Genomic origin and EGFR-TKI treatments of pulmonary adenosquamous carcinoma.

BACKGROUND: Adenosquamous carcinoma (ASC) of the lung is a heterogeneous disease that is composed of both adenocarcinoma components (ACC) and squamous cell carcinoma components (SCCC). Their genomic profile, genetic origin, and clinical management remain controversial. PATIENTS AND METHODS: Resected ASC and metastatic tumor in regional lymph nodes (LNs) were collected. The ACC and SCCC were separated by microdissection of primary tumor. The 1021 cancer-related genes were evaluated by next-generation sequencing independently in ACC and SCCC and LNs. Shared and private alterations in the two components were investigated. In addition, genomic profiles of independent cohorts of adenocarcinomas and squamous cell carcinomas were examined for comparison. We have also carried out a retrospective study of ASCs with known EGFR mutation status from 11 hospitals in China for their clinical outcomes. RESULTS: The most frequent alterations in 28 surgically resected ASCs include EGFR (79%), TP53 (68%), MAP3K1 (14%) mutations, EGFR amplifications (32%), and MDM2 amplifications (18%). Twenty-seven patients (96%) had shared variations between ACC and SCCC, and pure SCCC metastases were not found in metastatic LNs among these patients. Only one patient with geographically separated ACC and SCCC had no shared mutations. Inter-component heterogeneity was a common genetic event of ACC and SCCC. The genomic profile of ASC was similar to that of 170 adenocarcinomas, but different from that of 62 squamous cell carcinomas. The incidence of EGFR mutations in the retrospective analysis of 517 ASCs was 51.8%. Among the 129 EGFR-positive patients who received EGFR-TKIs, the objective response rate was 56.6% and the median progression-free survival was 10.1 months (95% confidence interval: 9.0-11.2). CONCLUSIONS: The ACC and SCCC share a monoclonal origin, a majority with genetically inter-component heterogeneity. ASC may represent a subtype of adenocarcinoma with EGFR mutation being the most common genomic anomaly and sharing similar efficacy to EGFR TKI.

Immune focusing and enhanced neutralization induced by HIV-1 gp140 chemical cross-linking.

Experimental vaccine antigens based upon the HIV-1 envelope glycoproteins (Env) have failed to induce neutralizing antibodies (NAbs) against the majority of circulating viral strains as a result of antibody evasion mechanisms, including amino acid variability and conformational instability. A potential vaccine design strategy is to stabilize Env, thereby focusing antibody responses on constitutively exposed, conserved surfaces, such as the CD4 binding site (CD4bs). Here, we show that a largely trimeric form of soluble Env can be stably cross-linked with glutaraldehyde (GLA) without global modification of antigenicity. Cross-linking largely conserved binding of all potent broadly neutralizing antibodies (bNAbs) tested, including CD4bs-specific VRC01 and HJ16, but reduced binding of several non- or weakly neutralizing antibodies and soluble CD4 (sCD4). Adjuvanted administration of cross-linked or unmodified gp140 to rabbits generated indistinguishable total gp140-specific serum IgG binding titers. However, sera from animals receiving cross-linked gp140 showed significantly increased CD4bs-specific antibody binding compared to animals receiving unmodified gp140. Moreover, peptide mapping of sera from animals receiving cross-linked gp140 revealed increased binding to gp120 C1 and V1V2 regions. Finally, neutralization titers were significantly elevated in sera from animals receiving cross-linked gp140 rather than unmodified gp140. We conclude that cross-linking favors antigen stability, imparts antigenic modifications that selectively refocus antibody specificity and improves induction of NAbs, and might be a useful strategy for future vaccine design.

Müllerian adenosarcoma: a review of cases and literature.

OBJECTIVE: Mullerian adenosarcoma usually originates in the endometrium and grows as a polypoid mass in post-menopausal women presenting as abnormal vaginal bleeding. This report reviewed Miillerian adenosarcoma cases to clarify the clinical and pathologic characteristics. MATERIALS AND METHODS: Fifteen cases ofMiillerian adenosarcoma in two medical centers covering a 15-year period were reviewed. Their clinical characteristics, pathologic findings, treatment, and outcomes were compared. RESULTS: Of the 15 cases, three originated from the endometrium, six arose from uterine adenomyosis, three from the adnexa, and three from the cervix. There was only one post-menopausal case. One case was of breast cancer with tamoxifen (TMX) therapy. There were four Miillerian adenosarcoma with sarcomatous overgrowth (MASO) cases, three of which died within one year after surgery. Only the focal MASO case survived. CONCLUSION: The rare variant of MASO is very aggressive and associated with poor prognosis.

Drug-targeting strategies in cancer therapy.

Genetic changes in cell-cycle, apoptotic, and survival pathways cause tumorigenesis, leading to significant phenotypic changes in transformed cells. These changes in the tumor environment - elevated expression of surface proteases, increased angiogenesis and glucuronidase activity - can be taken advantage of to improve the therapeutic index of existing cancer therapies. Targeting cytotoxics to tumor cells by enzymatic activation is a promising strategy for improving chemotherapeutics.

Protein domains governing interactions between E2F, the retinoblastoma gene product, and human papillomavirus type 16 E7 protein.

Human papillomaviruses (HPVs) are the etiological agents for genital warts and contribute to the development of cervical cancer in humans. The HPV E7 gene product is expressed in these diseases, and the E7 genes from HPV types 16 and 18 contribute to transformation in mammalian cells. Mutation and deletion analysis of this gene suggests that the transforming activity of the protein product resides in the same domain as that which is directly involved in complex formation with the retinoblastoma gene product (pRB). This domain is one of two conserved regions (designated CRI and CRII) shared by E7 and other viral oncoproteins which bind pRB, including adenovirus E1A protein. Binding of HPV type 16 E7 protein to pRB has previously been shown to affect pRB's ability to bind DNA and to form complexes with other cellular proteins. In the current study, we map the functional interaction between E7 protein and pRB by monitoring the association between a 60-kDa version of the pRB, pRB60, and the cellular transcription factor E2F. We observe that CRII of E7 (amino acids 20 to 29), which completely blocks binding of full-length E7 protein, is necessary but not sufficient to inhibit E2F/pRB60 complex formation. While CRI of E1A (amino acids 37 to 55) appears to be sufficient to compete with E2F for binding to pRB60, the equivalent region of E7 is neither necessary nor sufficient. Only E7 fragments that contained both CRII and at least a portion of the zinc-binding domain (amino acids 60 to 98) inhibited E2F/pRB60 complex formation. These results suggest that pRB60 associates with E7 and E2F through overlapping but distinct domains.

Reactive atmospheric pressure plasma for highly efficient removal of structure-directing agents from zeolite thin films.

We present a novel approach to remove the structure-directing agent (SDA) from as-synthesized zeolites using an atmospheric pressure plasma jet (APPJ). This reduces the time required to less than 60 seconds as compared to the existing thermal calcination, whose durations range from hours to days. The highly reactive plasma also results in a pronounced Q(3)-to-Q(4) transformation in the pure-silica zeolite MFI.

An SH3 domain is required for the mitogenic activity of microinjected phospholipase C-gamma 1.

Phospholipase activity is elevated in dividing cells. In response to growth factor stimulation, phospholipase C-gamma (PLC-gamma) binds to activated tyrosine kinase receptors via SH2 binding domains, resulting in phosphorylation of PLC-gamma and activation of its enzyme activity. These observations suggest that PLC-gamma participates in the signal transduction pathway employed by growth factors to promote mitogenesis. Consistent with this hypothesis, microinjection of purified bovine PLC-gamma into quiescent fibroblasts has been previously reported to initiate a mitogenic response [Smith et al. (1989) Proc. Natl. Acad. Sci. 86, 3659]. We have reproduced this result using recombinant rat PLC-gamma protein. Surprisingly, however, a catalytically inactive mutant of PLC-gamma, H335Q, also elicited a full mitogenic response. The capacity to induce mitogenesis by microinjection of PLC-gamma was mapped to the 'Z' domain of the protein, which contains PLC-gamma's SH2 and SH3 motifs. Inactivation of the phosphorylated tyrosine binding properties of both SH2 domains had no effect on the mitogenic activity of the Z-domain peptide. However, deletion of the SH3 domain resulted in a complete loss of activity. These results suggest that PLC-gamma's mitogenic properties do not require the enzyme's phospholipase activity, but are instead mediated by a novel pathway for mitogenic stimulation which is dependent upon an intact SH3 domain.

Malignant carcinoid tumor metastatic to the dura mater simulating a meningioma.

A 50-year-old man, transferred from another hospital, was admitted because of adult onset seizures. Nine months earlier, he had undergone an esophagogastrectomy; the lesion was confirmed to be a carcinoid tumor. Laboratory tests, chest x-rays, and electrocardiogram were normal. A second liver and spleen scan was performed. A computed tomographic scan revealed a well-circumscribed homogeneous enhancement of a lesion in the left frontal superficial area. On the 10th day, the patient underwent a left frontal parietal craniotomy. Postoperatively, he manifested no residual neurological deficits and was discharged on the 6th postoperative day. A week later, he was readmitted for treatment of aphasia and right hemiparesis; he was treated and discharged. The patient survived 16 more months. The occurrence of central nervous system metastasis from carcinoid tumor is rare. This tumor resembled, in many respects, a parasagittal meningioma. Radiological findings on the computed tomographic scan were typical of these tumors. This patient was diagnosed as having metastatic disease just 9 months after the diagnosis of the primary tumor and 13 months from the onset of any symptoms. This is a short period of time compared with that reported in other cases.

Klippel-Feil syndrome in association with a craniocervical dermoid cyst presenting as aseptic meningitis in an adult: case report.

Intracranial tumors associated with Klippel-Feil syndrome usually occur in children, with spinal tumors being more common in adults affected by the syndrome. A rare case of a dermoid cyst at the craniocervical junction presenting as aseptic meningitis in an adult with Klippel-Feil syndrome is described. A review of the literature on tumors associated with this syndrome is also presented.

Automatic gait recognition via statistical approaches for extended template features.

A gait recognition system using extended template features is presented. A proposed statistical approach is applied for feature extraction from spatial and temporal templates. This method can be used to reduce data dimensionality and to optimize the class separability of different gait sequences simultaneously. Dimensionality reduction is achieved by template extraction followed by principal component analysis. Gait recognition is achieved in the canonical space using a measure of accumulated distance as the metric. By incorporating spatial and temporal information into an extended feature, gait recognition becomes more robust and accurate than using spatial or temporal features alone.

[Immunohistochemical and cytochemical analysis of lymphocytes and plasma cells in nasopharyngeal carcinoma].

Cytoplasmic IgA+, IgG+ and IgM+ in plasma cells, present in biopsy tissue of 68 patients with nasopharyngeal carcinoma (NPC) and 40 patients with chronic nasopharyngitis (CN), were studied by immunoperoxidase (PAP) technique. EB virus VCA-IgA serum antibody in all these patients was determined. At the same time, the activity of T lymphocytes of 41 patients (23 with NPC and 18 with CN) was investigated by alpha-naphthyl acetate esterase (ANAE) method. The number of T lymphocytes in NPC was far less than that in CN. This suggests that the impediment or deficiency in cellular immunity may promote the development and growth of tumor. The number of IgA+ plasma cells in NPC was obviously more than that in CN. As the increase in the level of VCA-IgA serum antibody in NPC patients corresponded to the increase in the number of IgA+ plasma cells in the tumor tissue, it was presumed that part of the IgA+ plasma cells might participate in the production and introduction of VCA-IgA antibody. We suggest that the examination of VCA-IgA serum antibody be a reliable screening test for NPC. No significant difference was found in the numbers of IgG+ plasma cells between NPC and CN. IgM+ plasma cells were rare in both.

Blood alcohol concentration among injured drivers in Hualien County.

Vehicle-related injuries are the major cause of death and injuries in Hualien County. Driving under the influence of alcohol plays a major role in such crashes. From December 1997 to May 1998, we determined the blood alcohol concentrations of 750 injured drivers from vehicle crashes, visiting the two emergency rooms of teaching hospitals in Hualien. The objectives of this study were to investigate the incidence of alcohol used in vehicle crashes, to identify the prevalence groups for prevention and to discuss alcohol testing at emergency services. Sixty-four percent were male; 27.5% were aborigines. The mean age was 36.5 +/- 16.5 years. About 54.1% tested positive for blood alcohol concentration (BAC), which ranged from 13 to 611 mg/dL; 38.6% had BAC levels exceeding 50 mg/dL and 21.1% exceeding 200 mg/dL. The mean BAC was 85.9 mg/dL (+/- 118.5). Middle-aged males and aborigines were more likely to drive under the influence of alcohol. We recommend blood alcohol testing to be mandatory at the emergency service and to be used as evidence for prosecution in a court of law. Preventing drunk driving through community programs is imperative, especially in the aboriginal communities.

[The establishment of genomic DNA libraries for the human malaria parasite Plasmodium falciparum].

The DNA of Plasmodium falciparum has been purified and fragmented with restriction endonuclease BamHI. The fragments have been incorporated in vitro into derivatives of bacteriophage lambda EMBL4 digested with BamHI and Sal I. The recombinant mixture has been ligated and packaged in vitro. The recombinant phages have been identified in E. coli L95 host cell and the libraries have been established in which most of the parasite DNA is represented. The ligation proportion of vector to insert is 3:1. The recombinant phages of 4 x 10(5) have been obtained. By plaque hybridization, we have been able to recover from these libraries specific clones containing repetitive DNA sequences.

Regulated expression of neuronal SIRT1 and related genes by aging and neuronal ß2-containing nicotinic cholinergic receptors.

Longevity genes attenuate the aging process, but their expression in the brain during aging remains unknown. Loss of the majority of heteromeric brain nicotinic acetylcholine receptors (nAChRs) results in premature brain aging, and altered regulation of longevity genes could be involved. Using in situ hybridization, the expression of SIRT1, Ku70, Nampt, p53, forkhead Box O3 (FoxO3), and mitochondria uncoupling protein 5 (UCP5) was determined in neocortex and hippocampus of young adult 3-month and middle-aged 18-month-old wild-type (WT), and age-matched mice lacking ß2* heteromeric nAChRs (ß2-/-). Age-related structural changes were detected in WT mice. In particular, cortical thickness was decreased but neuronal density increased, and hippocampal volume increased with age. In contrast, young ß2-/- mice exhibited increased cortical neuronal density, and with age, cortical thickness decreased more dramatically, and hippocampal volume did not increase. Thus, young ß2-/- mice exhibited cortical signs of aging, and aging was accelerated at 18 months. The longevity genes probed exhibited similar expression patterns in frontal brain structures, with strong expression in hippocampus, medial habenula (MHb), and cortex. In WT mice, age significantly decreased expression of all genes except SIRT1 in cortical structures, and a similar pattern was detected in the MHb. Genotype had no effect on expression in young adults in either cortex or MHb, but increased mRNA expression of SIRT1, Nampt, and Ku70 was detected in cortex, hippocampus, and MHb of aged ß2-/- mice compared with WT mice. This is the first study to determine age-related expression of survival genes in forebrain areas. Although, structural changes indicative of accelerated aging are evident in young ß2-/- mice, the data suggest that nAChRs do not directly regulate expression of survival genes. However, loss of ß2* nAChRs could result in augmented cellular stress, which indirectly increases expression of SIRT1, Nampt, and Ku70 as an adaptive response to provide protection against neurodegeneration.

Use of thin films for high-sensitivity angle measurement.

We propose to use thin films to provide a drastic improvement of measurement sensitivity in the recently developed small-angle measurement method, namely, angle measurement based on the internal-reflection effect. By designing the thin films (single layer or multiple layers) so that they provide an antireflection effect in the vicinity of the critical angle, we show that the sensitivity of angle measurement can be increased exponentially with the increase of the number of thin-film layers. This method provides a new means of designing angle sensors with increased sensitivities without having to increase the number of reflections and therefore the physical size and the required fabrication accuracy of the reflection prisms. We describe the design of the thin films for this particular application and the analysis of measurement sensitivity and range as determined by the material and the number of layers of the thin films. Selection of the optimal initial angle for high linearity performance is also discussed.