Galactorrhoea of the neck following pectoralis major reconstruction of a pharyngeal defect.

We describe a case of postoperative galactorrhea following the use of a pedicled pectoralis major myocutaneous flap for reconstruction of a pharyngolaryngeal defect in a woman with squamous cell carcinoma. We believe this to be unique in the literature, and an important complication to be reported, due to the similarities in appearance of galactorrhoea and postoperative aerodigestive tract/cutaneous fistula.

Immunization of mice with Plasmodium TCTP delays establishment of Plasmodium infection.

Translationally controlled tumour protein (TCTP) may play an important role in the establishment or maintenance of parasitemia in a malarial infection. In this study, the potential of TCTP as a malaria vaccine was investigated in two trials. In the initial vaccine trial, Plasmodium falciparum TCTP (PfTCTP) was expressed in Saccharomyces cerevisiae and used to immunize BALB/c mice. Following challenge with Plasmodium yoelii YM, parasitemia was significantly reduced during the early stages of infection. In the second vaccine trial, the TCTP from P. yoelii and P. berghei was expressed in Escherichia coli and used in several mouse malaria models. A significant reduction in parasitemia in the early stages of infection was observed in BALB/c mice challenged with P. yoelii YM. A significantly reduced parasitemia at each day leading up to a delayed and reduced peak parasitemia was also observed in BALB/c mice challenged with the nonlethal Plasmodium chabaudi (P.c.) chabaudi AS. These results suggest that TCTP has an important role for parasite establishment and may be important for pathogenesis.

Cervical metastases of squamous cell carcinoma of the maxilla: a retrospective study of 25 years.

OBJECTIVES: Squamous cell carcinomas (SCCs) of the maxilla are relatively rare; therefore, only little data is available regarding the frequency of cervical metastasis (CM) and therapy strategies. Most authors only undertake clinical observation of the lymph nodes. The aim of this retrospective study was to evaluate the manner of metastasis in SCC of the maxilla. MATERIALS AND METHODS: All patient records from 1987 to 2011 were scanned for SCC of the maxilla. Patients with SCC limited to the maxilla were comprised. The cases were analyzed regarding tumor node metastasis staging system and any special occurrences in the follow-up time such as tumor recurrence, metastasis, and exitus letalis. Classification and staging were performed according to the 2003 UICC system. RESULTS: One hundred thirty-eight patients were comprised of 36 % females and 64 % males (average age, 66 years; women, 71 years; men, 63 years). The average follow-up time was 43 months (range, 0-195). Fifty-eight percent smoked or declared regular consumption of alcohol. About 50 % of the patients had an advanced tumor stage (III-IV). At the time of the primary diagnosis, 38 % of the patients had CM. There is an increased risk for CM occurrence with increasing tumor size and grading and a tumor localized in the postcanine region. Contralateral CM arises frequently in T4 tumors and tumors localized in the postcanine region. CONCLUSION: The data exhibit aggressive regional metastatic behavior of SCC of the maxilla. CLINICAL RELEVANCE: Therefore, surgical treatment of the draining lymphatic system as a primary management strategy is recommended for patients with SCC of the maxilla.

In situ quantification of HER2-protein tyrosine kinase 6 (PTK6) protein-protein complexes in paraffin sections from breast cancer tissues.

BACKGROUND: Protein tyrosine kinase 6 (PTK6; breast tumour kinase) is overexpressed in up to 86% of the invasive breast cancers, and its association with the oncoprotein human epidermal growth factor receptor 2 (HER2) was shown in vitro by co-precipitation. Furthermore, expression of PTK6 in tumours is linked with the expression of HER2. METHOD AND RESULTS: In this study, we used the proximity ligation assay (PLA) technique on formalin-fixed paraffin sections from eighty invasive breast carcinoma tissue specimens to locate PTK6-HER2 protein-protein complexes. Proximity ligation assay signals from protein complexes were assessed quantitatively, and expression levels showed a statistically significant association with tumour size (P=0.015) and course of the cancer disease (P=0.012). CONCLUSION: Protein tyrosine kinase 6 forms protein complexes with HER2 in primary breast cancer tissues, which can be visualised by use of the PLA technique. Human epidermal growth factor receptor 2-PTK6 complexes are of prognostic relevance.

WWOX mRNA expression profile in epithelial ovarian cancer supports the role of WWOX variant 1 as a tumour suppressor, although the role of variant 4 remains unclear.

WWOX is a candidate tumour suppressor gene that exhibits LOH or homozygous deletion in several tumour types. As well as the predominant full-length transcript (variant 1) there also exist alternatively spliced transcripts found previously only in malignant tissue. It has been suggested that proteins encoded by these variants may interfere with normal WWOX function in a dominant negative fashion. The most prevalent alternate transcript demonstrated in ovarian cancer is variant 4, which lacks exons 6-8. Here, we report the first comparison of the mRNA expression of WWOX variants 1 and 4 in human ovarian tumours and normal ovaries, and correlate expression with clinical data. We demonstrate significantly lower WWOX variant 1 expression in tumours than in normal ovaries. This reduction was not associated with any specific clinical subgroup. Variant 4 was expressed at low levels, and significantly associated with high grade and advanced stage ovarian cancer. Furthermore, tumours co-expressing variant 4 and relatively high levels of variant 1 showed significantly worse survival than tumours expressing variant 1 alone. However, variant 4 was also frequently identified in non-malignant ovarian tissue. These results support the role of WWOX variant 1 as a suppressor of ovarian tumourigenesis, but the role of variant 4 remains speculative.

WWOX: a candidate tumor suppressor gene involved in multiple tumor types.

We previously reported the construction of a P1-derived artificial chromosome (PAC) contig encompassing a set of homozygous deletions of chromosome 16q23-24.1 found in primary ovarian tumor material and several tumor cell lines. Using these PAC clones in a cDNA selection experiment, we have isolated a Sau3A fragment homologous to the WWOX transcript (GenBank accession no. ) from normal human ovarian surface epithelial (HOSE) cells. We demonstrate the homozygous deletion of WWOX exons from ovarian cancer cells and three different tumor cell lines. We also identify an internally deleted WWOX transcript from a further primary ovarian tumor. In three of these samples the deletions result in frameshifts, and in each case the resulting WWOX transcripts lack part, or all, of the short chain dehydrogenase domain and the putative mitochondrial localization signal. Sequencing revealed several missense polymorphisms in tumor cell lines and identified a high level of single nucleotide polymorphism (SNP) within the WWOX gene. This evidence strengthens the case for WWOX as a tumor suppressor gene in ovarian cancer and other tumor types.

A hydroxychalcone derived from cinnamon functions as a mimetic for insulin in 3T3-L1 adipocytes.

OBJECTIVES: These studies investigated the ability of a hydroxychalcone from cinnamon to function as an insulin mimetic in 3T3-LI adipocytes. METHODS: Comparative experiments were performed with the cinnamon methylhydroxychalcone polymer and insulin with regard to glucose uptake, glycogen synthesis. phosphatidylinositol-3-kinase dependency, glycogen synthase activation and glycogen synthase kinase-3beta activity. The phosphorylation state of the insulin receptor was also investigated. RESULTS: MHCP treatment stimulated glucose uptake and glycogen synthesis to a similar level as insulin. Glycogen synthesis was inhibited by both wortmannin and LY294002, inhibitors directed against the PI-3-kinase. In addition, MHCP treatment activated glycogen synthase and inhibited glycogen synthase kinase-3beta activities, known effects of insulin treatment. Analysis of the insulin receptor demonstrated that the receptor was phosphorylated upon exposure to the MHCP. This supports that the insulin cascade was triggered by MHCP. Along with comparing MHCP to insulin, experiments were done with MHCP and insulin combined. The responses observed using the dual treatment were greater than additive, indicating synergism between the two compounds. CONCLUSION: Together, these results demonstrate that the MHCP is an effective mimetic of insulin. MHCP may be useful in the treatment of insulin resistance and in the study of the pathways leading to glucose utilization in cells.

Cancer screening in a high risk population: a clinical trial.

The purpose of this paper is to report the results of screening high risk women for ovarian cancer using endovaginal ultrasound (EVUS), color flow Doppler and cancer antigen (CA) 125. A total of 252 women were recruited with a family history of ovarian cancer in at least 1 first-degree relative. All women underwent a pelvic examination and EVUS twice during the first year and annually thereafter. Of 210 premenopausal women in the study, 14 underwent surgery. Of these, 2 were based on the endovaginal ultrasound (US) results and proved to be false-positive. There were 48 postmenopausal women who underwent 9 operations. Of these, 2 were stimulated by finding a thickened stripe on EVUS, and proved to be endometrial carcinomas. There were 2 ovarian cancers, both advanced, 1 colon cancer and 1 renal cell cancer. Of the women, 6 had a history of breast cancer and 11 more developed it during the study. This high-risk population has a very high prevalence of breast cancer, and mammography must be a part of ovarian cancer screening programs.

Involving nursing students in continuous improvement projects.

Reducing medication errors is a topic of national concern and action. Nursing students participated in a healthcare organization's continuous quality improvement project targeting patient safety. Students were actively involved in chart review and became acutely aware of safety issues related to medication administration, order transcription and implementation, and documentation. Both the students and the hospital realized expected and unexpected benefits.

Sonography of palpable breast cancer.

PURPOSE: Because of the increasing use of sonography to rule out cancer in women with palpable breast abnormalities, this study was performed to determine the rate of sonographically occult malignancy in this clinical setting. METHODS: Women who were recommended for biopsy based on mammographic and/or clinical findings underwent breast sonography. This study retrospectively analyzed the subset of patients with palpable malignant lesions. Lesions were classified as visible or occult on mammography and sonography. Patients without a tissue diagnosis of tumor were excluded. RESULTS: Of 1,346 masses that underwent biopsy or aspiration, 616 lesions were palpable, and of these, 293 were malignant. Sonography detected all 293 palpable malignant lesions (95% confidence interval for sensitivity, 99-100%). Eighteen lesions were mammographically occult. The median lesion size as determined by sonography was 1.8 cm; for the lesions that were mammographically occult, the median size was 1.6 cm. The most common histopathologic diagnosis for both groups of lesions was infiltrating ductal carcinoma. CONCLUSIONS: All palpable malignant breast lesions were visible by sonography in patients in whom a biopsy was recommended. However, we caution that until the false-negative rate of sonography for equivocal palpable abnormalities is determined prospectively, sonography cannot be accurately applied to rule out malignancy in this setting.

A 700-kb physical map of a region of 16q23.2 homozygously deleted in multiple cancers and spanning the common fragile site FRA16D.

We have identified a >600-kb region at 16q23.2 that is homozygously deleted from malignant ovarian ascites using representational difference analysis. Overlapping homozygous deletions were also observed in the colon carcinoma cell line HCT116 and a xenograft established from the small cell lung cancer cell line WX330. This region coincides with that described previously by others as showing loss of heterozygosity in prostate and breast cancers (C. Li et al., Genes Chromosomes Cancer, 24: 175-182, 1999; A. Latil et al., Cancer Res., 57: 1058-1062, 1997; K. Driouch et al., Genes Chromosomes Cancer, 19: 185-191, 1997; A. Iida et al., Br. J. Cancer, 75: 264-267, 1997). In addition, the minimally deleted region spans the common fragile site FRA16D. We have constructed a 700-kb physical map encompassing the deleted region. By fluorescence in situ hybridization of aphidicolin-induced metaphase chromosomes, we have preliminary data to suggest that P1-derived bacterial artificial chromosome clones from the contig lie on both sides of FRA16D. This is confirmed by extensive fluorescence in situ hybridization analysis of the region reported in the accompanying article (M. Mangelsdorf et al., Cancer Res., 60: 1683-1689, 2000) and is consistent with an involvement of this common fragile site in the loss of 16q23.2 material in various cancer types. The minimally deleted region of approximately 210 kb has been characterized using our own markers and public domain markers. Eleven distinct expressed sequences mapped to the region, providing a basis for identifying the predicted tumor suppressor gene in this region.

Contrast-enhanced ultrasound for guidance of local tumor ablation.

The objective was to assess the efficacy of sonography with and without contrast medium enhancement in guiding and monitoring percutaneous ethanol ablation of tumors in an animal model. VX-2 carcinoma was implanted into the thighs of New Zealand white rabbits and examined by grey-scale ultrasound, color, power, and pulse Doppler, before and after injection of 95% ethanol into the tumors. Injections of ethanol were guided by ultrasound to sites of tumor vascularity, until all tumor vascularity had been obliterated. Microbubble contrast medium or saline was injected i.v. prior to each of the ultrasonic interrogations. Arteriography was performed before and after ablation. Selected tumor samples were submitted for histologic examination. Contrast enhanced tumor vascularity over saline controls in all cases. In some, incompletely ablated foci of tumor could only be identified with contrast medium enhancement. Arteriography showed complete ablation of all but 1 tumor. We conclude that ultrasound enhanced by contrast better shows the presence or absence of tumor vascularity. Ultrasound enhanced by contrast might offer an accurate means of guiding and monitoring percutaneous ethanol injection for tumor ablation.

Identification of a region of frequent loss of heterozygosity at 11q24 in colorectal cancer.

Loss of heterozygosity (LOH) at 11q23-qter occurs frequently in ovarian and other cancers, but for colorectal cancer, the evidence is conflicting. Seven polymorphic loci were analyzed between D11S897 and D11S969 in 50 colorectal tumors. Two distinct LOH regions were detected, suggesting possible sites for tumor-suppressor genes involved in colorectal neoplasia: a large centromeric region between D11S897 and D11S925, and a telomeric 4.9-Mb region between D11S912 and D11S969. There was no correlation with clinicopathological features. This analysis describes a region of LOH in the region 11q23.3-24.3 for the first time in colorectal cancer and provides complementary evidence for the ongoing effort to identify the gene(s) involved.

Identification and characterization of a homozygous deletion found in ovarian ascites by representational difference analysis.

We have performed representational difference analysis (RDA) on DNA from tumor cells and normal fibroblasts isolated from the ascites of a patient with ovarian cancer. Five of six products of the RDA were homozygously deleted from the tumor DNA. One of these products has been characterized and identifies a homozygous deletion of approximately 6.9 Mb at chromosome 9p21 in the original ovarian tumor material. This deletion encompasses CDKN2A (p16), CDKN2B (p15), and IFN-alpha. PCR analysis of other tumor cell lines using the novel STS based on the RDA product has shown it to lie between IFN-alpha and p16, and to identify the distal extent of a homozygous deletion in another ovarian cancer cell line. These data provide further evidence for a tumor suppressor locus distinct from, but mapping close to, p16 on 9p21. Cytogenetic analysis using comparative genomic hybridization (CGH) performed on the same primary tumor confirmed a loss of material from chromosome 9p. However, the CGH technique had neither the resolution nor the sensitivity to define a subregion of homozygous loss.

Cloning of mnuA, a membrane nuclease gene of Mycoplasma pulmonis, and analysis of its expression in Escherichia coli.

Membrane nucleases of mycoplasmas are believed to play important roles in growth and pathogenesis, although no clear evidence for their importance has yet been obtained. As a first step in defining the function of this unusual membrane activity, studies were undertaken to clone and analyze one of the membrane nuclease genes from Mycoplasma pulmonis. A novel screening strategy was used to identify a recombinant lambda phage expressing nuclease activity, and its cloned fragment was analyzed. Transposon mutagenesis was used to identify an open reading frame of 1,410 bp, which coded for nuclease activity in Escherichia coli. This gene coded for a 470-amino-acid polypeptide of 53,739 Da and was designated mnuA (for "membrane nuclease"). The MnuA protein contained a prolipoprotein signal peptidase II recognition sequence along with an extensive hydrophobic region near the amino terminus, suggesting that the protein may be lipid modified or that it is anchored in the membrane by this membrane-spanning region. Antisera raised against two MnuA peptide sequences identified an M. pulmonis membrane protein of approximately 42 kDa by immunoblotting, which corresponded to a trypsin-sensitive nucleolytic band of the same size. Maxicell experiments with E. coli confirmed that mnuA coded for a nuclease of unknown specificity. Hybridization studies showed that mnuA sequences are found in few Mycoplasma species, suggesting that mycoplasma membrane nucleases display significant sequence variation within the genus Mycoplasma.

Lower extremity volumetric arterial blood flow in normal subjects.

The objective of this clinical study was to establish normal values for volumetric blood flow in the leg at rest using Doppler ultrasound, and to determine what biophysical factors influence resting volumetric flow. Arterial blood flow was measured at four sites in the legs of 40 healthy subjects using an ATL Ultramark 9 HDI system. All subjects were nonhypertensive nonsmokers with ankle brachial index values greater than 1 and no history of vascular disease. The subjects, 20 of each gender, in age ranging from 20 to 64 y were examined. Blood flow was calculated from the time-averaged, intensity-weighted mean velocity Doppler waveforms and vessel cross-sectional area at the same site. Thigh and calf circumference measurements were used to estimate muscle masses. The mean flow and standard error measured in four arteries in the leg were: 284+/-21 mL/min in the common femoral (CFA); 152+/-10 mL/min in the superficial femoral (SFA); 72+/-5 mL/min in the popliteal; and 3+/-1 mL/min in the dorsalis pedis. Although women tended to have higher time-averaged mean velocities in the CFA and SFA than men (t-test, p < 0.008), their arterial cross-sectional areas tended to be smaller (t-test, p < 0.004) and no statistically significant difference was found between men and women in volumetric flow at any site. No correlation was found between age, weight, height, muscle mass and volumetric flow at all four sites. These estimates of lower extremity volumetric flow in healthy subjects provide a baseline for future studies of flow rates in patients with vascular disease.