Novel regulators of the humoral immune response.

Activation of mature B cells to proliferation and terminal differentiation is a multistep process controlled mainly by macrophages and T cells. However, there is growing evidence that B cells and other factors can also regulate the humoral immune response. Here, Ferenc Uher, Eva Rajnavölgyi and Anna Erdei describe their work on the emerging role of regulatory interactions between subsets of B cells and their soluble products.


Functional studies on Fc receptors.

Fc receptors (FcRs) are cell surface molecules that recognize and bind to the constant domains of immunoglobulins. In doing so, they enable antibodies to perform several biological functions, by forming a link between specific antigen recognition and FcR-bearing cells. Here, Gabriella Sármay provides an overview of recent studies on FcRs in Hungarian laboratories, concentrating on their role in selected biological systems.


Bcl-2: a repressor of lymphocyte death.

The genes and mechanisms that control programmed cell death are currently the subject of intense study. The bcl-2 gene, a repressor of lymphocyte death, is perhaps the best understood of the programmed cell death associated genes. Here, Stanley Korsmeyer provides a brief overview of bcl-2, concentrating on its roles in B- and T-cell development and in oncogenesis.


Endothelial cell binding of NAP-1/IL-8: role in neutrophil emigration.

In this article Antal Rot calls into question the generally accepted notion that gradients of soluble chemotactic factors are responsible for leukocyte emigration from the circulation into sites of inflammation. He presents an alternative model in which chemoattractants bound to the surface of endothelial cells promote neutrophil adhesion and emigration, while soluble blood-borne attractants inhibit adhesion and emigration.


VDJ recombination.

The ability of lymphocyte receptor V, D and J gene segments to rearrange generates much of the receptor diversity that is the hallmark of the immune system. Naturally, the mechanisms of immunoglobulin and T-cell receptor gene recombination are of enormous interest. Here, Fred Alt and colleagues review current understanding of the process and speculate on future findings.


The origins and early years of IUIS.

The International Union of Immunological Societies (IUIS) was born of the global linkages between scientists involved in an exciting intellectual journey of discovery and search for a conceptual framework for regulation, specificity and memory of the immune response. Starting in the 1950s, a dozen of us discussed the need for an international organization; we founded it in 1969.


The antagonistic effects of tetrahydroprotoberberines on dopamine receptors: electrophysiological studies.

Tetrahydroprotoberberines (THPBs), including (-)-stepholidine ((-)-SPD), (-)-tetrahydropalmatine ((-)-THP) and tetrahydroberberine (THB), have been demonstrated to be a new class of DA antagonists in biochemical and neuropharmacological studies. In this paper, the antagonistic action of THPBs was examined by means of single unit recording from nigral DA neuron in chloral hydrate-anesthetized and gallamine-paralyzed rats. Intravenous injection of these compounds could promptly and completely reverse the inhibition of the spontaneous firing induced by DA agonist apomorphine (APO) in a dose-dependent way. Pretreatment with (-)-SPD, (-)-THP or THB could significantly reduce the inhibitory effect of APO and shift the dose-action curve to the right. Besides, the compounds could increase the spontaneous firing of DA neurons. The above results not only strongly support the conclusion that (-)-SPD, (-)-THP and THB are DA antagonists, but also demonstrate that one of their blocking sites is at somatodendritic DA autoreceptors (D-2 receptors). In other words, (-)-SPD did not exhibit any DA agonistic action in this acute electrophysiological study, although its DA agonistic action can be demonstrated in rotational behavior of 6-OHDA-lesioned rats. The dual actions of (-)-SPD, dependent upon different experimental conditions, are discussed.


The role of iron in oxygen-mediated toxicities.

The transition metal iron is capable of catalyzing redox reactions between biomolecules and oxygen that would not occur if catalytically active iron were not present. Although these biological oxidations (which are known collectively as "oxidative stress") have been implicated in numerous toxicities, the exact role of the iron catalyst remains to be elucidated. This review focuses on our current understanding of the role of iron in oxidative stress, discussing biologically relevant sources, biochemical forms, and reaction mechanisms of iron as a catalyst of biomolecular oxidations. Specific toxicities in which alterations in normal iron metabolism is thought to overwhelm the body's antioxidant defense system are presented, and future treatment regimens involving novel antioxidant drugs are discussed.


Assessment of the carcinogenicity of the nonnutritive sweetener cyclamate.

The weight of the evidence from metabolic studies, short-term tests, animal bioassays, and epidemiological studies indicates that cyclamate (CHS) is not carcinogenic by itself; however, there is evidence from in vitro and in vivo studies in animals that implies it may have cancer-promoting or cocarcinogenic activity. Epidemiological studies indicate that the use of nonnutritive sweeteners (CHS and saccharin) has not resulted in a measurable overall increase in the risk of bladder cancer in individuals who have ever used these products. No epidemiological information exists on the possible associations of these sweeteners and cancers other than those of the urinary tract. It is recommended that (1) no further studies on the metabolism of CHS to evaluate its carcinogenicity are required since no potentially hazardous metabolites have been appreciably detected in humans; (2) no further animal bioassays to test for the carcinogenicity of CHS by itself are necessary; (3) the studies in rodents that suggest a promotional or cocarcinogenic effect of CHS should be repeated because they cannot be ruled out; (4) because the significance to human health of a positive outcome of such studies is uncertain, additional research aimed at understanding the predictive value for human health of such results and more generic studies to develop well-validated systems that can be relied on in the assessment of cancer-promoting agents are recommended; (5) in populations where CHS continues to be used, epidemiological monitoring should be continued to determine whether there is an increased risk of cancer in humans who are heavy or long-term users or for those observed long after first exposure. In such monitoring, other cancer sites--in addition to the bladder--should be considered.


[Determinants of the smoking habit in the city of Pelotas, Brazil].

Although the harmful effects of smoking on health are well documented, this habit continues to be highly prevalent in Latin America. The objective of this study is to evaluate the frequency of this habit and its possible determinants in the adult population of the city of Pelotas, state of Rio Grande do Sul, Brazil, using a simplified methodology. With the use of a cross-sectional design, a probabilistic sample of 1,401 adults was studied; the no-response rate was 11%. Of those interviewed, 54% had never smoked, 14% were former smokers, and 33% were smokers at the time of the study; 16% smoked 20 or more cigarettes daily. Smoking was more frequent in men (44%) than in women (23%). In relation to educational level, the highest prevalence of smoking was among individuals with five to eight years of schooling (39.7%). The study showed that it is possible with a limited investment of time and resources, to evaluate the frequency of smoking in a representative sample of a medium-sized city, yielding data that will be highly useful for preventive campaigns.


[Prevalence of mental disorders in adults in Subtiava, León, Nicaragua].

The objective of this study was to estimate the prevalence of mental disorders in the Subtiava district of León, Nicaragua. Two-stage cluster sampling was used to select 219 families (n = 584 persons). The study was carried out in two phases. In the first, probable mental disorder cases were screened using the Self-Reporting Questionnaire. In the second, the diagnoses were confirmed through the Present State Examination questionnaire. The validity and reliability of the questionnaires had been confirmed in a pilot study. The present study was conducted in October and November 1987, when Nicaragua was still immersed in armed conflict. Construction of a family map made it possible to analyze, through the multiple imputation method, the characteristics of persons who were absent at the time of the interviews or who did not respond. The estimated prevalence percentages and their respective 95% confidence intervals were as follows: neurosis, 7.5% (5.2%-9.8%); depression, 6.2% (2.6%-8.1%); reactive crisis, 3.3% (2.6-6.5%); alcoholism, 5.8% (3.9%-8.9%); organic brain syndrome, 3.9% (1.7%-5.2%); psychosis, 0.5% (0.2%-1.6%); and other disorders, 0.7% (0.2%-1.6%). The estimated overall prevalence of mental disorders in the study population was 27.9%. Disorders were more prevalent among men (30.8%) than women (26.3%). It is concluded that these high estimated prevalences are associated with stress caused by the war.


Purification and characterization of an extracellular thiol-containing serine proteinase from Thermomyces lanuginosus.

An extracellular protease produced by the filamentous fungus Thermomyces lanuginosus has been purified and characterized. The results indicate that the enzyme, which we have called humicolin, is a thiol-containing serine protease with a molecular mass of 38,000 kilodaltons. Secretion of humicolin, which is glycosylated, is tightly regulated by protein substrates. Kinetic characterization has revealed that humicolin activity is highly dependent upon the deprotonation of a group with a pKa of 6.6 and that the enzyme has a specificity for phenylalanine in the P1 position of the substrate.


Purification of sn-glycerol-3-phosphate dehydrogenase from Trypanosoma brucei brucei.

A protein has been purified from the membranes of bloodstream forms of Trypanosoma brucei brucei. The purified material contained a single polypeptide chain of molecular mass 67 kilodaltons as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; under "native" conditions it migrated through a Sephacryl S-300 column with a similar molecular mass. The purified protein catalysed electron transfer from sn-glycerol 3-phosphate to oxygen with the subsequent formation of water. Electron transfer by the purified enzyme to O2 was dependent on the presence of low concentrations of the mediator phenazine methosulfate. This protein is clearly the major membrane-bound sn-glycerol-3-phosphate dehydrogenase, but it also has some characteristics suggestive of the trypanosome alternative oxidase activities.


Expression of c-fos and c-myc genes in uterus and nonreproductive organs during the implantation period in the rat.

The c-fos and c-myc mRNA levels were investigated in the uterus and nonreproductive organs of rats during the implantation period (from day 5 to 7 of pregnancy). They were determined by densitometric analysis of slot blots and the mean values (n = 4) at a defined age of pregnancy were compared with those observed in nonpregnant control rats (NP group). No significant variations of c-fos level were observed in the liver and brain of pregnant rats. In the uterus, the c-fos level decreased, reached a minimum on day 6 (4-fold decrease compared with the NP group), and then increased. The c-fos level significantly increased in the kidney (2- and 2.5-fold on days 6 and 7), and in the lung (2.5-fold on day 7). The c-myc level did not significantly vary in the uterus, liver, and lung. It significantly increased in the kidney (3-fold on day 7) and in the brain (4.1-, 3.8-, and 6-fold, on days 5, 6, and 7). The changes in c-fos and (or) c-myc gene expression in the uterus and nonreproductive organs could be due to sexual steroids and (or) systemic factors from uterine cells or blastocysts.


Developmentally regulated troponin C mRNAs of chicken skeletal muscle.

Fast and slow/cardiac troponin C (TnC) are the two different isoforms of TnC. Expression of these isoforms is developmentally regulated in vertebrate skeletal muscle. Therefore, in our studies, the pattern of their expression was analyzed by determining the steady-state levels of both TnC mRNAs. It was also examined if mRNAs for both isoforms of TnC were efficiently translated during chicken skeletal muscle development. We have used different methods to determine the steady-state levels of TnC mRNAs. First, probes specific for the fast and slow TnC mRNAs were developed using a 390 base pair (bp) and a 255 bp long fragment, of the full-length chicken fast and slow TnC cDNA clones, respectively. Our analyses using RNA-blot technique showed that fast TnC mRNA was the predominant isoform in embryonic chicken skeletal muscle. Following hatching, a significant amount of slow TnC mRNA began to accumulate in the skeletal (pectoralis) muscle. At 43 weeks posthatching, the slow TnC mRNA was nearly as abundant as the fast isoform. Furthermore, a majority of both slow and fast TnC mRNAs was found to be translationally active. A second method allowed a more reliable measure of the relative abundance of slow and fast TnC mRNAs in chicken skeletal muscle. We used a common highly conserved 18-nucleotide-long sequence towards the 5'-end of these mRNAs to perform primer extension analysis of both mRNAs in a single reaction. The result of these analyses confirmed the predominance of fast TnC mRNA in the embryonic skeletal muscle, while significant accumulation of slow TnC mRNA was observed in chicken breast (pectoralis) muscle following hatching. In addition to primer extension analysis, polymerase chain reaction was used to amplify the fast and slow TnC mRNAs from cardiac and skeletal muscle. Analysis of the amplified products demonstrated the presence of significant amounts of slow TnC mRNA in the adult skeletal muscle.


Characterization of two forms of cationic peroxidase from cultured peanut cells.

Two forms of cationic peroxidase from peanut cells were differentiated by concanavalin A affinity chromatography. They differed in molecular mass as well as concanavalin A binding, leading to the initial suggestion that they represented two isozymes of peroxidase. However, similar values for the specific activity, Soret absorption, calcium content, and peptide molecular mass were observed for each of the forms. Therefore, the binding and nonbinding fractions most likely represent two molecular forms of cationic peanut peroxidase, rather than two distinct cationic isozymes. The difference between these two forms is discussed in terms of glycosylation. Through the amino acid sequence analysis of the formic acid treated peptide, the cationic isozyme has been shown to be identical in amino acid sequence to the cDNA clone PNC1.


Formation of C19 11-hydroxysteroids by porcine Leydig cells.

In a previous study, we reported the presence of 11 beta-hydroxyandrostenedione and 11 beta-hydroxytestosterone in testicular vein blood from mature male pigs. Since C19 steroids with an oxygen function at C11 have not been recorded as products of steroid biosynthesis in normal mammalian testes, we have examined their possible production in purified preparations of porcine Leydig cells. Both androstenedione and cortisol were added as substrates in studies using cell incubations of Leydig cells from mature boars (greater than 8 months old). Steroids were recovered from media by solid-phase extraction and separated by reversed-phase high performance liquid chromatography. Peaks corresponding to retention times of authentic standard steroids were seen for both 11 beta-hydroxyandrostenedione and 11 beta-hydroxytestosterone from each substrate. Generally, lesser amounts of C19 11-oxosteroids were noted also. Definitive confirmation was made by gas chromatography - mass spectrometry for 11 beta-hydroxyandrostenedione in the media.


Characterization and localization of large sulfated glycoproteins in the extracellular matrix of the developing asteroid Pisaster ochraceus.

In this study techniques commonly used to extract and purify proteoglycans of vertebrates were applied to the embryo of the asteroid Pisaster ochraceus at the early bipinnaria larva stage, a stage in which extensive cell migration is occurring within the extracellular matrix of the blastocoel. Several large sulfated glycoproteins were isolated and shown to consist of protein cores covalently bound to sulfated polysaccharide chains. The polysaccharide chains consisted primarily of neutral sugars and were not susceptible to glycosaminoglycan-degrading enzymes, suggesting that these were not glycosaminoglycans. The sulfated glycoproteins could be fractionated by electrophoresis on sodium dodecyl sulfate-agarose-acrylamide composite gels. Two types of monoclonal antibodies prepared against isolated extracellular matrix of these embryos reacted with two subsets of bands on Western blots of the composite gels. Staining of sections of the embryos with the antibodies showed that the epitopes that they recognized were located throughout the extracellular matrices of the embryos. That these high molecular weight glycoproteins were located within the extracellular matrix of the embryos suggests that they may be involved in the control of morphogenesis and cellular movement.


Control of O-glycan synthesis: specificity and inhibition of O-glycan core 1 UDP-galactose:N-acetylgalactosamine-alpha-R beta 3-galactosyltransferase from rat liver.

The specificity of glycosyltransferases is a major control factor in the biosynthesis of O-glycans. The enzyme that synthesizes O-glycan core 1, i.e., UDP-galactose:N-acetylgalactosamine-alpha-R beta 3-galactosyltransferase (beta 3-Gal-T; EC 2.4.1.122), was partially purified from rat liver. The enzyme preparation, free of pyrophosphatases, beta 4-galactosyltransferase, beta-galactosidase, and N-acetylglucosaminyltransferase I, was used to study the specificity and inhibition of the beta 3-Gal-T. beta 3-Gal-T activity is sensitive to changes in the R-group of the GalNAc alpha-R acceptor substrate and is stimulated when the R-group is a peptide or an aromatic group. Derivatives of GalNAc alpha-benzyl were synthesized and tested as potential substrates and inhibitors. Removal or substitution of the 3-hydroxyl or removal of the 4-hydroxyl of GalNAc abolished beta 3-Gal-T activity. Compounds with modifications of the 3- or 4-hydroxyl of GalNAc alpha-benzyl did not show significant inhibition. Removal or substitution of the 6-hydroxyl of GalNAc reduced activity slightly and these derivatives acted as competitive substrates. derivatives with epoxide groups attached to the 6-position of GalNAc acted as substrates and not as inhibitors, with the exception of the photosensitive 6-O-(4,4-azo)pentyl-GalNAc alpha-benzyl, which inhibited Gal incorporation into GalNAc alpha-benzyl. The results indicate that the enzyme does not require the 6-hydroxyl of GalNAc, but needs the 3- and the axial 4-hydroxyl as essential requirements for binding and activity. In the usual biochemical O-glycan pathway, core 2 (GlcNAc beta 6[Gal beta 3] GalNAc alpha-) is formed from core 1 (Gal beta 3GalNAc-R). We have now demonstrated an alternate pathway that may be of importance in human tissues.


[Spectrum analysis of the r-r interval using a PC].

Spectral analysis of r-r variability has been recently proposed as a clinical tool to assess the autonomic nervous system function. In this article we present the results obtained using an equipment and an analysis software (based on Maximum Entropy Method) developed in our laboratory. Analyzing the tachograms derived from prolonged ECG registrations of 12 young healthy subjects, 24 to 36 years old (mean 31 +/- 4), we observed the two classic components of the signal: a low frequency component (0.7 +/- 0.2 Hz) and a high frequency component (0.21 +/- 16.6 Hz). As expected, standing, a simple manoeuvre augmenting sympathetic activity, caused a stronger predominance of the low frequency component. We conclude that our method is reliable to evaluate, by means of spectral analysis, rhythmical oscillations of r-r variability.


[Effect of insulin-like growth factor I (IGF-I) on Sertoli cell metabolism in the pubescent rat].

The influence of Insulin-like Growth Factor I (IGF-I) on some metabolic functions of Sertoli cells from peripubertal rats was investigated. Sertoli cells were isolated from the testes of 24-day-old animals and cultured at 32 degrees C in Eagle's MEM with or without 1 nM IGF-I. Sertoli cells cultured in the presence of IGF-I showed increased nuclear RNA polymerase activity (+80%) and augmented protein synthesis (+50%).


[Chemoprevention of genotoxic damage in lung cells of rats exposed to cigarette smoke].

Male Sprague-Dawley rats were exposed to cigarette smoke (CS) according to a complex protocol which lasted 40 days. Some of the groups were pre-treated with N-acetyl-L-cysteine (NAC) by gavage (1 g/kg b.w.) 5 h before each exposure. Bronchoalveolar lavage was performed in each animal at the end of each exposure period in order to recover pulmonary alveolar macrophages (PAM). Cells were identified and counted under the microscope, and the number of micronucleated (MN) and binucleated (BN) PAM was registered. The results showed an increase in the number of MN PAM, which was already evident after 8 days of CS exposure; this increase remained constant after 28 and 40 days of exposure. A significant decrease in the number of MN PAM was observed in the animals pre-treated with NAC. BN PAM were significantly increased after 28 and 40 days of exposure; again, a slight yet not significant decrease was detected in NAC-pre-treated animals. On the whole, this study demonstrates that CS is clearly clastogenic to alveolar macrophages and that NAC can efficiently prevent this cytogenetic damage.


[Facilitating and inhibiting effects of the orientation of attention using peripheral cues].

The attentional field resulting from the presentation of peripheral cues; which were either informative or non informative about the position of the imperative stimulus, was studied. Different time intervals between cue and stimulus (120, 300, 600 ms) were used. The results showed facilitation of the response with the informative cue and inhibition with the non informative cue. This happened for the longest cue-stimulus intervals and when the position of the cue and the position of the stimulus were congruent. Also order of cue presentation (i.e., either informative followed by the non informative cue or vice versa) proved important in producing facilitatory and inhibitory effects.


[Morphological characteristics of mastocytes in the infertile human testicle].

Mast cells, basophilic elements of the connective tissue, have been studied in numerous researches carried out both in man and animals. Previous studies showed that mast cells have been found in increased number in testis affected by pathology. In the present research morphologic characteristics of the mast cells in human infertile testis have been studied. Testicular biopsies obtained from 49 subjects, aged from 21 to 61 years, have been treated according to the current techniques for transmission electron microscopy. Results showed that it is possible to distinguish at least two types of mast cells: 1) "Interstitial" mast cells, large, round shaped, with great content of large characteristic granules, in relation with the loose connective tissue surrounding capillary vessels and Leydig cells; 2) "Peritubular" mast cells, flattened and relatively poor in granules, trapped in the conspicuous peritubular collagenic layers. These two types of mast cells differ from one another as concerns not only localization, but also general morphological characteristics and number and ultrastructure of cytoplasmic granules. Probably it is possible to establish a relationship between peritubular mast cells trapping and the increased deposition of fibrous connective tissue in the peritubular layers; such a deposition characterizes a lot of testicular pathologies.


Calcium oxalate smear layer: mineralogical and crystallographic study.

Different methods to reduce dentine hypersensitivity have been tested by means of oxalate treatments. Natural material (20 teeth) and commercial oxalates have been employed. As a result the dentinal tubules are filled by small crystals preventing dental disease. Permeability measurements have been performed in order to quantify results. The composition of the synthetic smear layer produced after oxalate treatment have been investigated by powder diffraction method suggesting calcium oxalate formation.


Nonhuman primate models for HIV vaccine development.

Animal models will be essential in developing a vaccine to protect against human immunodeficiency virus (HIV) infection. While HIV-1 infects great apes, it induces no disease in these species. Some simian immunodeficiency virus (SIV) isolates cause an AIDS-like disease in macaque monkeys. The SIV/macaque model has proven a valuable system for exploring AIDS pathogenesis and assessing strategies for HIV vaccination. Studies in nonhuman primates have shown that a variety of vaccine approaches transiently elicit immunity which can protect against a very small intravenous challenge of cell-free virus. Longstanding immunity which protects against cell-associated or mucosal virus challenge has not yet been achieved. Nonhuman primate models will be essential for achieving this goal.


Extraplacental human fetal tissues express mRNA transcripts encoding the human chorionic gonadotropin-beta subunit protein.

The glycoprotein hormone human chorionic gonadotropin (hCG) is synthesized in large quantities by the developing placenta, reaching peak concentrations in maternal blood during the late first trimester and early midtrimester of pregnancy. In general it is believed that the alpha-subunit of this dimeric hormone is expressed in pituitary gonadotropes, thyrotropes, and trophoblasts, while the beta-subunit is expressed exclusively by trophoblasts. Studies from our laboratory and other laboratories have shown that some midtrimester human fetal tissues, in addition to the placenta, can synthesize proteins that appear to be very similar to the beta-subunit of hCG. To define precisely the nature of this putative hCG-beta-subunit in extraplacental fetal tissues, we have examined the mRNA from a variety of human fetal and adult tissues using nucleic acid hybridization and reverse transcription-polymerase chain reaction (PCR) methods. Our results demonstrate that midtrimester fetal kidney and adrenal tissues contain hCG-beta mRNA transcripts at concentrations comparable to that of placenta, while fetal lung, brain, muscle, and adult adrenal contain only trace to undetectable levels of hCG-beta mRNA. By restriction endonuclease mapping of PCR fragments from fetal tissue cDNAs, we show that the hCG-beta transcript expressed in midtrimester human fetal organs is a bone fide copy of hCG-beta gene No. 5 of the beta-subunit gene family located on chromosome 19.


Characterization of low Mr zona pellucida binding proteins from boar spermatozoa and seminal plasma.

A group of low Mr (16 kDa-23 kDa) glycoproteins on ejaculated boar spermatozoa have been shown to have high affinity for homologous zona pellucida glycoproteins (ZPGPs). These ZPGP binding proteins are derived from seminal plasma as shown by their absence from epididymal spermatozoa and their presence in seminal plasma as identified by N-terminal amino acid sequence analysis. They bind to ZPGPs by a polysulphate recognition mechanism similar to that found for proacrosin-ZPGP interactions. The haemagglutination activity of boar seminal plasma is also associated with these low Mr glycoproteins. It is suggested that they play a role in regulating the rate of sperm capacitation and survival in the female reproductive tract.


Purification and characterization of vitellogenin and lipovitellins of the sand crab Emerita asiatica: molecular aspects of crab yolk proteins.

In the mole crab Emerita asiatica, the main yolk proteins consist of two slow moving lipovitellins (Lv I and Lv II) of glycolipoprotein nature. Lv I cleaves into subunits (MW: 109,000 and 105,000) and Lv II gives rise to six subunits (MW: 65,000, 54,000, 50,000, 47,000, 44,000, and 42,000) in SDS-PAGE (with beta-mercaptoethanol). In order to observe the stability of Lv II as well as to achieve better resolution of the proteins, two different buffer systems (Phosphate buffered saline and tris-buffered saline), 40% sucrose, and glass distilled water were used as homogenizing media. Among them, better resolution was achieved with tris-buffered saline and 40% sucrose, and tris-buffered saline seems to be the ideal medium for elution of Lv II. The analysis of biochemical constituents of the major Lv II reveals a percentage composition of 69.325, 27.927, and 2.753 respectively for protein, lipid, and bound sugars. In the I stage embryo, protein comprises about 67.276%, lipid 29.65%, and bound sugars 3.015%. Vitellogenin (Vg) electrophoretically corresponding to the Lv I and Lv II was present in the female haemolymph during the entire period of embryogenesis. The number of subunits (8) of Vg in all stages remained unaltered and their approximate molecular weights were Vg1, 91,000; Vg2, 87,000; Vg3, 83,000; Vg4, 61,000; Vg5, 58,000; Vg6, 45,000; Vg7, 42,000; and Vg8, 38,000. Different proteins present in the embryos (I and IV stage) and the serum obtained from the animal carrying the I stage embryo were separated by gel-filtration in high performance liquid chromatography (HPLC). Sephadex (G-200) gel filtration chromatography was used to purify the Lv II in large quantity. Total lipid extracted from Lv II as well as the embryos belonging to different stages of development were separated into their constituent neutral, glycolipids, and phospholipids, using silicic acid column chromatography. Thin layer chromatography (TLC) was used to isolate the different phospholipids purified from various stages of embryos and Lv II. As many as seven different phospholipids were separated from Lv II and I and IX stage embryos; and whereas thin layer chromatogram of V and VI stage embryos showed six different phospholipids, embryos of VII and VIII stage contained four phospholipid species. Cholesterol, glycolipids, and individual phospholipids isolated from the Lv II and I stage embryo were quantified spectrophotometrically and the results were discussed.


Reinvestigation of the ultrastructure of spermiogenesis and the spermatozoon of Hymenolepis nana (Cestoda, Cyclophyllidea), parasite of the small intestine of Rattus rattus.

Spermiogenesis in Hymenolepsis nana begins with the formation of a differentiation zone. This is limited at the front by arched membranes, is surrounded by cortical microtubules associated with 12 crested-like bodies, and contains a single centriole made up of doublets. The distal centriole gives rise to a flagellum that grows at the same pace as the cortical microtubules. Migration of the nucleus takes place after the formation of the flagellum. It is followed by the separation of the old spermatid from the residual cytoplasm. The mature H. nana spermatozoon is filiform and lacks mitochondria. The axoneme, of the 9 + "1" pattern of the Platyhelminthes, does not reach the extremities of the spermatozoon. The nucleus is electron dense and is in close contact with the axoneme around which it coils in a spiral making an angle of 10 degrees to 15 degrees with the spermatozoon axis. The cortical microtubules follow a 10 degrees to 15 degrees helicoidal path along almost their whole length, except at their posterior extremity, where they are parallel to the spermatozoon axis. H. nana is distinguished by the early development of 12 crested-like bodies of different lengths and by the existence of a single centriole in the differentiation zone. Such a high number of crested-like bodies had never previously been reported in a cestode.


Recovery of penetration ability in protease-treated zona-free mouse eggs occurs coincident with recovery of a cell surface 94 kD protein.

Previous studies have demonstrated that protease treatment of zona-free mouse eggs impairs sperm-egg interaction (Boldt et al.: Biol Reprod 39:19-27, 1988) and causes modification of a 94 kD egg plasma membrane protein (Boldt et al., Gamete Res 23:91-101, 1989). In this report, the ability of eggs to recover penetration ability following protease treatment was examined. Zona-free mouse eggs were isolated and treated with either trypsin or chymotrypsin (1 mg/ml, 20 min), then cultured for 0, 3, or 6 hr before insemination. Eggs cultured for 3 or 6 hr displayed significantly higher penetration levels than eggs inseminated immediately after protease treatment, indicating a recovery of penetration ability during the 3 or 6 hr incubation period. The recovery of penetration ability was not blocked by inclusion of cyclohexamide (50 micrograms/ml) during the 3 or 6 hr culture period, indicating that protein synthesis was not required for recovery of fusion ability. Cell surface radiolabeling studies with 125I revealed that a 94 kD cell surface protein was lost immediately following trypsin or chymotrypsin treatment but was found on the egg surface after the 3 or 6 hr recovery period. Recovery of the 94 kD egg surface protein occurred in the presence of cyclohexamide, and metabolic radiolabeling studies with 35S-methionine confirmed that synthesis of a 94 kD protein was blocked by cyclohexamide. These results suggest that the recovery of penetration ability after protease treatment of zona-free eggs is due to recovery of the 94 kD cell surface protein, providing further evidence for the involvement of the 94 kD protein in sperm-egg interaction.


The kinetics of oocyte activation and polar body formation in bovine embryo clones.

The kinetics of polar body formation were examined in parthenogenetically activated, in vitro matured and aged bovine oocytes. Subsequently, the presence or absence of polar body formation was determined in bovine embryo clones. Polar body formation, defined as telophase II, occurred by 1 (13/40, 43%) and 2 h (15/21, 71%) postparthenogenetic activation of metaphase II stage oocytes. Parthenogenetically activated oocytes readily formed pronuclei by 4 h. Some oocytes had chromatin in a highly condensed state at 1, 2, and 4 h postactivation (13/72, 18%). These oocytes often (10/13, 77%) appeared to be "self-enucleated," as the condensed chromatin was found in a membrane-bound extrusion. The phenomenon was most prevalent when oocytes were handled at room temperature (25-27 degrees C). Nuclear transfer procedures were established to bring about synchronous blastomere fusion and oocyte activation conditions. Synchronous conditions were achieved only when oocytes were handled and manipulated at 37-39 degrees C. Embryo clones examined 2 h postfusion did not form a polar body. Conversely, nucleate demi-oocyte controls were at the late telophase II stage of meiosis. The results are discussed in relation to cell cycle effects on bovine nuclear transfer.


Effects of protein synthesis on maturation, sperm penetration, and pronuclear development in porcine oocytes.

In vitro matured porcine oocytes were used to test the importance of protein synthesis for sperm penetration, the second meiotic division, and pronuclear development. Experiments were carried out to measure rates of protein synthesis in the presence of protein synthesis inhibitors (35 microM or 350 microM cycloheximide or a combination of inhibitors) (study 1); to test for sperm penetration and pronuclear development when protein synthesis was inhibited during fertilization (study 2); to test for oocyte meiosis, sperm penetration, and female and male pronuclear development when protein synthesis was inhibited during maturation (oocyte maturation in vitro with addition of inhibitor at 0, 24, or 36 hr of culture) (study 3); and to analyze the changes in the pattern of protein synthesis during these phases. Sperm penetration, oocyte meiosis, and female pronuclear development were not affected by the total inhibition of protein synthesis during fertilization. By contrast, inhibiting protein synthesis during maturation severely impaired the completion of meiosis and pronuclear development. Although inhibition of protein synthesis after 36 hr of maturation culture did not totally block male pronuclear development (MPN), the rate of MPN formation was lower than for controls (52% vs. 72%, P less than 0.05). However, protein synthesis was absolutely essential between 24 and 36 hr for the formation of MPN after decondensation. This period of maturation coincided with the dominant phase of protein reprogramming in the oocyte.


Cauda epididymal sperm interactions with seminal vesicle fluid.

The interaction of rat cauda epididymal sperm cAMP-dependent protein kinase (PKA) with seminal vesicle fluid (SVF) proteins was examined. Specific proteins in SVF act as substrates for the sperm cell PKA. The apparent molecular weights of these proteins are 45.0, 31.5, 17.2, 14.7, and 13.3 kDa. The phosphorylation of one low-molecular-weight cauda sperm protein is blocked in the presence of SVF. There is no PKA enzyme activity in SVF. The presence of phosphate transfer activity between the sperm cell enzyme and the SVF proteins is species dependent. For example, mouse and rat SVF proteins are efficient phosphate acceptors, but there is no phosphorylation activity when hamster SVF is used as the enzyme substrate. The sperm cell samples were also assessed for membrane integrity. Specifically, cauda sperm cells used in these assays were judged to be intact when examined microscopically using the fluorescent vital dye carboxyfluorodiacetate. Although there was enzyme activity in the supernatants of the rat sperm cell samples, in the protein kinase assay it required three times as much supernatant volume (compared with intact cell sample volume) to measure the activity. Supernatant enzyme activity did not increase with washing, indicating that the cells were not damaged by this procedure. The enzyme itself does not adhere to the sperm cells, so the PKA enzyme activity is most likely oriented on the external surface of the sperm cell.


Functional properties of caltrin proteins from seminal vesicle of the guinea pig.

Caltrin proteins from seminal vesicle content of the guinea pig bind with great specificity to different regions of the spermatozoa. Indirect immunofluorescence studies with polyclonal antibodies showed that caltrin I binds to the head, on the acrosomal cup, while caltrin II binds on the principal tail and the neck. No fluorescence was detected either in the midpiece or in the post-acrosomal area of the head when sperm were exposed to either of the caltrins. Calcium-induced hyaluronidase release, which occurs during the acrosomal reaction, was dramatically inhibited by caltrin I (approximately 85% inhibition). Caltrin II was less effective in preventing the enzyme release (approximately 50% inhibition). Chemical modification of the structure modified the biological activity of the two caltrins. Reduction and carboxymethylation of the cysteine residues diminished the inhibitory activity on 45Ca2+ uptake and reduced the ability of the proteins to react with their antibodies. Removal of the carbohydrate portion by chemical deglycosylation transformed the inhibitor proteins into enhancers of calcium uptake into the spermatozoa. Caltrin proteins from the guinea pig appear to play the same physiological role as bovine caltrin, regulating specifically calcium transport across the spermatozoal membranes related with the acrosome reaction and hyperactivation process. The dual behavior of caltrins to inhibit or enhance Ca2+ uptake enables them to fulfill this function. Nevertheless, molecular mechanisms different from those described for bovine caltrin seem to be involved in the control of the functional activity of the guinea pig caltrins.


Subzonal insemination of a single mouse spermatozoon with a personal computer-controlled micromanipulation system.

A personal computer-controlled micromanipulation system was developed for automatic injection of spermatozoa into the perivitelline space of mouse ova. A pair of three-dimensional hydraulic micromanipulators driven by pulse motors was used for this automatic system. The pulse signals that regulate the motors are initiated by the computer program, and these signals cause the micromanipulator to move the microtool precisely. The computer program was designed to perform the most effective movements of the sperm injection needle used during manual micromanipulation. Prior to the manipulation, the computer locates the tip of the injection needle and the end of the egg-holding pipette in the microscope field using image processing. The trajectory of the injection needle is determined according to these initial positions. Using this robotic system, subzonal insemination with a single mouse spermatozoon was attempted in a total of 143 ova. The sperm insertion was successfully completed in all cases without damaging any of the ova. Spermatozoa treated with ionophore A23187 and those without the treatment were used. The fertilization rate (68.8%) of the ova inseminated with treated sperm was significantly higher than that (37.5%) obtained with the nontreated sperm (P less than 0.05). These findings suggest the feasibility and potential for further applications of a robotic microinsemination system and, in addition, that a higher fertility rate in the subzonal insemination of mouse ova can be achieved with the ionophore treatment of spermatozoa.


Structural organization of surface domains of sperm mitochondria.

Sperm mitochondria are assembled into an organized sheath surrounding the outer dense fibers and axoneme of the flagellar midpiece. Each mitochondrion is arranged so that its surface faces three different cellular organelles including the plasma membrane, neighboring mitochondria and the outer dense fiber-axoneme complex. In this manuscript we present data on structural differentiations of these three different surface domains of the outer mitochondrial membrane. We demonstrate that the apposed surfaces of adjacent mitochondria are joined by a two dimensional network of studs unique to this domain. By contrast, the surface domain facing the outer dense fiber-axoneme complex exhibits a different, but highly ordered structural organization, evident as a parcrystalline network of parallel stripes; this domain is further distinguished by its exclusive association with a midpiece-specific cytoskeletal complex. These differentiations are not seen on the surface domain of mitochondria which faces the plasma membrane. The implications of the mosaic composition of the outer mitochondrial membrane in the assembly and function of the mitochondrial sheath are discussed.


F-actin in acrosome-reacted boar spermatozoa.

Biochemical and immunoelectron microscopic methods have been used to analyze the distribution of actin in boar spermatozoa and its state of aggregation before and after acrosome reaction. F-actin was detected on sperm head and tail by electron microscopy using an improved phalloidin probe: incubation with a fluorescein-phalloidin complex and an anti-fluorescein antibody, followed by labeling with protein A-gold complex. Gold particles, indicating the presence of F-actin, were localized on the sperm surface of the acrosome-reacted spermatozoa. Specific labeling was localized (1) between the outer acrosomal membrane and the plasma membrane in the equatorial region, (2) between the outer surface of the fibrous sheath and the plasma membrane in the postacrosomal region, (3) around the connecting piece and the neck region, and (4) on the external surface of the fibrous sheath in the principal piece of the tail. Furthermore, after NP-40 extraction, the SDS-PAGE revealed a difference in solubility between reacted and unreacted boar spermatozoa, reflecting actin polymerization. We conclude that most actin in the acrosome reacted boar spermatozoa is polymeric.


Multiplane transesophageal echocardiography: latest evolution in an imaging revolution.

Multiplane imaging with a rotating phased-array transducer from within the esophagus represents the latest development in transesophageal cardiac ultrasound. Transverse, longitudinal, and all possible intermediate oblique planes are easily obtained from the same transducer with minimal probe manipulation. Three-dimensional conceptualization of complex structures and pathologic conditions is facilitated. The major advantages are a simplified examination procedure and much less patient discomfort than monoplane and biplane probe imaging.


The role of transesophageal echocardiography in critical care: UCSF experience.

The applications of transesophageal echocardiography (TEE) now encompass the arena of critical care. We reviewed the indications, findings, and outcome of TEE in the critical care setting in a tertiary care hospital and report our experience. Eighty-three transesophageal echocardiograms were performed in 69 critically ill patients for the following indications: endocarditis (43%), embolic source (13%), hypotension (10%), mitral regurgitation (10%), left ventricular function (6%), aortic dissection (4%), prosthetic valve dissection (4%), and other (10%). Findings were unexpected in 21 of 83 studies and led to a change in management in 17%. On the basis of the results of TEE, 22% of patients had further evaluation by a more invasive technique, and 19% of patients had surgical intervention without further study. No significant complications attributable to TEE were noted. We conclude that TEE is a safe and useful diagnostic technique with fairly broad applications in the critical care setting.


Sensory blockade for difficult passage of transesophageal echocardiography probes.

During transesophageal echocardiography probe passage, airway reflexes are usually obtunded with topical local anesthetics. This technique meets with varying degrees of success. Even partially intact airway reflexes result in coughing, retching, and withdrawal, which may prevent transesophageal echocardiography examination or predispose to life-threatening tachycardia and hypertension. Proper preparation of the patient enhances comfort and helps protect against tachycardia and hypertension as well as reducing the time required for examination. This article outlines specific monitoring issues and offers precautions that are critical to transesophageal echocardiography probe passage. It suggests a premedication regimen and describes methods of sedation. Furthermore, it outlines the afferent innervation of the upper airway and specific techniques of temporary reflex interruption.


Intravascular ultrasound imaging of blood: the effect of hematocrit and flow on backscatter.

This study evaluates two key parameters influencing the ultrasonic backscatter from blood--hematocrit and flow rate--at 30 MHz in an in vitro flow system. A range of hematocrits from 0 to 50% was studied at a constant flow rate; various flow rates between stagnation and physiologic levels were studied at a constant hematocrit. The relation between backscatter intensity and hematocrit was a convex function with a maximum between a hematocrit of 10% and 20%. In the flow rate studies, the blood backscatter intensity was a maximum at a flow rate of 0 and rapidly decreased at higher flow rates. These in vitro results suggest that blood backscatter intensity is minimally dependent on hematocrit in the physiologic range. However, a dramatic increase in backscatter intensity occurs with stagnant flow, presumably the result of red blood cell aggregation. Clinically, blood backscatter intensity may provide an index for risk of thrombus formation.


Two-dimensional color Doppler flow velocity profiles can be time corrected with an external ECG-delay device.

Although two-dimensional ultrasound color flow imaging is often considered to be a real-time technique, the acquisition time for two-dimensional color images may be up to 200 msec. Time correction is therefore necessary to obtain correct flow velocity profiles. We have developed a time-correction method in which a specially designed unit detects the QRS complex from the patient and creates a trig pulse that is delayed incrementally in relation to the QRS complex. This trig pulse controls the acquisition of the ultrasound images. A number of consecutively delayed images, with known incremental delay between the sweeps, can thus be stored in the memory of the echocardiograph and transferred digitally to a computer. The time-corrected flow velocity profile is obtained by interpolation of data from the time-delayed profiles. The system was evaluated in a Doppler string phantom test. With this technique it is possible to study time-corrected flow velocity profiles without the need to alter existing ultrasound Doppler equipment.


Beat-by-beat aortic valve area measurements indicate constant orifice area in aortic stenosis: analysis of Doppler data with varying RR intervals.

Gorlin formula calculation of aortic valve area suggests that orifice area increases in patients with aortic stenosis with rising cardiac output. Evidence that aortic orifice area varies was sought in patients with aortic stenosis by analyzing Doppler data beat by beat versus RR interval in 22 patients with spontaneous RR variability. Stroke volume increased in all patients from minimum to maximum RR interval by 129% +/- 19%. Over the same range of RR intervals, assessment of aortic valve area by (A) simultaneous inner and outer continuous wave Doppler signals, or (B) nonsimultaneous RR-matched pulsed wave Doppler from the left ventricular outflow tract and continuous wave Doppler from the aortic valve failed to suggest an increase in aortic valve area. A positive relationship between aortic valve area and RR interval was not consistently observed with the exception of seven out of eight patients with mild to moderate (pulsed wave Doppler/continuous wave Doppler time velocity integral ratio of 0.3 to 0.7) aortic stenosis (p less than 0.05). Beat-by-beat measurements of aortic valve orifice area using Doppler techniques do not suggest that the aortic stenosis orifice varies over a wide range of RR intervals and stroke volumes.


Utility of transesophageal echocardiography in the diagnosis of aortic conduit endocarditis in patients who have undergone the Cabrol procedure.

The Cabrol procedure is being increasingly performed in patients with aortic pathologic conditions. Transesophageal echocardiography can help in the prompt diagnosis of endocarditis in these patients. In this report we present transesophageal echocardiographic features of two patients in whom endocarditis developed after they underwent the Cabrol procedure for ascending aortic aneurysm caused by Marfan's syndrome.


Variation in the color Doppler area of a regurgitant jet with changes in the absolute chamber pressure: an in vitro study.

The color Doppler appearance of a regurgitant jet depends on jet momentum, determined in part by the pressure difference between the two chambers. However, it is not clear if absolute chamber pressure has an independent effect on jet area. To test this question, an in vitro experiment was performed in which dynamically decaying jets were created with identical initial pressure gradients but five different levels of absolute chamber pressures. At every level of chamber pressure, color Doppler images were recorded with two different transducers (3.5 and 5.0 MHz) and jet areas were measured at four different flow rates (0 to 9.9 cm3/sec). A multilinear regression model was created with jet area as the dependent variable and jet flow rate, transducer frequency, and absolute chamber pressure as independent parameters. Jet area was most strongly predicted by flow rate (univariate r = 0.90) and transducer frequency (r = 0.32). Even after adjusting for these effects, however, a small but significant (p less than 0.0001) effect of absolute chamber pressure on jet area was seen with jet area rising by 0.89 cm2 for each 10 mm Hg increase in absolute chamber pressure (multivariate r = 0.96, p less than 0.0001). We conclude that the color Doppler area of a regurgitant jet is dependent not only on the relative pressure and flow between the two chambers but also on the absolute chamber pressure.


Diagnosis of tumor embolus to the pulmonary artery by transesophageal echocardiography.

This article describes a patient with renal cell carcinoma and massive tumor extension into the inferior vena cava who experienced sudden tachypnea and subsequent cardiopulmonary arrest. Transesophageal echocardiography was used to diagnose a massive tumor embolus to the pulmonary artery that was subsequently confirmed at autopsy. Transesophageal echocardiography provided a rapid bedside diagnosis of this unusual entity in an acutely ill patient. This technique may prove more widely applicable in the evaluation of patients with suspected pulmonary embolus, whether neoplastic or thromboembolic.


Misplaced Greenfield filter: diagnosis by transesophageal echocardiography.

A case of accidental placement of a Greenfield filter in the right atrium is described. Transthoracic echocardiography demonstrated the filter but was unable to provide details regarding the exact location of the filter. Transesophageal echocardiography showed the filter to be fixed to the tricuspid anulus. This finding, which was confirmed during surgery, was used to guide the subsequent surgical management.


On the process for evaluating proposed new diagnostic EEG tests.

Assessment of new technology is an important part of the evolving art of medicine. For a variety of reasons, it is appropriate to restrict clinical applications of new technology to those tests which have been shown to be safe and effective. Efficacy of new EEG technology can be assessed through a variety of standard procedures, generally based on controlled, well organized clinical studies. Scientific reports using new EEG technology all too often fail to meet the standards traditionally expected for such clinical trials. Many such studies were never designed to be clinical trials. Misunderstandings occur when reports of scientific studies and informal clinical series become confused with formal clinical trials of efficacy. Based upon examples given during the 1991 ISBET meeting symposium on discriminant analysis, examples are discussed regarding how well individual kinds of presentations can be used to help clarify the generic clinical efficacy of the presented diagnostic tests.


Embolic occlusion of a patent foramen ovale: a cause of false negative contrast echocardiogram.

A 74-year-old man who had dyspnea and cyanosis demonstrated venous thrombosis and multiple lung ventilation-perfusion mismatches suggesting pulmonary emboli. Transthoracic echocardiography demonstrated a right atrial mass and a microbubble contrast study did not reveal right-to-left shunt. Transesophageal echocardiography revealed an embolus entrapped in the foramen ovale projecting into the left atrium that was subsequently removed. Embolic occlusion of patent foramen ovale is an important cause of false negative microbubble contrast study.


Echocardiographic demonstration of an unusual pulmonary venous pathway in a patient with left ventricular inflow obstruction.

A newborn infant with mitral atresia and a restrictive patent foramen ovale had normal pulmonary venous connection to the left atrium demonstrated by echocardiography. However, pulmonary venous blood then coursed through an unusual pathway in the roof of the left atrium into the right atrium. This case illustrates the need for the echocardiographer to be reminded of and then search for unusual drainage pathways in cases of left ventricular inflow obstruction, even when the pulmonary veins connect normally.


QEEG profiles of psychiatric disorders.

While reports of EEG correlates of psychiatric disorders date back five decades, clinical sensitivity of the EEG to psychiatric disorders has been greatly enhanced with the advent of quantitative methods of analysis (QEEG). Using a QEEG methodology known as neurometrics we have identified distinctive electrophysiological profiles associated with different psychiatric disorders. With this method quantitative features are extracted from 2 minutes of artifact- free eyes closed resting EEG data, log transformed to obtain Gaussianity, age-regressed, and Z-transformed relative to population norms. Using small subsets of neurometric features, multiple stepwise discriminant analyses were used to construct mathematical classifier functions, the values of which are different for members of different a priori defined diagnostic groups. Using this approach, we have demonstrated high discriminant accuracy in independent replications separating many populations of psychiatric patients from normal as well as from each other, including major affective disorder, schizophrenia, dementia, alcoholism, and learning disabilities, as well as high accuracy of discrimination between known subtypes of depression (unipolar vs bipolar). The use of classification accuracy curves (CACs) which allow one to assess the sensitivity and specificity achieved by the discriminant functions is discussed. In addition, using cluster analysis, neurometric subtypes can be identified in several clinically homogenous populations. Preliminary results suggest that baseline membership in some neurometric subtypes may be highly correlated with response to treatment.


QEEG in a public health system.

For the past decade the Cuban Neuroscience Center has organized on behalf of the Ministry of Public Health of the Republic, a nationwide Program for the introduction of quantitative EEG (qEEG). This Program has involved a) development of standardized equipment for "paperless" EEG, qEEG and brain topography; b) establishment of a network of 21 laboratories of clinical neurophysiology; c) creation of the specialty of clinical neurophysiology which trains physicians from all provinces in both traditional and quantitative electrophysiological methods; d) introduction of standardized protocols for the collection of clinical and electrophysiological information; e) organization of a national normative and neuropsychiatric database; f) establishment of normative regression equations. Among the special issues discussed are: 1) relationship between traditional and quantitative methods; 2) evaluation of the effectiveness of the technology introduced; 3) use of qEEG in the early detection of brain dysfunction.


Aortopulmonary fistula detected by echocardiography 12 years after aortoplasty for correction of ascending aorta aneurysm.

We describe a case of aortopulmonary fistula in which the correct diagnosis was made by transthoracic echocardiography. The transesophageal approach, because of severe aortic dilatation, failed to provide the correct diagnosis, underlining the importance of complete transthoracic and transesophageal studies in the evaluation of aortic aneurysms.


Measurement processes and spatial principal components analysis.

Spatial principal components analysis (SPCA) applied to the ongoing EEG yields factor loadings which, when mapped, consistently reveal symmetrical patterns resembling the spherical harmonics. In this paper, we consider the mechanisms responsible for these characteristic patterns. In doing so, we demonstrate that volume conduction is one of a family of processes capable of generating such patterns with SPCA. It is shown that any series of measurements on a sphere in which the covariance is only a function of measurement site angular separation (shift invariant processes) will yield the spherical harmonics as the eigenvectors or factor loadings of the covariance matrix. Simulations further indicate that this effect is robust and not determined by the geometry of the measurement sites. In situations where shift invariant signals coexist with those generated at specific sites (anatomically specific processes), such as evoked potentials and some artifacts, it is shown that the anatomically specific signals do not influence the eigenvectors of the covariance matrix in a uniform or random fashion. The factors most influenced are those whose symmetry is similar to that of the site specific signal.


Subclavian artery stenosis masquerading as prosthetic aortic stenosis: a potential source of confusion in Doppler evaluation of aortic valve disease.

In this case report we describe a patient with a prosthetic aortic valve in whom a high-velocity signal from a right subclavian artery stenosis initially was mistaken for the aortic jet signal. Differences in the shapes of the jets obtained from an apical and right supraclavicular position suggested different origins of these two high-velocity systolic signals. Correct identification of the origin of each signal was possible with pulsed Doppler recordings of the subclavian artery and high pulse-repetition-frequency pulsed Doppler interrogation of the aortic valve.


A five-chambered heart: rare coronary artery anomaly with fistula draining into a separate interventricular cavity.

The combination of color flow Doppler imaging, contrast echocardiography, magnetic resonance imaging, and coronary arteriography documented the presence of a right coronary artery fistula that opened into a separate cavity in the anterior part of the interventricular septum in a 32-year-old man. Although a traumatic origin could not be excluded, the abnormality was most likely a congenital anomaly. Because there were no significant symptoms and no signs of hemodynamic burden on the heart, a conservative management was decided and no complications occurred during 3 years of follow-up. So far the literature includes only one similar case described by autopsy findings.


Neurometrics in cerebral ischemia and uremic encephalopathy.

The neurometric method as introduced by John was used to study three groups of patients with cerebral ischemia, three groups of patients with renal disease and an additional normal control group. The traditional neurometric approach was slightly modified: relative band power values were not expressed as a percentage of the total power per derivation but as a percentage of the "global power"; frequency matrices were used in addition to power matrices. From the study of the three groups of patients with one-sided supratentorial ischemia it appeared that sensitivity and specificity are completely satisfactory when using neurometrics in patients with severe ischemia in the middle cerebral artery territory studied within 48 hours of the onset of the stroke. However, in ischemia patients with less pronounced clinical signs and especially in patients without persistent neurological deficit the sensitivity is much lower. In studying dialysed and non-dialysed renal patients signs of an (often subclinical) encephalopathy could be detected in approximately 37% of all patients. Follow-up studies of the ischemia patients and the renal patients over a period of several years revealed a parallelism between clinical scores and qEEG scores in the ischemia patients; almost all qEEG improvement occurred in the first three months after the stroke. The qEEG profile of the groups of dialysed patients tended to be more or less stable over a period of several years.


Quantitative EEG in elderly depressives.

Thirty-one elderly depressive patients were evaluated with topographic, quantitative EEG using relative measures, absolute measures, and computations of interhemispheric asymmetry and interhemispheric coherence in four frequency bands: delta, theta, alpha and beta. Patients were found to have lower than normal delta, higher than normal theta, higher than normal alpha and lower than normal beta values. EEG values were greater over the left than the right hemisphere in theta, alpha and beta bands. Lower than normal anterior interhemispheric coherence was found in all four frequency bands.


Unrestricted principal components analysis of brain electrical activity: issues of data dimensionality, artifact, and utility.

Principal components analysis (PCA) was performed on the 1536 spectral and 2944 evoked potential (EP) variables generated by neurophysiologic paradigms including flash VER, click AER, and eyes open and closed spectral EEG from 202 healthy subjects aged 30 to 80. In each case data dimensionality of 1500 to 3000 was substantially reduced using PCA by magnitudes of 20 to over 200. Just 20 PCA factors accounted for 70% to 85% of the variance. Visual inspection of the topographic distribution of factor loading scores revealed complex loadings across multiple data dimensions (time-space and frequency-space). Forty-two non-artifactual factors were successful in classifying age, gender, and a separate group of 60 demented patients by linear discriminant analysis. Discrimination of age and gender primarily involved EP derived factors, whereas dementia primarily involved EEG derived factors. Thirty-eight artifactual factors were identified which, alone, could not discriminate age but were relatively successful in discriminating gender and dementia. The need to parsimoniously develop real neurophysiologic measures and to objectively exclude artifact are discussed. Unrestricted PCA is suggested as a step in this direction.


Frequency domain models of the EEG.

The structure of the normal resting EEG crosspectrum SVV(omega) is analyzed using complex multivariate statistics. Exploratory data analysis with Principal Component Analysis (PCA) is followed by hypothesis testing and computer simulations related to possible neural generators. The SVV(omega) of 211 normal individuals (ages 5 to 97) may be decomposed into two types of processes: the xi process with spatial isotropicity reflecting diffuse, correlated cortical generators with radial symmetry, and processes that seem to be generated by more spatially concentrated, correlated sources. The latter are reflected as spectral peaks such as the process. The eigenvectors of the xi process are the Spherical Harmonic Functions which explains the recurring pattern of maps characteristic of the spatial PCA of qEEG data. A new method for estimating sources in the frequency domain which fits dipoles to the whole crosspectrum is applied to explain the characteristics of the localized sources.


Subtyping of psychiatric patients by cluster analysis of QEEG.

We have previously reported successful classification of patients with a variety of psychiatric disorders, using multiple discriminant functions based upon selected neurometric QEEG variables. In independent replications, these functions accurately separate patients with different DSM-III-R diagnoses from one another and from normals. This capability demonstrates that distinctive and replicable patterns of neurometric abnormalities are correlated with the clinical symptom clusters upon which DSM-III-R diagnostic criteria are based. However, patients with the same clinical diagnoses often respond very differently to the same treatments. Similar symptoms may arise from different pathophysiology. This study explored the 'natural structure' of a population of psychiatric patients in 8 diagnostic categories, using uninformed cluster analysis based upon the same set of neurometric variables found useful in separating each of these categories from normal. This preliminary numerical taxonomic approach reveals that groups of patients in each of these DSM-III-R categories contain subtypes with markedly different pathophysiology; further, patients in different DSM-III-R categories were aggregated together within each cluster, displaying similar pathophysiological profiles. Objective classification based on such physiological measurements may add information useful to improve treatment outcomes.


Autoregulation of prostaglandin E2-induced Ca2+ influx in osteoblast-like cells: inhibition by self-induced activation of protein kinase C.

In cloned osteoblast-like MC3T3-E1 cells, prostaglandin E2 (PGE2) stimulated 45Ca2+ influx even in the presence of nifedipine, a Ca2+ antagonist that inhibits voltage-dependent Ca2+ channel, in a dose-dependent manner, attaining a maximum at 0.5 microM. Dose of PGE2 above 0.5 microM caused less than maximal stimulation. While PGE2 stimulated the formation of inositol trisphosphate dose dependently in the range between 1 nM and 10 microM. 12-O-Tetradecanoylphorbol-13-acetate (TPA), a protein kinase C (PKC)-activating phorbol ester, which by itself had little effect on 45Ca2+ influx, significantly suppressed the 45Ca2+ influx induced by PGE2 in a dose-dependent manner between 1 nM and 1 microM. 4 alpha-Phorbol 12,13-didecanoate, a phorbol ester which is inactive for PKC, showed little effect in this capacity. Staurosporine, a PKC inhibitor, enhanced the PGE2-induced 45Ca2+ influx. On the other hand, dibutyryl cAMP had little effect on the 45Ca2+ influx induced by PGE2. Our data suggest that PGE2 regulates Ca2+ influx through self-induced activation of PKC. These results indicate that there is an autoregulatory mechanism in signal transduction by PGE2, and PGE2 modulates osteoblast functions through the interaction between Ca2+ influx and phosphoinositide hydrolysis in osteoblast-like cells.


v-Src induces elevated levels of diglyceride by stimulation of phosphatidylcholine hydrolysis.

When Rat-1 cells bearing the ts LA29 mutant of Rous sarcoma virus (Rat1 LA29) are shifted from restrictive to permissive temperature, the pp60v-Src tyrosine kinase is activated and there is an increase in the cellular level of sn1,2-diacylglycerol (DRG) within 30 min which is not accompanied by increased inositol phospholipid hydrolysis. Temperature shift also increases the hydrolysis of phosphatidylcholine (PC), as determined by an increase in the generation of water soluble choline metabolites. Transphosphatidylation studies have shown that this occurs at least in part via a phospholipase D (PLD) catalysed pathway.


Mitogen stimulation of T-cells increases c-Fos and c-Jun protein levels, AP-1 binding and AP-1 transcriptional activity.

We have analysed the effect of mitogenic lectins on c-Fos and c-Jun protein levels as well as on activator protein-1 (AP-1) binding and enhancer activity in Jurkat T-cells. Both c-Fos and c-Jun protein levels were increased after Con A and PHA stimulation. Since T-cell stimulation increases both intracellular Ca2+ and cAMP levels and activates protein kinase C (PKC), the possible involvement of these intracellular messengers in c-Fos and c-Jun induction was tested. PMA, which directly activates PKC, mimicked the effect of the lectins on c-Fos and c-Jun, but elevation of either intracellular Ca2+ or cAMP levels had little or no effect. The mitogen-induced increase of c-Fos and c-Jun immunoreactivity was inhibited by H-7, a kinase inhibitor with relatively high specificity for PKC, and less efficiently by H-8, a structurally related kinase inhibitor less active on PKC, but more active on cyclic nucleotide-dependent kinases. Con A stimulation was found to increase both binding of AP-1 to the AP-1 consensus sequence, TRE, and AP-1 enhancer activity, in Jurkat cells. PMA was also found to increase the AP-1 enhancer activity, whereas elevation of Ca2+ or cAMP had only minor effects. We conclude that stimulation with mitogenic lectins is sufficient to increase both c-Fos and c-Jun protein levels, AP-1 binding and AP-1 enhancer activity in Jurkat cells and that they act via mechanisms that could involve the activation of PKC.


Increased phosphorylation of nuclear substrates for rat brain protein kinase C in regenerating rat liver nuclei.

Protein phosphorylation catalysed by rat brain protein kinase C (PKC) has been studied in nuclei isolated from normal and regenerating rat liver. Histone H1 and a 40,000 molecular weight protein were hyperphosphorylated at all the explored regeneration times, ranging from 3 to 22 h after partial hepatectomy. Phosphorylation of the two substrates was totally dependent on calcium and lipids and was abolished by low concentration of staurosporine. The observed early change of phosphate content of histone H1 and of the 40,000 molecular weight protein on the time scale of liver regeneration suggests that PKC might be involved in the initial nuclear events leading to cell proliferation.


Prostaglandin E2 can bimodally inhibit and stimulate the epididymal adipocyte adenylyl cyclase activity.

Measurements of prostaglandin E2 (PGE2)-induced adenylyl cyclase activity in membranes isolated from epididymal rat adipocytes revealed inhibition of cAMP production at low concentrations of PGE2 (less than 10 mM) and stimulation at higher concentrations. This biphasic effect of PGE2 was obtained when adenylyl cyclase was stimulated with GTP or NaF. In the presence of forskolin only the inhibitory phase by PGE2 was observed. Sulprostone, a PGE2 analogue, did not affect cAMP synthesis in the presence of either GTP or NaF; however, in the presence of forskolin, it inhibited cAMP production similarly to PGE2. Treatment of the membranes with cholera or pertussis toxin did not alter the biphasic effect of PGE2 on cAMP production. These findings raise the possibility that PGE2 acts through several receptor subtypes which are coupled to GTP binding proteins different from the classical Gi or Gs proteins.


The clinical use of thermocouple thermometry.

The development of a clinical thermometry system entails solving both clinical and technical problems. A thermometry system will, in general, be comprised of a temperature transducing element, a mechanism for interrogating the transducer temperature, and a means of interpreting, reporting and recording the measured temperature. Thermocouples are thermometers which report their own temperature, while the parameter of interest is the temperature of the medium. The two key questions which arise are then: (1) is the thermometer correctly reporting its own temperature, and (2) what is the relationship between the medium temperature and the thermometer temperature? In this work some of the important parameters influencing the temperature of the thermometer and the thermal relationship between the thermometer and the medium are investigated.


Electrical impedance tomography: prospects for non-invasive control of deep hyperthermia treatments.

Electrical Impedance Tomography (EIT) is assessed as a means of monitoring deep hyperthermia treatments in both phantom and in vivo studies. EIT measurements during heating of an inhomogeneous leg phantom show good correlation with internal temperatures measured with invasive probes. Measurements whilst altering the temperature of the stomach contents in volunteers demonstrate the capability of EIT for imaging temperature change at depth in the torso. Significant problems remain to be overcome before EIT can be used in the clinic; however, the technique may be an important additional control factor as a complement to invasive monitoring.


Non-invasive thermometry with multi-frequency microwave radiometry.

The present status of the development of a non-invasive thermometer based on microwave radiometry at our laboratory is reported. We have developed a model fitting technique combined with a Monte Carlo technique to retrieve temperature-depth profiles from multi (4-6)-frequency-band microwave radiometric data along with confidence intervals (2-sigma) of tissue temperatures as a function of depth. In order to make the radiometric technique compatible with the heating, brightness temperatures are measured through a 1 cm thick water bolus. Results of phantom experiments are presented to demonstrate the above capabilities of the method. Numerical simulation studies have shown that 2-sigma intervals would be 1.0 K or less over a 0-4 cm range and 1.4 K at 5 cm from the surface with using a six-band, 1-5 GHz radiometer having brightness temperature resolution of 0.03 K (3 s integration time). The six-band instrument is currently being assembled at our laboratory.


Simultaneous estimation of blood flow rate and tissue temperature.

A new method for simultaneously estimating blood flow rate and tissue temperature under a RF hyperthermia treatment is presented. In this method the finite element method is used to solve the two-dimensional Laplace equation for a specific absorption rate (SAR) distribution and the two-dimensional bioheat transfer equation for a tissue temperature distribution from the SAR pattern over a cross-section of a patient-specific model input from computer tomography images. Blood flow rates used in the temperature distribution computation are determined from basal values given for respective types of tissues and tissue temperatures through an assumed function that is intended to represent the temperature dependence of the blood flow rate. The tissue temperature computation procedure is iterated until the estimated temperature at a given sampling point matches a measured value at that point. This iterative procedure allows one to simultaneously estimate blood flow rate and tissue temperature. This paper describes the numerical method along with results of rudimentary in vivo measurements made in a human brain, where blood flow rate was measured by the single photon emission computed tomography (SPECT) imaging technique and tissue temperature was measured by an invasive probe technique.


Comparison of normal respiratory sounds recorded from the chest and trachea at various respiratory air flow levels.

Respiratory sounds (RS)s were recorded from the trachea and chest of 10 normal adult subjects at respiratory air flow levels of 1.6, 2.1 and 2.6 l/s using an oral flow transducer, and at approximately 2.1 l/s without the flow transducer. Tracheal RS (TRS) and chest RS (CRS) frequency spectra were generated using Fast Fourier Transform, and the peak, mean and maximum frequency parameters were derived from each spectra. Parametric analysis showed: (i) all three parameters for TRS spectra are significantly higher than those for CRS spectra; (ii) TRSs are on average eight times louder than CRSs; (iii) both TRSs and CRSs are air-flow independent over the flow range, though TRSs are significantly modified by the flow transducer while CRSs are not; and (iv) though of similar loudness, inspired and expired RSs (both TRSs and CRSs) have some significant spectral differences. To compare the complex shapes of RS spectra, each spectra was divided into narrow frequency bands (to create a feature set) and principal component analysis was performed on all spectral feature sets. TRSs and CRSs were shown to be independent biological signals with little overlap in their respective spectral characteristics.


Energetics of the fibrillating ventricle.

We have proposed a new mechanical index, equivalent pressure-volume area (ePVA), as a measure of the total mechanical energy of a fibrillating ventricle. ePVA is an analogue of the pressure-volume area (PVA) of a contracting ventricle and the specific area surrounded by the horizontal pressure-volume line at the pressure of ventricular fibrillation (VF) and the end-systolic and end-diastolic pressure-volume relations in the beating state in the pressure-volume diagram. In the isolated, cross-circulated heart preparation, we obtained myocardial oxygen consumption (VO2) during VF and ePVA at various left ventricular volumes in the control, epinephrine, propranolol and hypothermia runs. ePVA was highly linearly correlated with VO2 in all runs (r = 0.95, 0.98, 0.96 and 0.90, respectively). We also determined equivalent heart rate (eHR) as an estimate of the contraction frequency of individual myocytes in a fibrillating ventricle from mechanically unloaded VO2 in beating and fibrillating states. Using both ePVA and eHR, VO2 during VF was estimated and correlated with directly measured VO2. Estimated VO2 almost agreed with measured VO2 in all runs. We conclude that ePVA is a primary determinant of VO2 during VF, and that VO2 of a fibrillating ventricle can be reasonably accounted for by the combination of ePVA and eHR. This paper is a review of our previous studies on the energetics of a fibrillating ventricle.


Guidelines for thermometry in clinical hyperthermia.

High quality thermometry is needed in hyperthermia to (1) ensure safe delivery of adequate therapy and (2) provide the quantitative information needed to develop prognostic parameters which will aid research into planning and dosimetry. Currently, only invasive thermometry which is subject to several sources of artifact is available to address these requirements. This paper summarizes techniques which have been developed to reduce these artifacts to acceptable levels, outlines quality assurance requirements ensuring good practice in the use of invasive thermometry during clinical treatments and ends with a brief discussion of thermometry requirements for control of various hyperthermia systems.


Pharmacokinetics of mitomycin C following hepatic arterial chemoembolization with gelfoam.

Twelve mongrel dogs were randomly allocated into two groups using matched paired-design. Catheters were inserted into the hepatic artery, hepatic vein and the femoral vein, respectively. In the first group, gelfoam supplemented with mitomycin C (MMC) was injected into the hepatic artery, whereas the second group received a hepatic arterial injection of MMC solution alone. Simultaneous blood sampling from the hepatic and femoral vein at regular intervals was performed. MMC concentrations in plasma was determined using high performance liquid chromatography (HPLC) and the pharmacokinetics of MMC were determined. MMC concentrations in hepatic and femoral veins did not differ and no significant difference in pharmacokinetics was found when comparing MMC administration into the hepatic artery with or without gelfoam supplementation. Thus, our results revealed that gelfoam could not delay the clearance of MMC from the liver.


Choledochoplasty by vein grafts in iatrogenic bile duct injuries.

The quality of immediate repair of common bile duct injuries with or without tissue loss occurring during elective cholecystectomy is crucial and maybe the sole factor behind future stricture formation with its considerable morbidity and mortality. Successful repair of iatrogenic common bile duct injuries has been achieved by immediate saphenous vein grafts in two patients with cystic duct avulsion, in one patient whose duct was split by a balloon catheter, and in one patient where a segment of the duct was resected. Follow-up for 5 years demonstrated that the grafting remained sound and produced no complications. Consequently, the immediate repair of iatrogenic bile duct injuries using vein grafts deserves consideration.


A case of primary squamous cell carcinoma of the liver arising in a solitary cyst.

A case of primary squamous cell carcinoma in a pre-existing hepatic cyst is presented. A review of the literature suggests that this rare type of liver tumor tends to arise from solitary, nonparasitic cysts, lined with squamous epithelium. Effective therapy is not available, the prognosis is grave.


Tuberculous pancreatic abscess: a rare condition mimicking carcinoma.

We report a rare case of obstructive jaundice caused by a tuberculous abscess in the head of pancreas, mimicking carcinoma. The case was successfully treated by pylorus-preserving proximal pancreatoduodenectomy and antituberculous drugs.


Oral cancer diagnosis and delays in referral.

Delays at various levels in the management pathway of oral cancer have previously been reviewed. It is a clinical impression that advanced tumours are being misdiagnosed by practitioners and consequently delays in referral are still prevalent. A study was conducted to compare the diagnosis and referral patterns of medical and dental practitioners. The results indicated that despite seeing similar populations of patients with oral cancer, the medical practitioner was better at diagnosing and referring cases early, and noting the relevance of lymphadenopathy. A high index of suspicion is a prerequisite for early diagnosis and referral of patients with oral cancer. The findings of this study highlight the importance of correct diagnosis, and support recent recommendations regarding practitioner education.


A three-dimensional soft tissue analysis of 16 skeletal class III patients following bimaxillary surgery.

A three-dimensional soft tissue study of the results of surgery in a group of 16 skeletal Class III adult patients following orthognathic surgery was carried out using laser scans (Arridge et al., 1985). The patient group was compared to a control group of the same population. Laser scans were taken prior to surgery, 3 months post-surgery, and at least 1 year after retention. Preoperative comparison to the control groups revealed that the facial disproportion related to both the maxilla and the mandible. Le Fort I advancements resulted in broadening of the lateral aspects of the nose, advancement of the dorsum, and overcorrection of the alar bases. There was a degree of change over the cheeks bilaterally, because of alterations in the general drape of the soft tissues. There was a degree of overcorrection in the female group following mandibular set back but the male group were still more prognathic, when compared to the control group. There was a marked degree of relapse in the mandible from 3 months to 1 year postoperatively, with a resultant anterior movement of the maxillary arch. Laser scanning has proved to be a simple non-invasive method of measuring three-dimensionally, and is a very useful tool in auditing surgical outcome and measuring surgical relapse.


Reproducibility of three-dimensional CT-assisted model production in the maxillofacial area.

In the mid-eighties, computerised tomography (CT) assisted three-dimensional imaging, and modelling of skull structures was introduced into preoperative diagnosis in maxillofacial surgery. This new method is already being used in the preoperative planning of corrections of post-traumatic defects and craniofacial deformities as well as in tumour surgery and implantology. The aim of the present study was to collect information on the reproducibility of a skull model milled from hardened polyurethane foam, and based on the CT data of a real skull. Thirty one measurements were carried out on the model and on the original skull, leading to the result that the model showed a mean inaccuracy of 1.5 mm. The deviations ranged between 0.0 and 3.6 mm. Generally, the model structures, however, tended to be larger than those of the original. As far as the total of all measurements is concerned, the model deviated from the original skull by 1.6%. A convincing aspect of the model, which cannot be obtained by any other method, is its plasticity and the possibility of 3-D orientation on a lifesize model. However, the study indicates that surgeons using this technique should consider the possible discrepancies between the model and the real skull when making preoperative assessments.


An update on recent advances in the understanding of non-neoplastic diseases of the salivary glands.

The recent advances in the understanding of the non-neoplastic disorders of salivary glands are summarised in this paper. The common infections are discussed and newer aspects of juvenile recurrent parotitis outlined. The benign lympho-epithelial lesion is reviewed in relation to Sjogren's syndrome and the salivary gland pathology seen in relation to infection with human immunodeficiency virus. A range of unusual conditions that may affect the minor salivary glands in particular, and can cause diagnosis dilemmas, is summarised.


An investigation of the effect of midazolam on the pain experience.

In a randomised, double-blind, cross-over study on 20 healthy human volunteers, the effect of intravenous midazolam on pain tolerance using the 'submaximum effort tourniquet technique' is described. Intravenous midazolam administered in conscious sedation doses was found to significantly reduce the affective and motivational component of the pain experience.


Concentration of cefuroxime in mandibular alveolar bone following a single preoperative dose.

A pilot study was undertaken on 12 patients undergoing the bilateral removal of impacted mandibular wisdom teeth to determine the level of cefuroxime sodium within the alveolar bone adjacent to the third molar teeth following a single preoperative intravenous administration. The results indicated that a single 1.5 g intravenous bolus injection provided satisfactory alveolar bone concentrations for routine antibiotic prophylaxis at the time of wisdom tooth removal.


Blank paper or printed set task forms for retrievable patient notes?

Patients' clinical notes made at review appointments following removal of third molars under a general anaesthetic at five Mersey Hospitals have been compared with those formed from the recording of the findings of preselected tasks. Consistent high quality retrievable records were achieved using the preselected task forms, superior to those formed on blank paper.


Infantile fibromatosis with involvement of the mandible.

Two unusual cases of infantile fibromatosis involving the mandible occurring in 2-year-old children are described. These tumours, though initially highly aggressive, underwent spontaneous regression in the absence of definitive treatment.


Obstructive sleep apnoea syndrome presenting as lingual ulceration.

Two patients with obstructive sleep apnoea syndrome are reported where the initial presenting complaint was of lingual ulceration. This unusual presentation has not been reported previously. Both patients experienced frequent apnoeic episodes during sleep with a profound fall in the arterial oxygen saturation. It is postulated that the lingual ulceration resulted from repeated trauma to the tongue by the teeth as the patient made violent inspiratory efforts at the termination of an apnoeic episode. The diagnosis of sleep apnoea syndrome was based upon suggestive symptoms of snoring, morning fatigue and day-time somnolence plus a minimum of 15 apnoeic episodes per hour of sleep. The first-line investigations of this condition are available in all district general hospitals and a diagnosis of sleep apnoea syndrome obtained. Referral to a regional sleep study centre may be appropriate prior to the commencement of therapy. Management is predominantly medical, consisting of weight loss and the administration of nocturnal nasal continuous positive airways pressure.


Liposarcoma of the tongue: a case report and review of the literature.

A case of the rare intraoral liposarcoma which presented as a lump on the tongue of 5 years duration is reported. The difficulty of accurate clinical and histopathological diagnosis of this lesion is outlined.


Papillary cystadenocarcinoma of the sublingual gland presenting as a ranula.

A case is reported of a papillary cystadenocarcinoma of the sublingual gland which presented as a ranula. This is both rare in site and mode of presentation.


Sialolithiasis of minor salivary glands.

Sialolithiasis of minor salivary glands is generally considered to be extremely rare. Reports of seven cases are presented. A review of the literature suggests that the condition is not as uncommon as previously considered, and that definitive histopathological diagnosis of lip swellings should be obtained due to the incidence of labial pleomorphic adenoma and the possibility of the clinical misdiagnosis with sialolithiasis.


Prediction of structural and functional features of protein and nucleic acid sequences by artificial neural networks.

The applications of artificial neural networks to the prediction of structural and functional features of protein and nucleic acid sequences are reviewed. A brief introduction to neural networks is given, including a discussion of learning algorithms and sequence encoding. The protein applications mostly involve the prediction of secondary and tertiary structure from sequence. The problems in nucleic acid analysis tackled by neural networks are the prediction of translation initiation sites in Escherichia coli, the recognition of splice junctions in human mRNA, and the prediction of promoter sites in E. coli. The performance of the approach is compared with other current statistical methods.


Functional topology of a surface loop shielding the catalytic center in lipoprotein lipase.

Lipoprotein lipase (LPL), hepatic lipase, and pancreatic lipase show high sequence homology to one another. The crystal structure of pancreatic lipase suggests that it contains a trypsin-like Asp-His-Ser catalytic triad at the active center, which is shielded by a disulfide bridge-bounded surface loop that must be repositioned before the substrate can gain access to the catalytic residues. By sequence alignment, the homologous catalytic triad in LPL corresponds to Asp156-His241-Ser132, absolutely conserved residues, and the homologous surface loop to residues 217-238, a poorly conserved region. To verify these assignments, we expressed in vitro wild-type LPL and mutant LPLs having single amino acid mutations involving residue Asp156 (to His, Ser, Asn, Ala, Glu, or Gly), His241 (to Asn, Ala, Arg, Gln, or Trp), or Ser132 (to Gly, Ala, Thu, or Asp) individually. All 15 mutant LPLs were totally devoid of enzyme activity, while wild-type LPL and other mutant LPLs containing substitutions in other positions were fully active. We further replaced the 22-residue LPL loop which shields the catalytic center either partially (replacing 6 of 22 residues) or completely with the corresponding hepatic lipase loop. The partial loop-replacement chimeric LPL was found to be fully active, and the complete loop-replacement mutant had approximately 60% activity, although the primary sequence of the hepatic lipase loop is quite different. In contrast, replacement with the pancreatic lipase loop completely inactivated the enzyme. Our results are consistent with Asp156-His241-Ser132 being the catalytic triad in lipoprotein lipase.(ABSTRACT TRUNCATED AT 250 WORDS)


Denaturation of human Cu/Zn superoxide dismutase by guanidine hydrochloride: a dynamic fluorescence study.

The unfolding of holo and apo forms of human Cu/Zn superoxide dismutase by guanidine hydrochloride has been investigated by steady-state and dynamic fluorescence. In agreement with previous observations, a stabilizing effect of the metal ions on the protein tertiary structure was apparent from comparison of apo- and holoproteins, which both showed a sharp sigmoidal transition though at different denaturant concentrations. The transition was also followed by circular dichroism to check the extent of secondary structure present at each denaturant concentration. The results are incompatible with a simple two-state mechanism for denaturation. The occurrence of a more complicated process is supported by the emission decay properties of the single tryptophanyl residue at different denaturant concentrations. A complex decay function, namely, two discrete exponentials or a continuous distribution of lifetimes, was always required to fit the data. In particular, the width of the lifetime distribution, which is maximum at the transition midpoint, reflects heterogeneity of the tryptophan microenvironment and thus the presence of different species along the denaturation pathway. In the unfolded state, the width of the lifetime distribution is broader than in the folded state probably because the tryptophan residue is affected by a larger number of local conformations. The dissociation of the dimer was also studied by varying the protein concentration at different denaturant concentrations. This process affects primarly the surface of the protein rather than its secondary structure as shown by a comparison between the tryptophan emission decay and circular dichroism data under the same conditions. Another consequence of dissociation is a greater instability in the structure of the monomers, which are more easily unfolded.(ABSTRACT TRUNCATED AT 250 WORDS)


Kinetic analysis of folding and unfolding the 56 amino acid IgG-binding domain of streptococcal protein G.

The 56 amino acid B domain of protein G (GB) is a stable globular folding unit with no disulfide cross-links. The physical properties of GB offer extraordinary flexibility for evaluating the energetics of the folding reaction. The protein is monomeric and very soluble in both folded and unfolded forms. The folding reaction has been previously examined by differential scanning calorimetry (Alexander et al., 1992) and found to exhibit two-state unfolding behavior over a wide pH range with an unfolding transition near 90 degrees C (GB1) at neutral pH. Here, the kinetics of folding and unfolding two naturally occurring versions of GB have been measured using stopped-flow mixing methods and analyzed according to transition-state theory. GB contains no prolines, and the kinetics of folding and unfolding can be fit to a single, first-order rate constant over the temperature range of 5-35 degrees C. The major thermodynamic changes going from the unfolded state to the transition state are (1) a large decrease in heat capacity (delta Cp), indicating that the transition state is compact and solvent inaccessible relative to the unfolded state; (2) a large loss of entropy; and (3) a small increase in enthalpy. The most surprising feature of the folding of GB compared to that of previously studied proteins is that its folding approximates a rapid diffusion controlled process with little increase in enthalpy going from the unfolded to the transition state.