled in All Care Hospice's home care program were given the option to receive acupuncture to supplement usual care offered by the hospice team. Treatment was provided by licensed acupuncturists in the patient's place of residence. RESULTS: Seven out of nine symptoms were significantly (P < .001) improved with acupuncture, the exceptions being drowsiness and appetite. Although the ESAS scale demonstrated a reduction in symptom severity post-treatment for both drowsiness and appetite, this reduction was not found to be significant. CONCLUSION: Acupuncture was found to be effective for the reduction and relief of symptoms that commonly affect patient QOL. Acupuncture effectively reduced symptoms of pain, tiredness, nausea, depression, anxiety, and shortness of breath, and enhanced feelings of well-being. More research is required to assess the long-term benefits and symptom reduction of acupuncture in a palliative care setting.

Acupuncture in Uganda: a research letter on the integration of acupuncture with conventional biomedical treatments.

The PanAfrican Acupuncture Project (PAAP) was created in 2001 and is a volunteer-based not-for-profit training organization. It encourages community empowerment and wellness through training local healthcare providers how to use simple, effective acupuncture protocols to treat the symptoms of HIV/ AIDS, malaria, tuberculosis, and other chronic conditions. A manual explains the theory of acupuncture and the techniques associated with it in clear and simple terms with specific acupuncture protocols associated with the signs and symptoms of illnesses commonly seen in Uganda. PAAP maintains contact with the trainees to provide ongoing support through: (1) three week-long meetings per year, (2) ongoing contact with the PAAP local coordinator, and (3) electronic communications including email, text messaging, and Facebook communication. A primary focus is to help integrate acupuncture into the local context and current public-health system. This is accomplished by working with local governmental and nongovernmental agencies and healthcare facilities to improve access to and effectiveness of care through collaboration with Western and traditional healthcare modalities. The integration of acupuncture and other traditional complementary and alternative medicines (TCAMs) in the international public health sector at the level of local health workers can increase access to treatment in a sustainable manner while also building the confidence and self-worth of the public health-care practitioners.

Spectral detection of micro-metastases in lymph node histo-pathology.

The first detection of breast cancer micrometastases in lymph nodes by infrared spectral imaging and methods of multivariate analysis is reported. Micrometastases are indicators of early spread of cancer from the organ originally affected by disease, and their detection is of prime importance for the staging and treatment of cancer. Infrared spectral imaging, at a spatial resolution of ca. 10-12 mum, can detect small metastases down to the level of a few cancerous cells. The results presented here add to a rapidly growing database of infrared spectral imaging results for cancer diagnostics.

Chapter 10: Infrared and Raman microscopy in cell biology.

This chapter presents novel microscopic methods to monitor cell biological processes of live or fixed cells without the use of any dye, stains, or other contrast agent. These methods are based on spectral techniques that detect inherent spectroscopic properties of biochemical constituents of cells, or parts thereof. Two different modalities have been developed for this task. One of them is infrared micro-spectroscopy, in which an average snapshot of a cell's biochemical composition is collected at a spatial resolution of typically 25 mum. This technique, which is extremely sensitive and can collect such a snapshot in fractions of a second, is particularly suited for studying gross biochemical changes. The other technique, Raman microscopy (also known as Raman micro-spectroscopy), is ideally suited to study variations of cellular composition on the scale of subcellular organelles, since its spatial resolution is as good as that of fluorescence microscopy. Both techniques exhibit the fingerprint sensitivity of vibrational spectroscopy toward biochemical composition, and can be used to follow a variety of cellular processes.

FTIR analysis and monitoring of synthetic aviation engine oils.

Synthetic turbine oils from military aircraft engines were analysed for antioxidant content and total acid number using infrared (IR) spectroscopy. Two-dimensional IR correlation analysis was employed to investigate and interpret observed trends in the spectra, as acid was formed and antioxidant species were depleted in the oils, as a function of aging and engine wear. Principal components and partial least squares algorithms were used and compared for the development of calibration and prediction models. Transmission IR spectrometry is demonstrated to be effective for the analysis and monitoring of synthetic aviation turbine engine oils and shown to provide rapid and accurate information as compared with traditional analytical techniques and methods.

Mie-type scattering and non-Beer-Lambert absorption behavior of human cells in infrared microspectroscopy.

We report infrared microspectral features of nuclei in a completely inactive and contracted (pyknotic) state, and of nuclei of actively dividing cells. For pyknotic nuclei, the very high local concentration of DNA leads to opaqueness of the chromatin and, consequently, the absence of DNA signals in the IR spectra of very small nuclei. However, these nuclei can be detected by their scattering properties, which can be described by the Mie theory of scattering from dielectric spheres. This scattering depends on the size of the nucleus; consequently, quite different scattering cross-sections are calculated and observed for pyknotic and mitotic nuclei.

Infrared microspectroscopy of individual human cervical cancer (HeLa) cells.

We report infrared (IR) spectra observed for individual, cultured human cervical cancer (HeLa) cells. Spectra were collected microscopically, in reflection/absorption modes, from cells deposited and dried on microscope slides or from cells grown directly on slides. Within the spectra of the dried cells, significant spectral heterogeneity exists that was previously attributed to different stages of the cell cycle [Boydston-White. S., et al., Biospectroscopy 1999, 5, 219-227; Holman, H. Y., et al., Biopolymers Biospectrosc 2000, 57, 329-335; Tobin, M. J., et al., in First BASIE Workshop, Karlsruhe, Germany, 2003]. The results reported here confirm earlier findings and present the possibility of determining the abundance of cells within each stage of the cycle from the IR spectra. In an accompanying paper, we show that the spectra of cells in suspension exhibit spectral intensity distributions that are different from that of the dried cells. This result has far-reaching implications for the use of infrared microspectroscopy to screen dried cell preparations for the presence of abnormal cells.

Infrared microspectroscopy of individual human cervical cancer (HeLa) cells suspended in growth medium.

We report for the first time the infrared spectra of individual human cervical cancer (HeLa) cells suspended in buffer or cell culture medium. Although we did not establish whether these cells were viable at the time of spectral data acquisition, we believe that the methodology used is applicable to the study of live cells. Data were collected either from entire cells, using 25- to 40-microm apertures, or via an imaging approach, where pixels measuring 6.25 x 6.25 microm were assembled to form a map of a cell in suspension. Measurements were carried out both in reflection/absorption and in transmission modes. The results reported here might have far-reaching implications for the use of infrared microspectroscopy to monitor cell proliferation, drug response, and other cell biological parameters in live cells.

Influence of benign cellular changes in diagnosis of cervical cancer using IR microspectroscopy.

IR spectroscopy and principal components analysis (PCA) of endocervical cells and smears diagnosed with benign cellular changes were investigated to determine the influence of these potential confounding variables in the diagnosis of cervical cancer. Spectral differences in all cell and diagnostic types investigated were found in the phosphodiester and carbohydrate regions. However, the spectral differences in other bands were not distinct enough to allow differentiation between groups. The PCA was successfully used to obtain a separation of normal ectocervical smears from normal endocervical cells and smears diagnosed with inflammation, Candida albicans, and bacterial vaginosis. A separation with a slight overlap of abnormal ectocervical smears from normal endocervical cells, inflammation, and bacterial vaginosis was obtained with PCA. Candida was not separated from abnormal ectocervical smears with any success.

Removal of blood components from cervical smears: implications for cancer diagnosis using FTIR spectroscopy.

Red and white cell lysis buffers were investigated to determine their ability to remove blood components from cervical smears prior to IR microspectroscopy. A white cell lysis buffer was effective in increasing the spectral reproducibility and sample homogeneity and in reducing the presence of inflammatory exudate, particularly leukocytes, from cervical smears. The reduction of leukocytes appeared to cause abnormal samples to be grouped with normal samples, resulting in poor discrimination. Despite differences between the cytological and histological diagnoses of cervical abnormalities, the results indicate that the differences seen in the spectra of exfoliated malignant and nonmalignant cervical cells could be due to the presence of inflammation.