Despite the efficient HIV-1 replication blockage achieved with current HAART therapies, HIV-1 persists in the body and survives in a latent state that can last for the entire life of the patient. A long-lived reservoir of latently infected CD4+ memory T-cells represents the most important sanctuary for the virus and the greatest obstacle for viral eradication. In this work, we present an initial step towards a gene therapy approach aimed at the activation of latent provirus to induce the death of latently infected T-cells. Latent HIV-1 infection is characterized by the failure of viral-gene expression as a consequence of uninitiated or aborted transcription. We have constructed an HIV-1-based lentiviral vector (p5p53RTAT3) that expresses the viral transactivating protein Tat in a drug-regulated manner and p53 in a Rev-dependent manner. We have demonstrated that the Tat-expressed protein from p5p53RTAT3 vector reactivates latent HIV-1 proviruses in J1.1 and ACH-2 cell lines and promotes p53-induced apoptosis in the presence of Rev. Our system was able to trigger the transactivation of the provirus 5'LTR, stimulate the expression of the Rev protein from a tat-defective provirus, and provoke apoptosis selectively in the cells transfected with a tat-defective HIV-1 provirus in contrast to those with no HIV-1 provirus. However, the Rev-dependent-p53 killing of latently infected cells was not effective enough for complete elimination of the awakened HIV-1 viruses. In summary, we have developed a vector system that is quite efficient at activating latent HIV-1 proviruses but that needs further improvement to kill infected cells.

Many wrasses on coral reefs exhibit daily spawning that peaks around daytime high tides. In this study, we examined tidal-related ovarian development in the threespot wrasse, Halichoeres trimaculatus, a species common on coral reefs in the Indo-Pacific Ocean. When the fish were collected in the morning at different tidal phases, the gonadosomatic index (GSI) and ovarian histology changed; concomitant with increases in GSI towards high tide, a clutch of the most advanced oocytes developed from vitellogenic to maturation stages. Ovulated eggs and post-ovulated follicles (POF) existed in most ovaries around high tide, but only POF remained around ebb tide, suggesting that spawning occurred during or after high tide. We noticed that tidal-related spawning was considerable in the morning and that most ovaries collected on the afternoon high tide exhibited post-spawning features. This suggests that certain labrid species possess plasticity with regard to their spawning time and utilize potent environmental cues to ensure their reproductive success. When pieces of ovary were incubated with precursor steroids, high conversion of testosterone to 17beta-estradiol occurred during high and ebb tides, while that of 17alpha-hydroxyprogesterone to 17alpha,20beta-dihydroxy-4-pregnen-3-one and 17alpha,20beta,21-trihydroxy-4-pregnen-3-one was observed during low and flood tides. Incubation of pieces of ovary with human chorionic gonadotropin resulted in similar fluctuations in the steroid hormones with tidal phase. Production of these steroid hormones correlated with oocyte development in the ovaries and was probably regulated by gonadotropin. These results demonstrate that the daily cycle is fundamental for oocyte development, and that the tidal cycle is superimposed on this process.

The aim of this study was to evaluate the usefulness of targeted intra-arterial carboplatin chemoradiotherapy in allowing less invasive surgery for patients with oral and oropharyngeal squamous cell carcinoma. Twenty patients with previously untreated squamous cell carcinoma of the oral cavity and oropharynx (T4; 8, T2; 12 patients) were treated with targeted transfemoral intra-arterial carboplatin infusion with concurrent hyperfractionated radiotherapy and the administration of tegafur/uracil (UFT). Of 20 patients, 15 underwent surgery after completion of one course of targeted chemoradiotherapy, and five were given another course or radiotherapy only. Eighteen (90%) of 20 patients had a clinically complete response at the primary site and two (10%) had a partial response. Of the 15 patients who underwent tumor resection, 11 (73%) showed histopathological disappearance of cancer cells at the primary site. Sixteen (80 %) of 20 tumors were controlled at the primary site within a mean follow-up of 30 months. Adverse effects were relatively mild. Targeted intra-arterial chemoradiotherapy caused a down-staging of tumors and facilitated the use of less invasive surgery in patients with squamous cell carcinoma of the oral cavity and oropharynx as a result of its favorable anti-tumor effect.

Our aim was first to evaluate the association between blood vessel density (BVD) and free platinum concentration in experimentally induced tumours in rabbits. We also investigated the association between tumour BVD and the clinical response of patients who had undergone targeted carboplatin intra-arterial (i.a.) chemoradiotherapy. VX2 carcinoma cells were transplanted into 46 inbred female Japanese white rabbits. In the i.a. group, carboplatin was infused into the lingual artery, and in the intravenous (i.v.) group, carboplatin was infused through the auricular vein. In the clinical study, we evaluated 19 patients with squamous cell carcinomas of the oral cavity and oropharynx, who had undergone targeted carboplatin i.a. chemoradiotherapy and had been administered i.a. tegafur/uracil chemotherapy before surgery. We quantified angiogenesis in both studies. Increased BVD was associated with a higher free platinum concentration in the tumour region in the i.a. group of rabbits. In the clinical study, using multivariate logistic regression analysis, only the BVD was related independently to the treatment effect. Therefore, BVD is a valid predictor of the effects of i.a. targeted carboplatin chemotherapy and concurrent radiotherapy for treating human oral and oropharyngeal squamous cell carcinomas.British Journal of Cancer advance online publication, 2 May 2006; doi:10.1038/sj.bjc.6603138 www.bjcancer.com.

Tumour angiogenesis is mediated by increased levels of vascular endothelial growth factor (VEGF). We have studied the mechanism by which endogenous activation of Rho oncoproteins regulates VEGF expression in COS-7 and NIH3T3 cells. We carried out transient and stable transfection with constitutively activated rhoA, rac1, and cdc42 mutants in COS-7 and NIH3T3 cells, respectively in the absence of external stimuli. Western blot and inmunohistochemistry assays of those cells revealed increased VEGF protein expression. Cotransfection with constitutively activated rhoA, rac1, and cdc42 mutants and a VEGF promoter-reporter construct showed an increase in VEGF promoter transcriptional activity induced by Rho oncoproteins in COS-7 and NIH3T3. c-Jun kinase had been described as a MAPK involved in Rho oncoproteins pathways. Interestingly, we found that c-Jun kinase chemical inhibition as well as transient transactivation assays using dominant negative c-Jun kinase mutant abolished the VEGF promoter transcriptional induction by Rac1 and Cdc42 but not by RhoA. These findings indicate that Rho oncoprotein endogenously activated regulates VEGF expression through a transcriptional mechanism, and that the c-Jun kinase activity is a mediator in the expression of VEGF induced by Rac1 and Cdc42 oncoproteins, but not of that induced by RhoA. J. Cell. Biochem.(c) 2006 Wiley-Liss, Inc.

This study was performed to analyze treatment of fractures of the edentulous mandible and to discuss this method in relation to the mandibular height at the fracture site. Fifteen fracture sites in 11 patients with an edentulous mandible were retrospectively examined. These fractures were located: nine fractures in the mandibular body, three in the paramedian region, and three in the mandibular angle. Fractures in a mandible measuring more than 10 mm in the vertical height were treated with one miniplate. Fractures in an extremely atrophic mandible with 10 mm or less were treated using one or two miniplates, also using a modified Champy plate with 1.3 mm in thickness. A mandibular fracture with a height of 5 mm was treated with a combination of a microplate on the buccal side and a miniplate on the inferior border of the mandible with additional direct circumferential wiring. Oblique or splitting fractures were treated with direct circumferential wiring or a Herbert screw, at one fracture site each, respectively. Complications, including infection, fibrous union, nonunion and trismus, were not seen. In one patient, hypesthesia of the lower lip was, however, persistent 1 month after surgery. Miniplate osteosynthesis is the less invasive treatment, and it is suitable for fractures of the atrophic edentulous mandible, except for comminuted or defect fractures. To obtain stable fixation in severely atrophic mandibles, we need to consider the use of two miniplates or a combination with microplates.

At present, virus replication is quite efficiently blocked by highly active antiretroviral therapy (HAART). However current HAART therapies still have important limitations that compromise in the long term the health of the patient. This failure is due to the existence of viral sanctuaries of replication-competent HIV-1, which can persists for the whole life. Two main strategies of HIV-1 gene therapy have been tried. One is based in the rationale of killing the infected cells before the virus can produce infective particles and the other strategy is to protect the cell from the virus. The first strategy uses lethal genes that are expressed in response to the expression of a HIV-1 provirus. Although technical limitations and safety concerns have limited the use of this approach in the last years, recent development of efficient lentiviral vectors and the need of the elimination of the latently infected cells are reasons to reconsider and come back to use this approach. Elimination of latently infected cells would also represent a perfect complement of the HAART therapy and may represent a hope in the long term for the development of a real cure of this infection.

Recent advances in orthognatic surgery have been remarkable. Among the surgical procedures, sagital ramus osteotomy has been often used for mandibular protrusion. Causes of dentofacial deformities include not only congenital but also acquired ones, which are caused by trauma or resection of tumor in the maxillofacial region. In all cases, functional and psychological disorders would often occur in the patient with these dentofacial deformities. We report a favorable result of anterior alveolar osteotomy of the mandible, which was applied to a patient complaining of malocculusion and facial deformity developed from a healed fracture of the mandible treated at another hospital.

A 73-year-old man presented with well-differentiated squamous cell carcinoma of the tongue (T3N0N0). After the completion of external beam radiotherapy (32 Gy) and oral tegafur-uracil (UFT) administration, the tumor had clinically disappeared. Two years later, a recurrent lesion was confirmed histologically. Without any specific therapy, the tumor gradually and spontaneously regressed, from 4 months after the recurrence was diagnosed, and it had completely disappeared both clinically and by computed tomography imaging at 1 year and 4 months after recurrence. There was no evidence suggesting regrowth of the disease in the patient at 3 years and 7 months after the histological confirmation of tumor recurrence.

We investigated whether intra-arterial administration of carboplatin using Calvert's formula is useful for avoiding thrombocytopenia in targeted chemoradiotherapy in patients with squamous cell cancer of the oral cavity and oropharynx. Carboplatin was infused intra-arterially under digital subtraction angiography in 28 patients. In the first group of patients, the dose of carboplatin was calculated according to the body surface area (BS group). In the second group, the dose was calculated using Calvert's formula (AUC group). The value for AUC (area under concentration vs time curve; mg ml(-1) min(-1)) in the formula was set at 4.5. All patients received concurrent radiotherapy (30 Gy) and were given oral tegafur-uracil (UFT((R)), 400-600 mg day(-1)). The AUC group showed a significantly lower percentage platelet reduction than the BS group (49.0+/-22.0 vs 65.1+/-23.2%; P=0.045) and also tended to have a higher platelet nadir count (10.9+/-4.2 vs 8.4+/-5.8 x 10(4); P=0.27) without reducing the antitumour effect. The value of 4.5 for target AUC is recommended clinically. However, AUC of Calvert's formula could not predict thrombocytopenia associated with intra-arterial chemoradiotherapy due to the variability of the actual AUC.British Journal of Cancer (2004) 90, 2062-2066. doi:10.1038/sj.bjc.6601818 www.bjcancer.com Published online 20 April 2004