J.H. Price

A bioadhesive patch cervical drug delivery system for the administration of 5-fluorouracil to cervical tissue

Abstract A novel bioadhesive cervical patch drug delivery is described, containing 5-fluorouracil for the treatment of cervical intraepithelial neoplasia (CIN). The patch was of bilaminar design, with a drug-loaded bioadhesive film cast from a gel containing 2% (w/w) Carbopol® 981 plasticised with 1% (w/w) glycerin. The casting solvent was ethanol/water 30:70, chosen to give a non-fissuring film with an even particle size distribution. The film, which was mechanically stable on storage under ambient conditions, was bonded directly to a backing layer formed from thermally-cured polyvinyl chloride emulsion. Bioadhesive strength was independent of drug loading in the bioadhesive matrix over the range investigated but was influenced by both the plasticiser concentration in the casting gel and the thickness of the final film. Release of 5-fluorouracil from the bioadhesive layer into an aqueous sink was rapid but was controlled down to an undetectable level through the backing layer. The latter characteristic was desirable to prevent drug spill from the device onto vaginal epithelium in vivo. Despite the relatively hydrophilic nature of 5-fluorouracil, substantial drug release through human cervical tissue samples was observed over approximately 20 h. Drug release, which was clearly tissue rather than device dependent, may have been aided by a shunt diffusion route through aqueous pores in the tissue. The bioadhesive and drug release characteristics of the 5-fluorouracil cervical patch indicated that it would be suitable for further clinical investigation as a drug treatment for CIN.

Menopause, hormone replacement and gynaecological cancers

Approximately 18,000 women are diagnosed with a gynaecological cancer in the UK each year. Predisposing risk factors for some of these gynaecological cancers include an early menarche/late menopause and hormone replacement therapy (HRT). Furthermore, treatment of gynaecological malignancies often induces an iatrogenic menopause, which may be more severe than a natural onset. HRT is an extremely effective treatment that may dramatically improve physical and psychological symptoms and ultimately quality of life in patients with cancer. However, the safety of using HRT in patients with gynaecological cancer is a controversial issue and not entirely clear. The main concern is the theoretical risk of the stimulation of residual cancer cells by estrogen replacement. The review of the evidence in this article found that for most gynaecological cancers this hypothesis was not proven. No study to date has found HRT to have a detrimental effect on survival in patients with early stage endometrial cancer, epithelial ovarian cancer, cervical cancer and vulval tumours. HRT is only an absolute contraindication in low-grade endometrial stromal sarcomas and is best avoided in granulosa cell ovarian tumours. Therefore, HRT should not be withheld in the majority of patients with gynaecological cancer. If quality of life is being adversely affected by symptoms of the menopause, then patients with cancer should be counselled regarding the known risks and benefits of HRT to enable them to make an informed decision on their treatment.

Serum CA19.9 levels are commonly elevated in primary ovarian mucinous tumours but cannot be used to predict the histological subtype

Aims CA19.9 is a monosialoganglioside secreted by mucinous tumours of the gastrointestinal tract, including the pancreas and biliary tree. Limited studies have shown that this tumour marker may also be elevated in primary ovarian mucinous neoplasms, but no study has assessed whether serum CA19.9 levels can be used to predict if a primary ovarian mucinous tumour is benign, borderline or malignant. The aim of this study was to correlate the serum CA19.9 level with the histological features in a large series of primary ovarian mucinous neoplasms. Methods 144 cases of primary ovarian mucinous neoplasm (79 benign, 45 borderline and 20 malignant) were identified in which a preoperative serum CA19.9 level had been performed. The association between the serum levels and the histological subtype and a variety of other parameters was investigated. In a subset of cases, immunohistochemical staining for CA19.9 was performed on tumour blocks. Results Serum CA19.9 levels were elevated in 27%, 38% and 40% of mucinous cystadenomas, borderline mucinous tumours and mucinous carcinomas, respectively. Markedly elevated levels of serum CA19.9 were observed in each group, with the highest serum CA19.9 measurements being noted in borderline mucinous tumours. There was no relationship between the serum CA19.9 level and whether the tumours were benign, borderline or malignant (Kruskal–Wallis test p value=0.32). A weak but statistically significant correlation was found between tumour maximum dimension and CA19.9 level (Spearmans rank correlation coefficient=0.17, p=0.04). In those cases in which CA19.9 immunohistochemistry was performed, all tumours showed positive staining for CA19.9, with 60% of these cases being associated with an elevated serum CA19.9 level. Conclusion Preoperative CA19.9 levels cannot be used to predict whether a suspected ovarian mucinous tumour is benign, borderline or malignant. Markedly elevated serum levels (>1000 U/ml) may be found in benign mucinous neoplasms as well as in borderline and malignant tumours.

Is laparoscopically assisted radical vaginal hysterectomy for cervical carcinoma safe? A case control study with follow up

Objective  To compare a new surgical approach, laparoscopically assisted radical vaginal hysterectomy (LARVH) with open radical hysterectomy in women with cervical cancer. Can selected women benefit from the minimally invasive approach without compromising safety (recurrence rate) and morbidity (complications)?

Successful photodynamic therapy of vulval Paget's disease using a novel patch‐based delivery system containing 5‐aminolevulinic acid

Case report A 66 year old Caucasian female attended for treatment of vulval Paget’s disease. She had an eight year history of persistent vulval itch, which had been treated unsuccessfully with antifungal, antibiotic and steroid creams. Biopsy at a district hospital revealed groups of large cells with atypical nuclei located within the epidermis with abundant clear to eosinophilic cytoplasm (Fig. 1A). These cells were positive with cytokeratin 7 and negative with cytokeratin 20. A diagnosis of Paget’s disease was made. There was no evidence of dermal invasion by tumour. Mammography, colonoscopy and computed tomography of the abdomen were all negative. Clinical examination showed a 4 3 cm, purple-red, well-demarcated, moist lesion on the left labia majora. Itch was graded on the visual analogue scale (0 to 10) at 8, becoming worse during the night and interrupting her sleep pattern. The patient was reluctant to undergo surgical excision and an experimental treatment using 5-aminolevulinic acid (ALA)-based photodynamic therapy was offered. Informed consent, as part of a trial protocol, was obtained. The treatment comprised part of an ongoing trial using photodynamic therapy for the treatment of a range of vulval lesions. The trial was performed under local Ethical Committee approval. A bioadhesive patch, which was developed in the School of Pharmacy, Queen’s University Belfast, containing 38 mg cm 2 ALA (Crawford Pharmaceuticals, Milton

An immunohistochemical and morphological analysis of post-chemotherapy ovarian carcinoma

Aims: Traditional management of advanced ovarian carcinoma is surgical debulking followed by chemotherapy; however, there is an increasing tendency for neoadjuvant chemotherapy followed by surgery. The morphology of ovarian carcinoma following chemotherapy often differs markedly from native tumour. This study aimed to compare the immunophenotype of post-chemotherapy ovarian carcinomas with that of untreated tumour. Methods: Post-chemotherapy ovarian carcinomas (n = 16) were stained with a range of antibodies. In six cases, pre-chemotherapy core biopsies were also stained; all were high-grade serous carcinomas. Antibodies used in the study were CK7, CA125, WT1, ER, PR, p53, p16, p63 and MIB1. Results: In eight post-treatment cases, there was minimal or no morphological response to chemotherapy, and in eight there was a significant response (in two additional cases, no residual tumour was identified). All pre-chemotherapy biopsies showed diffuse positivity of the tumour cells with CK7, CA125 and WT1. ER, p53 and p16 were diffusely positive in five, four and three cases respectively. One case was focally PR positive, and all were p63 negative. MIB1 staining was high; all but one case exhibited a proliferation index of >60%. Post-chemotherapy tumours exhibited a similar immunophenotype: diffuse positivity with CK7 in all cases and with CA125, WT1, ER, p53 and p16 in the majority, an immunophenotype in keeping with a serous carcinoma. All were negative with p63, and all but two with PR. The MIB1 proliferation index was lower in those cases exhibiting a significant morphological response, and p53 was less likely to be positive in cases with minimal or no response. Conclusions: The immunophenotype of post-chemotherapy ovarian carcinomas is very similar to that of native untreated tumours, illustrating that CK7, CA125, WT1, ER, p53 and p16 may be of value in identifying residual tumour cells and in subtyping the neoplasm if a pre-chemotherapy biopsy has not been obtained.

Response of vulval lichen sclerosus and squamous hyperplasia to photodynamic treatment using sustained topical delivery of aminolevulinic acid from a novel bioadhesive patch system.

This study evaluated the clinical and histopathological responses of vulval lichen sclerosus (LS) and squamous hyperplasia (SH) to photodynamic therapy (PDT). A novel bioadhesive patch containing aminolevulinic acid (ALA) at a dose of (38 mg/cm2) was used to treat 10 patients before irradiation with light of 630 nm. Clinical, histopathological and pathological responses to treatment were assessed at 6 weeks post‐treatment. After 17 cycles of PDT, six patients reported significant symptomatic relief and no cutaneous photosensitivity. Histopathological differences were not demonstrated, but statistically significant induction of apoptosis was seen. It can be concluded that ALA‐PDT patch‐based formulation is pragmatic and primarily offers symptomatic management of vulval LS and SH.

Clinical and immunohistochemical assessment of vulval intraepithelial neoplasia following photodynamic therapy using a novel bioadhesive patch-type system loaded with 5-aminolevulinic acid

BACKGROUND The work in this study appraised photodynamic treatment (PDT) as a treatment method for vulval intraepithelial neoplasia (VIN) using a novel bioadhesive patch to deliver aminolevulinic acid. An analysis of changes in expression of apoptotic and cell cycle proteins (p53, p21, Mdm2, Blc-2, Bax, Ki-67) in response to PDT was evaluated. METHODS PDT was performed using non-laser light, either as a one or two-cycle treatment, with clinical and pathological assessment following after 6 weeks. Twenty-three patients with 25 VIN lesions underwent 49 cycles of PDT. Patches were designed to conform to uneven vulval skin and contained 38 mg cm(-2) aminolevulinic acid. Assessment was carried out at 6 weeks post-treatment. Patient-based treatment assessment, along with clinical and pathological changes, were monitored. Immunohistochemical staining was used to elucidate a possible biomolecular basis for induced cellular changes. RESULTS Most patients (52%) reported a symptomatic response, with normal pathology restored in 38% of lesions. The patch was easy to apply and remove, causing minimal discomfort. Fluorescence inspection confirmed protoporphyrin accumulation. Pain during implementation of PDT was problematic, necessitating some form of local analgesia. Changes in expression of cell cycle and apoptotic-related proteins suggested involvement of apoptotic pathways. Down regulation of p21 and inverse changes in Bcl-2 and Bax were key findings. CONCLUSION Treatment of VIN lesions using a novel bioadhesive patch induced changes in cell cycle and apoptotic proteins in response to PDT with possible utilisation of apoptotic pathways. The efficacy of PDT in treating VIN could be improved by a better understanding of these apoptotic mechanisms, the influence of factors, such as HPV status, and of the need for effective pain management.

Expression of the SEPT9_i4 isoform confers resistance to microtubule-interacting drugs

BackgroundThe evolutionarily conserved septin family of genes encode GTP binding proteins involved in a variety of cellular functions including cytokinesis, apoptosis, membrane dynamics and vesicle trafficking. Septin proteins can form hetero-oligomeric complexes and interact with other proteins including actin and tubulin. The human SEPT9 gene on chromosome 17q25.3 has a complex genomic architecture with 18 different transcripts that can encode 15 distinct polypeptides. Two distinct transcripts with unique 5′ ends (SEPT9_v4 and SEPT9_v4*) encode the same protein. In tumours the ratio of these transcripts changes with elevated levels of SEPT9_v4* mRNA, a transcript that is translated with enhanced efficiency leading to increased SEPT9_i4 protein.MethodsWe have examined the effect of over-expression of SEPT9_i4 on the dynamics of microtubule polymer mass in cultured cells.ResultsWe show that the microtubule network in SEPT9_i4 over-expressing cells resists disruption by paclitaxel or cold incubation but also repolymerises tubulin more slowly after microtubule depolymerisation. Finally we show that SEPT9_i4 over-expressing cells have enhanced survival in the presence of clinically relevant microtubule acting drugs but not after treatment with DNAinteracting agents.ConclusionsGiven that SEPT9 over-expression is seen in diverse tumours and in particular ovarian and breast cancer, such data indicate that SEPT9_v4 expression may be clinically relevant and contribute to some forms of drug resistance.

Phototoxicity of 5-aminolevulinic acid in the HeLa cell line as an indicative measure of photodynamic effect after topical administration to gynecological lesions of intraepithelial form.

AbstractPurpose. The depth-resolved pattern of aminolevulinic acid (ALA) concentration in excised vaginal tissue was determined after in vitro application of an ALA-loaded bioadhesive patch. From this data, the tissue concentration of ALA achievable at a specified depth from the surface could be related to the concentration needed to elicit a photodynamic effect in a model gynecological tumor cell line (HeLa). Methods. Excised vaginal tissue was mounted in a modified Franz diffusion cell and exposed to a water-soluble, ALA-loaded, bioadhesive patch. After a period of time, the tissue was cryostatically sectioned and the stratal concentration of radiolabeled ALA determined using scintillation spectroscopy. HeLa cells were cultured in media containing specific concentrations of ALA and exposed to standard photodynamic protocols of light exposure. Results. An ALA concentration of 65.6 mM was achievable at 2.375 mm from the tissue surface after application of ALA-loaded patch. The photodynamic effectiveness of this concentration was demonstrated in HeLa with exposure to concentrations exceeding 1.0 mM ALA bringing about reductions in viable cell numbers by 90%. An enhancement of PpIX production using adjunctive EDTA over the clinically relevant 4 h application time interval was shown to be minimal in HeLa. Instead, PpIX production was more closely correlated with ALA concentration, with 100 mM ALA producing approximately 3100 ng PpIX mg-1 protein in the same time period. Conclusions. Given that vaginal intraepithelial neoplasias can extend to 2.0 mm from the lesion surface, the ALA permeability derived from a bioadhesive patch is sufficient to induce photosensitization suitable for light induced destruction at deep sites of this type of lesion.