Metastable oxidation states of tetrathiafulvalenes on the surface of liposomes.

Redox-active liposomes are prepared by the incorporation of tetrathiafulvalene-cholesterol conjugate 1 in phospholipid vesicles. The oxidation of tetrathiafulvalene (TTF) on the surface of the liposomes in aqueous solution is monitored by UV-vis spectroscopy. It is shown that metastable (TTF+˙)2π-dimers of the mono-oxidized cation radical are formed due to the high local concentration of TTF groups in the lipid membrane. These dimers can be further stabilized by the addition of cucurbit[8]uril or by reduction of the lateral mobility in the membrane by variation of the lipid composition.

Hyperthermia sensitization and proton beam triggered liposomal drug release for targeted tumor therapy.

PURPOSE: The objectives of this study were to: 1) determine if mild hyperthermia (40-42°C) can sensitize tumor cells for more effective proton beam radiotherapy (PBRT); 2) characterize the survival fraction of cells exposed to PBRT; and 3) characterize release of the drug doxorubicin (Dox) from low temperature sensitive liposomes (LTSLs) without exposure to mild hyperthermia in combination with PBRT. METHODS: Dox was actively loaded in LTSLs. A549 monolayer cells were incubated with 100-200 nM of Dox-LTSL (±mild hyperthermia). Cell irradiation (0-6 Gy) was performed by placing the cell culture plates inside a solid water phantom and using a clinical proton treatment beam with energy of 150 MeV. End points were survival fraction, radiation-mediated Dox release, and reactive oxygen species (ROS) production. RESULTS: Hyperthermia effectively sensitized cells for PBRT and lowered the cell survival fraction (SF) by an average of 9.5%. The combination of 100 nM Dox-LTSL and PBRT (1-6 Gy) achieved additive to synergistic response at various dose combinations. At higher radiation doses (>3 Gy), the SF in the Dox and Dox-LTSL groups was similar (~20%), even in the absence of hyperthermia. In addition, 30% of the Dox was released from LTSLs and a 1.3-1.6 fold increase in ROS level occurred compared to LTSL alone therapy. CONCLUSIONS: The combination of LTSLs and PBRT achieves additive to synergistic effect at various dose combinations in vitro. Concurrent PBRT and Dox-LTSL treatment significantly improved the cytotoxic outcomes of the treatment compared to PBRT and Dox chemotherapy without LTSLs. We hypothesize that PBRT may induce drug release from LTSL in the absence of hyperthermia.

Inhibition of the bacterial lectins of Pseudomonas aeruginosa with monosaccharides and peptides.

Pseudomonas aeruginosa (PA) can cause infections in compromised hosts by interacting with the glycocalyx of host epithelial cells. It binds to glycostructures on mucosal surfaces via two lectins, which are carbohydrate-binding proteins, named PA-IL and PA-IIL, and blocking this interaction is, thus, an attractive anti-adhesive strategy. The aim of this study was to determine by ciliary beat frequency (CBF) analysis whether monosaccharides or peptides mimicking glycostructures represent blockers of PA lectin binding to human airway cilia. The treatment with monosaccharides and peptides alone did not change the CBF compared to controls and the tested compounds did not influence the cell morphology or survival, with the exception of peptide pOM3. PA-IL caused a decrease of the CBF within 24 h. D-galactose as well as the peptides mimicking HNK-1, polysialic acid and fucose compensated the CBF-modulating effect of PA-IL with different affinities. PA-IIL also bound to the human airway cilia in cell culture and resulted in a decrease of the CBF within 24 h. L(-)-fucose and pHNK-1 blocked the CBF-decreasing effect of PA-IIL. The HNK-1-specific glycomimetic peptide had a high affinity for binding to both PA-IL and PA-IIL, and inhibited the ciliotoxic effect of both lectins, thus, making it a strong candidate for a therapeutic anti-adhesive drug.

Transrectal ultrasound-integrated spectral optical tomography of hypoxic progression of a regressing tumor in a canine prostate.

The objective of this study was to evaluate if transrectal optical tomography implemented at three wavelength bands for spectral detection could monitor changes of the hemoglobin oxygen saturation (StO2) in addition to those of the total hemoglobin concentration ([HbT]) in lesions of a canine prostate, including an induced tumor modeling canine prostate cancer. Near-infrared (NIR) optical tomography was integrated with ultrasound (US) for transrectal imaging. Multi-spectral detection at 705_nm, 785_nm and 808_nm rendered measurements of [HbT] and StO2. Canine transmissible venereal tumor (TVT) cells were injected into the right lobe of a dog's prostate gland, which had a pre-existing cyst in the left lobe. Longitudinal assessments of the prostate were performed weekly over a 63-day duration by NIR imaging concurrent with grey-scale and Doppler US. Ultrasonography revealed a bi-lobular tumor-mass regressing from day-49 to day-63. At day-49 this tumor-mass developed a hypoxic core that became larger and more intense by day-56 and expanded further by day-63. The tumor-mass presented a strong hyper-[HbT] feature on day-56 that was inconsistent with US-visualized blood flow. Histology confirmed two necrotic TVT foci within this tumor-mass. The cyst appeared to have a large anoxic-like interior that was greater in size than its ultrasonographically delineated lesion, and a weak lesional elevation of [HbT]. On day-56, the cyst presented a strong hyper-[HbT] feature consistent with US-resolved blood flow. Histology revealed acute and chronic hemorrhage in the periphery of the cyst. The NIR imaging features of two other TVT nodules and a metastatic lymph node were evaluated retrospectively. Transrectal US-integrated spectral optical tomography seems to enable longitudinal monitoring of intra-lesional oxygenation dynamics in addition to the hemoglobin content of lesions in the canine prostate.

Trans-rectal ultrasound-coupled spectral optical tomography of total hemoglobin concentration enhances assessment of the laterality and progression of a transmissible venereal tumor in canine prostate.

OBJECTIVES: To evaluate whether trans-rectal spectral optical tomography of total hemoglobin concentration (HbT) can image longitudinal and lateral developments of a canine transmissible venereal tumor (TVT) in a canine prostate. METHODS: A near-infrared (NIR) applicator was integrated with a trans-rectal ultrasound (TRUS) transducer to perform ultrasound (US)-coupled optical tomography of the canine prostate. Spectral detection at 785 and 830 nm enabled quantitation of HbT. Canine TVT cells were injected into the right lobe of a dog's prostate gland. Longitudinal imaging assessment of the post-injection prostate was performed by coupled US/NIR imaging over a 45-day duration. RESULTS: By day 7, NIR indicated TVT infiltration in the noninjected left prostatic lobe with the gray-scale US indistinct. By day 31, both NIR and gray-scale US revealed more widespread TVT involvement in the left than in the right lobe, as well as an extensive TVT mass in the caudal aspect of the gland, of which the peak HbT increased 3-fold and the mass volume grew exponentially over the 45-day duration. Increased blood supply to the mass was also observed on Doppler US. CONCLUSIONS: TRUS-coupled spectral optical tomography enhances assessment of the laterality and progression of prostate tumor compared with using gray-scale and Doppler TRUS.

Tissue levels, histologic changes and plasma pharmacokinetics of meta-Tetra (hydroxyphenyl) chlorin (mTHPC) in the cat.

The biodistribution and pharmacokinetics of meta-tetra(hydroxyphenyl)chlorin (mTHPC)(1) have been documented in humans, rats, dogs and rabbits. It has been demonstrated to be an effective photodynamic therapy agent for treatment of squamous cell carcinoma. Squamous cell carcinoma is a common feline neoplasm, causing significant morbidity and mortality in the feline population. The association between ultraviolet radiation exposure and occurrence of this neoplasm in the cat provides a useful model for the study of human cutaneous squamous cell carcinoma. In this study, we document the biodistribution, pharmacokinetics and toxicity of mTHPC in a group of normal cats. Four groups of cats were given the drug intravenously at dosages of 0, 0.15, 0.30 and 0.60 mg/kg. mTHPC levels were measured in plasma and tissues at 0, 24, 48, 72, 96 and 336 h after drug administration. Additionally, plasma samples were collected at 1 and 6 h post-injection and analysed. Biodistribution and pharmacokinetics of mTHPC in cats mirrors that in other animal species. There were no clinical or pathological changes associated with administration of the drug. The biodistribution and pharmacokinetics of mTHPC in cats mirrors that in other species studied. There were no clinical or pathological changes attributable to administration of the drug at the doses administered. mTHPC may be a useful photodynamic therapy drug in cats.

Failure of holmium:yttrium-aluminum-garnet laser lithotripsy in two horses with calculi in the urinary bladder.

Holmium:yttrium-aluminum-garnet (Ho:YAG) laser lithotripsy was attempted in a mare and a gelding with calculi in the urinary bladder. The procedure was unsuccessful in producing adequate fragmentation of the calculi. In the gelding, pulsed dye laser lithotripsy was subsequently used to fragment the urolith. Manual removal of the urolith via the urethra was performed in the mare.

Laser lithotripsy of a urethral calculus via ischial urethrotomy in a steer.

A steer examined because of obstructive urolithiasis and urethral rupture underwent laser lithotripsy, using a chromium-thulium-holmium:yttrium-aluminum-garnet (Ho:YAG) laser inserted through an ischial urethrotomy. Procedures were performed with caudal epidural anesthesia. Six months after surgery, the urethra was patent with no clinical evidence of urethral stricture or fistula. Ischial urethrotomy provided rapid access to the bladder for catheterization and to the obstructive urolith for lithotripsy. Laser lithotripsy was a rapid and effective means of urolith removal in this steer.

Evaluation of carbon dioxide laser and conventional incisional techniques for resection of soft palates in brachycephalic dogs.

OBJECTIVE: To compare clinical outcome, healing, and effect of tracheostomy in conventional incisional and carbon dioxide (CO2) laser techniques for resection of soft palates in brachycephalic dogs. DESIGN: Prospective randomized trial. ANIMALS: 20 adult brachycephalic dogs. METHODS: Dogs were randomly allocated into 4 groups, and 1 of the following was performed: palate resection by use of a CO2 laser; incisional palate resection and closure with suture; and palate resection by use of a C02 laser or incision with tracheostomy. A clinical score for respiratory function was assigned to each dog at 0, 2, 8, 16, and 24 hours. Biopsy specimens of incision sites obtained at days 0, 3, 7, and 14 were examined. Data were analyzed to determine the effects of technique on clinical and histologic outcome. RESULTS: Mean surgical time for laser (309 seconds) was significantly shorter than for sharp dissection (744 seconds). Surgical technique significantly affected clinical scores at 3 of the 5 postoperative time points, but differences were not clinically apparent. Tracheostomy significantly affected clinical scores at 3 of 5 postoperative time points. After tracheostomy tube removal, clinical scores were similar to those of dogs without tracheostomies. Inflammation, necrosis, and ulceration were evident in all groups at day 3; these lesions had almost resolved by day 14. Most complications were associated with tracheostomy. CONCLUSIONS AND CLINICAL RELEVANCE: Clinical outcomes appear to be similar with the laser and incisional techniques. Regarding surgical time and ease, laser resection of the soft palate appears advantageous. Tracheostomy is not warranted in dogs that have uncomplicated surgeries and recoveries.

Urinary bladder transitional cell papilloma in a shorthorn heifer.

A 12-month-old shorthorn heifer was presented for pollakiuria of 4 months' duration. Urinary bladder transitional cell papilloma was diagnosed. The heifer had no exposure to bracken fern and no papillomavirus or bacterium was demonstrated. Laser surgery was used in an attempt to debulk the mass.

First experiences in screening women at high risk for breast cancer with MR imaging.

Women with a genetic predisposition for breast cancer are often advised surveillance with physical examination twice a year and mammography once a year from 25 years onwards. However, the sensitivity of the mammography decreases when breast tissue is dense and this is seen in 40-50% of women under 50 years. We therefore investigated whether magnetic resonance imaging (MRI) in addition to the normal surveillance could detect cancers otherwise missed. In 109 women with over 25% risk of breast cancer, MRI was performed because over 50% dense breast tissue was seen at mammography and no suspect lesion was seen at the previous screening. MRI detected breast cancers in three patients (2.8%) occult at mammography and with no new palpable tumor, twice at stage T1bN0 and T1cN0 once. Two cancers were expected. MRI was false positive in six women, resulting in two benign local excisions because ultrasound or fine needle examination confirmed suspicion. We had no false negative MRI results. MRI proved true benign in four BRCA 1/2 gene mutation carriers at histologic examination. Preoperative wire localization of the malignancies detected at MRI proved necessary as the tumor was not palpable in the lumpectomy specimen nor visible at specimen radiology. The extra cost of breast MRI in addition to mammography and physical examination was [symbol: see text]uro13.930 per detected cancer. The cost of the detection of one breast cancer patient in our national screening program is [symbol: see text]uro9000. During follow-up of patients with a familial risk in whom the first breast cancer was detected at MRI, MRI detected two recurrent cancers in stage T1bN0 and T1cN0 and one contralateral cancer T1aNo. Breast MRI is promising in screening young women at high risk for breast cancer, as it can advance the detection of cancers still occult at mammography and physical examination; but the cost may be considerable.

Ovarian cancer surgery: a clinical pathway.

Clinical pathways are useful tools in providing quality care while decreasing the cost of that care. Pathways help to facilitate managed care, identify patient and family educational needs, encourage multidisciplinary communication, and expedite patient discharge. This article discusses a clinical pathway for ovarian cancer surgery.

Cloning of a new Kunitz-type protease inhibitor with a putative transmembrane domain overexpressed in pancreatic cancer.

In a previous large scale screen for differentially expressed genes in pancreatic cancer, we identified a gene highly overexpressed in cancer encoding a novel putative transmembrane protein with two Kunitz-type serine protease inhibitor domains. The identified gene named kop (Kunitz domain containing protein overexpressed in pancreatic cancer) was assigned to chromosome 19 in the region 19q13.1. Kop was detected at high levels in pancreatic cancer cell lines and was overexpressed in pancreatic cancer tissues as compared to both, normal pancreas and chronic pancreatitis tissues. Being a member of the Kunitz-type serine protease inhibitor family, this new gene may participate in tumour cell invasion and metastasis and in the development of the marked desmoplastic reaction typical for human pancreatic cancer tissues. In this context, the fact that kop has a putative transmembrane domain may have functional implications of particular interest.

Patient use of the symptom reporting tool.

Patients frequently report symptoms to nurses without reporting them to their physicians. A variety of reasons are cited for this occurrence. The Symptom Reporting Tool was developed so that patients and family members could document their symptoms at home. By bringing the tool with them to appointments, patients found it easier to discuss problems with their physicians. Because the results of attempts to relieve these symptoms also were documented, the effectiveness of the symptom management plan was evaluated easily and a more positive patient outcome resulted.

Comparison of a high power diode laser with the Nd:YAG laser using in situ wound strength analysis of healing cutaneous incisions.

BACKGROUND AND OBJECTIVE: The laser-tissue interaction of a high power semiconductor diode laser was compared to the continuous wave neodymium yttrium aluminum garnet (Nd:YAG) laser by evaluating primary wound healing of cutaneous incisions in rats. STUDY DESIGN/MATERIALS AND METHODS: Full thickness incisions were made in rat skin using a diode laser (805 nm, 10 W, contact mode), an Nd:YAG laser (1,064 nm, 10 W, contact mode), and a stainless steel scalpel blade (control). In situ wound breaking strength measurements were obtained at 7, 14, and 21 days using a specially designed tensiometer. Cross sectional area of non-disrupted wounds was calculated in two groups prior to testing to allow for calculation of tensile strength. Blinded histopathologic analysis was also performed. RESULTS: Analysis of variance (P < or = 0.05) was used to determine differences in breaking strengths and tensile strengths due to incision method. There was no significant difference in the breaking strengths (group 1) or tensile strengths (groups 2 and 3) of the diode and Nd:YAG laser incisions. As predicted, breaking strengths and tensile strengths of scalpel blade incisions were significantly greater than those of incisions made with laser energy. Histopathologic evaluation revealed that through day 14, the degree of inflammation and collagen production was similar for diode and Nd:YAG laser incisions. Laser incisions had greater inflammation and a lag in fibroblast invasion and collagen production compared with scalpel incisions. By day 21, all incisions were similar in fibroblast population and collagen production, but laser incisions had slightly more inflammation than scalpel incisions. CONCLUSION: In the primary wound healing model described, the tissue effect, cellular response, and development of wound strength were essentially the same for the high power diode laser at 10 W and the Nd:YAG laser at 10 W.

Anomalous Dispersion with Edges in the Soft X-ray Region: First Results of Diffraction from Single Crystals of Trypsin Near the K-Absorption Edge of Sulfur.

Anomalous dispersion of X-ray diffraction at wavelengths near the X-ray K-absorption edge of sulfur at wavelengths around 5 A has been applied to single crystals of trypsin obtained from an ammonium sulfate solution. The multiwavelength anomalous-dispersion method based on 775 unique reflections (+183 Bijvoet mates) measured at three wavelengths near the K-absorption edge of sulfur in trypsin (two methionines and disulfide bridges of six cystines) reproduces the known features of the trypsin structure of a resolution of 4 A. It appears that there is anisotropic anomalous scattering from the disulfide bridges of cystine. The multiwavelength anomalous solvent contrast shows up at wavelengths near the K-absorption edge of the sulfate ions, which is shifted by 10 eV to higher energies with respect to that of sulfur in trypsin. The influence of the complex contrast of trypsin in 2.5 M ammonium sulfate on the dispersion of a low-order reflection is analyzed. The measurement of anomalous dispersion of X-ray diffraction at long wavelengths beyond 5 A requires a special diffractometer, the features of which are presented. An outstanding one is a detector system consisting of four multiwire proportional counters. Its efficiency is compared with that of imaging plates. The influence of radiation damage with soft X-ray diffraction from single crystals of trypsin is presented and possible remedies are discussed.

Use of the holmium yttrium aluminum garnet laser for percutaneous thoracolumbar intervertebral disk ablation in dogs.

OBJECTIVE-To evaluate a laser ablation-technique for treatment of thoracolumbar intervertebral disk disease in dogs. DESIGN-Prospective case series. ANIMALS-33 dogs with signs of recurrent back pain associated with intervertebral disk disease after the initial episode had been managed conservatively for a minimum of 2 weeks. PROCEDURE-Spinal needles were placed percutaneously through the annulus fibroses to permit delivery of an optical fiber into the nucleus pulposes of thoracolumbar intervertebral disks T10-11 through L3-4. Fluoroscopy was used to guide needle placement. Holmium yttrium aluminum garnet laser energy then was used to ablate the contents of each selected intervertebral disk. Intervals from time of treatment to time of assessment ranged from 3 to 114 weeks. RESULTS-All dogs recovered without complication. Results of follow-up radiography and histologic evaluation indicated that percutaneous holmium yttrium aluminum garnet laser ablation reduces the volume of nucleus pulposus in treated disks. CLINICAL IMPLICATIONS-Used as a clinical treatment and prophylactically, this minimally invasive procedure should prevent further extrusion of partially herniated disks and should reduce the chances of subsequent herniation of disks at other treatment sites.

Photothermal effects on murine mammary tumors using indocyanine green and an 808-nm diode laser: an in vivo efficacy study.

Murine mammary tumors were treated using indocyanine green and an 808 nm diode laser, and the in vivo chromophore-enhanced photothermal effects on the tumor burden and on tumor rat survival were investigated. The power of the laser was selected in the range of 5-10 W, and irradiation duration 3-5 min. One percent aqueous indocyanine green solution in a volume of 100-200 microliters was administered in situ, either acutely or 24 h prior to the treatment. The photothermal interaction was apparent under all our treatment conditions with a well-defined spatial containment in this study and the tumor growth was slowed after treatment. The post-treatment observation showed tumor recurrence and metastasis; no long-term survival was achieved with the single application of laser in conjunction with indocyanine green. Our results pose a question on the efficacy of the photothermal interaction even though tumor cell destruction can be achieved in a large and controlled scale. However, this highly selective photothermal impact on the tumor tissue did suggest that this method be applied repeatedly to be more effective and be used as the precursor of other modalities, such as chemotherapy, radiation therapy, immunotherapy, and surgery.