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Background: Development of an efficacious egg-free mock-up H5N1 vaccine is key to our preparedness against pandemic avian flu. Methods: This is a single-center, randomized, observer-blinded phase I clinical trial evaluating the safety and immunogenicity of an alum-adjuvanted Madin–Darby canine kidney (MDCK)-derived inactivated whole-virion H5N1 influenza vaccine in healthy adults. Hemagglutination inhibition (HAI) and neutralizing antibody titers were measured using horse and turkey red blood cells (RBCs). Results: Thirty-six adult subjects were randomized to receive two doses of 0.5 mL of the MDCK-derived H5N1 alum-adjuvanted vaccine containing 7.5, 15, or 30 μg of hemagglutinin (HA) 21 days apart. The candidate vaccine was well tolerated and safe across the three dosing groups. The most frequent adverse event was injection site pain (46.5%). Both HAI and neutralizing antibody titers increased after each vaccination in all three dosing groups. The best HAI responses, namely a seroconversion rate of 91.7% and a geometric mean ratio of 9.51 were achieved with the HA dose of 30 μg assayed using horse RBCs at day 42. HAI titers against H5N1 avian influenza virus was significantly higher when measured using horse RBCs compared with turkey RBCs. Conclusions: This Phase I trial showed the MDCK-derived H5N1 candidate vaccine is safe and immunogenic. The source of RBCs has a significant impact on the measurement of HAI titers (ClinicalTrials.gov number: NCT01675284.). Keywords: Influenza A, Avian influenza, H5N1, Vaccination, Immune response

Influenza is a contagious acute respiratory disease caused by the influenza virus infection. Hemagglutinin (HA) is an important target in the therapeutic treatment and diagnostic detection of the influenza virus. Influenza A virus encompasses several different HA subtypes with different strains, which are constantly changing. In this study, we identified a fully human H1N1 neutralizing antibody (32D6) via an Epstein-Barr virus-immortalized B cell-based technology. 32D6 specifically neutralizes the clinically isolated H1N1 strains after the 2009 pandemic but not the earlier strains. The epitope was identified through X-ray crystallographic analysis of the 32D6-Fab/HA1 complex structure, which revealed a unique loop conformation located on the top surface of HA. The major region is composed of two peptide segments (residues 172–177 and 206–213), which form an abreast loop conformation. The residue T262 between the two loops forms a conformational epitope for recognition by 32D6. Three water molecules were observed at the interface of HA and the heavy chain, and they may constitute a stabilizing element for the 32D6-HA association. In addition, each 32D6-Fab is likely capable of blocking one HA trimer. This study provides important information on the strain specificity of 32D6 for the therapeutic treatment and detection of viral infection.

The telomerase-specific replication-competent oncolytic adenovirus, Telomelysin, was developed for virus-mediated preferential lysis of tumor cells. Its selectivity is derived from a human telomerase reverse transcriptase (hTERT) promoter-driven active viral replication, which occurs in cancer cells with high telomerase activity but not in normal cells lacking such activity. Because the TERT activity is elevated in most cases of hepatocellular carcinoma (HCC), the current study aims to investigate whether Telomelysin can be used for treatment of HCC. The oncolytic effect of Telomelysin has been investigated both in vitro using cell culture and in vivo using an immunocompetent in situ orthotopic HCC model. In this model, HCC developed spontaneously in the liver of HBx transgenic mice, which is pathologically and genetically similar to human HCC. In cell culture assay, Telomelysin lyses HCC cell lines at a low multiplicity of infection (MOI), ranging 0.77-6.35 (MOI [PFU/cell]). In the orthotopic HCC model, Telomelysin showed a potent oncolytic effect on HCC but spared normal liver tissue. Dose escalation analysis identified a safety dose of 1.25 × 10(8) PFU for this model. The effect of multiple injections of Telomelysin was also evaluated in this immunocompetent HCC model. We found that the virus replicates in HCC after a second intratumoral injection despite an immune response induced by the previous injection. This preclinical study shows that Telomelysin can be used for treatment of human HCC at an appropriate dosage and that its tumor-killing activity persists after multiple injections.

Cordyceps sinensis (C. sinensis) is one of the well known fungi used in traditional Chinese medicine for treatment asthma and bronchial and lung inflammation. In this study, effects of C. sinensis methanolic extracts on bronchoalveolar lavage fluids (BALF) cells proliferation, inflammatory cytokines production, and genes expression were evaluated. The proliferative response of BALF cells to lipopolysaccharide (LPS) was determined by the tritiated thymidine uptake method. The cell-free supernatants were harvested then tested for interlukin-1beta (IL-1beta), interlukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), interleukin-10 (IL-10), interleukin-12 (IL-12), and interferon-gamma (IFN-gamma) by the enzyme immunoassay. The results indicated that the CS-19-22 fraction dose dependently suppressed BALF cells proliferation activated by LPS. The CS-19-22 fraction also reduced IL-1beta, IL-6, IL-8, IL-10 and TNF-alpha production in LPS activated BALF cell cultures. Furthermore, the IL-12 and IFN-gamma production in activated BALF cells were enhanced by CS-19-22 treatment. The CS-19-22 fraction did not affect IL-1beta, IL-6, TNF-alpha, and IL-8 mRNAs expression in BALF cells detected by reverse transcription-polymerase chain reaction (RT-PCR). By contrast, the CS-19-22 fraction increased IL-12 and IFN-gamma mRNAs expression and decreased IL-10 mRNA expression in the BALF cells activated with LPS. These results indicated the CS-19-22 fraction suppressed IL-1beta, IL-6, TNF-alpha, and IL-8 cytokines production in BALF cells through other than inhibition of mRNAs expression pathway. These results also demonstrate that the therapeutic activity of C. sinensis in Chinese medicine may be related to modulation of TH1 and TH2 cells functions in bronchial airway.

A panel of urologists experienced in interstitial laser coagulation (ILC) gathered to formulate practice guidelines for ILC. The aim was to avoid abuse or inappropriate applications leading to problems that may jeopardise the welfare of patients, and to formulate a common rationale for treatment to facilitate reimbursement for benign prostatic hyperplasia (BPH) treatment in Asia-Pacific countries and elsewhere. The authors reviewed data from the literature and accumulated experience from over 1000 patients who have undergone ILC since 1991. ILC for BPH treatment is a safe procedure with a low complication profile and its effectiveness is comparable to that of transurethral resection of the prostate (TURP). Temporary voiding irritation for 2–4 weeks and a catheterisation period of 5–7 days are the major disadvantages seen in those who had eight punctures or more for their BPH. Despite the seemingly higher direct cost arising from investment in laser unit and fibre-optic equipment, other benefits of ILC — s...

BACKGROUND Photodynamic therapy (PDT) is an experimental approach for treating prostate cancer localized to the gland that does not involve surgery or irradiation. Second-generation photosensitizers 5-aminolevulinic acid (ALA) and aluminum disulfonated phthalocyanine (A1S2Pc) were studied in the normal canine prostate. METHODS Tissue biodistribution of photosensitizers on serial biopsies was examined using fluorescence microscopy. Photodynamic therapy was done by delivering red light interstitially at 100mW through fibers placed under transrectal ultrasound guidance. RESULTS Peak levels of A1S2Pc apppeared at 5-24 hr and at 3 hr for ALA. Macroscopic PDT lesions were up to 12 mm in diameter using A1S2Pc, but only 1-2 mm with ALA. Light at 300 mW caused thermal lesions. At 28 days, damaged glands remained atrophic, but the interlobular supporting stroma was well preserved. Urethral lesions healed by 28 days without functional impairment. CONCLUSIONS Although the results with ALA were disappointing, PDT using A1S2Pc looks like a promising modality for treatment of localized prostate cancer. Prostate 32:89–98, 1997. © 1997 Wiley-Liss, Inc.

Background and Objective Photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) for sensitization is a promising treatment for carcinoma in situ and diffuse premalignant changes of the bladder. We studied the biodistribution of PpIX in a range of tissues with oral and intravesical routes of administration of ALA and compared the photodynamic effects on bladder and skin. Study Design/Materials and Methods: Normal Wistar rats were given oral or intravesical ALA and PpIX levels in the liver, kidney, skin, and bladder measured by fluorescence microscopy on tissue sections. At the time of maximum PpIX levels, the bladder and skin on the back were illuminated with light at 630 nm and the PDT effects compared. Results PpIX fluorescence in the urothelium after 200 mg/kg given intravesically was comparable to that found after 100 mg/kg orally. The ratio of PpIX levels between the urothelium and the underlying muscle was the same for both routes of administration, although there appeared to be more selectivity of urothelial PDT necrosis after intravesical administration. Skin photosensitization was greater after oral ALA, the epidermal PpIX level being three times higher than after intravesical administration for comparable urothelial levels and the PDT effect being more marked. Conclusions Intravesical instillation is preferable to oral administration of ALA for PDT ablation of the urothelium of the rat bladder without damage to the underlying tissue layers and for minimizing skin photosensitivity. The technique is now ready for clinical trials. Lasers Surg. Medicine 20:254–264, 1997. © 1997 Wiley-Liss, Inc.

Photodynamic therapy (PDT) produces localised necrosis with light after prior administration of a photosensitising drug. Although the technique is promising for small tumours of hollow organs, little work has been done on solid organs like the prostate. We studied the tissue biodistribution and photodynamic effects of meso-tetra-(m-hydroxyphenyl) chlorin (mTHPC), a potent second-generation photosensitiser, on normal canine prostate in vivo. Using quantitative fluorescence microscopy, the highest concentration of mTHPC in the prostate was seen 24-72 hr after intravenous administration. For PDT, red light (650 nm) was delivered to the prostate by laser fibres inserted via the transurethral or transperineal route under transrectal ultrasound guidance. PDT lesions up to 40 mm in diameter (using 4 fibre sites) were produced, characterised by swelling, inflammatory response and extensive glandular destruction. There was persistent glandular atrophy at 90 days, but no disruption of the main stroma and no change in the ultimate size or shape of the gland. Urethral damage sometimes caused temporary urinary retention, but this resolved by 7 days, and no animal became incontinent. Occasional small lesions were seen in the rectum, but these healed without sequelae and there were no fistulae. Since cancer and normal prostate are likely to respond similarly, PDT has considerable promise for treating cancer confined to the gland as large areas of glandular tissue can be necrosed with safe healing. Because the structural integrity of the gland is maintained, PDT is unlikely to be of value in the management of benign prostatic hypertrophy.

Purpose: Photodynamic therapy (PDT) has considerable potential for the treatment of superficial bladder neoplasia. Problems of detrusor muscle scarring and prolonged cutaneous photosensitivity with current photosensitizers may be reduced by using the new photosensitizer precursor, 5-aminolevulinic acid (ALA). We studied the fluorescence distribution of protoporphyrin IX (PpIX, the active derivative of ALA) in rat urinary bladder after intravesical administration of ALA as the first step to undertaking PDT using locally administered ALA.Materials and Methods: Solutions of varying concentrations of ALA and pH were given intravesically to Wistar rats. The bladder was removed 1 to 24 hours later for measurement of the tissue levels of PpIX by fluorescence microscopy in the layers of the bladder wall.Results: The stability of the ALA solution steadily decreased as the pH was increased from 2.1 to 7. The best value was pH 5.5, at which the stability of the solution and the urothelial tolerance were both...

Purpose: Photodynamic therapy (PDT) has the potential to treat extensive premalignant lesions and microinvasive tumors in the bladder, but its use has been hampered by the risk of detrusor muscle damage and prolonged skin photosensitivity. We have shown that the rat urothelium can be sensitized by selectively using a 10 percent solution of 5-aminolevulinic acid (ALA) at pH 5.5 administered intravesically. This paper evaluates the photodynamic effects on sensitized bladders.Materials and Methods: The bladders of Wistar rats were instilled with ALA solutions of different concentrations at pH 5.5 and subsequently treated with laser light at 630 nm. Bladders were harvested 1 to 7 days after PDT for histological assessment.Results: Under optimum conditions (10 percent intralipid diffusion medium, light dose 50J) uniform urothelial necrosis was seen after 1 to 2 days; it healed in 7 days without damage to the underlying muscle layer although some increase in collagen was seen in the lamina propria. Over...

In an attempt to assess the usefulness of a ureteral stent in facilitating disintegration of obstructed proximal ureteral stone with extracorporeal shock wave lithotripsy (ESWL), a prospective randomized study on 120 patients with ultrasonic evidence of hydronephrosis whose stone sizes were greater than 1.0 x 0.6 cm on KUB film and located above the level of L5 were allocated into three distinct treatment strategies: group 1: in situ without stone manipulation; group 2: bypass stones with stent, and group 3: placement of a catheter below stone with continuous saline irrigation. Of 77 patients, 26, 27 and 24 in each group, followed up for a period of 6 months with sufficient data for analysis, the stone-free rates were 76.9, 59.3 and 62.5%, respectively. The average number of shock waves delivered to accomplish a stone-free state were 3,245, 3,250 and 3,880 for individual groups and the comparisons among them were statistically insignificant (p's > 0.1). ESWL retreatment was common, being 33.3, 29.4 and 43.8%, respectively. Hydronephrosis has a major impact on the outcome of ESWL treatment, as more stone-free cases were found in the mildly hydronephrotic category than in the severer counterparts. About 90% of cases who finally were stone-free, passed all their stone fragments within 28 days. Ancillary procedures were needed in 13 (16.9%), 7 received ureterolithotomy, whilst 6 were removed of their stones by ureteroscopy.

The management of posterior urethral obliteration secondary to urethral disruption associated with pelvic fracture has been one of the greatest challenges to a urologist. Although different surgical procedures are available, a recent prevailing notion has been that if feasible, any completely obliterated urethra may be tried out with an optic urethrotomy to regain the alignment. A decisive incision by the cold knife over the site of urethral obliteration into the urinary bladder could be guided precisely by linear transrectal ultrasound used concomitantly with an optic urethrotomy. In a period of 2 years, transrectal ultrasound-guided optic urethrotomy was conducted successfully in five patients with traumatic complete posterior urethral obliteration and two with complete bladder neck obstruction. Although as many as six subsequent optic urethrotomies have been required for extremely long segment obliterations, there has been no impotence or, urinary incontinence over a period of 12 to 30 months. Optic ur...

Background/Aims Hepatocellular carcinoma recurrence after curative treatment adversely influences clinical outcome. It is important to explore adjuvant therapies. This phase II/stage 1 multi-center, randomized trial investigated the safety, optimal dosage and preliminary efficacy of PI-88, a novel heparanase inhibitor, in the setting of post-operative recurrence of HCC according to a Simon's 2-stage design. Methods Three groups were included: one untreated arm (Group A) and two PI-88 arms (Group B: 160mg/day; Group C: 250mg/day). Treatment groups received PI-88 over nine 4-week treatment cycles, followed by a 12-week treatment-free period. Safety and optimal dosage were assessed. Results Overall, 172 patients were randomized and 168 were included in the intention-to-treat (ITT) population. Treatment-related adverse effects included cytopenia, injection site hemorrhage, PT prolongation, etc. Four serious adverse events were possibly related to PI-88 treatment. One (1.8%) group B patients and six (10.5%) group C had hepatotoxicity-related withdrawals. Among the ITT population, 29 patients (50%) in Group A, 35 (63%) in Group B, and 22 (41%) in Group C remained recurrence-free at completion. Calculated T 1 value suggested 160mg/day treatment satisfied the criteria for the next stage of the trial. Conclusions PI-88 at 160mg/day is optimal and safe, and shows preliminary efficacy as an adjunct therapy for post-operative HCC.

AIM: To demonstrate that administering heparanase inhibitor PI-88 at 160 mg/d is safe and promising in reducing hepatocellular carcinoma (HCC) recurrence for up to 3 year following curative resection. METHODS: A total of 143 patients (83.1% of the 172 participants in the phase II study) participated in the follow-up study. Of these patients, 50 had received no treatment, 48 had received 160 mg/d PI-88, and 45 had received 250 mg/d PI-88 during the phase II trial. Safety parameters and the following efficacy endpoints were investigated: (1) time to recurrence; (2) disease-free survival; and (3) overall survival. RESULTS: PI-88 at 160 mg/d delayed the onset and frequency of HCC recurrence, and provided a clinically significant survival advantage for up to 3 years after treatment compared with those of the control group: (1) the recurrence-free rate increased from 50% to 63%, and (2) time to recurrence at the 36th percentile was postponed by 78%. The efficacy of administering PI-88 at 250 mg/d was confounded by a high dropout rate (11 out of 54 patients). Additionally, subgroup analyses of patients with (1) multiple tumors or a single tumor ≥ 2 cm; and (2) hepatitis B or C revealed that administering PI-88 at 160 mg/d conferred the most significant survival advantage (56.8% improvement in disease-free survival, P = 0.045) for patients with both risk factors for recurrence. CONCLUSION: Administering PI-88 at 160 mg/d is a safe and well-tolerated dosage that may confer significant clinical benefits for patients with HCC.

Photodynamic therapy (PDT) is proved to have potential for managing various malignancies. We investigated tissue biodistribution and photodynamic effects on a canine model in vivo using second generation photosensitizers, meso-tetra(m-hydroxyphenyl)chlorin (mTHPC) and 5-aminolaevulinic acid (ALA) to evaluate the feasibility and possible future application of PDT on the prostate. Using fluorescence microscopy, the optimal sensitization time of the prostate was between 24 - 72 hours with mTHPC and, 3 hours with ALA. After optimum time of sensitization, prostates of mature beagle were treated with laser at various sites by placing fiber interstitially under the guidance of transrectal ultrasound. The light dose for each treatment site was 100 J (100 mW for 1,000 seconds at the wavelength of 650 and 630 nm, respectively). With mTHPC, single laser fiber was able to induce organ confined PDT lesion as large as 20 by 18 by 18 mm in size. However, the PDT lesion with ALA was negligible 3 days after treatment. Physical distress manifested as urinary retention, poor appetite and body weigh loss, was more prominent with increasing number of treatment sites as a result of extensive prostatic swelling and urethral damages. However, these problems usually alleviated spontaneously 7 to 10 days after PDT. The characteristic histological changes were hemorrhagic necrosis and glandular destruction with preservation of interlobular collagen fibers. Urethral damage seen at the early stage healed by regeneration of urothelium in 4 weeks. We conclude that interstitial PDT with mTHPC is technically possible to produce extensive glandular necrosis in the normal prostate which heals safely and does not change the prostatic architecture. ALA, although it seems promising for bladder tumors, is much less effective than mTHPC on the prostate. With mTHPC, it might have the potential for treating prostate cancers localized in the periphery of the gland.© (1996) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Photodynamic therapy (PDT) has considerable potential for the treatment of superficial bladder neoplasia. Complications such as scarring of the detrusor muscle and prolonged cutaneous photosensitivity may be reduced by using the new photosensitizer precursor, 5- aminolaevulinic acid (ALA). After instillation of ALA, the concentration, pH, and time of bladder retention of ALA solution were found to be the key factors to a satisfactory PpIX buildup in the mucosa. The optimum PpIX fluorescence intensity ratio between mucosa and muscle layer is 10 to 1 with a pH 5.5, 1% ALA solution retained for 5 hours. Higher concentration resulted in more mucosal PpIX formation, but less selectivity. Unbuffered ALA was unsuitable for bladder instillation. Two days after laser treatment with 25 J/cm2 at 630 nm with optimal sensitization, typical histological findings were urothelial sloughing and lamina propria edema without obvious muscle damage. After 7 days, recovery of the urothelium was almost complete and fibroblast infiltration was minimal. ALA induced PpIX after bladder instillation may be an appropriate photosensitizer for future management of superficial bladder cancer.© (1994) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

5-aminolaevulinic acid(ALA) induced protoporphyrin IX(PpIX) has been proved to be a useful photosensitizer forphotodynamic therapy(PDT). In living cells, the conversion of PpIX to photoinactive haem is catalysed by ferrochelatase inthe presence of tissue iron and inhibition of this final committed step results in increased accumulation of PpIX. The invivo effect of a new iron chelator, 1 ,2-diethyl-3-hydroxypyridin-4-one (CP94) on the buildup of PpIX in different bladder layers was evaluated. In CP94 treated rats, 5-7 hours after intravesical instillation of ALA solution, the fluorescence intensity of PpIX in the urothelium was doubled whilst in the muscle layer it remanied low at a similar level to those seenwithout the iron chelator. With CP94, further reduction of skin photosensitisation is possible as similar photodynamiceffect on the bladder could be achieved at lower ALA concentration. The addition of CP94 seems an effective and convenientway to potentiate ALA induced PpIX tissue selectivity.Keywords: 5-aminolaevulinic acid(ALA), 3-hydroxypyridin-4-ones(HPOs), 1 ,2-diethyl-3-hydroxypyridin-4-one(CP94),bladder instillation, tissue selectivity, Desferrioxamine(DFO), photodynamic therapy(PDT)

In a series of 229 lower ureteral calculi manipulated with either extracorporeal shock wave lithotripsy (ESWL) or ureteroscopy the stone clearance rate 7 days after treatment was 40.6% and 90%, respectively. Only 17 of 32 patients (53.1%) treated with ESWL were rendered stone-free at 4-week followup. In addition to concern about stone clearance, other medical and social factors, and the availability of health insurance coverage must be considered. Since the differences of living standards between highly developed and developing countries are great, the selection of ESWL or ureteroscopy as primary treatment of lower ureteral calculi must be based on the socioeconomical status of a specific country. Because ESWL devices are not readily available in most developing countries and ureteroscopy is cost-effective, highly efficacious (greater than 92%) and involves minimal morbidity (less than 10%) for calculi removal in experienced hands, we recommend ureteroscopy as the treatment of choice for lower ureteral stones, with ESWL as the alternative, in developing countries.

Transrectal ultrasonic sagittal transducer is an excellent modality to image the bladderand urethra in dynamic change, In female patients, we found it is also helpful for the diagnosis of urinary stress incontinence. The posterior urethrovesical (PUV) angle is measured with the transrectal sonoprobe under strain and non-strain conditions. We compared the results of sonographic cystourethrogram with the radiographic chain cystourethrogram. The sonographic cystourethrography is superior to the radiograph, The former may estimate the PUV angle accurately and differentiate between the patients with and without stress urinary incontinence. Furthermore, we also use the transrectal sagittal probe intraoperatively to adjust the suspension force as well as PUV angle in patients who underwent vesical neck suspension for stress urinary incontinence (SUI).