Targeting Semaphorin 3C in Prostate Cancer With Small Molecules.

Despite the amenability of early-stage prostate cancer to surgery and radiation therapy, locally advanced and metastatic prostate cancer is clinically problematic. Chemical castration is often used as a first-line therapy for advanced disease, but progression to the castration-resistant prostate cancer phase occurs with dependable frequency, largely through mutations to the androgen receptor (AR), aberrant AR signaling, and AR-independent mechanisms, among other causes. Semaphorin 3C (SEMA3C) is a secreted signaling protein that is essential for cardiac and neuronal development and has been shown to be regulated by the AR, to drive epithelial-to-mesenchymal transition and stem features in prostate cells, to activate receptor tyrosine kinases, and to promote cancer progression. Given that SEMA3C is linked to several key aspects of prostate cancer progression, we set out to explore SEMA3C inhibition by small molecules as a prospective cancer therapy. A homology-based SEMA3C protein structure was created, and its interaction with the neuropilin (NRP)-1 receptor was modeled to guide the development of the corresponding disrupting compounds. Experimental screening of 146 in silico‒identified molecules from the National Cancer Institute library led to the discovery of four promising candidates that effectively bind to SEMA3C, inhibit its association with NRP1, and attenuate prostate cancer growth. These findings provide proof of concept for the feasibility of inhibiting SEMA3C with small molecules as a therapeutic approach for prostate cancer.

High-level dosimetry at the demagnetization experiments of permanent magnets.

The measurements of high-energy and high dose mixed radiation from high-energy electron accelerator are carried out using a radiation damage monitor. It consists of two Radiation-Sensing Field-Effect Transistors (RADFETs) for total absorbed dose from mainly gamma ray and other charged particles and a Si PIN diode for neutron fluence. This is a part of the demagnetization study of rare earth permanent magnet irradiated by 2.5-GeV electron beam. The sensitivities of damage detectors are measured using 65-MeV quasi-monoenergic neutron, 14-MeV D-T neutron, (252)Cf neutron for Si PIN diode and (60)Co and (137)Cs gamma ray for RADFETs. Measured sensitivities are in acceptable range in the comparison of producer's proposed values. The dose and fluence measurements are carried out for the same target condition, Cu and Ta, as that for the demagnetization study. The 5 x 5 cm(2) cross-sectional and 5.5-cm-thick Pb target is also used for the general comparison with photoneutron yields. All measured dose and fluence are compared with the calculated results using the FLUKA code and agree well each other. The application of this kind of radiation damage monitor to high-level dosimetry at high-energy electron accelerator has been discussed.

Financial burden of national health insurance for treating patients with transfusion-dependent thalassemia in Taiwan.

The thalassemias are a heterogeneous group of inherited hypochromic anemias of varying severity. The mainstay of supportive treatment is regular blood transfusion accompanied by iron-chelating therapy. Hematopoietic stem cell transplantation (HSCT) provides an alternative option when curative therapy is considered. More than 400 patients in Taiwan have beta-thalassemia major or other transfusion-dependent thalassemias, and their treatment costs account for a considerable percentage of the National Health Insurance expenditure. In this report, we estimated the treatment costs of conventional therapy (regular blood transfusion accompanied by iron-chelating agents) and HSCT. The undiscounted medical cost of 20 years of follow-up (20 years from diagnosis) and the undiscounted total lifetime cost were NT$ 4 739 888 (NT$ means New Taiwan Dollars)/US$ 149 288 and NT$ 11 529 990/US$ 363 149, respectively, for patients undergoing conventional therapy, and NT$ 2 639 982/US$ 83 149 and NT$ 3 511 172/US$ 110 588, respectively, for those undergoing successful HSCT. Comparisons of treatment costs and other parameters between these two modalities can add to the information base on which policy is made by health authorities or clinicians.

Inactivation of farnesyltransferase and geranylgeranyltransferase I by caspase-3: cleavage of the common alpha subunit during apoptosis.

Caspase plays an important role in apoptosis. We report here that farnesyltransferase/geranylgeranyltransferase (FTase/GGTase)-alpha, a common subunit of FTase (alpha/beta(FTase)) and GGTase I (alpha/beta(GGTase)), was cleaved by caspase-3 during apoptosis. FTase/GGTase-alpha (49 kDa) was cleaved to 35 kDa (p35) in the Rat-2/H-ras, W4 and Rat-1 cells treated with FTase inhibitor (LB42708), anti-Fas antibody and etoposide, respectively. This cleavage was inhibited by caspase-inhibitors (YVAD-cmk, DEVD-cho). Serial N-terminal deletions and site-directed mutagenesis showed that Asp59 of FTase/GGTase-alpha was cleaved by caspase-3. The common FTase/GGTase-alpha subunit, but not the beta subunits, of the FTase or GGTase I protein complexes purified from baculovirus-infected SF-9 cells was cleaved to be inactivated by purified caspase-3. In contrast, FTase mutant protein complex [(D(59)A)alpha/beta(FTase)] was resistant to caspase-3. Expression of either the cleavage product (60-379) or anti-sense of FTase/GGTase-alpha induced cell death in Rat-2/H-ras cells. Furthermore, expression of (D(59)A)FTase/GGTase-alpha mutant significantly desensitized cells to etoposide-induced death. Taken together, we suggest that cleavage of prenyltransferase by caspase contributes to the progression of apoptosis.

Selectivity and antagonism of chemokine receptors.

The chemokine superfamily can be subdivided into two groups based on their amino terminal cysteine spacing. The CXC chemokines are primarily involved in neutrophil-mediated inflammation and, so far, two human receptors have been cloned. The CC chemokines tend to be involved in chronic inflammation, and recently we have cloned a fourth leukocyte receptor for this group of ligands. Understanding what makes one receptor bind its range of agonists is important if we are to develop potent selective antagonist. We have started to investigate the molecular basis of this receptor selectivity by looking at why CC chemokines do not bind to the CXC receptors in several ways. First, we looked at the role of the three-dimensional structure of the ligand, and have solved the three dimensional structure of RANTES using nuclear magnetic resonance spectroscopy. The structure is similar to that already determined for the CC chemokine macrophage inflammatory protein-1 beta, and it has a completely different dimer interface to that of the CXC chemokine interleukin-8 (IL-8). However, the monomer structures of all the chemokines are very similar, and at physiological concentrations the proteins are likely to be monomeric. Second, by examining all the known CC and CXC chemokines, we have found a region that differs between the two subfamilies. Mutations of one of the residues in this region, Leu-25 in IL-8, to tyrosine (which is conserved at this position in CC chemokines) enables the mutant IL-8 to bind CC chemokine receptor-1 (CC-CKR-1) and introduces monocyte chemoattractant activity. Using other mutations in this region, we can show a direct interaction with the N-terminus of CC-CKR-1. Third, we have found that modification of the amino terminus of RANTES by addition of one amino acid makes it into an antagonist with nanomolar potency. Taken together, this data suggests a two-site model for receptor activation and for selectivity between CC and CXC chemokines, with an initial receptor contact provided by the main body of the chemokine, and activation provided by the amino terminal region.

Feasibility study of nitrogen removal with the mecellulose wasted liquor as an external carbon source in the two-stage denitrification process.

The utilization of mecellulose wasted liquor (MWL) as an external carbon source was investigated to find an alternative for methanol in the two-stage denitrification pilot process. The pilot plant was supplied with the raw water from the J-Municipal Sewage Treatment Plant (J-MSTP) in Korea. The raw water of J-Municipal Sewage Treatment Plant contains low and high concentration of biodegradable organics and nitrogen source, respectively, due to the inflow of industrial wastewater and landfill leachate. Methanol was fed to provide external carbon source for high concentration of nitrogen source removal by denitrification in this J-Municipal Sewage Treatment Plant, and thus this study was performed to test effects to the effluent quality and efficiencies of nitrogen source removal with an alternative carbon source for the cost reduction. The 6.5mg 1(-1) and 5.7mg l(-1) of total nitrogen (TN) concentration in the effluent were achieved with mecellulose and methanol, whereas mecellulose and methanol were fed to give the same ratio of gCODgNO,-N(-1), respectively. The 60% of COD in mecellulose wasted liquor was used as a carbon source for denitrification and the stable denitrification rate was earned when one half of the required total carbon source for denitrification was fed to pre-anoxic tank in the pilot plant. The required gCODgNO,-N(-1) ratio with mecellulose wasted liquor was 1.4 times higher than with methanol. Mecellulose wasted liquor is feasible to be used as external carbon source for organic loading, nitrogen and phosphorus removal. If mecellulose wasted liquor is considered as an alternative external carbon source to substitute methanol 26-28m3 mecellulose wasted liquor per 1 m3 methanol will be required. However, to meet with the effluent standard (10 mg BOD l(-1)) for J-Municipal Sewage Treatment Plant, the feed concentration of mecellulose wasted liquor should be recommended to be lower than 200 mgl(-1).

Sensitization potentials of methyl, ethyl, and n-butly methacrylates and mutual cross-sensitivity in guinea pigs.

Guinea pigs could be strongly sensitized to methyl, ethyl, and n-butyl methacrylates in ethanol or olive oil by the topical route, or in saline by the intradermal route. For elicitation of skin reactons, topical challenge with the compounds in olive oil or intradermal challenge with saline as the solvent was neccessary. Topical challenge with the methacrylates in ethanol failed to elicit any allergic skin reactions because of their volatility. All sensitized animals responded strongly not only to the inducing methacrylate but also to the other methacrylates, showing that mutual cross-sensitivity had occurred. Since methyl methacrylate has been reported to be a potent sensitizer in humans, the guinea-pig model described here may be useful for screening products before marketing.

Sensitization potentials and immunologic specificities of neomycins.

The use of sensitization indices for expressing allergenic skin reactions in guinea pigs is described. The method is convenient for comparing allergens and cross-reacting substances and permits the use of both irritating and nonirritating challenge concentrations of allergens. It also permits determination of both optimal reading time and challenge concentrations for each experiment. By this technique commercial neomycin complex, neamine (neomycin A), neomycin B, neomycin C, and streptomycin were found to be allergenic in guinea pigs via intradermal (id) and foot-pad (fp) immunizations. The immunizing emulsion consisted of an allergen and an adjuvant containing Mycobacterium butyricum (MB) or Mycobacterium tuberculosis H37Ra (Ra). The adjuvant MB was as effective as Ra by the id route, but inferior to Ra by the fp route. The cross-reactivity of neomycin C was generally greater than neomycin B in guinea pigs sensitized to neamine, neomycin B, neomycin C, or streptomycin. In guinea pigs sensitized to neomycin complex by repeated immunizations, neomycins A, B, and C were effective elicitors of skin reactions, whereas the N-acetylated derivatives of the components failed to cause reactions. This finding is interpreted to mean that the amino groups of the aminoglycosides are the coupling sites to host proteins in the processes of sensitization and elicitation of skin reactions in vivo.

Peptidergic innervation of the primate meibomian gland.

PURPOSE: To localize and characterize nerves in primate meibomian glands using immunohistochemical staining for neuropeptides and neuronal enzymes. METHODS: Upper eyelids were obtained from seven rhesus monkeys (Macaca mulatta) and one cynomolgous monkey (Macaca fascicularis). The tissues were fixed either by immersion in Zamboni's fixative or by transcardiac perfusion with paraformaldehyde and glutaraldehyde and were then postfixed. Cryostat tissue sections of the lids were stained by immunohistochemistry using rabbit antisera to neuron-specific enolase (NSE), tyrosine hydroxylase (TH), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), and substance P (SP), followed by a fluorescence visualization system. RESULTS: Used as a marker for the overall nerve distribution, NSE antibodies revealed abundant smooth and varicose nerve fibers closely apposed to the basement membranes of acini of the meibomian glands. Numerous nerve fibers near the meibomian gland acini were immunoreactive for NPY and VIP, but nerve fibers containing TH, CGRP, and SP were more sparse in the meibomian glands. Nerve fibers also were visualized in other eyelid structures, including conjunctiva, epidermis, hair follicles, and subconjunctival lymphoid follicles. CONCLUSIONS: The meibomian glands of rhesus and cynomolgous monkeys are richly innervated by diverse nerve fiber types. The immunohistochemical staining suggests a largely parasympathetic origin for this innervation, with relatively smaller contributions from sympathetic and sensory sources. These findings also suggest that meibomian gland secretion is under the control of diverse neurotransmitter-neuromodulator mechanisms.

Pulmonary artery aneurysm presenting as a lung mass.

We describe a case of pulmonary artery aneurysm in which clinical clues and conventional imaging suggested a lung tumor, and the actual nature of the lesion was discovered at the time of thoracotomy. This case shows the importance of an awareness of this condition in the formulation of a differential diagnosis for a lung mass.

Method of determining kidney position as guide for percutaneous nephrostomy.

Computed tomography scans were used to assess the distance between the skin and the collecting system in 256 kidneys in 128 adult patients of various heights and weights. Body surface area correlated well with kidney depth. A graph is presented that can be used to calculate kidney depth.

Cross-sensitivity of common aminoglycoside antibiotics.

Guinea pigs were sensitized to neomycin (A, B, or C), paromomycin, gentamicin, kanamycin, streptomycin, and dihydrostreptomycin via intradermal or foot-pad injection with an adjuvant containing killed Mycobacterium butyricum or M tuberculosis H37Ra (Ra). These antibiotics produced greater cross-sensitization with an increase in the number of immunizations and chemical structural similarities. After repeated intradermal injections (adjuvant Ra) of neomycin, guinea pigs showed cross-sensitization to paromomycin, kanamycin, and streptomycin. A single intradermal injection of one of these antibiotics produced stronger reactions to the most closely related antibiotics, with no meaningful sensitization to the least-related allergens. Streptomycin-sensitized guinea pigs seldom showed a meaningful cross-sensitization to dihydrostreptomycin or the other antibiotics (except neomycin C); however, guinea pigs sensitized to dihydrostreptomycin or the other antibiotics often showed strong cross-sensitization to streptomycin.

The effect of the nasopharyngeal air cavity on x-ray interface doses.

We investigated the impact of air cavities in head and neck cancer patients treated by photon beams based on clinical set-ups. The phantom for investigation was constructed with a cubic air cavity of 4 x 4 x 4 cm3 located at the centre of a 30 x 30 x 16 cm3 solid water slab. The cavity cube was used to resemble an extreme case for the nasal cavity. Apart from measuring the dose profiles and central axis percentage depth dose distribution, the dose values in 0.25 x 0.25 x 0.25 cm3 voxels at regions around the air cavity were obtained by Monte Carlo simulations. A mean dose value was taken over the voxels of interest at each depth for evaluation. Single-field results were added to study parallel opposed field effects. For 10 x 10 cm2 parallel opposed fields at 4, 6 and 8 MV, the mean dose at regions near the lateral interfaces of the cavity cube were decreased by 1 to 2% due to the lack of lateral scatter, while the mean dose near the proximal and distal interfaces was increased by 2 to 4% due to the greater transmission through air. Secondary build-up effects at points immediately beyond the air cavity cube are negligible using field sizes greater than 4 x 4 cm2. For most head and neck treatment, the field sizes are usually 6 x 6 cm2 or greater, and most cavity volumes are smaller than our chosen dimensions. Therefore, the influence of closed air cavities on photon interface doses is not significant in clinical treatment set-ups.

Human immunodeficiency virus p24 antigen testing in cornea donors.

PURPOSE: Testing for the p24 antigen of the human immunodeficiency virus (HIV) may detect early HIV infection in the seronegative window; however, falsely reactive results may occur in cadaver specimens. Although neither the Food and Drug Administration (FDA) nor the Eye Bank Association of America requires p24 testing of cornea donors, many tissue banks using other organs from cornea donors do perform this assay, and the FDA requires that eye banks reject corneal tissue if a reactive p24 assay is reported. We investigated the impact of p24 testing on eye banking and corneal transplantation. METHODS: Two clinical cases and records from the Lions Eye Bank of Delaware Valley (LEBDV) were reviewed retrospectively. RESULTS: Two corneas from the LEBDV were transplanted before the reporting of p24 reactivity by other tissue banks. In one case, because of the young age of the recipient, the surgeon elected to replace the cornea with new tissue hours after the original transplant, and later polymerase chain reaction (PCR) testing was negative. In the other case, there was not enough specimen to perform Western blot or PCR confirmatory testing. The patient was followed with periodic serologic testing for HIV and has remained seronegative. To avoid such problems in the future, the LEBDV initiated testing of all donors with p24 and other nonrequired screening tests. Over a 2-month period, 22 corneas (from 11 donors) were discarded because of these tests: 4 donors had reactive p24 tests, 6 were reactive for antibody to hepatitis B core antigen, and 1 had a reactive syphilis test. CONCLUSIONS: Results from p24 assays by other tissue banks may cause difficult clinical situations when the results are received after transplantation of the tissue, but the use of the p24 assay in the screening of cornea donors may result in excessive waste of donor tissue. Further guidance is needed regarding the management of positive results from this and other nonrequired screening tests.

Penetrating keratoplasty in iridocorneal endothelial syndrome.

PURPOSE: To evaluate the clinical outcome of penetrating keratoplasty (PK) in iridocorneal endothelial (ICE) syndrome. METHODS: Clinical charts of patients who underwent penetrating keratoplasty for ICE syndrome between 1985 and 1999 were reviewed retrospectively. Glaucoma control, best corrected visual acuity pre- and post-PK, graft clarity, graft rejection episodes, improvement in pain, and additional procedures were analyzed. RESULTS: Fourteen cases were reviewed with an average follow-up of 58 months after PK. Initial grafts failed in seven patients (50%), in six cases because of rejection, and one owing to endothelial failure without signs of rejection. Repeat PKs were performed in six patients. At final follow-up, 12 grafts were clear. Glaucoma was controlled pre- and post-PK (average intraocular pressure, 16 mmHg for both eyes). Pre-PK, eight patients were using glaucoma medicines and nine had had glaucoma surgery. At the end of the follow-up, seven patients were using glaucoma medicines; six patients required glaucoma surgery after their initial PK. At the final follow-up visit, visual acuity in three patients (21%) was 20/40 or better, it ranged from 20/50 to 20/100 in four patients (29%) and 20/200 to 20/400 in five patients (36%), and in two patients with failed grafts (14%) it was counting fingers or worse. CONCLUSION: Clear grafts were achieved in 12 cases, although six patients (43%) underwent repeat PKs. All patients had glaucoma, which was controlled before and after PK by medical treatment and surgical procedures. Favorable outcomes can be achieved in patients with ICE syndrome but may require multiple corneal and glaucoma procedures.

A case of Leigh's disease with initial manifestation of dystonia.

A case of Leigh's disease (subacute necrotizing encephalomyelopathy) is reported with such noteworthy features as early onset, dystonia, paraparesis the presence of low attenuation areas in both basal ganglias on computerized tomography of the brain and the presence of a high signal intensity in both basal ganglias in T2 weighted image by MR. The electron microscopic findings of muscle biopsy are suggestive of pleoconial mitochondrial myopathy.

Laparoscopic splenectomy for immune thrombocytopenic purpura--long-term result of 40 laparoscopic splenectomies.

Laparoscopic surgery has recently extended its indications and it has also become an acceptable surgical approach for splenectomy. In the last five years, we have performed 40 laparoscopic splenectomies for immune thrombocytopenic purpura. Thirty-five patients were female and 5 patients were male. The mean age was 34, varying from 17 to 56. After learning to perform laparoscopic splenectomy with five ports, we are now usually using three or four ports in a right lateral kidney position. There was no case of conversion to exploratory laparotomy. The mean hospital stay was 7 days. There was no perioperative mortality; but in 2 cases we had postoperative subphrenic abscesses which were successfully managed by catheter drainage. Since undergoing laparoscopic splenectomy, 28 patients (70%) were weaned effectively from their steroid medications. Eight patients (20%) have been on small doses of steroid, and 4 patients (10%) have been on the same doses of steroid with no response. The patient group with rapidly increasing platelet count after splenectomy showed a statistically significant relation with the complete response group (p < 0.001). Laparoscopic splenectomy is a safe and reasonable operative procedure for patients with immune thrombocytopenic purpura.

Comparison of balloon dacryocystoplasty to probing as the primary treatment of congenital nasolacrimal duct obstruction.

PURPOSE: In children older than 18 months, primary probing procedures for congenital nasolacrimal duct obstruction (CNLDO) are thought to have lower rates of success. This study compares the results of primary probing to balloon dacryocystoplasty (DCP) in children stratified by age category. METHODS: In a retrospective chart review, 29 eyes with CNLDO that underwent balloon DCP in children older than 18 months were identified and age-matched to 29 eyes that underwent probing. The eyes were divided into 3 age categories: category 1 (18-24 months), category 2 (24-36 months), and category 3 (>36 months). RESULTS: Of the 29 eyes treated with balloon DCP (mean age, 37.1 months), 26 were successfully treated. Twenty-five of the 29 matched probed eyes (mean age, 31.1 months) were successfully treated, resulting in an overall success rate of 90% for balloon DCP and 86% for primary probing. Within each age category, the success rate varied but did not show an advantage to balloon DCP. The presence of crusting and expressible discharge from the puncta during preoperative evaluation predicted a successful probing (OR, 16; 95% CI, 1.3-192). CONCLUSION: Overall, balloon DCP did not appear to present an advantage as compared with primary probing as the initial treatment in these children. Primary probing has an impressive overall success rate that did not diminish in the children older than 36 months.

Development of a robotic navigation system for neurosurgery.

This paper presents a robotic navigation system for image-guided neurosurgery, which can be applied to the treatment of Parkinson's disease and biopsy of brain tumor. The system integrates a computer for real-time display of brain anatomy, a magnetic tracking device for measuring the positions and orientations of surgical instruments, and a robot manipulator for guiding surgical instruments to the preplanned positions and orientations. The computer display of brain anatomy offers a convenient tool for surgeons to diagnose brain disease and to plan safe surgical paths; while the tracking device assists the robot manipulator to automatically guide surgical instruments to the preplanned direction. The registrations among the tracking device, the image system, and the robot are completed on the base of coordination mappings of external markers. An experiment of using a skull model for simulating a robotic biopsy of brain tumor has been done to verify the performance of the navigation system. The result shows that the system can accomplish a positioning accuracy around 2 mm.