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Brazil has the greatest plant diversity in the world. Many species exhibit a wide range of phytochemical compounds which can be exploited in food, agronomic, pharmacological and medicinal plant industries. Therefore, the chemical composition and in vitro bioactivities of volatile oil from Psidium firmum fresh leaves (PfVO) were investigated for the first time. GC-FID and GC-MS analyses revealed 28 compounds in PfVO. The major ones were α-selinene (20.8%), β-caryophyllene (16.5%) and nerolidol (10.4%). Results showed that PfVO affected the growth of Leishmania amazonensis promastigote forms in a dose-dependent manner; its IC50 value was 14.05 µg/mL. PfVO also exhibited antibacterial activity against Salmonella enteritidis, Yersinia enterocolitica, Staphylococcus aureus, Pseudomonas aeruginosa and Listeria monocytogenes; MIC values ranged from 25 µg/mL to 250 µg/mL. Moreover, PfVO promoted normal cell growth inhibition at 61.02 ± 1.97 µg/mL. Antiproliferative activity was observed against human tumor cell lines; IC50 values of MCF-7 cells, HeLa cells and M059J cells were 47.91 µg/mL, 73.78 µg/mL and 41.94 µg/mL, respectively. Results provided strong evidence of the promising potential of PfVO as a nature-based antileishmanial, antibacterial and antiproliferative agent.

SUMMARYRecently, organic farming systems have attracted the attention of consumers because of their low environmental impact. Organic agriculture is a valid alternative to conventional farming and ancient wheat, such as KAMUT® khorasan wheat (T. turgidum ssp. turanicum), has emerged as an industry leader for its nutritional and functional properties (anti-inflammatory, antioxidant and prebiotic).The aim of the present study was to evaluate environmental effects on the quality of KAMUT® khorasan grains harvested in the last two decades (1989–2012) on one farm in Montana (USA), through the evaluation of phytochemical accumulation. Results revealed high variability in the amounts of macronutrients and nutraceuticals. In particular, from 1989 to 2012, there was a decreasing trend in starch content (ranging from 70·87 to 50·54/100 g) and amylose (from 41·48 to 31·46% of total starch) with a slight increase of insoluble dietary fibre (from 12·14 to 17·75/100 g). The soluble dietary fibre content varied among the years of cultivation even if the general trend remained constant (4·57–2·82/100 g). High variability of total polyphenols content was observed with the free soluble fraction present at lower levels than bound polyphenols (BP). Moreover, an inverse correlation between free and BP was observed. The results obtained in the present study show that the influence of environmental conditions plays a fundamental role in the accumulation of primary and secondary metabolites in wheat kernels and strongly modulate the nutritional and nutraceutical value of flour.

Background/objectives Khorasan wheat is an ancient grain with widely acclaimed beneficial effects on human health. The objective of the study was to examine the effect of a Khorasan-based diet on the wellbeing and inflammatory profile of young athletes. Results We conducted a randomized, single-blinded crossover trial involving 20 male young athletes. The participants were randomly assigned to consume products (pasta, bread, biscuits and crackers) made either with Khorasan (KAMUT® brand) or modern semi-whole-grain wheat for 4-weeks with a 4-week washout period before the crossover. Laboratory analyses and fitness tests were performed both at the beginning and end of each diet period. The consumption of Khorasan products was associated with a significant reduction of monocyte chemoattractant protein-1 (MCP-1; mean reduction: -36.15 pg/mL; -25.67%) while the consumption of modern wheat was not associated with significant differences in Interleukin-8 (IL-8) or Interleukin-1 receptor antagonist (IL-1ra). The consumption of the Khorasan-based diet also resulted in a significant improvement in self-rated health status. No statistically significant differences in any athletic performance parameter were observed between the two diets. Conclusion The present results suggest that a Khorasan-based diet could be effective in reducing the inflammatory status in young athletes. © 2019 Society of Chemical Industry.

Legionella spp are the causative agents of Legionnaires&rsquo, diseases, which is a pneumonia of important public health concern. Ubiquitous freshwater and soil inhabitants can reach man-made water systems and cause illness. Legionella enumeration and quantification in water systems is crucial for risk assessment and culture examination is the gold standard method. In this study, Legionella recovery from potable water samples, at presumably a low concentration of interfering microorganisms, was compared by plating on buffered charcoal yeast extract (BCYE) and glycine, vancomycin, polymyxin B, cycloheximide (GVPC) Legionella agar media, according to the International Standard Organization (ISO) 11731: 2017. Overall, 556 potable water samples were analyzed and 151 (27.1%) were positive for Legionella. Legionella grew on both BCYE and GVPC agar plates in 85/151 (56.3%) water samples, in 65/151 (43%) on only GVPC agar plates, and in 1/151 (0.7%) on only BCYE agar plates. In addition, GVPC medium identified Legionella species other than pneumophila in six more samples as compared with the culture on BCYE. Although the medians of colony forming units per liter (CFU/L) detected on the BCYE and GVPC agar plates were 2500 and 1350, respectively (p-value &lt, 0.0001), the difference did not exceed one logarithm, and therefore is not relevant for Legionella risk assessment. These results make questionable the need to utilize BCYE agar plates to analyze potable water samples.

Background Performing accurate anatomical reconstruction is a challenging task in the treatment of internal orbital floor fractures. Compared with traditional transcutaneous incisions, endoscopic transmaxillary approaches have the advantage of avoiding complications related to external scars, and provide direct access to the orbital floor. Autogenous bone provides the ideal material for defect reconstruction, but determination of the correct size and shape of the graft is crucial for a stable support. This study introduces a new protocol for the treatment of internal orbital floor fractures that combines endoscopy, virtual reality, and 3D printing. The authors also investigated the impact of computer-aided surgery (CAS) on the overall accuracy of reconstruction in aiming to achieve the triple objective of restoring anatomy, volume, and function. Materials and methods Fourteen patients with orbital floor fractures were recruited for this study. High-resolution CT scans provided appropriate imaging for detailed orbital floor defect visualization. A virtual reconstruction of the orbital floor defect was developed and a 3D printed template was fabricated to provide intraoperative guidance in the graft harvesting phase, according to the orbital defect. Virtual analyses were conducted to evaluate the accuracy of reconstruction both in terms of graft size and graft orientation. Results Postoperative CT scans showed that in all cases orbital floor reconstruction was successfully performed, resulting in restoration of the correct globe position. No intraoperative complications occurred. Correspondence of graft size was evaluated using color-coded maps and RMSE, while comparison of angular measurements allowed the authors to relate simulated and actual reconstruction. Conclusions Orbital floor reconstruction performed via transmaxillary endoscopy is a safe technique, which allows for detailed visualization of the fracture rim, avoids external scars, and permits an easier reduction of the prolapsed orbital content into the overlying orbital cavity. Virtual planning plays an important role in defining the appropriate geometry of the bone graft and establishing the optimal reconstruction strategy. Our preliminary results indicate that virtual planning and 3D printing should become part of an integrated protocol for the endoscopic treatment of orbital floor fractures.

Made available in DSpace on 2018-11-12T17:26:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-08-01. Added 1 bitstream(s) on 2018-11-12T17:34:52Z : No. of bitstreams: 1 S1516-18462018000400442.pdf: 323574 bytes, checksum: 8ced7f7ee3c535c02f7c6feff0967e91 (MD5) RESUMOObjetivo: descrever e comparar os resultados das provas oculares (sacádicos, rastreio, e optocinético) da vectoeletronistagmografia entre os grupos com dislexia, transtorno de aprendizagem e controle.Métodos: participaram deste estudo 28 estudantes do sexo masculino e feminino, com idade entre 8 e 11 anos. Foram divididos em três grupos, sendo Grupo I: Dez escolares com Dislexia, Grupo II: Nove escolares com Transtornos de Aprendizagem e Grupo III: Nove escolares sem dificuldades de aprendizagem. Nesta pesquisa foi realizada avaliação da vectoeletronistagmografia digital: provas oculares (sacádicos, rastreio, e optocinético).Resultados: verificou-se que as provas de movimento sacádico, nistagmo optocinético e rastreio pendular mostraram diferenças sutis entre os três grupos. Na comparação das provas de movimentos sacádicos dos olhos e pesquisa do nistagmo optocinético foi observada que a velocidade do movimento do olho esquerdo que se mostrou mais lento para o Grupo I, e ainda mais lentos para o Grupo II. Foi observado também que os grupos GI e GII foram mais lentos para o rastreio em pêndulo do estímulo luminoso em relação ao grupo controle.Conclusão: de modo geral, foi observada diferença entre os grupos para a medida da vectoeletronistagmografia, que indicam rastreios e reflexo vestibular-ocular mais lentos nas crianças com dislexia e transtornos de aprendizagem. ABSTRACTObjective: to describe and compare the results of ocular (saccadic, screening, and optokinetic) tests of vectoelectronystagmography among the groups with dyslexia, learning disorder and control.Methods: 28 male and female students aged 8 to 11 years participated in this study. They were divided into three groups: Group I, 10 students with dyslexia, Group II, 9 students with learning disorders and Group III, 9 students with no learning disorders. In this research, digital vectoelectronystagmography - ocular test - was performed.Results: saccadic movement, optokinetic nystagmus, and pendular tracking tests were found to show subtle differences among the three groups. Comparing the saccadic eye movements and the optokinetic nystagmus tests, it was observed that the movement in the left eye was slower for Group I, and even slower for Group II. It was also observed that GI and GII were slower for the pendular tracking of luminous stimulus in relation to the control group.Conclusion: in general, there are differences among the groups in the vectoelectronystagmography, which indicated slower tracking and vestibulo-ocular reflex in children with dyslexia and learning disorders. Universidade de São Paulo Faculdade de Medicina UNESP Faculdade de Filosofia e Ciências Universidade Estadual de Campinas UNESP Faculdade de Filosofia e Ciências

Made available in DSpace on 2018-11-12T17:26:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-12-01. Added 1 bitstream(s) on 2018-11-12T17:34:47Z : No. of bitstreams: 1 S1516-18462015000801838.pdf: 319362 bytes, checksum: 36aada8f8b2887b240182cad220b8cf2 (MD5) RESUMO:Objetivo: descrever os achados dos exames eletrofisiológicos de indivíduos com gagueira e comparar com indivíduos com desenvolvimento típico.Métodos: participaram desta pesquisa 34 indivíduos, de ambos aos gêneros, com idade entre 7 e 31 anos. O grupo pesquisa foi constituído por 13 crianças (G1a) e 4 adultos (G1b) diagnosticadas com gagueira e o grupo controle por 13 crianças (G2a) com bom desempenho acadêmico e 4 adultos (G2b) com desenvolvimento típico. Foi realizada a avaliação dos potenciais auditivos, na varredura de frequência e de duração.Resultado: quando comparados os grupos de crianças gagas e controle, foi observado que as crianças gagas apresentaram na varredura de frequência, aumento da latência dos componentes P2, N2 em Cz na orelha direita e N2 e P3 em Fz na orelha esquerda, e diferença na amplitude de P2 e P3 em Cz na orelha direita. Na varredura de duração, houve redução da amplitude dos componentes N2 em Cz e P3 em Fz na orelha direita. Já no grupo de adultos gagos, observou-se na varredura de frequência, aumento da latência no componente P3 em Cz e Fz na orelha direita e amplitude reduzida no P3 em Cz na orelha esquerda, e na varredura de duração, aumento de latência de N2 e P3 em Cz na orelha direita e N2 em Cz na orelha esquerda, quando comparados ao grupo controle.Conclusão: há diferenças entre os exames eletrofisiológicos de indivíduos com gagueira comparados aqueles com desenvolvimento típico. No entanto, novos estudos neste âmbito ainda são necessários para que os achados aqui apresentados possam ser confirmados. ABSTRACT:Purpose: to describe the findings of electrophysiological examinations of individuals who stutter and compare with typically developing individuals.Methods: 34 subjects participated in this study, both genders, aged between 7 and 31 years. The research group consisted of 13 children (G1a) and 4 adults (G1b) diagnosed with stuttering and the control group of 13 children (G2a) with good academic performance and 4 fluent adults (G2b). The auditory potentials assessment was performed on frequency and duration discrimination tests.Result: when compared groups of stuttering children and control group, it was observed that the stuttering children had the scan frequency, increased latencies of P2, N2 components at Cz in the right ear and N2 and P3 at Fz in the left ear, and the difference amplitude P2 and P3 in the right ear Cz. In the scan duration, decreased the amplitude of the N2 and P3 components in Cz Fz in the right ear. In the group of adult stutterers was observed in the frequency sweep, increased latency P3 component at Cz and Fz in the right ear and reduced P3 amplitude at Cz in the left ear, and scan duration, increased latency of N2 Cz and P3 in the right ear and N2 at Cz in the left ear, when compared to the control group.Conclusion: there are differences between the electrophysiological examinations of individuals who stutter compared with those with typical development. However, further studies in this area are still needed for the findings presented here can be confirmed. Universidade Estadual Paulista Júlio de Mesquita Filho Universidade Estadual Paulista Júlio de Mesquita Filho

An investigation was carried out into the recovery from calf faeces of Bacillus coagulans spores added to the feed as probiotic. For this purpose, Bacillus coagulans spores (9 log₁₀ CFU g⁻¹) were given daily to 10 calves during the whole farming periods; another 10 calves acted as controls. Throughout the trial the faecal spore counts were significantly (P < 0.01) higher in the treated group than in the controls (averaging 2.1 x 10⁵ vs 3.7 x 10⁴ CFU g⁻¹). Bacterial cells were recovered from faecal samples and ribotyping matched the strain isolated from faecal sample to the clone administered to the animals. In addition, the recovered cells were found to maintain their functionality aspects of acid production, survival in artificial gastric juice and in the presence of bile, and attachment to human intestinal epithelial cells. The results further elucidate the fate of spore formers administered to calves, and this will help in the development of new species-specific nutritional strategies.

A PCR-RFLP technique has been applied on 13 species of Bifidobacterium in order to update a previous study carried out by Baffoni et al. [1]. This method is based on the restriction endonuclease activity of HaeIII on the PCR-amplified hsp60 partial gene sequence, and allows a rapid and efficient identification of Bifidobacterium spp. strains at species and subspecies level.

The halogenated compounds aeruginosin and cyanopeptolin (protease inhibitors) are well documented in several cyanobacteria. In this work, the presence of genetic coding for aeruginosin and cyanopeptolin in Sphaerocavum genus and their characterization by mass spectrometry and potential toxic effects were investigated. Three strains of Sphaerocavum brasiliense (CCIBt 3094, CCIBt 3096 and CCIBt 3316) isolated from different reservoirs in Sao Paulo State, and one (CCIBt 3179) from Rio Grande do Norte State were selected for this study. Fragments of aerA-aerB and mcnC-mcnE gene regions (aeruginosin and cyanopeptolin, respectively) were amplified by polymerase chain reaction (PCR) and sequenced. Compounds were characterized by mass spectrometry. All the studied strains presented aeruginosin and cyanopeptolin synthetase genes. Mass spectrometry showed the presence of several aeruginosin and cyanopeptolin variants. The Allium test showed that the ethanolic extract promoted toxic and cytotoxic actions in acute and chronic treatments. These naturally occurring protease inhibitors can play an important role in the physiopathology of human diseases.

Xylella fastidiosa is a plant pathogen that threatens a US$ 4.6 billion worldwide wine and citrus industry. Monitoring its presence and distribution in plants and vectors is crucial for designing control strategies, as well as for understanding its ecological role and fate. We developed two fluorescent oligonucleotide probes complementary to different regions of the 16S rRNA gene of X. fastidiosa. The specificity of the newly designed probes S-S-X.fas-0067-a-A-18 and S-S-X.fas-1439-a-A-18 was demonstrated using fluorescence in situ hybridization (FISH) for 12 Xylella isolates, 15 closely related microorganisms and three plant endophytes. These probes were used to detect and quantify X. fastidiosa in plant sap (average value of 2.9 +/- 0.3 x 10(6) cells ml(-1)) from three different citrus orchards. In a second experiment, cells were quantified in honeydew (2.2 +/- 0.2 x 10(4) cells ml(-1)) collected from the insect vector Bucephalogonia xanthophis during the acquisition access period on an infected plant. The number of pathogen cells retained or digested by the insect is 10,000 times greater than the estimated minimum value to ensure an efficient transmission. Polymerase chain reaction (PCR) amplification using specific primers with plant sap and honeydew samples, followed by sequencing, confirmed the presence of the plant pathogen. This is the first demonstration of FISH being used for environmental samples, such as plant sap and insect honeydew, to estimate the abundance of a plant pathogen during infection.

Made available in DSpace on 2022-04-29T07:13:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-01-01 Cyanobacteria are an important source of structurally bioactive metabolites, with cytotoxic, antiviral, anticancer, antimitotic, antimicrobial, specific enzyme inhibition and immunosuppressive activities. This study focused on the antitumor and antimicrobial activities of intra and extracellular cyanobacterial extracts. A total of 411 cyanobacterial strains were screened for antimicrobial activity using a subset of pathogenic bacteria as target. The in vitro antitumor assays were performed with extracts of 24 strains tested against two murine cancer cell lines (colon carcinoma CT-26 and lung cancer 3LL). Intracellular extracts inhibited 49 and 35% of Gram-negative and Gram-positive pathogenic bacterial growth, respectively. Furthermore, the methanolic intracellular extract of Cylindrospermopsis raciborskii CYP011K and Nostoc sp. CENA69 showed inhibitory activity against the cancer cell lines. The extracellular extract from Fischerella sp. CENA213 and M. aeruginosa NPJB-1 exhibited inhibitory activity against 3LL lung cancer cells at 0.8 μg ml-1 and Oxynema sp. CENA135, Cyanobium sp. CENA154, M. aeruginosa NPJB-1 and M. aeruginosa NPLJ-4 presented inhibitory activity against CT26 colon cancer cells at 0.8 μg ml-1. Other extracts were able to inhibit 3LL cell-growth at higher concentrations (20 μg ml-1) such as Nostoc sp. CENA67, Cyanobium sp. CENA154 and M. aeruginosa NPLJ-4, while CT26 cells were inhibited at the same concentration by Nostoc sp. CENA67 and Fischerella sp. CENA213. These extracts presented very low inhibitory activity on human peripheral blood lymphocytes. The results showed that some cyanobacterial strains are a rich source of natural products with potential for pharmacological and biotechnological applications. © 2013 Bentham Science Publishers. University of São Paulo Center for Nuclear Energy in Agriculture Laboratory of Molecular Ecology of Cyanobacteria, 13400-970 Piracicaba, SP São Paulo State University Dept. Microbiology and Immunology Institute of Biosciences, 18618-970 Botucatu, SP São Paulo State University Dept. Microbiology and Immunology Institute of Biosciences, 18618-970 Botucatu, SP

Cyanobacteria possess the ability to produce compounds with remarkable biological activity, and have thus attracted the attention of the pharmaceutical industry. Cyanopeptides acting as protease inhibitors have shown potential in the field of pharmacotherapy through regulation of abnormal physiological processes in the human body. Despite the already described cyanopeptide protease inhibitors, the search for new congeners is of considerable interest which may pave the way for more efficient molecules. In this study, the presence of the protease inhibitors aeruginosin and cyanopeptolin with non-, mono- and dichlorination and also genes coding for their synthetases was investigated in 90 cyanobacterial strains. Mass spectrometry analyses highlighted production of 91, 19 and 3 non-, mono- and dichlorinated congeners, respectively. The purified extract of Microcystis botrys SPC759 inhibited 61% of pepsin protease. PCR amplifications of aeruginosin and cyanopeptolin synthetase gene regions were observed in 41 and 28% of evaluated strains, respectively. The sequences obtained for the aerA-aerB (aeruginosin) and mcnC-mcnE (cyanopeptolin) gene regions grouped together with their homologues found in other cyanobacterial strains in the phylogenetic analyses with high bootstrap support. Antimicrobial activity assays performed using all intracellular extracts inhibited 31 and 26% of Gram-negative and Gram-positive pathogenic bacterial growth, respectively. The results of this study showed the production of aeruginosin and cyanopeptolin and the presence of their genes in several cyanobacterial genera for the first time besides the discovery of novel congeners.

Water pollution caused by toxic cyanobacteria is a problem worldwide, increasing with eutrophication. Due to its biological significance, genotoxicity should be a focus for biomonitoring pollution owing to the increasing complexity of the toxicological environment in which organisms are exposed. Cyanobacteria produce a large number of bioactive compounds, most of which lack toxicological data. Microcystins comprise a class of potent cyclic heptapeptide toxins produced mainly by Microcystis aeruginosa. Other natural products can also be synthesized by cyanobacteria, such as the protease inhibitor, aeruginosin. The hepatotoxicity of microcystins has been well documented, but information on the genotoxic effects of aeruginosins is relatively scarce. In this study, the genotoxicity and ecotoxicity of methanolic extracts from two strains of M. aeruginosa NPLJ-4, containing high levels of microcystin, and M. aeruginosa NPCD-1, with high levels of aeruginosin, were evaluated. Four endpoints, using plant assays in Allium cepa were applied: rootlet growth inhibition, chromosomal aberrations, mitotic divisions, and micronucleus assays. The microcystin content of M. aeruginosa NPLJ-4 was confirmed through ELISA, while M. aeruginosa NPCD-1 did not produce microcystins. The extracts of M. aeruginosa NPLJ-4 were diluted at 0.01, 0.1, 1 and 10 ppb of microcystins; the same procedure was used to dilute M. aeruginosa NPCD-1 used as a parameter for comparison, and water was used as the control. The results demonstrated that both strains inhibited root growth and induced rootlet abnormalities. The strain rich in aeruginosin was more genotoxic, altering the cell cycle, while microcystins were more mitogenic. These findings indicate the need for future research on non-microcystin producing cyanobacterial strains. Understanding the genotoxicity of M. aeruginosa extracts can help determine a possible link between contamination by aquatic cyanobacteria and high risk of primary liver cancer found in some areas as well as establish water level limits for compounds not yet studied.

Cyanobacterial strains isolated from terrestrial and freshwater habitats in Brazil were evaluated for their antimicrobial and siderophore activities. Metabolites of fifty isolates were extracted from the supernatant culture media and cells using ethyl acetate and methanol, respectively. The extracts of 24 isolates showed antimicrobial activity against several pathogenic bacteria and one yeast. These active extracts were characterized by Q-TOF/MS. The cyanobacterial strains Cylindrospermopsis raciborskii 339-T3, Synechococcus elongatus PCC7942, Microcystis aeruginosa NPCD-1, M. panniformis SCP702 and Fischerella sp. CENA19 provided the most active extracts. The 50 cyanobacterial strains were also screened for the presence of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) genes and microcystin production. Putative fragment genes coding for NRPS adenylation domains and PKS keto-synthase domains were successfully PCR amplified from 92% and 80% of cyanobacterial strains, respectively. The potential therapeutical compounds siderophores were detected in five cyanobacterial isolates. Microcystin production was detected by ELISA test in 26% of the isolates. Further a protease inhibitor substance was detected by LC-MS/MS in the M. aeruginosa NPLJ-4 extract and the presence of aeruginosin and cyanopeptolin was confirmed by PCR amplification using specific primers, and sequenced. This screening study showed that Brazilian cyanobacterial isolates are a rich source of natural products with potential for pharmacological and biotechnological applications.

Xylella fastidiosa is a pathogenic bacterium found in several plants. These bacteria secrete extracellular proteases into the culture broth as visualized in sodium-dodecyl-sulfate polyacrylamide activity gels containing gelatin as a copolymerized substrate. Three major protein bands were produced by the citrus strain with molar masses (MM) of 122, 84 and 65 kDa. Grape strain 9,713 produced two bands of approximately 84 and 64 kDa. These organisms produced zones of hydrolysis in agar plates amended with gelatin, casein and hemoglobin. Gelatin was the best substrate for these proteases. Sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) activity gel indicated that the protease of Xylella fastidiosa from citrus and grape were completely inhibited by PMSF and partially inhibited by EDTA. The optimal temperature for protease activity was 30 degrees C with an optimal pH of 7.0. Among the proteolytic enzymes secreted by the phytopathogen, chitinase and beta-1,3-glucanase activities were also detected in cultures of Xylella fastidiosa (citrus). From these results, it is suggested that proteases produced by strains of Xylella fastidiosa from citrus and grape, belong to the serine- and metallo-protease group, respectively.

Synonymous codon usage varies considerably among Caenorhabditis elegans genes. Multivariate statistical analyses reveal a single major trend among genes. At one end of the trend lie genes with relatively unbiased codon usage. These genes appear to be lowly expressed, and their patterns of codon usage are consistent with mutational biases influenced by the neighbouring nucleotide. At the other extreme lie genes with extremely biased codon usage. These genes appear to be highly expressed, and their codon usage seems to have been shaped by selection favouring a limited number of translationally optimal codons. Thus, the frequency of these optimal codons in a gene appears to be correlated with the level of gene expression, and may be a useful indicator in the case of genes (or open reading frames) whose expression levels (or even function) are unknown. A second, relatively minor trend among genes is correlated with the frequency of G at synonymously variable sites. It is not yet clear whether this trend reflects variation in base composition (or mutational biases) among regions of the C. elegans genome, or some other factor. Sequence divergence between C. elegans and C. briggsae has also been studied.

Serological markers of hepatitis delta virus (HDV) infection were found in 18 (12%) of 146 consecutive patients with chronic hepatitis B, and the characteristics of patients who had antibody to HDV (anti-HDV-positive) were analyzed. During one to 15 years of follow-up, histological deterioration was documented in 77% of anti-HDV-positive patients; however, in hepatitis B surface antigen (HBsAg) carriers without HDV infection, histology deteriorated in 30% but improved or remained unchanged in the majority of patients (P less than .01). In seven (70%) of the 10 anti-HDV-positive patients who showed transition from chronic active hepatitis to cirrhosis, this event was observed within the first two years of follow-up. The probability of evolution to cirrhosis was significantly higher in anti-HDV-positive patients than in patients without antibody to HDV (P less than .001). These findings indicate that HDV infection in patients with chronic hepatitis B is associated with a more-rapid progression to cirrhosis compared with HBsAg carriers with chronic hepatitis and no evidence of HDV infection.

A family with a peculiar defect of factor VII is described. The propositi, a brother and sister, were born of a nonconsanguineous marriage and presented a mild to moderate bleeding tendency since childhood (epistaxis, excessive bleeding after tooth extraction). The coagulation pattern was characterized by a mildly prolonged prothrombin time, normal partial thromboplastin time, and normal Stypven-cephalin clotting time. Prothrombin consumption and thromboplastin generation were normal. Factor VII activity was 20%-25% of normal but factor VII antigen was about 50% of normal. The father and other members of his family had factor VII activity of 36%-50% of normal but the factor VII antigen level was 100%. The mother had factor VII activity and antigen levels of about 50% of normal. It appeared that the propositi had inherited an abnormal factor VII from the father and a heterozygous factor VII deficiency from the mother. Therefore, the defect would seem to result from a double heterozygous state. The brother of the propositi had 65% factor VII activity and a normal factor VII antigen, probably the result of the interaction of a paternal abnormal factor VII gene and a normal maternal gene. The condition is referred to as factor VII Verona.

Glucosylgalactosylhydroxylysine (GGHYL), galactosylhydroxylysine (GHYL), pyridinoline (PYD) and deoxypyridinoline (DPD) were measured in the urine (6 h serial specimens over 96 and 24 h urine specimens for 4 days) collected from four adult Sprague Dawley rats and in the femoral and tibia] bone as well as in the dorsal skin of the same rats. No significant daily variations were found in the urine excretion of GGHYL, GHYL, PYD and DPD but significant diurnal variations. The GGHYL/GHYL ratio in rat urine (0.46 +/- 0.1) reflected neither the bone collagen ratio (1.9 to 2.4) nor the skin collagen ratio (1.22 +/- 1.07), a finding that may reflect GGHYL conversion into GHYL. The content of both pyridinolines was very low in the skin and high in the bone collagen and the urinary PYD/DPD ratio (1.46 +/- 0.15) reflected essentially the bone collagen ratio (0.8-3.0). These results suggest the usefulness of measuring GGHYL, GHYL, PYD and DPD in 24 h urine specimen and, based on the inter-animal variations, the necessity to consider each animal as its own control when bone turnover needs to be monitored.

To investigate the possibility that hemopoietic cells may become infected with hepatitis B virus (HBV), viral DNA was studied by molecular hybridization in bone marrow aspirates of 51 children with leukemia. HBV-DNA was found in the bone marrow of eight children (15%) and Southern blot analysis revealed the presence of free, monomeric viral sequences. Only one of the eight children with HBV-DNA in bone marrow cells was HBsAg-positive in serum, whereas two additional patients were transiently HBsAg-positive in serum during follow-up, but were negative at the time HBV-DNA was found in bone marrow. Four other cases developed antibodies to HBV. Cases of myeloid leukemia were more frequently positive for HBV-DNA in bone marrow (55%), compared with cases of lymphoid leukemia (7%). These results indicate that hemopoietic cells are susceptible to infection with hepatitis B virus and stimulate new interest into the relation of HBV infection to the development of some forms of leukemia, as four of eight cases of myeloid leukemia were HBV-DNA positive in bone marrow aspirates at diagnosis, prior to receiving any transfusion therapy.

In an attempt to evaluate a possible correlation between cryptogenic chronic liver disease and a present or past hepatitis B virus (HBV) infection, we studied 17 patients with hepatitis B surface antigen (HBsAg)-negative, nonalcoholic chronic liver disease; 9 of them were positive for serum HBsAg detected by a solid-phase enzyme immunoassay with monoclonal antibody (M-EIA) and 8 were negative for the same marker. Liver hepatitis B core antigen (HBcAg), studied by an indirect immunofluorescence technique, was present in 55.5% of the patients positive for serum HBsAg by M-EIA. In the same group of patients, liver HBV-DNA was found in 66.6% of the patients. On the other hand, only 1 patient without serum positivity for HBsAg by M-EIA was positive for liver HBcAg and HBV-DNA. None of our patients showed serum positivity for HBV-DNA sequences. We conclude that HBV infection may be a possible cause of cryptogenic chronic liver disease; this HBV-related, HBsAg-negative chronic liver disease seems to have no viral replication or undetectable levels of HBV-DNA in serum. HBsAg, detected by a monoclonal assay, seems to be a suitable marker to identify this subgroup of patients with HBsAg-negative chronic liver disease.