Safety assessment of Asterarcys quadricellulare, a microalga, with applications in poultry and livestock feed.

Asterarcys quadricellulare (AQ) is a microalgal species with potential applications in improving the quality of animal feed, and safety studies on this species are lacking. Therefore, this study presents safety data on an industrially cultivated strain of AQ tested using the following Organisation for Economic Co-operation and Development (OECD) guidelines: acute skin irritation in rabbits; skin sensitisation in guinea pigs; acute eye irritation in rabbits; acute oral fixed-dose procedure in rats; and bacterial reverse mutation using the B.N. Ames technique. Results showed that AQ is non-irritant and non-sensitising to skin. AQ caused transient conjunctival lacrimation and redness; however, the scores for these clinical signs translated into low ocular irritation indices and classification of AQ as non-irritant to the eyes. An acute oral dose of AQ (2000 mg/kg) did not cause mortality, change in body weight gain, or any general, functional, and neurobehavioral clinical signs. In five strains of Salmonella typhimurium bacteria, treatment with AQ did not cause biologically or statistically significant changes in the number of revertant colonies, indicating that AQ does not cause mutagenic toxicity. This study demonstrates the safety of a heterotrophically-produced strain of AQ and supports its use as a safe and non-toxic feed ingredient.

Toxicological evaluation of a glycan preparation from an enzymatic hydrolysis of Saccharomyces cerevisiae.

The aim of this paper was to provide a comprehensive toxicological and safety evaluation of a yeast cell wall preparation (YCWP) for use as an animal feed ingredient. The following toxicological assessments were carried out: the mutagenic activity was tested using the Ames' Test in five Salmonella typhimurium strains; clastogenic activity was investigated using the mammalian micronucleus test in Swiss ICO OF1 (IOPS Caw) mice; genotoxic activity was assessed using the in vitro mammalian chromosomal aberration test in human lymphocytes; acute oral toxicity was tested by administration of a single dose of 2000 mg/kg BW. Eye and skin irritation were assessed in rabbits according to OECD guidelines; skin sensitivity was established in guinea pigs by means of the Buehler test, while acute dermal and inhalation studies in rats were further completed, also according to OECD guidelines. All conducted tests were considered valid under the experimental conditions. No significant mutagenic activity or genotoxic activity was observed, and it was concluded that the test article did not induce any clastogenic effect. YCWP was found to be mildly irritating to the eye, slightly irritating to the skin but was found to be non-sensitizing in the guinea pig. The acute oral, dermal and inhalation studies did not yield any evidence of gross toxicity or pharmacological effects.

Heterotrophic Aurantiochytrium sp. supplementation to layer diets sustainably increases the omega-3 concentration of eggs.

1. The consumption of adequate amounts of the long-chain polyunsaturated omega-3 fatty acids (n-3 LC-PUFA) has been associated with beneficial effects on human health. Eggs are commonly consumed worldwide, and their omega-3 content can be easily altered by changing the diets of laying hens and so represent an important target for enrichment. 2. In this study, the effect of supplementing laying hens with DHA-rich, Aurantiochytrium limacinum at three different inclusion levels was investigated over a 24-week period. 3. Significant increases in egg DHA concentrations were observed after four weeks and were maintained for the duration of the 24-week study. The supplemented eggs in the current study had a DHA content of 82, 101, and 129 mg/yolk when supplemented with 0.25%, 0.5% and 1% treatments, respectively, which meets the EU criteria to be considered 'high in omega-3'. 4. Using the sustainably grown protist Aurantiochytrium limacinum to supplement layer diets increased the egg DHA concentration and decreased the n-6/n-3 ratio, improving the nutritional value of the eggs for human consumers.

Redressing the balance: including DHA-rich Aurantiochytrium limacinum in broiler diets increases tissue omega-3 fatty acid content and lowers the n-6:n-3 ratio.

1. The consumption of sufficient quantities of long chain omega-3 fatty acids (n-3 LCPUFA) from meat and other animal products can lead to a variety of health benefits in humans. The fatty acid content of poultry meat can be increased by feeding birds ingredients that are rich in n-3 LCFUFA 2. The effect of feeding a docosahexaenoic acid (DHA) rich Aurantiochytrium limacinum biomass (AURA) on the fatty acid content of breast and thigh tissues was investigated in a feeding trial with 2880 male Ross 308 broilers. The broiler diets were supplemented with either 0, 0.25, 0.5 or 1% AURA from day 21 to 42 of age. 3. Supplementation significantly increased the DHA content of both breast and thigh meat at an inclusion rate of 1% in the diet, leading to a total of 42 and 46 mg DHA/100 g of fresh breast or thigh tissue respectively. Significant increases in the tissue eicosapentaenoic acid (EPA) concentration were seen alongside a reduced omega-6/omega-3 ratio, improving the nutritional value of the meat for consumers and identifying supplementation of broiler diets with A. limacinum as an effective and sustainable method to increase n-3 LCPUFA consumption in the human population.

The effect of dietary supplementation with Aurantiochytrium limacinum on lactating dairy cows in terms of animal health, productivity and milk composition.

The aim of this study was to investigate the effect of dietary supplementation with the docosahexaenoic acid (DHA)-rich microalgae, Aurantiochytrium limacinum (AURA), on a variety of health and productivity parameters in lactating cows. Twenty-four cows were blocked by parity and number of days in milk and then randomly assigned to a control (CON; n = 12) group with no algal supplementation, or a treatment group (AURA; n = 12) provided with 100 g AURA cow-1  day-1 or 16 g DHA cow-1  day-1 . A variety of health and productivity measurements were taken, and results indicated that supplementation had no negative effects on animal health in terms of somatic cell count, haematological and biochemical blood parameters, while body condition was marginally improved by algal supplementation. No differences were found for the various production parameters measured; however, a tendency towards increased milk production was observed for the AURA group during the final stage of the study (+4.5 kg cow-1  day-1 , day 78-84). The fatty acid profile of milk was improved by supplementation, with significantly lower saturated fatty acids, significantly higher omega-3 fatty acids and an improved omega-3/omega-6 ratio observed when compared to the control group. The amount of DHA in the milk of cows provided 105 g AURA head-1  day-1 was 4.7 mg/100 g milk with a peak transfer efficiency from feed to milk at day 49 of 8.3%. These results indicate that supplementation with 105 g AURA head-1  day-1 resulted in the successful enrichment of milk with DHA without negatively impacting the health or productivity of the animals.

Effects of a DHA-rich unextracted microalgae as a dietary supplement on performance, carcass traits and meat fatty acid profile in growing-finishing pigs.

Two 125-day experiments of the same design were conducted to evaluate the effects of a heterotrophically grown microalgae (AURA) containing docosahexaenoic acid (DHA) on pig performance, carcass traits and the fatty acid composition of lean and adipose tissue. In each experiment, 144 Hypor pigs were blocked by sex, allocated to three treatment groups, and fed 0, 0.25% or 0.50% AURA in isonutritive, isocaloric diets. Pigs were weighed on days 0, 28, 56, 84 and 112. Feed and water intakes were measured every 28 days. Pigs were slaughtered on day 125. Data from the two studies were analysed as a single data set. Performance and carcass traits did not differ between treatments. Both microalgae treatment levels enriched (p < .05) Longissimus lumborum (LL) and backfat in DHA and improved (p < .05) their ratios of n-6 to n-3 fatty acids.

Dietary safety of a dual-enzyme preparation for animal feed: Acute and subchronic oral toxicity and genotoxicity studies.

Animal feed is routinely supplemented with exogenous enzymes to improve nutrient utilization, such as proteases to enhance protein hydrolysis in vivo and xylanases to alleviate feed related anti-nutritional factors. The present studies were conducted to evaluate the potential oral toxicity and genotoxicity of a dual-enzyme preparation, Vegpro® concentrate (VPr-C). Acute oral toxicity studies were conducted in adult male and female Sprague-Dawley Crl CD rats and CHS Swiss ICO:OFI (IOPS Caw) mice. Thirteen week preliminary and final subchronic oral toxicity studies were conducted in male and female rats. Genotoxicity was evaluated through a bacterial reverse mutation test (Ames test), an in-vitro mammalian chromosomal aberration test, and a mammalian micronucleus test. The LD50 was >2000 mg/kg of BW in mice and rats. In the 13-week oral toxicity study, the No Observed Adverse Effects Level (NOAEL) was 1000 mg/kg BW per day for females and 300 mg/kg BW per day for males. VPr-C showed no mutagenic activity in Salmonella typhimurium, did not induce significant chromosomal aberrations in cultured human lymphocytes, and did not increase the frequency or proportion of micronucleated immature erythrocytes in mice. There was no evidence of acute or subchronic toxicity or genotoxicity associated with the test article at these test dosages.

Performance and environmental impact of egg production in response to dietary supplementation of mannan oligosaccharide in laying hens: a meta-analysis.

A meta-analysis was conducted to examine the effect of supplementing mannan oligosaccharide (MOS; Bio-Mos, Alltech Inc., Nicholasville, KY) in the diets of laying hens on the performance and environmental impact of egg production. Data on production performance (feed intake, hen-day production [HDP], feed conversion ratio [FCR], and mortality) and egg quality attributes (egg weight, egg mass, and eggshell thickness) were extracted from 18 studies to build a database of comparisons between nonsupplemented diets (control) and diets supplemented with MOS. A total of 4,664 laying hens were involved in the comparisons and the average MOS dosage and age of hens were 0.97 kg/ton and 44 wk, respectively. The dataset was analyzed using the random-effects model to estimate the effect size of MOS supplementation on production performance and egg quality attributes. The impact of feeding MOS on the carbon footprint (feed and total emission intensities) of egg production was evaluated by using the meta-analysis results of production performance to develop a scenario simulation that was analyzed by a life cycle assessment (LCA) model. Overall pooled effect size (raw mean difference) indicated that MOS supplementation did not affect feed intake. In contrast, HDP increased by +1.76% and, FCR and mortality reduced by -26.64 g feed/kg egg and -2.39%, respectively. Dietary MOS did not influence egg weight while egg mass increased (P < 0.01) by +0.95 g/day/hen and eggshell thickness tended to increase (P = 0.07) by +0.05 mm. Subgroup analysis indicated that dietary MOS exhibited consistent improvement on HDP and FCR under several study factors (age of hens, number of hens, production challenges, MOS dosage, and study duration). Additionally, the simulated LCA revealed that supplementing MOS decreased feed and total emission intensities of egg production by -1.3 and -1.5%, respectively. Overall, dietary supplementation of MOS at 1.0 kg/ton improved the production performance of laying hens and reduced the carbon footprint and, therefore, can enhance the sustainability credentials of egg production.

Technical Innovation for Visual Assessment of Preterm Newborns in a Neonatal Intensive Care Unit: Exploratory Study.

OBJECTIVE: The present study is aimed at assessing heart rate variability (HRV) and its correlation with visual acuity (VA) assessment of preterm newborns (PTNB) in neonatal intensive care units. METHOD: Cross-sectional study analyzing HRV during assessment of VA with the aid of a Polar RS800CX heart rate monitor (Polar Electro Oy, Finland). HRV was analyzed according to time and frequency domains and the chaos domain used the autocorrelation coefficient and entropy. The sample consisted of hospitalized PTNB, and static analysis included simple regression diagnosis. RESULTS: A total of 14 PTNB were included in the sample. VA varied between 0.23 and 1.60 cpd, and only five PTNB obtained below-expected values for age. Statistical analysis demonstrated a negative correlation between VA and time domain (SDDN and SD2) and a positive correlation between frequency domain (heart rate and hertz), but in simple linear regression analysis, these variables did not influence VA. CONCLUSION: The results of the study demonstrate that visual acuity was inversely correlated with SDNN and SD2 and during stimulation, showing that the higher the visual performance, the lower the autonomic modulation response.

Toxicological evaluation of an unextracted Aurantiochytrium limacinum biomass, a novel docosahexaenoic acid rich feed ingredient.

The purpose of this paper was to evaluate the toxicological potential of a heterotrophically grown unextracted Aurantiochytrium limacinum biomass (AURA) when used as a food additive. The following toxicological assessments were conducted on this novel docosahexaenoic acid rich feed ingredient: Mutagenic activity was tested by means of the Ames' test using five Salmonella typhimurium strains; clastogenic activity was investigated using the micronucleus test in male and female Sprague Dawley rats; genotoxic activity was assessed by means of the in vitro metaphase analysis tests in human lymphocytes; oral toxicity was tested by administration of AURA at various concentrations; eye and skin irritation was assessed in rabbits according to OECD guidelines; skin sensitivity was established in guinea pigs by means of the Buehler test. All conducted tests were considered valid under the experimental conditions. No significant mutagenic activity or clastogenic activity was observed. Genotoxic activity in human lymphocytes was not induced. Oral administration of 276 mg AURA/kg bw1 and 2000 mg AURA/kg bw resulted in no mortality or signs of acute toxicity. Daily administration of 1000 mg AURA/kg bw caused no mortality or biologically relevant signs of toxicity and was established as the No Observable Adverse Effect Level. AURA was also found to be a non-irritant for the eye and skin of the rabbit and was non-sensitizing to guinea pig skin.

Primary peritoneal mesotheliomas in children: a clinicopathological and immunohistochemical study of eight cases.

AIMS: To present eight cases of primary diffuse peritoneal malignant mesothelioma in children <15 years old, with a discussion of the pitfalls of this diagnosis in the paediatric age group. METHODS AND RESULTS: The cases were selected based on the following criteria: (i) primary peritoneal neoplasms confined grossly or radiographically to the abdominal cavity; (ii) negative history of previous or another associated malignancy; (iii) histopathological confirmation. All patients (five female, three male) presented clinically with symptoms of abdominal pain, distention and ascites. Grossly, the tumours showed multiple, diffuse peritoneal nodules. Histologically, seven cases corresponded to epithelioid mesotheliomas and one case displayed biphasic (epithelioid and spindle) cellular proliferation. Immunohistochemical studies for cytokeratin (CK) 5/6, calretinin and low-molecular-weight CK (CAM5.2) showed strong cytoplasmic positivity in the neoplastic cells. Three patients were treated by chemotherapy. On clinical follow-up, four patients with epithelioid mesotheliomas were alive and well from 12 to 18 months after initial diagnosis; one patient with a mixed (biphasic epithelioid/sarcomatoid) mesothelioma died of tumour 24 months after diagnosis. CONCLUSIONS: Peritoneal malignant mesothelioma in children is a rare condition that can introduce difficulties in histopathological diagnosis.

Antigen binding properties of highly purified bispecific antibodies.

A panel of three bispecific monoclonal antibodies (bsMAbs) binding to follitropin (FSH) and to beta-galactosidase have been prepared by fusion of hybridoma cell lines resistant to oubain and neomycin. One of these bispecific antibodies contains heavy chains of the same IgG subclass, and two are composed of heavy chains of different IgG subclasses. We have investigated methods for the purification of bispecific antibodies from hybrid hybridoma supernatant fluids grown in serum-free medium. Following ammonium sulfate precipitation, bispecific antibodies can be purified in a single step by mixed mode ion-exchange HPLC on Bakerbond Abx columns. In one case, three species were resolved by ion-exchange HPLC and functional analysis showed that two peaks contained parental antibodies, and the third contained the bispecific. Ion-exchange HPLC purification of serum-free preparations from two other hybrid hybridomas resolved seven protein-containing peaks, only one of which was active in a bispecific ELISA. The equilibrium affinity constants for each of the parental antibodies for both FSH and beta-galactosidase were determined and found to be similar to those of the purified bsMAbs. Further, the association of FSH to one binding site on a bispecific antibody was shown to have no effect on the equilibrium binding constant for beta-galactosidase binding to the other site. Our results suggest that bsMAbs can be readily purified from hybrid hybridomas by a simple and rapid method, and the binding of antigen to one binding site on a bsMAb is independent of antigen binding to the second site.

Primary thymic epithelial neoplasms showing combined features of thymoma and thymic carcinoma. A clinicopathologic study of 22 cases.

Thymic epithelial neoplasms are unusual tumors that may span the gamut from clinically benign, well-differentiated lesions (encapsulated thymoma) to overtly malignant, poorly differentiated neoplasms (thymic carcinoma). It has been commonly believed that lesions displaying obvious cytologic features of malignancy (i.e., thymic carcinoma) represent a unique and separate group that is histogenetically distinct from thymoma. We have studied 22 cases of thymic epithelial neoplasms characterized by the admixture of areas displaying conventional features of thymoma with areas showing features of thymic carcinoma. The tumors occurred in six women and 16 men whose ages ranged from 23 to 83 years (median, 53). The lesions presented in eight patients with symptoms of chest discomfort resulting from the involvement of surrounding structures; in 14 patients, they were asymptomatic and discovered incidentally on routine chest radiographs. Histologically, most tumors showed a combination of conventional thymomatous elements with well-differentiated squamous-cell carcinoma (10 cases), followed by thymoma and poorly-differentiated squamous carcinoma (seven cases) and spindle-cell thymoma with poorly-differentiated squamous carcinoma (five cases). Areas of transition between the two different components could be identified in most cases. In five cases, areas showing the features of clear-cell carcinoma could be seen either arising from squamous carcinomatous elements or within the thymomatous component, and in one case transitions between lymphoepithelioma-like carcinoma and anaplastic carcinoma could be observed. Two patients had a history of myasthenia gravis with biopsy-proven thymomas in whom the tumors had been monitored without treatment for 10 and 14 years before the sudden enlargement of the mass. The resected specimens in both patients showed the emergence of a carcinoma arising from a thymoma. The present cases appear to support the existence of a continuum in the spectrum of differentiation between thymoma and thymic carcinoma, suggesting a close histogenetic relationship between these two conditions. Such findings are important not only for our understanding of these tumors but may also play a significant role in the assessment of the biologic behavior and management of these lesions.

Mucoepidermoid carcinomas of the thymus. A clinicopathologic study of six cases.

Six cases of mucoepidermoid carcinoma of the thymus are presented. The patients were two men and four women aged 17 to 66 years (median age, 34.5). Clinically, three patients had symptoms of chest discomfort and three were asymptomatic. Grossly, the tumors in three patients were described as cystic structures varying in size from 7 to 8 cm in greatest dimension, with focal areas of induration within the walls of the cyst that averaged from 1.5 to 3 cm. The other three cases had grossly and radiographically well-circumscribed, homogeneous tumor masses. Histologically, the lesions showed a spectrum of features that ranged from those of well-differentiated, to moderately well-differentiated, to poorly differentiated mucoepidermoid carcinoma, with sheets and solid islands of squamoid cells admixed with mucin-secreting epithelium lining gland-like spaces. In four cases, the tumor was histologically seen in continuity with the epithelial lining of multilocular cystic structures; the nonneoplastic components of the cysts contained abundant inflammation and showed the features of otherwise conventional acquired multilocular thymic cysts. Clinical follow-up showed that the two patients with intermediate and high-grade tumors died within 2 and 7 months after initial diagnosis. One of these patients showed at autopsy residual tumor limited to the mediastinum, whereas the other patient died with metastases to pericardium and myocardium despite postoperative radiation therapy. Two patients with low-grade tumors were alive and well with no residual disease 2 and 3 years after surgery, and the other two were lost to follow-up. Mucoepidermoid carcinoma of the thymus should be included in the differential diagnosis of cystic neoplasms of the thymus. As with their counterparts at other sites, the biologic behavior of these tumors closely correlates with their degree of differentiation and amount of cytologic atypia.

Mediastinal seminomas with prominent cystic changes. A clinicopathologic study of 10 cases.

We present 10 cases of thymic seminomas associated with prominent cystic changes. All patients were males, aged 16 to 79 years (median, 23.5). Clinically, two patients presented with chest pain/four were asymptomatic and the tumors were discovered on routine chest radiographs; one tumor was discovered incidentally at autopsy; and in three patients no clinical information was obtained. Grossly, the tumors were described as multilocular cystic lesions that ranged in size from 7 to 19 cm in greatest dimension, showing small focal areas of induration within the cyst walls. Histologically, the lesions were characterized by cystic spaces lined by squamous or cuboidal epithelium showing severe chronic inflammatory changes with areas of cholesterol cleft granulomas, lymphoid follicular hyperplasia, and scattered foci of residual thymic parenchyma within the walls of the cysts, resulting in a picture indistinguishable from acquired multilocular thymic cysts. Careful examination, however, revealed microscopic foci composed of a neoplastic proliferation of large polygonal cells with slightly eosinophilic to clear cytoplasm and large nuclei with prominent nucleoli. The atypical cells were admixed with an inflammatory background and were often accompanied by a florid granulomatous reaction. Periodic acid-Schiff histochemical reaction with diastase revealed moderate amounts of glycogen within the cytoplasm of the tumor cells. Immunohistochemical studies in five cases showed positive labeling of the tumor cells with placental alkaline phosphatase. Nine patients were treated by complete surgical excision of the mass, and additional postoperative radiation therapy was given to two patients. Follow-up information available for five patients showed all to be alive and well from 2 to 19 years after diagnosis (mean follow-up, 9 years). Four of the patients were lost to follow-up. The pathogenesis of the cystic process in these cases remained unsettled but may represent a reactive change secondary to epithelial hyperplasia of thymic epithelium. Thymic seminoma should be considered in the differential diagnosis of cystic lesions of the anterior mediastinum; extensive sampling of such lesions is therefore recommended for proper evaluation.

Pleomorphic large cell lymphomas of the mediastinum.

Nine cases of primary non-lymphoblastic, non-Hodgkin's large cell lymphomas of the mediastinum characterized by a highly pleomorphic histologic appearance are described. The patients, four women and five men, were aged 30 to 65 years. All patients presented with symptoms referable to their tumors, including cough, chest pain, dyspnea, pleural effusion, and superior vena cava syndrome. Clinical and pathologic staging in all patients showed that the bulk of the tumor was confined to the chest cavity at the time of initial diagnosis, with local infiltration into the neck, lung hilum, and surrounding mediastinal structures. Three different histological growth patterns were observed: one composed of a diffuse proliferation of pleomorphic, highly atypical cells with bizarre nuclear features that closely resembled a high grade sarcoma; another one composed of sheets of large, epithelial-appearing atypical cells suggestive of anaplastic carcinoma; and another pattern characterized by a pleomorphic proliferation of large lymphoid cells admixed with numerous scattered Reed-Sternberg-like cells reminiscent of the lymphocyte-depleted variant of Hodgkin's disease. Immunohistochemical studies on paraffin-embedded tissue sections in all cases showed positive staining of the tumor cells with CD20 and CD45 antibodies and negative staining with a large panel of markers, including broad-spectrum keratin, CAM 5.2, carcinoembryonic antigen, epithelial membrane antigen, vimentin, actin, desmin, HMB 45, S-100 protein, CD3, CD15, CD30, and CD45RO. Because of their location restricted to the anterior mediastinum, frequent lack of recognizable lymph node architecture, and bizarre cytologic features, the present group of lesions posed difficulties for diagnosis, their correct identification was achieved through the application of a panel of immunohistochemical markers. An awareness of these unusual histologic appearances of primary large cell lymphoma in the mediastinum and inclusion of a broad panel of lymphoid markers are therefore recommended for the evaluation of pleomorphic, undifferentiated malignant neoplasms of this anatomic region.

Primary mediastinal choriocarcinomas: a clinicopathologic and immunohistochemical study of eight cases.

Primary choriocarcinoma of the anterior mediastinum is by far the rarest and most controversial form of extragonadal germ cell tumor. A clinicopathologic study of eight primary mediastinal neoplasms bearing the histopathologic and immunohistochemical features of choriocarcinoma is presented. The patients were all men between the ages of 21 and 63 years (mean, 42 years). Clinical symptoms included shortness of breath, chest pain, cough, and superior vena cava syndrome; one patient also had gynecomastia. All patients presented with large anterior mediastinal masses on chest radiographs that measured an average of 10 cm in greatest diameter. Grossly, the tumors were described as large, soft, extensively hemorrhagic, and with foci of necrosis. Histologically, they were characterized by a dual cell population composed of cytotrophoblastic cells with uniform, round nuclei, clear cytoplasm, and prominent nucleoli admixed with large, multinucleated syncytiotrophoblastic cells with bizarre nuclei, prominent nucleoli, and abundant eosinophilic cytoplasm. Immunohistochemically, the tumors were notable for strong keratin and beta-human chorionic gonadotropin (HCG) positivity. Seven patients presented at the time of diagnosis with thoracic and extrathoracic (liver, adrenal, kidney, and spleen) metastases. In one case, the tumor was entirely confined to the mediastinum. All patients died over a period of 1 to 2 months. Complete autopsies were performed in all cases; none of the patients showed evidence of a testicular tumor or scar after thorough examination of the testes on serial sectioning. The present cases demonstrate the widespread distribution of germ cells in the human body and lend further support to the existence of primary extragonadal choriocarcinoma arising in the thymic region.

Yolk sac tumors of the mediastinum with prominent spindle cell features: a clinicopathologic study of three cases.

Three cases of primary mediastinal yolk sac tumors with prominent spindle cell features are presented. The patients were three men 24-34 years of age (mean 29). Clinically, two patients presented with symptoms of chest pain and cough; no clinical information was provided for the third patient. Grossly, the tumors were described as large mediastinal masses, with a hemorrhagic and necrotic cut surface. Histologically, the tumors were characterized by a predominantly atypical spindle cell proliferation admixed with areas that showed focally the characteristic reticular growth pattern of yolk sac tumors, with the presence of Schiller-Duval bodies and intra- and extracellular hyaline globules. Immunohistochemical studies performed in one case showed positive staining for keratin and alpha-fetoprotein in both the spindle cell and reticular components of the tumor. Follow-up information was obtained in two patients; they both died of tumor with metastases to the lungs 1 year after initial diagnosis. The present cases expand the spectrum of histopathologic growth patterns that may be observed in yolk sac tumors of the mediastinum and stress the issue of careful sampling and evaluation of mediastinal neoplasms for arriving at the correct diagnosis.

Primary melanoma of the lung: a clinicopathologic and immunohistochemical study of eight cases.

Primary malignant melanoma of the lung (PMML) is an uncommon neoplasm that may be confused with more conventional types of lung cancer. Although the previously proposed criteria for diagnosis, including the presence of an in situ component, are often difficult to satisfy, this lesion is characterized by a poor prognosis, ultimately leading to patient death. We report eight cases of PMML that presented as solitary, central endobronchial neoplasms, resulting in a picture that closely resembled carcinoid tumor or poorly differentiated non-small-cell carcinoma of the lung. The mean age at diagnosis was 51 years (range 45-71). The patients included one woman and seven men. The histologic growth pattern varied from organoid to fascicular and included epithelioid to spindled cells with hyperchromatic to vesicular nuclei, prominent eosinophilic nucleoli, and abundant eosinophilic to clear cytoplasm with occasional intranuclear cytoplasmic inclusions. A bronchial in situ component was present in four cases. Initial interpretations included carcinoid tumor, non-small-cell carcinoma, and malignant melanoma. Melanin was present in all neoplasms on hematoxylin and eosin staining, although very focally in one case, and was Fontana-Masson positive in all cases. Immunohistochemically, diffuse strong positivity for S-100, HMB-45, and vimentin was present in all seven tumors tested. All seven tumors were negative for cytokeratin, CAM 5.2, and chromogranin. Ultrastructural examination of the eighth case showed dysmorphic premelanosomes but no neurosecretory granules. None of the patients had disseminated disease at presentation, and all patients underwent surgical resection (seven lobectomies and one excision). In this series, primary malignant melanoma of the lung was characterized by an aggressive postoperative course, with five patients dying of metastatic disease from 4 to 32 months after resection (median 14 months). Two patients are alive with metastatic disease at 4 and 30 months after surgery, and the eighth patient is alive with no evidence of disease 108 months after surgery at last follow-up. Metastatic melanoma was identified in various sites, including the lungs, adrenal glands, liver, mesentery, brain, and bone. The cases herein presented indicate that PMML should be included in the differential diagnosis of primary bronchial tumors.

Hepatoid yolk sac tumors of the mediastinum: a clinicopathologic and immunohistochemical study of four cases.

Four cases of primary hepatoid yolk sac tumors of the anterior mediastinum are described. The patients were all men between the ages of 26 and 40 years (median 33). Clinically, they all presented with a history of shortness of breath and chest pain of several weeks' duration. None of the patients had a history of germ cell tumor elsewhere or evidence of any hepatic abnormality. Grossly, all the tumors were described as large mediastinal masses that impinged on adjacent structures. Histologically, they were characterized by sheets of medium-sized, round to polygonal neoplastic cells with moderate amounts of eosinophilic cytoplasm and round to oval nuclei with prominent nucleoli. The cellular proliferation was homogeneous and displayed moderate cellular atypia and scattered mitotic activity. All the tumors showed focally the presence of more conventional areas of yolk sac tumor, with islands of tumor cells showing a reticular pattern of growth admixed with scattered intra- and extracellular hyaline globules and occasional Schiller-Duval bodies. Immunohistochemical studies showed strong positivity of the tumor cells for alpha-fetoprotein in both components of the lesions. Follow-up information was available in three patients, all of whom developed lung metastases within a year after initial diagnosis. Two of these patients died of tumor within the same period, whereas a third patient has been lost to follow-up. The present cases illustrate an unusual histologic pattern of yolk sac tumor in the mediastinum and highlight the importance of considering this tumor in the differential diagnosis of lesions showing a hepatoid pattern of growth in the mediastinal area.