Springer Science and Business Media LLC

Objective: This study was designed to analyze expression patterns of estrogen receptor (ER), human epidermal growth factor receptor-2 (HER2/ERBB2), and nonmetastatic protein 23 (NM23-H1/NME1) proteins in patients with invasive ductal carcinoma and different menopausal status to identify their relationships with axillary lymph node metastasis. Materials and Methods: 213 pre-menopausal and 177 post-menopausal women diagnosed with invasive ductal carcinoma were evaluated for ER, HER2, and NM23-H1 protein expression by immunohistochemistry. When HER2 immunoreactivity was equivocal (category 2+), specimens were confirmed by fluorescence in situ hybridization. Results: ER expression showed no correlation with menopausal status or lymph node metastasis (each p > 0.05). However, expression of ER was associated with negative expression of HER2 (r =−0.214, p < 0.05) and positive expression of NM23-H1 (r = 0.137, p < 0.05) in the pre-menopausal group. Over-expression of HER2 was correlated with menopausal status (r =−0.107, p < 0.05) and lymph node metastasis in the ER-negative post-menopausal group (r = 0.222, p < 0.05). NM23-H1 was associated with less lymph node metastasis in the ER-positive pre-menopausal group (r = −0.237, p < 0.05). Conclusion: Our results indicated that expression patterns of ER, NM23-H1, and HER2 in primary breast cancer lesions warn that cells might have metastatic potential, which could assist clinicians to provide a more accurate prognosis and tailor therapeutic management for individual patients.

The majority of patients with JAK2 V617F-negative essential thrombocythemia or primary myelofibrosis harbor mutations involving the calreticulin (CALR) gene. These mutations are located in CALR exon 9 and lead to a frameshift with subsequent alteration of the CALR protein C-terminus. They have emerged as valuable molecular markers for the diagnosis of clonal myeloproliferative diseases. Although a variety of CALR mutations have been described, two mutations, denoted type 1 and type 2, account for around 85 % of cases. The type 1 mutation encompasses a 52 bp deletion and the type 2 mutation a 5 bp TTGTC insertion. This work describes the development and testing of quantitative real-time PCRs (qPCRs) for detecting these two mutations. The final type 1 CALR qPCR displayed a sensitivity of <0.1 % mutant alleles and the type 2 CALR qPCR had a sensitivity of <0.01 % mutant alleles. Additionally, two new CALR mutations are reported. These sensitive and specific qPCRs should be helpful in establishing the diagnosis and in monitoring minimal residual disease in patients during or after therapy.

Background: Interferon-β (IFNβ) has proven to be an important advance in the therapy of multiple sclerosis (MS), but optimal markers for bioactivity have not been identified. To accurately measure bioactivity in MS patients treated with IFNβ, we developed and tested a real-time reverse transcriptase (RT)-PCR assay for gene expression of MxA, an IFNβ-induced gene in the peripheral blood of patients treated with IFNβ. Methods: We compared IFNβ-treated patients with MS to controls in expression of MxA relative to the house-keeping gene, GAPDH. 2′-5′oligoadenylate synthetase (OAS) gene expression was also tested by real-time RT-PCR on RNA from the same patient specimens. Anti-IFNβ antibody was measured by ELISA and a cytopathic effect assay. Results: Seven of 54 patients were found to have complete loss of bioactivity. MxA expression correlated well with OAS expression. All patients with lost bioactivity had high levels of binding antibodies or neutralizing antibodies. Conclusions: This is the first demonstration that a real-time RT-PCR assay can be used to monitor therapy with interferons. These data identify MxA mRNA as an excellent biomarker for INFβ action on the IFN receptor, and clarify the relationship between anti-IFNβ antibodies and bioactivity in patients with MS treated with IFNβ.

Skin wound healing is a crucial process for regenerating healthy skin and avoiding the undesired consequences associated with open skin wounds. For epidermolysis bullosa (EB), a debilitating group of fragile skin disorders currently without a cure, skin blistering can often be severe and heal poorly, increasing susceptibility to life-threatening complications. To prevent these, investigational therapies have been exploring the use of tissue-engineered skin substitutes (TESSs) aimed at replacing damaged skin and promoting long-term wound closure. These products have either been developed in house or commercially sourced and are composed of allogeneic or autologous human skin cells, often with some form of bioscaffolding. They can be broadly classified based on their cellular composition: keratinocytes (epidermal substitutes), fibroblasts (dermal substitutes) or a combination of both (composite substitutes). Encouraging long-term wound healing has been achieved with epidermal substitutes. However, these substitutes have not demonstrated the same efficacy for all patients, which may be due to the molecular heterogeneity observed between EB subtypes. Autologous composite TESSs, which more closely resemble native human skin, are therefore being investigated and may hold promise for treating an extended range of patients. Additionally, future TESSs for EB are focused on using gene-corrected patient skin cells, which have already demonstrated remarkable long-term wound healing capabilities. In this review, we provide an overview of the different TESSs that have been investigated in clinical studies to treat patients with EB, as well as their long-term wound healing results. Where available, we describe the methods used to develop these products to inform future efforts in this field.

Hexaminolevulinate (HAL) is an ester derivative of 5-aminolevulinate, and is used in conjunction with blue light (fluorescence) cystoscopy (BLC) to detect bladder cancer. In various studies, HAL-BLC was generally a better detection method than the gold standard method of white light cystoscopy (WLC), as assessed by endpoints including additional lesions (at both the lesion and patient levels) not detected by WLC, higher detection rates, and more complete treatment decisions as a result of improved detection. HAL-BLC in addition to WLC was associated with reduced tumor recurrence rates compared with WLC alone in randomized trials with up to 2 years’ follow-up. HAL-BLC was generally well tolerated, and the most common adverse events (i.e. hematuria, dysuria, and bladder spasm) were also the most common adverse events in the WLC alone group, and were expected as a result of the resection procedure. Although its impact on disease progression and survival rates is not yet known, HAL-BLC is a valuable addition to WLC for the diagnosis of bladder cancer in patients who are suitable candidates for its use.

Osteonecrosis is a disabling complication of treatment for pediatric acute lymphoblastic leukemia, and much effort has been made to predict which patients are prone to develop this disease. Multiple clinical and genetic factors have already been identified as being associated with osteonecrosis; however, a prediction model that combines pretreatment genetic biomarkers and clinical factors has not yet been designed. Such a prediction model can only be developed with continuing international collaborations and research efforts, including large genome-wide association studies.