bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood culture bottles for transportation and recovery of anaerobic bacteria from non‐blood samples
Authors: Melhus Åsa , Tjernberg Ingela
Published: October 2003

Using bacterial suspensions as simulated non‐blood specimens, the capacity of three different BacT/Alert blood culture bottles for the transportation and recovery of anaerobic bacteria with different sensitivity to air was evaluated. To better assess the performance of the BacT/Alert bottles, three other liquid media specially designed for anaerobes were included in the study. Attention was paid to recovery rates in relation to species, initial bacterial concentration, and time needed for detection. Of the BacT/Alert blood culture bottles, the anaerobic FAN bottle yielded the highest recovery rates, but its performance was limited compared with chopped meat broth in tubes. This broth allowed detection of all the tested species within 48 h. Since collection and transportation of anaerobic bacteria are of major importance for a reliable culture result, improvements are necessary.
The role of cytokeratins 20 and 7 and estrogen receptor analysis in separation of metastatic lobular carcinoma of the breast and metastatic signet ring cell carcinoma of the gastrointestinal tract
Authors: Tot Tibor
Published: October 2003

Metastatic signet ring cell carcinomas of unknown primary site can represent a clinical problem. Gastrointestinal signet ring cell carcinomas and invasive lobular carcinomas of the breast are the most common sources of these metastases. Immunohistochemical algorithms have been successfully used in the search for the unknown primary adenocarcinomas. In the present study a series of primary invasive lobular breast carcinomas (79 cases) and their metastases and a series of gastrointestinal signet ring cell carcinomas (22 primary and 13 metastases) were stained with monoclonal antibodies for cytokeratin (CK) 20 and CK7 and for estrogen receptors (ER). The staining was evaluated as negative (no staining), focally (less than 10% of the tumor cells stained) or diffusely positive. All the primary and metastatic gastrointestinal signet ring cell carcinomas proved to be CK20 positive, while only 2/79 (3%) of the primary and 1/21 metastatic lobulr carcinomas (5%) stained positively for this CK. None of the gastrointestinal carcinomas and the majority of the lobular carcinomas expressed ER. The majority of the tumors were CK7+. Using CK20 alone, 33 of 34 metastases could be properly classified as gastrointestinal (CK20+) or mammary (CK20‐). ER identified 31/34 of breast cancer metastases. By combining the results of CK20 and ER staining all the metastases could be properly classified as the CK20+/ER‐ pattern identified all the gastrointestinal tumors.
Teratocarcinosarcoma of the nasal cavity Report of a case showing favorable prognosis
Authors: Fukuoka K. , Hirokawa M. , Shimizu M. , Shirabe T. , Manabe T. , Hirai M. , Akisada T.
Published: October 2003

We report a very rare case of teratocarcinosarcoma of the nasal cavity showing a favorable prognosis. The patient was a 66‐year‐old man with a mass completely obstructing the right nasal cavity. Subsequently, extirpation of the mass and Denker‐Watsuji operation were performed, and the patient was treated with a combination of radiation therapy and chemotherapy. Neither recurrence nor distant metastasis was observed during follow‐up lasting 30 months. Histologic examination of the resected mass revealed several tissue elements including columnar and squamous epithelia with atypia, smooth muscle cells with rare mitotic activity, and neuroectodermal tissue. The glandular epithelium and smooth muscle cells were reminiscent of a primitive intestinal organoid structure, suggestive of tera‐tomatous tumorigenesis. Our case and a review of the literature indicate that the absence of invasiveness to the stroma or surrounding tissue is closely related to a favorable prognosis.
Changed expression of leukocyte adhesion molecules and increased production of reactive oxygen species caused by Streptococcus pyogenes in human whole blood
Authors: Saetre TORUNN , HÖIby E. ARNE , KÄHler HANNE , Lyberg TORSTEIN
Published: October 2003

To elucidate the innate immune responses to group A streptococci (GAS) important in the pathophysiology of sepsis, flow cytometric techniques were applied to study the effects of live and heat‐inactivated GAS, including their particulate and soluble components, on the expression of leukocyte adhesion molecules CDllb (Mac‐1) and CD62L (L‐selectin), and leukocyte production of reactive oxygen species (ROS) in human whole blood. GAS caused marked time‐ and concentration‐dependent increases in CD11b and ROS, while CD62L was downregulated. Live and heat‐inactivated GAS induced similar changes in leukocyte adhesion molecules, whereas ROS production induced by heat‐inactivated GAS (and its particulate fraction) was 4 (2.5)‐fold higher than with live GAS. Leukocyte nitric oxide production (24 h) was not enhanced. Although GAS proved a more potent inducer of ROS production, leukocyte responses to GAS were similar to those reported for lipolysaccharides, indicating that Gram‐positive and Gram‐negative bacteria activate common pathways in the inflammatory response. High ROS production may contribute to tissue damage caused by GAS.
Human exposure to endocrine disrupters: Standardisation of a marker of estrogenic exposure in adipose tissueNote
Authors: Rivas Ana , Fernandez Mariana F. , Cerrillo Isabel , Ibarluzea Jesús , Olea‐Serrano M. Fátima , Pedraza Vicente , Olea Nicolás
Published: October 2003

In many epidemiological studies based on the direct measurement of exposure to organochlorines, the chemicals of concern are determined directly from adipose tissue samples. Although the measurement of all possible organochlorines, their metabolites, isomers and congeners may be desirable, it is expensive and time‐consuming and many chemicals with hormonal activity may not yet have been identified. Testing systems are therefore required to screen for estrogenicity and to identify appropriate biomarkers of human exposure. To address this issue, we developed and standardised a method to assess the total estrogenic xenobiotic burden in human adipose tissue. The method extracts and separates the more lipophilic xenoestrogens from ovarian estrogens, with a subsequent bioassay determination of the cumulative effect of the xenoestrogens. It was applied to 400 women, using 200 mg of adipose tissue: 65% of samples showed measurable estrogenicity in the fraction where most non‐polar xenoestrogens eluted, and 76% of fractions where ovarian estrogens eluted were positive for estrogenicity. Residues of 16 organochlorine pesticides were determined. No correlation was found between pesticide content and estrogenicity of the samples. The high percentage of positive samples suggests that the method is sensitive enough to be used as a biomarker of human exposure to estrogenic xenobiotics and can be applied in epidemiological studies.
Identification of enterococcal isolates by temperature gradient gel electrophoresis and partial sequence analysis of PCR‐amplified 16S rDNA variable V6 regionsNote
Authors: Monstein Hans‐Jürg , Ahrné Siv , Molin Göran , Nikpour‐Badr Shohreh , Jonasson Jon
Published: October 2003

Based on partial sequence analysis of PCR‐amplified 16S rDNA variable V6 regions of 14 enterococcal type strains, Enterococcus faecalis, Enterococcus mundtii, Enterococcus gallinarum, Enterococcus avium, Enterococcus raffinosus and Enterococcus saccharolyticus showed characteristic sequence motifs which made it possible to separate them into six individual species lines. Furthermore, two species cluster groups could be identified, including (i) Enterococcus faecium, Enterococcus durans, Enterococcus hirae, Enterococcus malodoratus, and (ii) Enterococcus casseliflavus/Enterococcus flavescens, Enterococcus pseudoavium, Enterococcus dispar and Enterococcus sulfureus. There were identical DNA sequences in the V6 region within each group. Temporal temperature gradient gel electrophoresis (TTGE) of the PCR products from 16 type strains, 12 enterococcal reference strains and 8 clinical isolates revealed that a single nucleotide divergence in DNA sequences was sufficient for separation, identification and division of the studied enterococcal strains into corresponding TTGE profiles. It was concluded that partial DNA sequence analysis and TTGE profiling of PCR‐amplified 16S rDNA variable V6 regions provide useful tools for the identification of clinically important Enterococcus spp.
Performance characteristics and clinical utility of a hybrid ELISA for detection of ANANote
Authors: Ulvestad Elling
Published: October 2003

To investigate the clinical utility of a newly developed hybrid ELISA for antinuclear antibodies (ANA), a cross‐sectional study of patients admitted to the Section of Rheumatology was initiated. The ELISA was compared to indirect immunofluorescence (IIF) on HEp‐2 cells. Accuracy of tests was analyzed using receiver‐operating characteristic methodology (ROC). In addition, diagnostic sensitivity, specificity and predictive values were calculated for each assay. Results from the ROC analysis showed a slightly superior accuracy for IIF as compared to ELISA. Furthermore, IIF showed higher diagnostic sensitivity and positive predictive value for all combinations of patients and reference populations. This was due to enhanced detection by IIF, in contrast to ELISA, of diagnostically useful antibodies. IIF detected 87.4% and ELISA detected 84.2% of sera with antibodies against extractable nuclear antigens (ENA). In addition, IIF detected diagnostically important antibodies that are not included among the anti‐ENA. The hybrid ELISA either lacks or does not contain the relevant antigens in sufficient amount. Inclusion of these antigens may further enhance the performance characteristics of the ELISA.
Inverse relation between mRNA synthesis and secretion of parathyroid hormone in athymic mice grafted with human parathyroid tissueNote
Authors: Velin Åsa K. , Ander Stefan , Johansson Kenth J. M. , Trulsson Lena M. , Smeds Staffan
Published: October 2003

Parathyroid hormone (PTH) mRNA in original and transplanted human adenomatous parathyroid tissue and human serum intact PTH (S‐iPTH) was measured in athymic mice at 4, 7, 14, and 28 days after transplantation. Parathyroid tissue was obtained during surgery for hyperparathyroidism and implanted subcutaneously. PTH mRNA detection was done with RT‐PCR followed by membrane blot and hybridisation and S‐iPTH was analysed using a human specific immunoradiometric method. At 4 days, PTH mRNA was 79.6±5.3% (mean±SE) of that in original tissue whereas S‐iPTH was only 5.4 ng/l. At 28 days, PTH mRNA was significantly reduced to 60.7±4.1% whereas S‐iPTH was increased to 192 ng/l. The reduced PTH mRNA expression in the transplants at 28 days may be explained by an inhibited DNA transcription. The presence of human S‐iPTH in transplanted mice at 4 days may be due to cell disintegration and diffusion. The gradual increase in S‐iPTH during the experimental period probably reflects increased transplant cell volume and improved graft revascularisation.
Semen quality and male reproductive health: the controversy about human sperm concentration declineNote
Authors: Jouannet Pierre , Wang Christina , Eustache Florence , Kold‐Jensen Tina , Auger Jacques
Published: October 2003

Concern about the effect of environmental changes on male reproductive health has grown in recent years to become a major preoccupation in some developed countries. A possible decline in human sperm concentration was suggested in the early seventies following studies in the US. In 1992 a meta‐analysis of 61 articles published by Carlsen et al. concluded that the mean sperm count of healthy men had declined by 1% per year over the previous 50 years. From 1995 and onwards, some retrospective, longitudinal analyses of the sperm count of fertile or infertile men contradicted this while others did not. The demonstration of a geographical variation in sperm concentration, between and within countries or regions, appears to be less controversial. The amplitude of the difference observed cannot only be explained by methodological or confounding factors, and must to some extent be attributed to ethnic, genetic or environmental factors. As many of the published studies suffer from imprecision regarding the description of population characteristics and confounding factors, and were not designed with controlled and standardised methodology, the debate remains open. Prospective studies in well‐defined cohorts of men in various populations are required to evaluate the potential effect of external factors on male reproductive health. These studies should not be limited to the analysis of sperm concentration, as this may not be the best biomarker of testis function and human fertility.
Tissue‐specific expression pattern of estrogen receptors (ER): Quantification of ERα and ERβ mRNA with real‐time RT‐PCRNote
Authors: Pfaffl M. W. , Lange I. G. , Daxenberger A. , Meyer H. H. D.
Published: October 2003

We have examined the tissue‐specific mRNA expression of ERα and ERβ in various bovine tissues using real‐time RT‐PCR. The goal of this study was to evaluate the deviating tissue sensitivities and the influence of the estrogenic active preparation RALGRO on the tissue‐specific expression and regulation of both ER subtypes. RALGRO contains Zeranol (α‐Zearalanol), a derivative of the mycotoxin Zearalenon, shows strong estrogenic and anabolic effects, and exhibits all symptoms of hyperestrogenism, in particular reproductive and developmental disorders. Eight heifers were treated over 8 weeks with multiple‐dose implantations (0×, 1×, 3×, 10×) of Zeranol. Plasma Zeranol concentration, measured by enzyme immunoassay, of multiple treated heifers was elevated. To quantify ERα and ERβ transcripts also in low‐abundant tissues, sensitive and reliable real‐time RT‐PCR quantification methods were developed and validated on the LightCycler. Expression results indicate the existence of both ER subtypes in all 15 investigated tissues. All tissues exhibited a specific ERα and ERβ expression pattern and regulation. With increasing Zeranol concentrations, a significant downregulation of ERα mRNA expression could be observed in jejunum (p<0.001) and kidney medulla (p<0.05). These data support the hypothesis that ERβ may have different biological functions than ERα, especially in kidney and jejunum.
Uterotrophic effect of a saturated fatty acid 17‐ester of estradiol‐17β administered orally to juvenile ratsNote
Authors: Paris Alain , Goutal Isabelle , Richard Jacques , Bécret Annie , Guéraud Françoise
Published: October 2003

In comparison to estradiol‐17β, the naturally synthesized estradiol‐17β‐17‐fatty acid esters are potent estrogens when administered subcutaneously. A lipophilic character of estradiol‐17‐esters could partially protect them from metabolic inactivation. In order to compare their relative estrogenic potency when administered orally, the uterotrophic response to different dosages (0, 2.5, 25, 250 and 2500 nmol/kg BW/day) of estradiol‐17β and estradiol‐17β‐17‐stearate was assessed in juvenile Sprague‐Dawley female rats. Estrogens were administered by oral gavage once a day for 6 days. On the 7th day uterus and vagina were dissected, weighed, and examined microscopically. At 2.5 and 25 nmol/kg BW/day, no difference was detected in the uterus weight compared to control animals which received the vehicle alone (corn oil). At 250 nmol/kg BW/day, the uterotrophic response was maximal in estradiol‐17β‐17‐stearate‐treated animals (×2.40–2.70), whereas it was moderate in estradiol‐17β‐treated rats (×1.86) at the same dosage. This differential weight gain effect of estradiol‐17β‐17‐stearate was correlated with typical microscopic changes in uterus and vagina. The results are in favour of a stronger estrogenic effect of orally given lipoidal estrogens compared to estradiol‐17β. This could be explained by a slower but sustained absorption of estradiol‐17β released from estradiol‐17β‐17‐stearate by esterases and/or by a facilitated transfer of esters in the lymphatic circulation.
Conversion of Borrelia garinii cystic forms to motile spirochetes in vivoNote
Authors: Gruntar Igor , Malovrh Tadej , Murgia Rossella , Cinco Marina
Published: October 2003

Cystic forms (also called spheroplasts or starvation forms) and their ability to reconvert into normal motile spirochetes have already been demonstrated in the Borrelia burgdorferi sensu lato complex. The aim of this study was to determine whether motile B. garinii could develop from cystic forms, not only in vitro but also in vivo, in cyst‐inoculated mice. The cysts prepared in distilled water were able to reconvert into normal motile spirochetes at any time during in vitro experiments, lasting one month, even after freeze‐thawing of the cysts. Motile spirochetes were successfully isolated from 2 out of 15 mice inoculated intraperitoneally with cystic forms, showing the infectivity of the cysts. The demonstrated capacity of the cysts to reconvert into motile spirochetes in vivo and their surprising resistance to adverse environmental conditions should lead to further studies on the role and function of these forms in Lyme disease.
Acute viral lymphadenitis mimicking low‐grade peripheral T‐cell lymphoma A clinicopathological study of nine cases
Authors: Kojima Masaru , Nakamura Shigeo , Itoh Hideaki , Yoshida Katsue , Suchi Taizan , Masawa Nobuhide
Published: October 2003

Acute viral lymphadenitis, especially infectious mononucleosis (IM), often shows the presence of Reed‐Sternberg‐like cells, resulting in confusion with Hodgkin’s disease. However, acute viral lymphadenitis requiring differential diagnosis from non‐Hodgkin’s lymphoma is not widely recognized. We describe the clinicopathological and immunohistochemical features of lymph node lesions from nine such patients which pose serious problems of differential diagnosis from low‐grade peripheral T‐cell lymphoma. There were three males and six females with ages ranging from 21 to 44 years (median 25 years). All patients had “B” symptoms and multicentric lymphadenopathy. The clinical course was also self‐limiting. Each lymph node specimen showed an obvious expansion of an interfollicular area by pleomorphic and polymorphous infiltration with an increased number of arborizing postcapillary venules. The infiltrate was composed of variable numbers of small and medium‐sized lymphocytes, immunoblasts, plasma cells in various stage of maturation and occasional granulocytes. The small lymphocytes usually had regular round nuclei, whereas the medium‐sized lymphocytes occasionally showed nuclear pleomorphism. Hyperreactivity of B‐lymphocytes, including hyperplastic germinal centers and/or foci of monocytoid B‐cells, was seen in parts of the lesion. The majority of the interfollicular T‐lymphocytes, including T‐immunoblasts, expressed CD8 antigen. Various numbers of TIA‐1‐positive small and medium‐sized T‐cells were observed in the paracortical area. Despite these findings, the overall histological picture of this series posed serious difficulties when differentially diagnosing this condition from low‐grade peripheral T‐cell lymphomas such as angioimmunoblastic T‐cell (AILD) and T‐zone types, indicating that viral lymphadenitis occasionally presents with histological features of AILD and T‐zone lymphomas. To avoid overdiagnosis and overtreatment, we emphasize the need to pay careful attention to the clinical and laboratory findings as well as the morphological features.
Cloning, expression and antigenicity of the L. donovani reductase
Authors: Jensen Anja T. R. , Kemp Kåre , Theander Thor G. , Handman Emanuela
Published: October 2003

The protozoan parasite Leishmania undergoes a morphological and biochemical transformation from the promastigote to the amastigote form during its life cycle, which is reflected in the expression of stage‐specific proteins. One of these proteins shows homology to a superfamily of reductase proteins. We have cloned the reductase gene from L. donovani and have shown that it differs in only one nucleotide from the L. major homologue, resulting in one amino acid change. A cytosine (C) to guanine (G) transposition in the coding sequence leads to a nonconserved substitution of asparagine (N) for lysine (K). Only 2 of 22 plasma samples from patients with visceral leishmaniasis were found to have detectable anti‐reductase antibodies and peripheral blood mononuclear cells (PBMC) from one of three individuals previously infected with visceral leishmaniasis proliferated in the presence of recombinant reductase protein. Interestingly, 6 of 10 PBMC isolated from Danish controls proliferated in the presence of the reductase protein. Intracellular IFNγ was found in a significant percentage of cells in all the tested PBMC cultures from Danes, whereas IL4 was only found in a small proportion of cells, or not at all. The results indicate the presence of cross‐reacting CD45R0 memory T‐cells in individuals not exposed to Leishmania. Several previous studies have shown that T‐cells from nonexposed individuals often respond to crude Leishmania antigen preparations. The present study suggests that this reactivity is partly caused by T‐cells recognising L. donovani reductase.
Age of puberty: Data from the United States of AmericaNote
Authors: LEE PETER A. , GUO SHUMEI S. , KULIN HOWARD E.
Published: October 2003

In an attempt to determine whether the secular trend toward an earlier onset of puberty has continued over recent decades in the United States of America, published reports concerning the age of attainment of pubertal events have been reviewed. Such reports are very limited and vary in both design and inclusive ages of study subjects. Among females, two recent large cross‐sectional studies indicate that fifty percent of females in the United States attain Tanner breast stage 2 at 9.5 to 9.7 years of age. This is younger than previously thought, although adequate earlier studies of girls in the United States are not available for comparison. These two studies also indicate that about 14% of girls attain Tanner stage 2 while 8 years of age; one study extends earlier reporting that about 6% exhibit onset of breast development while 7 years of age. There is no evidence that the age of menarche or the attainment of adult (Tanner 5) breast development has decreased over the past 30 years. The data also suggest an earlier onset of Tanner stage 2 pubic hair but no change in attainment of stage 5. Among males, pubic hair may be appearing at younger ages, but data are inadequate or too inconsistent to allow firm interpretation. The lack of standardization of genital criteria of pubertal onset in the male makes any conclusions regarding secular trends impossible. In summary, earlier secular trends over recent decades related to better health, improved nutrition or socio‐economic status, or any putative influence by endocrine disrupters cannot be verified.
Genotoxic potential of xenobiotic growth promoters and their metabolitesNote
Authors: METZLER MANFRED , PFEIFFER ERIKA
Published: October 2003

This paper reviews data reported in the literature as well as recent and unpublished studies from our laboratory on the metabolism and genotoxicity of the xenobiotic growth promoters 17β‐trenbolone, melengestrol acetate and zeranol. In our metabolic study, the oxidative in vitro metabolites generated by hepatic microsomes from rats, bovine and humans were analyzed by HPLC and GC/MS. 17β‐Trenbolone gave rise to at least 13 monohydroxylated products, whereas 12 mono‐ and dihydroxylated metabolites were obtained with melengestrol acetate and at least 5 with zeranol. The genotoxic potential of the parent compounds was studied using the following endpoints: induction of HPRT mutations in cultured V79 cells and of lacI mutations in E. coli; induction of micronuclei in V79 cells; and formation of DNA adducts in cultured primary rat hepatocytes. Negative results were obtained in most of these assay systems. Only the micronucleus induction was marginally positive with 17β‐trenbolone and zeranol at near‐cytotoxic concentrations. Commercial melengestrol acetate was found to contain an impurity causing apoptosis in V79 cells. The genotoxic potential of the numerous oxidative metabolites of the xenobiotic growth promoters remains to be studied.
The birth rate of hypospadias in the Turku area in FinlandNote
Authors: VIRTANEN H. E. , KALEVA M. , HAAVISTO A.‐M. , SCHMIDT I. M. , CHELLAKOOTY M. , MAIN K. M. , SKAKKEBAEK N. E. , TOPPARI J.
Published: October 2003

Reports based on national registers of congenital malformations have suggested that the birth rate of hypospadias has increased during the last few decades. Register‐based information may, however, have pitfalls because of changes in diagnostics, reporting accuracy and registration system. The aim of this study was to determine the current birth rate of hypospadias in Turku University Central Hospital (TUCH) in Finland. This was a prospective study on live‐born boys born in TUCH from 1997 to 1999. In the total birth cohort (n=5,798) as well as in a special subcohort group (n=1,505) 0.3% of boys had hypospadias. Only one scrotal hypospadias was found in a boy who had a chromosomal anomaly. Other hypospadias were glandular or coronal. No increase was found in the birth rate of hypospadias when comparing our result with register‐based data of boys born in Finland during the years 1970 to 1986 and surgically treated for hypospadias by the age of 8 years. No difference was found either from malformation register‐based data concerning the nationwide birth rate of hypospadias during the years 1993 to 1998. Due to differences in national registration systems between countries, prospective studies with equal assessment criteria are needed in order to make reliable international comparisons.
Estrogenic activity of estradiol and its metabolites in the ER‐CALUX assay with human T47D breast cellsNote
Authors: HOOGENBOOM L. A. P. , DE HAAN L. , HOOIJERINK D. , BOR G. , MURK A. J. , BROUWER A.
Published: October 2003

A number of metabolites of 17β‐estradiol were tested for their estrogenic activity using the ER‐CALUX assay based on the increased expression of luciferase in exposed T47D breast cancer cells. E2β and estrone showed similar potencies in the test, whereas E2α was 100 times less active. Incubation of cells with estrone (0.35 μM) resulted in the formation of E2β, whereas the reverse reaction was observed for E2β. The resulting equilibrium may explain the similar estrogenic potency of estrone in the test. The synthetic 17‐hydroxy benzoate ester of E2β was 3 times less active than the parent compound. The 17‐hydroxy palmitate and oleate esters of E2β, were respectively 25 and 200 times less active than the parent compound. The 2‐hydroxy metabolites of E2β and estrone showed a 5,000 to 10,000 fold lower activity. The 4‐hydroxy metabolites were more potent than the 2‐hydroxy metabolites, showing only a 20–200 times lower activity. The 2‐ and 4‐methoxyesters of estrone were 700 times less active. It is concluded that the estrogenic potency of metabolites formed in cattle after treatment with E2β, like estrone, E2α and especially the esters of E2β, may be significant with respect to the potential risk of the use of estradiol for growth promotion in domestic animals in certain countries.
Predominant cultivable subgingival microbiota of healthy and HIV‐infected ethnic ChineseNote
Authors: TSANG C. S. P. , SAMARANAYAKE L. P.
Published: October 2003

Although clinically distinctive types of periodontal disease are known to be associated with HIV infection, the pathogenesis remains unclear. In the present study, the subgingival microflora of 21 HIV‐infected and 11 HIV‐free ethnic Chinese were studied using the direct microscopic and anaerobic culture methods. Motile curved rods were found to be three times higher in the HIV‐infected group under direct microscopy. Otherwise, there were no significant differences between the diseased and healthy groups when analyzed either in relation to the morphotype distribution or Gram stain morphology or oxygen tolerance. The most common bacteria isolated were Capnocytophaga species followed by Prevotella loescheii, Streptococcus sanguinis, Lactobacillus spp. and Fusobacterium spp. Although there were 20 bacterial species that were strictly limited to the HIV‐infected group, and 5 limited to the healthy group, none of the species was a predominant isolate in either group (p>0.05). These findings agree with previous studies which report that subgingival bacteria of HIV‐infected individuals (with or without periodontal disease) are similar to those found in healthy HIV‐free individuals (with or without periodontal disease). Further work examining the subgingival microflora from patients with specific and/or severe forms of HIV‐associated periodontal disease is required to shed light on the pathogenesis of this complex clinical entity.
The effect of oral bacteria on Candida albicans germ‐tube formationNote
Authors: NAIR RAJ G. , ANIL S. , SAMARANAYAKE LAKSHMAN P.
Published: October 2003

Abstract: A total of eight bacterial isolates belonging to six species, and a select group of 12 oral Candida albicans isolates, were used to study the effect of bacteria on germ‐tube formation. Briefly, each bacterial suspension (105–6 cells/ml) was mixed with a C. albicans suspension (107 cells/ml) and incubated at 37 °C for 90 min with bovine serum, and the percentage germ‐tube‐positive Candida cells was quantified using a haemocytometer, under light microscopy. In general, out of eight bacteria, Streptococcus sanguis SK21A, Streptococcus salivarius SK56, Escherichia coli ATCC 25922, and S. salivarius OBU3 suppressed germ‐tube formation to varying degrees, with different C. albicans isolates. Porphyromonas gingivalis Pg 50, Lactobacillus casei ATCC 7469 and Prevotella intermedia OBU4 elicited significant enhancement of germ‐tube formation, whereas S. sanguis OBU 2 had no effect. E. coli ATCC 25922 was the only organism to show statistically significant suppression of germ‐tube formation (p=0.0312). A significant increase in the germ tube production of C. albicans isolated from HIV‐infected compared with HIV‐free individuals was also noted. The current results tend to suggest that commensal and transient oral bacterial populations may selectively influence the differential expression of germ‐tube‐forming ability of C. albicans isolates.
Growth rate and biofilm thickness of Streptococcus sobrinus and Streptococcus mutans on hydroxapatiteNote
Authors: ROZEN RAMONA , BACHRACH GILAD , ZACHS BATIA , STEINBERG DORON
Published: October 2003

Bacteria in biofilm and planktonic bacteria exhibit different properties. The objective of the present study was to compare the growth rates of Streptococcus sobrinus and Streptococcus mutans on different types of biofilm with their planktonic growth rate. Our experimental model consisted of hydroxyapatite beads coated with human saliva (sHA). Glucans or fructans were synthesized in situ on sHA by immobilized cell‐free glucosyltransferase or fructosyltransferase isolated from oral bacteria. S. sobrinus or S. mutans was then adsorbed onto the glucan‐ or fructan‐coated sHA and incubated for different time intervals. The depth of the developing biofilm was measured. Our results show that growth rates of S. sobrinus and S. mutans on both fructan‐ and glucan‐coated sHA were similar during a 23 h period. In addition, the profile was similar to the growth profile of the same planktonic bacteria. The resemblance in growth rates between planktonic and biofilm bacteria may be attributed to the thin and non‐dense biofilm formed in the initial stages of the biofilm formation. The thin biofilm coat, reaching a maximal depth of 11 μm, has only imposed limited diffusion restrictions, thus not affecting the growth of the bacteria in the biofilm. Our study shows that growth of bacteria on surfaces may resemble their growth in suspension if the bacteria are not embedded in a thick dense biofilm.
From malformations to molecular mechanisms in the male: three decades of research on endocrine disruptersNote
Authors: McLachlan John A. , Newbold Retha R. , Burow Matthew E. , Li Shuan Fang
Published: October 2003

For three decades, we have known that estrogens alter the development of the mammalian reproductive system in predictable ways. In mice exposed prenatally to diethylstilbestrol (DES) or other estrogens, the male offspring exhibit structural malformations including cryptorchidism, epididymal cysts and retained Mullerian ducts. The estrogen‐associated alterations in the genital tract phenotype can be usefully considered as a model called Developmental Estrogenization Syndrome. While estrogen treatment during critical periods of morphogenesis of the male reproductive system has been associated with these changes, the mechanisms at the molecular level are still being discovered. Parallel findings on the hormones involved in Mullerian duct regression and testicular descent have helped guide research on the mechanisms of developmental estrogenization of the male. Cellular localization of molecular signals associated with key steps in genital tract development, use of mice with gene disruption, and knowledge of the mechanisms underlying persistent changes in gene expression are beginning to provide a blue print for both the physiological role and pathological effects of estrogens in reproductive tract development. Since many of the same biological principles underlie genital tract morphogenesis in mammals, one may expect some of the same changes in males of other species exposed to estrogen during the appropriate developmental periods.
Residues from anabolic preparations after good veterinary practiceNote*
Authors: Henricks D. M. , Gray S. L. , Owenby J. J. , Lackey B. R.
Published: October 2003

The purpose of this study was to determine the endogenous concentrations of estrogens, particularly estradiol‐17β (E2β), in edible tissues of beef cattle (females and intact and neutered males) and the concentrations of E2β, and trenbolone beta and alpha (βTb, αTb) after an E2β and/or trenbolone acetate (TA) ear implant. Radioimmunoassays were validated for quantitation of E2β (active isomer), E2α, estrone (E1), βTb and αTb for bovine muscle, liver, kidney and fat tissues. The criteria of accuracy, precision, specificity and sensitivity were applied according to the standards of the U.S. Food & Drug Administration. In steer tissues, endogenous E2β was <15 ppt, as was heifer muscle; but heifer liver and kidney were 3‐fold greater. An E2β implant in steers had no effect on muscle E2β concentration, but increased E2β in liver and fat 4‐ and 3‐fold, respectively, but by 24 h post‐implant removal, E2β had fallen by half. Tissue E1 concentrations in cyclic females were similar to E2β, but rose many fold greater than did E2β during gestation; E2β rose 3‐fold during gestation. After E2β/TA implant, steer tissues had E2β concentrations equal to (for muscle and fat) and one‐half (for liver) the E2β measured in E2β implant only steers; βTb was in a low range (250–380 ppt) in muscle, liver and fat and αTb was even lower, except in liver (800–1500 ppt). An implant of TA only (no E2β) resulted in βTb and αTb concentrations 2–3‐fold greater in liver, kidney and fat, but no greater in muscle than βTb in tissues of E2β/TA implant steers. In conclusion, anabolic implants in steers resulted in tissue E2β concentrations less than the FDA allowable increment and βTb in the lowest quartile (0.25) of a part per billion 30 days after implant.
Comparison of endobronchial and transbronchial biopsies with high‐resolution CT (HRCT) in the diagnosis of sarcoidosisNote
Authors: Halme M. , Piilonen A. , Taskinen E.
Published: October 2003

High‐resolution computed tomography (HRCT) has been shown to accurately visualise parenchymal infiltrates of sarcoidosis. The aim of this study was to compare the diagnostic yield (DY) of HRCT with that of endobronchial (EBB) and transbronchial (TBB) biopsies in establishing the diagnosis of sarcoidosis. Forty‐five patients referred to Helsinki University Central Hospital with a presumptive diagnosis of sarcoidosis underwent fiberoptic bronchoscopy (FOB) with bronchoalveolar lavage (BAL), EBB, TBB and HRCT. Thirty‐seven of the patients were diagnosed as having sarcoidosis, 34 of whom showed a parenchymal infiltrate in HRCT. One of the three patients with no parenchymal findings, had positive findings in TBB, and all had lymphocytosis in BAL. The DY for sarcoidosis by EBB and TBB was 24.3% and 50.0%, respectively. The addition of EBB to TBB improved the DY by 8.3%, whereas adding TBB to EBB improved the DY by 30.6%. There were no major complications after the FOB, which was always performed under fluoroscopic control. In conclusion, HRCT is a valuable tool in diagnosing sarcoidosis. However, the HRCT findings cannot be seen in all patients with positive findings in lung biopsy, nor is the parenchymal infiltrate specific for sarcoidosis. Attempting biopsy‐proven diagnosis of sarcoidosis is still recommended. Fiberoptic bronchoscopy with EBB and TBB under fluoroscopic control is a safe and well‐tolerated procedure.
Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human chondrosarcomasNote
Authors: Söderström M. , Aro H. T. , Ahonen M. , Johansson N. , Aho A. , Ekfors T. , Böhling T. , Kähäri V. M. , Vuorio E.
Published: October 2003

The aim of the present study was to characterise the ability of malignant chondrosarcomas to invade normal bone by analysing their production of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). For this purpose 12 chondrosarcomas were investigated for the expression of mRNAs for several MMPs and all 4 TIMPs by Northern hybridisation, and for immunohistochemical localisation of the proteins. A characteristic finding of these analyses was increased expression of MMP‐13, MMP‐14 and TIMP‐2 mRNAs in chondrosarcomas when compared with nonmalignant control samples. Individual chondrosarcomas also exhibited elevated levels of MMP‐1, MMP‐7 and MMP‐9 mRNAs. The results of Northern hybridisations were supported by immunohistochemical stainings of the corresponding tumour areas for MMP‐2, MMP‐14 and TIMP‐2, further suggesting that these may have prognostic value for determining whether individual chondrosarcomas are locally aggressive or have a probability of recurrence. Another finding of the present study was a marked heterogeneity in histologic appearance and gene expression of the chondrosarcomas, emphasising the importance of analysing several areas of these tumours to get representative results. These findings suggest that analysis of MMPs could be a useful diagnostic indicator in patients with cartilaginous tumours and could help in differentiating between a low‐grade malignant chondrosarcoma and a benign growing enchondroma.
Epithelial‐myoepithelial carcinoma harboring p53 mutationNote
Authors: Daa Tsutomu , Kashima Kenji , Gamachi Ayako , Nakayama Iwao , Yokoyama Shigeo
Published: October 2003

A case of epithelial‐myoepithelial carcinoma of the parotid gland harboring p53 mutation is reported. The tumor removed from a 67‐year‐old Japanese female was composed of an organoid biphasic population of cells: inner dark epithelial cells were surrounded by clear myoepithelial cells. The cells were immunopositive for EMA and smooth muscle actin, respectively. Some of the epithelial cells formed solid nests. Immunostaining for proliferating cell nuclear antigen (PCNA) resulted in a higher percentage of labeled cells in the solid epithelial region than in the region with the more general biphasic pattern. Genetic analysis, including polymerase chain reaction‐single strand conformational polymorphism (PCR‐SSCP) and nucleotide sequencing, revealed a mutation in codon 207 (aspartic acid to glycine) of the p53 tumor‐suppressor gene. To our knowledge, this is the first report of a mutation in the p53 gene in an epithelial‐myoepithelial carcinoma of the salivary gland.
In situ hybridisation for identification and differentiation of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae and Mycoplasma hyorhinis in formalin‐fixed porcine tissue sectionsNote
Authors: BOYE M. , JENSEN T. K. , AHRENS P. , HAGEDORN‐OLSEN T. , FRIIS N. F.
Published: October 2003

Oligonucleotide probes targeting 16S ribosomal RNA were designed for species‐specific identification of the porcine mycoplasmas Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma hyosynoviae using a fluorescent in situ hybridisation assay. The specificity of the probes was evaluated using pure cultures as well as porcine tissue sections with artificial presence of mycoplasma, and the probes were found specific for the target organisms. The assay was applied on sections of 28 tissue samples from pigs infected with one or more of the three Mycoplasma species as determined by cultivation. M. hyopneumoniae and M. hyorhinis were identified in accordance with cultivation in lung sections from nine pigs affected by catarrhal to purulent bronchopneumonia. Likewise, in eight cases of fibrinous pericarditis, M. hyopneumoniae, M. hyorhinis and M. hyosynoviae were the infectious agents according to cultivation and were correctly identified by in situ hybridisation. Out of 11 joints cultivation positive for M. hyosynoviae, the probe was only able to identify M. hyosynoviae in eight cases probably due to a low number of microorganisms in the tissue sections. The in situ hybridisation assay is well suited for use in diagnostic and experimental work as well as a tool for pathogenesis studies.
Histoplasma capsulatum var. capsulatum occurring in an HIV‐positive Ghanaian immigrant to Italy
Authors: RIVASI FRANCESCO , CASALI BRUNO , NANETTI ANNA , COLLINA GUIDO , MAZZONI ALDO
Published: October 2003

Histoplasmosis, which is highly endemic in the United States, is rare in Europe, usually imported but sometimes autochthonous. In Africa, histoplasmosis capsulati coexists with “African histoplasmosis”, a characteristic skin infection caused by H. capsulatum var. duboisii. Histoplamosis due to H. capsulatum is one of the 12 secondary infections listed in the surveillance definitions of AIDS. We report the case of a 36‐year‐old black man with acquired immunodeficiency syndrome (AIDS) who was living in Italy but originally came from Ghana. Histoplasmosis was disseminated with fever and cutaneous manifestations. The diagnosis was demonstrated morphologically based on the presence of yeast, observed by light microscopy, in skin lesions and by identification of H. capsulatum var. capsulatum DNA by nested PCR from a paraffin sample. No clinical reports of histoplamosis capsulati in Ghana have been published until now. The present case stresses the role of immigration of subjects from outside Europe who have been infected in their native country.
Proliferation conditions for human satellite cells The fractional content of satellite cells
Authors: GASTER M. , BECK‐NIELSEN H. , SCHRØDER H. D.
Published: October 2003

Primary satellite cell cultures have become an important tool as a model system for skeletal muscles. A common problem in human satellite cell culturing is fibroblast overgrowth. We combined N‐CAM (Leu19) immunocytochemical staining of satellite cells (Sc) with stereological methods to estimate the fraction of Sc in culture. Evaluation of different culture conditions allowed us to find proliferation conditions preferentially for Sc: a) Sc should be cultured on surfaces coated with ECM‐gel. b) Primary cell culture should be inoculated in DMEM supplemented with 10% fetal calf serum to increase cell adherence. c) Change of media to DMEM supplemented with 2% Ultroser‐G and 2% FCS after 24 h.d) Before subcultivation, cells should be preplated for 30 min. The fractional content of Sc in passage four when applying this method of cultivation was 0.82 +/‐ 0.07 (mean +/‐ SE, N=10). Our method enabled us to establish culture conditions which resulted in high Sc content despite several subcultivations. Estimation of the fractional cell content could be a useful tool for optimizing not only Sc‐culturing but all cultures initially containing more cell types.
Comparative evaluation of a commercial test for rapid identification of methicillin‐resistant Staphylococcus aureus
Authors: JUREEN R. , BOTTOLFSEN K. LILAND , GREWAL H. , DIGRANES A.
Published: October 2003

The performance and ease of use of the recently introduced MRSA screen test (Denka Seiken Co. Ltd., Japan) for the identification of methicillin‐resistant Staphylococcus aureus was evaluated in comparison with the BBL Crystal MRSA ID System (Becton Dickinson Europe, France). A total of 109 strains of S. aureus, consisting of 57 strains of MecA‐negative S. aureus and 52 strains of MecA‐positive S. aureus, were tested. With MecA PCR as the gold standard, the MRSA screen test had 98% sensitivity and 98% specificity, whereas the BBL Crystal MRSA ID System had 98% sensitivity and 95% specificity. The simplicity of use and rapid result make the MRSA screen test a valuable tool in the clinical microbiology laboratory pending demonstration of the MecA gene that should still always be done.
Benign solitary fibrous pleural tumour. Evidence of primitive features and complex genomic imbalances, including loss of 20qNote
Authors: AMO‐TAKYI B. K. , GÜNTHER K. , PETERS I. , MITTERMAYER C. , EBLENKAMP M. , TIETZE L.
Published: October 2003

Aims: Cytogenetic data on solitary fibrous tumours (SFT) are very limited. We studied a benign pleural SFT for its ultrastructural and immunohistochemical details, and made cytogenetic analyses for comparison with other genetic and ultrastructural studies of SFT. Results: Immunohistochemistry showed strong positivities for CD34 and vimentin, but no reactions with anti‐cytokeratins and epithelial membrane antigens. Electron microscopy revealed primitive desmosomes in our SFT. The results thus evinced fibroblast‐like cells with intermediate epithelial‐mesenchymal character. Comparative genomic hybridization of the tumour revealed losses of 1p33→pter, 17pter–q21, entire copies of chromosomes 19 and 22, and gains of 1p21–p22, 2q23–q32.3, 3p12–q13.2, 4p14–q28, 6p12–q21, 9p21→pter and 13q21–q31. Furthermore, there was loss of 20q, as was previously reported elsewhere in a case of benign and a case of malignant SFT. Conclusions: The results furnish further evidence of the involvement of −20q in SFT. In addition, they show that SFT may have complex genomic imbalances and primitive features, despite having a benign appearance.
Neutrophil free oxygen radical production and blood total antioxidant capacity in patients with coronary heart disease using various medicationsNote
Authors: MATTILA KIMMO , RISTOLA MATTI , REPO HEIKKI , ASIKAINEN SIRKKA , METSÄ‐KETELÄ TIMO
Published: October 2003

Despite convincing results of studies in vitro, less is known about the effects of antioxidants on in vivo redox balance in humans. We developed a novel parameter of in vivo redox balance, and studied it and its relation to dental infections in 51 patients on medication for coronary heart disease (CHD) and 39 random controls matched for age group, sex, social class and locality. In vivo redox balance was the ratio of plasma antioxidant capacity, as measured with radical‐trapping assay, to neutrophil respiratory burst capacity, as measured with whole blood chemiluminescence assay. Dental infections were quantitated with four rating scales. CHD patients had higher values than controls. Patients on acetosalicylic acid (ASA), diuretics or beta blockers, but not the ones on calcium channel blocker, had significantly higher redox balance than non‐users. Combination of calcium channel blockers and ASA was associated with redox balance similar to taking beta blockers or diuretics. Diuretics and ASA were independent determinants of redox balance in multivariate analyses. Redox balance did not correlate with severity of dental infections (Spearman's r 0.06 to 0.11). The results contrast experimental data indicating that calcium channel blockers are as antioxidants superior to other cardiovascular drugs. Total antioxidant capacity in parallel with oxygen species production capacity should be considered in attempts to solve the antioxidant paradox.