An unusual finding of an anaplastic meningioma in NF2-related schwannomatosis.

NF2-related schwannomatosis (NF2) is a rare autosomal-dominant genetic disorder characterized by bilateral vestibular schwannomas and multiple meningiomas. This case report presents the extremely rare occurrence of an anaplastic meningioma in a 12-year-old male with previously undiagnosed NF2. The patient presented with a history of abdominal pain and episodic emesis, gait unsteadiness, right upper and lower extremity weakness, and facial weakness. He had sensorineural hearing loss and wore bilateral hearing aids. MR imaging revealed a sizable left frontoparietal, dural-based meningioma with heterogeneous enhancement with mass effect on the brain and midline shift. Multiple additional CNS lesions were noted including a homogenous lesion at the level of T5 indicative of compression of the spinal cord. The patient underwent a frontotemporoparietal craniotomy for the removal of his large dural-based meningioma, utilizing neuronavigation and transdural ultrasonography for precise en bloc resection of the mass. Histopathology revealed an anaplastic meningioma, WHO grade 3, characterized by brisk mitotic activity, small-cell changes, high Ki-67 proliferation rate, and significant loss of P16. We report an anaplastic meningioma associated with an underlying diagnosis of NF2 for which we describe clinical and histopathological features.

Candida spondylodiscitis: a systematic review and meta-analysis of seventy two studies.

OBJECTIVES: Knowledge of Candida spondylodiscitis is limited to case reports and smaller case series. Controversy remains on the most effective diagnostical and therapeutical steps once Candida is suspected. This systematic review summarized all cases of Candida spondylodiscitis reported to date concerning baseline demographics, symptoms, treatment, and prognostic factors. METHODS: A PRISMA-based search of PubMed, Web of Science, Embase, Scopus, and OVID Medline was performed from database inception to November 30, 2022. Reported cases of Candida spondylodiscitis were included regardless of Candida strain or spinal levels involved. Based on these criteria, 656 studies were analyzed and 72 included for analysis. Kaplan-Meier curves, Fisher's exact, and Wilcoxon's rank sum tests were performed. RESULTS: In total, 89 patients (67% males) treated for Candida spondylodiscitis were included. Median age was 61 years, 23% were immunocompromised, and 15% IV drug users. Median length of antifungal treatment was six months, and fluconazole (68%) most commonly used. Thirteen percent underwent debridement, 34% discectomy with and 21% without additional instrumentation. Median follow-up was 12 months. The two year survivorship free of death was 80%. The two year survivorship free of revision was 94%. Younger age (p = 0.042) and longer length of antifungal treatment (p = 0.061) were predictive of survival. CONCLUSION: Most patients affected by Candida spondylodiscitis were males in their sixties, with one in four being immunocompromised. While one in five patients died within two years of diagnosis, younger age and prolonged antifungal treatment might play a protective role.

Ten-step minimally invasive slalom unilateral laminotomy for bilateral decompression (sULBD) with navigation.

BACKGROUND: Unilateral laminotomy for bilateral decompression (ULBD) is a MIS surgical technique that offers safe and effective decompression of lumbar spinal stenosis (LSS) with a long-term resolution of symptoms. Advantages over conventional open laminectomy include reduced expected blood loss, muscle damage, mechanical instability, and less postoperative pain. The slalom technique combined with navigation is used in multi-segmental LSS to improve the workflow and effectiveness of the procedure. METHODS: We outline ten technical steps to achieve a slalom unilateral laminotomy for bilateral decompression (sULBD) with navigation. In a retrospective case series, we included patients with multi-segmental LSS operated in our institution using the sULBD between 2020 and 2022. The primary outcome was a reduction in pain measured by Visual Analogue Scale (VAS) for back pain and leg pain and Oswestry Disability Index (ODI). RESULTS: In our case series (N = 7), all patients reported resolution of initial symptoms on an average follow-up of 20.71 ± 9 months. The average operative time and length of hospital stay were 196.14 min and 1.67 days, respectively. On average, VAS (back pain) was 4.71 pre-operatively and 1.50 on long-term follow-up of an average of 19.05 months. VAS (leg pain) decreased from 4.33 to 1.21. ODI was reported as 33% pre-operatively and 12% on long-term follow-up. CONCLUSION: The sULBD with navigation is a safe and effective MIS surgical procedure and achieves the resolution of symptoms in patients presenting with multi-segmental LSS. Herein, we demonstrate the ten key steps required to perform the sULBD technique. Compared to the standard sULBD technique, the incorporation of navigation provides anatomic localization without exposure to radiation to staff for a higher safety profile along with a fast and efficient workflow.

Atheroprotective effects of dietary L-arginine increase with age in cholesterol-fed rabbits.

NO has several putative atheroprotective properties but its precursor, L-arginine, and inhibitors of its synthesis have had inconsistent effects on the extent of experimental atherosclerosis in rabbits. The location and character of experimental atherosclerosis differ between immature and mature rabbits; both phenomena have been attributed to changes with age in the NO pathway. We investigated whether the influence of dietary L-arginine on experimental atherosclerosis is also age-related. The frequency of lesions was mapped in the descending thoracic and upper abdominal aorta of immature and mature rabbits fed 1 % cholesterol, with or without supplementary L-arginine, for 8 weeks. Consistent with earlier data, the distribution of lesions around the branch points changed with age in control rabbits. The mean frequency of lesions was essentially the same at both ages. L-Arginine supplements had no effect on the distribution of lesions at either age. They significantly reduced the mean frequency of lesions in mature animals but not in immature animals. Thus, the atheroprotective effect of dietary L-arginine in cholesterol-fed rabbits increases with age.

Prospective study of breast MRI in BRCA1 and BRCA2 mutation carriers: effect of mutation status on cancer incidence.

Annual MRI screening is recommended as an adjunct to mammography for BRCA1 and BRCA2 mutation carriers. Prophylactic oophorectomy has been shown to decrease breast cancer risk in BRCA1/2 mutation carriers. Here, we aimed to examine the combined effects of MRI and oophorectomy. For this purpose, 93 BRCA1/2 mutation carriers were screened with yearly mammograms and yearly MRI scans. Study endpoints were defined as date of breast cancer diagnosis, date of prophylactic mastectomy, or date of most recent contact. Of 93 women, with a median age of 47, 80 (86%) had prophylactic oophorectomy. Fifty-one women (55%) had BRCA1 mutations. A total of 283 MRI scans were performed. Eleven breast cancers (9 invasive, 2 ductal carcinoma in situ) were detected in 93 women (12%) with a median follow-up of 3.2 years (incidence 40 per 1,000 person-years). Six cancers were first detected on MRI, three were first detected by mammogram, and two were "interval cancers." All breast cancers occurred in BRCA1 mutation carriers (incidence 67 per 1,000 person-years). Apart from BRCA1 vs. BRCA2 mutation status, there were no other significant predictors of breast cancer incidence. Most invasive breast cancers were estrogen receptor negative (7 of 9) and lymph node negative (7 of 9). There have been no systemic recurrences with a median follow-up of 19 months after cancer diagnosis. Finally, it was concluded that all breast cancers occurred in BRCA1 mutation carriers, in most cases despite oophorectomy. These data suggest that surveillance and prevention strategies may have different outcomes in BRCA1 and BRCA2 mutation carriers.

Artificial neural network-based classification to screen for dysphonia using psychoacoustic scaling of acoustic voice features.

SUMMARY: For diagnosis and classification of dysphonia, voice specialists can choose from an array of diagnostic tools like perceptual tests or acoustic voice analysis. These methods have in common that they require a high level of specialized training and experience, and therefore are mostly reserved to specialized centers. We aimed at developing an acoustic voice analysis system that could be used as a screening device to monitor, document, and diagnose voice problems that are also encountered by non-voice specialists, such as anesthesiologists, head and neck surgeons, and general surgeons before surgery of the thyroid gland and the upper thoracic aperture. An acoustical feature extraction paradigm that focused on jitter, shimmer, standard deviation of fundamental frequency, and the glottal-to-noise excitation ratio was used to reanalyse 120 voice samples previously analyzed by Schönweiler et al (A Novel Approach to Acoustical Voice Analysis Using Artificial Neural Networks. JARO. 2000:1;270-282). An improved artificial neural network (ANN) was used for classification. Building on this preliminary work, we modified the mathematical algorithm to further improve classification accuracy. Eighty percent of all voice samples could be classified correctly as either healthy or hoarse (sensitivity: 63.0%; specificity: 93.9%; area under the curve: 0.854). The adaptation of the ANN-voice analysis system for mobile use may facilitate its use and acceptance by non-voice specialists for the discovery and documentation of preexisting voice disorders, and may thereby lead to a timely initiation of further diagnosis and therapy by voice specialists.

Biomedical vocabularies--the demand for differentiation.

The need of biomedical vocabularies is well known for various tasks, e.g., supporting structured data entry, decision support and electronic data exchange as well as retrieval and statistical evaluation of the data. Due to a considerable diversity of artifacts like interface terminologies, classifications and thesauri it seems to be reasonable to demand for a massive reduction and, finally, to end up with just one unique "multi-purpose world terminology". Concept-based reference terminologies like SNOMED CT might be candidates for that idea. Unfortunately, the above mentioned kinds of vocabularies cannot be replaced because of their specific purpose-dependent nature. Their mutual distinctive characteristics are outlined and compared with inherent purpose-less concept systems that are based on formal approaches like description logics. For supporting interoperability the different kinds of purpose-dependent vocabularies can and should be improved by mappings to machine-processible reference terminologies. As a side-effect, this paper may contribute to the meta-terminology in this field of medical terminology.

Sex-specific expression of gastrin-releasing peptide receptor: relationship to smoking history and risk of lung cancer.

BACKGROUND: Activation of gastrin-releasing peptide receptor (GRPR) in human airways has been associated with a proliferative response of bronchial cells to gastrin-releasing peptide and with long-term tobacco use. The GRPR gene is located on the X chromosome and escapes X-chromosome inactivation, which occurs in females. Increasing evidence demonstrates that women are more susceptible than men to tobacco carcinogenesis. We hypothesized that the susceptibility of women to the effects of tobacco may be associated with airway expression of GRPR. METHODS: We analyzed GRPR messenger RNA (mRNA) expression in lung tissues and cultured airway cells from 78 individuals (40 males and 38 females) and in lung fibroblasts exposed to nicotine in vitro. Nicotinic acetylcholine receptors in airway cells were assayed by use of radioactively labeled nicotine and nicotine antagonists. A polymorphism in exon 2 of the GRPR gene was used to detect allele-specific GRPR mRNA expression in some individuals. Statistical tests were two-sided. RESULTS: GRPR mRNA expression was detected in airway cells and tissues of more female than male nonsmokers (55% versus 0%) and short-term smokers (1-25 pack-years [pack-years = number of packs of cigarettes smoked per day multiplied by the number of years of smoking]) (75% versus 20%) (P =.018 for nonsmoking and short-term smoking females versus nonsmoking and short-term smoking males). Female smokers exhibited expression of GRPR mRNA at a lower mean pack-year exposure than male smokers (37.4 pack-years versus 56.3 pack-years; P =.037). Lung fibroblasts and bronchial epithelial cells exhibited high-affinity, saturable nicotinic acetylcholine-binding sites. Expression of GRPR mRNA in lung fibroblasts was elevated following exposure to nicotine. CONCLUSIONS: Our results suggest that the GRPR gene is expressed more frequently in women than in men in the absence of smoking and that expression of this gene is activated earlier in women in response to tobacco exposure. The presence of two expressed copies of the GRPR gene in females may be a factor in the increased susceptibility of women to tobacco-induced lung cancer.

Detection of human lung epithelial cell growth factors produced by a lung carcinoma cell line: use in culture of primary solid lung tumors.

Serum-free medium conditioned for 72 h by a human bronchioloalveolar carcinoma of lung, A549-1, stimulated the colony formation of normal human bronchial epithelial cells, newly cultured cells from human solid lung tumors, and established human lung tumor cell lines, including A549-1 cells themselves. This activity was concentration dependent and was stable to acid. Growth factors in A549-1 conditioned medium (CM) supported culture of solid lung tumors; primary cell cultures were obtained from nine of 10 solid lung tumors of non-small cell origin and from one small cell tumor using A549-1 CM. In addition, three cell lines have been established to date from these primary cultures. Gel filtration of concentrated A549-1 CM on Biogel P-10 separated the growth promoting activity into four regions of apparent Mr 70,000, 12,000, 8,000, and 6,000, and two broad regions of apparent Mr 3000-5000. All but the 12,000 Mr fraction contained activity which competed for specific binding of epidermal growth factor (EGF) to A431 cell membranes. CM was superior to both EGF and TGF alpha in stimulating growth of normal and neoplastic lung cells. EGF also was inhibitory to tumor cells while TGF alpha stimulated both normal and tumor cell growth. TGF beta was also found in CM but inhibited normal and neoplastic lung epithelial cell growth. Of other substances tested, ILGF-I stimulated colony formation. The results suggest that autocrine factors may be important in non-small cell lung tumor cell growth and that differences in response to EGF and TGF alpha may provide the basis for selective culturing of normal and neoplastic lung epithelial cells.

Chromosome abnormalities in human non-small cell lung cancer.

Clonal cytogenetic abnormalities found in 30 non-small cell lung carcinomas (NSCLC), including 28 newly diagnosed primary tumor specimens, are summarized. Multiple chromosome alterations were identified in every case, and 19 of 30 tumors had near-triploid or near-tetraploid karyotypes. Polysomy 7 and partial gains of 7p, including 7p11-p13 (site of the EGFR gene), were particularly frequent, occurring alone or in combination in 26 tumors. Recurrent losses involving 1p, 3p, 6q, 9p, 11p, 15p, and 17p (where the TP53 gene is located) were each seen in 16-25 cases. Five tumors exhibited double minutes, which were associated with amplified MYC1 (1 case) and EGFR (1 case), as determined by Southern analysis. The cytogenetic data were compiled from either short term cultures of tumor tissue harvested within 1-9 days (18 cases) or later harvests performed on long term cultures or cell lines (6 cases); in the other 6 cases results were obtained from both short term and long term cultures. Two studies were performed to validate the use of long term culture for cytogenetic analysis of solid lung tumors. First, in order to determine whether cytogenetic results from cultures are representative of the original tumor, the modal chromosome number of 13 specimens placed into culture was compared to the DNA index of the original tumor tissue, as measured by flow cytometry. The DNA indices of the solid tumor biopsies agreed with the degree of aneuploidy observed by cytogenetic analysis in every case. Second, in 6 cases we performed direct comparisons of karyotypes obtained from cells cultured by both methods. Identical chromosome abnormalities were detected in short term cultures and later harvests of the same specimen. Overall, our findings indicate that tumorigenesis in NSCLC is characterized by the accumulation of multiple chromosome alterations. Furthermore, these data demonstrate that recurrent cytogenetic changes can be identified in NSCLC and that detailed karyotypes from long term cultures are relevant to the original tumor. Chromosome abnormalities detected by these techniques may have clinical and biological significance. However, the complex pattern of karyotypic changes seen in newly diagnosed NSCLC emphasizes the need for future investigations of premalignant bronchial lesions in order to identify primary genetic changes important for early detection and intervention in this aggressive neoplasm.

Response of primary human lung carcinomas to autocrine growth factors produced by a lung carcinoma cell line.

Medium conditioned for 48 to 72 h by A549-1 lung carcinoma cells was used to culture primary solid lung tumors on feeder layers of inactivated Swiss 3T3 cells. Of 22 cases placed into culture, primary cultures of carcinoma cells were obtained in 20. Subcultures were obtained in 18 cases, and cell lines were established in nine cases. The neoplastic origin of the cultured cells was demonstrated by several criteria: tumorigenicity in athymic mice; anchorage-independent growth; expression of altered lactate dehydrogenase isoenzyme profiles; and expression of the lung tumor marker pregnancy-specific glycoprotein 1. The epithelial nature of cultured carcinoma cells was demonstrated by expression of keratin. These characteristics were compared to normal epithelial cells established in culture from bronchial explants from the same donors as tumor tissue, or other donors. The growth-stimulating effect of conditioned medium toward primary or newly cultured tumor cells was quantitated by clonal assays in soft agar and in monolayer culture. Growth response in clonal assays of newly cultured carcinoma cells to the purified growth factors transforming growth factor alpha and insulin-like growth factor 1, two known components of medium conditioned by A549-1 cells, was also demonstrated.

Effect of phorbol esters on clonal cultures of human, hamster, and rat respiratory epithelial cells.

The effect of the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) on the growth of epithelial cells from rat, hamster, and human respiratory tract has been measured by monitoring colony formation and cross-linked envelope formation in culture. TPA and its active derivatives stimulated colony formation of rat tracheal epithelial cells but did not stimulate cross-linked envelope formation. Tracheal epithelial cells from the hamster and human bronchial epithelial cells were inhibited from forming colonies by these agents. This inhibitory effect was also dependent on concentration. In the rat, the stimulation of cells to enter cell division induced by TPA decayed with time after removal of primary cells from the trachea, while in hamster and human cells, the inhibitory effect of TPA was independent of time. Although TPA inhibited colony formation in hamster and human cells, it did not elicit the same responses with respect to cross-linked envelopes. Hamster tracheal epithelial cells did not form cross-linked envelopes in response to TPA, whereas human bronchial cells did. A comparison was made of the response to TPA in cells from the human bronchi of 24 individuals; the extent of inhibition of colony formation induced by TPA varied by 130-fold. These results show that normal cells from these species vary in biological response to tumor promoters, implying that selective induction of terminal differentiation in normal cells may not be a universal mechanism of action of tumor promoters.

Alterations of human endometrial stromal cells produced by N-methyl-N'-nitro-N-nitrosoguanidine.

Stromal cell cultures obtained from human endometrium were treated repetitively with N-methyl-N'-nitro-N-nitrosoguanidine in vitro at concentrations ranging from 0.5 to 4.0 micrograms/ml, and alterations in growth potential and morphology were analyzed. A single exposure to the carcinogen resulted in morphological evidence of toxicity and reductions in growth rates, plating efficiency, and saturation density as compared to solvent-treated control cells. Cytotoxicity was reduced after additional exposures to the carcinogen. Following repetitive treatments with N-methyl-N'-nitro-N-nitrosoguanidine, human endometrial stromal cells developed enhanced growth potential, the capacity to form macroscopic colonies in soft agar, and elevated gamma-glutamyltranspeptidase activity. Carcinogen-treated cells displayed atypical morphology characterized by irregularities in cell and nuclear size and shape, large bizarre nucleoli, increased nuclear:cytoplasmic ratios, and cellular crowding. Control cells did not display altered morphology or growth parameters even following multiple exposures to solvent and repetitive subculturing. These alterations in growth potential and morphology suggest that the cells are progressing towards preneoplastic and perhaps neoplastic transformation in vitro.

Effects of anthracyclines on oxygenated and hypoxic tumor cells.

The cytotoxic effects of anthracyclines and other chemotherapeutic agents were examined in normally aerated and hypoxic Sarcoma 180 and EMT6 tumor cells in vitro. Adriamycin, daunomycin, and mitomycin C were selectively toxic to hypoxic Sarcoma 180 cells. The augmented sensitivity was not the result of an increase in susceptibility of oxygen-deprived cells toward antitumor agents in general. 1,3-Bis(2-chloroethyl)-1-nitrosourea, for example, exhibited equal cytotoxicity toward normally aerated and hypoxic cells, while streptonigrin was selectively toxic toward normally aerated cells. The cellular levels of [3H]daunomycin in both Sarcoma 180 and EMT6 cells were not different under the two conditions of oxygenation, and no greater production of either the alcohol or aglycone metabolites of daunomycin occurred in hypoxic cells, compared with their normally aerated counterparts. In addition, analysis of cellular pellets for residual drug remaining after exhaustive extraction showed no significant difference between normally aerated and hypoxic cells. The effects of reoxygenation of hypoxic cells on their sensitivity to mitomycin C and to Adriamycin were studied in both Sarcoma 180 and EMT6 cells. The enhanced efficacy of mitomycin C as a cytotoxic agent observed under hypoxia was reversed after a 2-hr reoxygenation. In contrast, the augmented toxicity of Adriamycin toward hypoxic cells was not reversible in either cell line after 2 or 4 hr of reoxygenation. The results suggest that neither the formation of a reactive oxygen species nor direct involvement of an alkylating agent generated by drug metabolism is an obligatory step in the cytotoxic action of these anthracyclines.

Metabolism of benzo(a)pyrene in monolayer cultures of human bronchial epithelial cells from a series of donors.

Benzo(a)pyrene [B(a)P] metabolism was measured in monolayer cultures of human bronchial epithelial cells derived from 18 specimens of explanted tissue. Bronchial epithelial cells converted B(a)P to dihydrodiols, phenols, quinone derivatives, and polyhydroxylated forms. Sulfate and glucuronide conjugates of B(a)P metabolites were also detected. Both total metabolism and distribution of metabolites showed a 10-fold or greater variation in cultures from different specimens. When the data were divided according to smoking status, however, no differences in total metabolism, extent of conjugation, or distribution of metabolites could be demonstrated between the two groups. Wide variation (over 1000-fold) in the cytotoxicity of B(a)P towards cells derived from different specimens was demonstrated but could not be directly correlated to the extent of metabolic activation. The results suggest that human bronchial epithelial cells which are newly grown from explanted tissue of smokers in culture do not demonstrate enzymatic induction. Variation among individuals observed in these studies probably represents basal differences in metabolic capability.

Correlation of model chromosome number of cultured non-small cell lung carcinomas with DNA index of solid tumor tissue.

The modal chromosome number of 13 non-small cell lung carcinomas placed into culture was compared to the DNA index of the tumor tissue as measured by flow cytometry in order to determine whether cytogenetic results from such cultures are representative of the original solid tumor. The modal chromosome number observed in culture, which ranged from 45-146, fell within the range of aneuploidy predicted from the DNA content of the original tissue in all 13 cases. In 7 cases, flow cytometry results showed that the aneuploid G1/G0 population of the tumor tissue (DNA index of 1.5 or higher) represented 11-76% of the cells present, while diploid cells (presumably normal tissue) made up the remainder of the population. In these 7 cases, modal chromosome numbers of 61-92 were found in tumor cells cultured from the tissue. In 3 cases, only a diploid or near-diploid population was found by flow cytometry, consistent with the near-diploid modal chromosome number of cultured cells observed (45-55). In 3 cases, the aneuploid G1/G0 population (DNA index of 1.5, 1.6, and 3.2) of the original tissue represented only a small fraction of the solid tumor (1-5% of cells). Modal chromosome number found in cells cultured from these 3 cases was 64-69, 62-68, and 136-146, which is in close agreement with the aneuploid peak observed in the tissue. Histological analysis of the tumor tissue in two of the latter cases showed large numbers of infiltrating lymphocytes and/or stromal tissue which could have dominated the measurement by flow cytometry. In the third case, tumor cells made up at least 75% of the specimen examined, implying that part of the population in the "diploid" peak contained tumor cells in this specimen. Only the aneuploid population was detected in culture of this tumor. Agreement between flow cytometry and cytogenetics was found in cases in which metaphase spreads were obtained within a few days of culture as well as after several months. These results indicate that highly aneuploid populations are found in many, but not all, non-small cell lung tumors. Although in some cases multiple populations may exist in the tumor which do not all proliferate in vitro, tumor cells which are found in culture of solid lung carcinomas are representative of the original tumor. Flow cytometry findings in the solid tumors confirmed the findings of aneuploidy observed by cytogenetic analysis.

Genetic instability of microsatellite sequences in many non-small cell lung carcinomas.

We have analyzed DNA obtained from 38 lung tumors and normal lung or blood DNA for microsatellite instability. Instability was examined at 10 different microsatellite loci on chromosome 3p, as well as loci on 3q, 11p, 11q, and 13q, and two on Xq. We observed microsatellite instability at one or more loci in 13 of the lung tumors analyzed, and this instability ranged from tumors showing instability in only a single microsatellite to two adenocarcinomas that had alterations in all 16 tested microsatellites. Microsatellite instability could therefore play a significant role in the development of a sizable portion of lung tumors.

Comparative genomic hybridization analysis detects frequent, often high-level, overrepresentation of DNA sequences at 3q, 5p, 7p, and 8q in human non-small cell lung carcinomas.

Comparative genomic hybridization analysis was used to identify chromosomal imbalances in 20 non-small cell lung carcinoma (NSCLC) biopsies and cell lines. The chromosome arms most often overrepresented were 3q (85%), 5p (70%), 7p (65%), and 8q (65%), which were observed at high copy numbers in many cases. Other common overrepresented sites were 1q, 2p, and 20p. DNA sequence amplification was often observed, with the most frequent site being 3q26 (six cases). Other recurrent sites of amplification included 8q24, 3q13, 3q28-qter, 7q11.2, 8p11-12, 12p12, and 19q13.1-13.2. The most frequent underrepresented segment was 3p21 (50%); other recurrent sites of autosomal loss included 8p21-pter, 15q11.2-13, 5q11.2-15, 9p, 13q12-14, 17p, and 18q21-qter. These regions of copy number decreases are also common sites of allelic loss, further implicating these sites as locations of tumor suppressor genes. Although some of the overrepresented segments harbor known or suspected oncogenes/growth-regulatory genes, we have identified 3q and 5p as new sites that are very frequently overrepresented in NSCLC. These findings could represent entry points for the identification of novel amplified DNA sequences that may contribute to the development or progression of NSCLC.

Recurrent deletions of specific chromosomal sites in 1p, 3p, 6q, and 9p in human malignant mesothelioma.

Detailed cytogenetic analyses were carried out on primary tumor specimens and cell lines from 23 patients with pleural malignant mesothelioma (MM). Clonal abnormalities were identified in 20 of 23 MM. In 3 cases, karyotypic data were compiled from harvests of both short-term cultures (1-3 days), and primary cultures grown on murine feeder layers for several weeks. The karyotypes obtained with these 2 different culture methods were very similar, although polyploid versions of abnormal clones were found only in the long-term cultures. In addition, while short-term cultures from 9 tumor biopsies usually exhibited near-diploid clones, cell lines derived from 11 tumors tended to have higher ploidies. Each of the cytogenetically abnormal MM displayed multiple clonal alterations. The 2 most frequent changes were chromosomal losses of specific regions in 1p (17 cases) and 9p (16 cases). The shortest regions of overlap of these losses were at 1p21-p22 and 9p21-p22, respectively. Other common abnormalities included losses of 3p21 (13 cases) and 6q15-q21 (9 cases), and numerical losses of chromosomes 14, 16, 18, and 22 (each observed in 10-13 tumors). In many of the MM examined, most or all of these recurrent changes occurred in combination, suggesting the involvement of a pathogenetic cascade in this cancer. The pattern of recurrent chromosomal losses suggests that these regions represent the locations of tumor suppressor genes whose loss/inactivation may have a pivotal role in MM tumorigenesis.

Association of immunoreactive hepatocyte growth factor with poor survival in resectable non-small cell lung cancer.

We have shown previously that hepatocyte growth factor (HGF), which is produced by lung fibroblasts, is a potent mitogen and motogen for both normal and neoplastic bronchial epithelium, and that expression of the HGF receptor, the c-met proto-oncogene protein, is uniformly found in the human bronchial epithelium and in non-small cell lung carcinomas (NSCLCs; P. Singh-Kaw et al., Am. J. Physiol., 268: L1012-L1020, 1995). Yamashita et al. have reported an association of HGF with poor survival in invasive ductal carcinoma of the breast (Cancer Res., 54: 1630-1633, 1994). There are few prognostic markers for lung cancer, and the high recurrence rate for stage I lung cancer suggests the frequent presence of undetectable tumor burden in such patients. Criteria are needed to evaluate these patients for risk of recurrence. We have now evaluated whether HGF present in resectable lung tumors has prognostic significance. In this study, 56 primary NSCLCs, mainly adenocarcinomas, were examined for presence of HGF by quantitative Western blot. These tumors consisted of tissue from 34 stage I patients, 9 stage II patients, and 13 stage IIIa patients who underwent curative resection for primary NSCLC. Extracts of whole tumor tissue were analyzed after separation of proteins by electrophoresis and transfer of proteins to nitrocellulose membranes. Immunoreactive (ir)-HGF was visualized by reaction with a polyclonal anti-HGF antiserum and quantitated by densitometry. Lung tumor content of ir-HGF varied widely among individuals. Median ir-HGF content in tumor extracts was 15.3 ng/40 microg of tumor protein; mean ir-HGF was 27.2 ng/40 microg of tumor protein. The median and mean ir-HGF were both significantly higher in tumor tissue from patients who suffered a recurrence during the follow-up period compared with those with no evidence or residual disease; this was true of all patients (P = 0.0001) and stage I patients analyzed separately (P = 0.002). Analysis of survival curves indicated that ir-HGF levels higher than the median were associated with poor overall survival (P < 0.03). Univariate analysis showed three factors related to poor overall survival in this set of patients: ir-HGF, tumor (T) status (a measure of primary tumor size and extent), and age. Nodal (N) status and stage were only marginally related to overall survival, most likely because the majority of the patients in the study were stage I. N status, stage, and T status were related to disease-free survival, however. Multivariate Cox analysis showed that ir-HGF, T status, and age independently had a negative impact on overall survival. ir-HGF was a strong independent negative prognostic indicator (P = 0.0001) with a relative risk of 1.022 per unit of ir-HGF (ng/40 microg of protein). This demonstrates that, in this group of patients, the relative risk of ir-HGF content increased continuously as ir-HGF increased, and exceeded 10 at units of ir-HGF of 100 or more. In comparison, in this group of patients, the relative risk of a T status greater than 1 was 4.75 and that of age greater than 65 was 3.95. The combined negative effect of a T status greater than 1 and elevated ir-HGF on survival was also highly pronounced (P < 0.005). In addition, elevated ir-HGF had a negative impact on survival when patients were stratified by stage or N status. Stage I patients with high ir-HGF values had a worse outcome than stage II or stage IIIa patients with low ir-HGF values. Elevated ir-HGF was strongly associated with poor outcome for resectable NSCLC patients as a group, and also identified stage I patients with poor outcome, indicating that it could be a useful indicator of risk of relapse and death in patients who have early lung cancer. The impact of elevated ir-HGF was especially prominent in patients whose T status was greater than 1, suggesting that patients with both risk factors who are stag