Inhalation anesthesia induced by isoflurane alters penicillin disposition in swine tissues.

Twelve healthy swine were dosed with penicillin G intramuscularly. Fluids and tissues samples were collected at the end of two periods of general anesthesia, performed 24 h apart. Tissue samples were collected by minimally invasive laparoscopy under general anesthesia at 8 and 28 h postdose. Four nonanesthetized, penicillin-treated pigs were euthanized at 8 h postdose, and a second set of four similarly treated control pigs were sacrificed 28 h postdose. Liver penicillin tissue concentrations from animals that underwent anesthesia and laparoscopic tissue collection had tissue concentrations that were higher than nonanesthetized pigs at both time points. Urine, plasma, kidney, skeletal, and cardiac muscle showed no differences between the two groups. Laparoscopic tissue collection under general anesthesia in swine induces physiological changes that cause alterations in tissue pharmacokinetics not seen in conscious animals.

Microbial Diversity in the Early In Vivo-Formed Dental Biofilm.

Although the mature dental biofilm composition is well studied, there is very little information on the earliest phase of in vivo tooth colonization. Progress in dental biofilm collection methodologies and techniques of large-scale microbial identification have made new studies in this field of oral biology feasible. The aim of this study was to characterize the temporal changes and diversity of the cultivable and noncultivable microbes in the early dental biofilm. Samples of early dental biofilm were collected from 11 healthy subjects at 0, 2, 4, and 6 h after removal of plaque and pellicle from tooth surfaces. With the semiquantitative Human Oral Microbiome Identification Microarray (HOMIM) technique, which is based on 16S rRNA sequence hybridizations, plaque samples were analyzed with the currently available 407 HOMIM microbial probes. This led to the identification of at least 92 species, with streptococci being the most abundant bacteria across all time points in all subjects. High-frequency detection was also made with Haemophilus parainfluenzae, Gemella haemolysans, Slackia exigua, and Rothia species. Abundance changes over time were noted for Streptococcus anginosus and Streptococcus intermedius (P = 0.02), Streptococcus mitis bv. 2 (P = 0.0002), Streptococcus oralis (P = 0.0002), Streptococcus cluster I (P = 0.003), G. haemolysans (P = 0.0005), and Stenotrophomonas maltophilia (P = 0.02). Among the currently uncultivable microbiota, eight phylotypes were detected in the early stages of biofilm formation, one belonging to the candidate bacterial division TM7, which has attracted attention due to its potential association with periodontal disease.

Interferon beta-1b-induced postmenopausal bleeding in a patient with multiple sclerosis.

OBJECTIVE: Postmenopausal bleeding must always be evaluated to rule out endometrial carcinoma, although there are many benign etiologies. There have been rare reports of premenopausal bleeding with interferon beta-1b, used to treat multiple sclerosis, but no prior reports in postmenopausal women. METHODS: Literature searches were performed using PubMed and Medline for articles with content related to premenopausal and postmenopausal bleeding while taking interferon beta-1b. The searches were restricted to the English language. Search terms included interferon beta-1b and/or uterine hemorrhage and/or vaginal bleeding and/or postmenopausal and/or menopause. RESULTS: The literature review found no related articles for postmenopausal bleeding while taking interferon beta-1b. We present a case of a patient with postmenopausal bleeding attributed to elevation of serum estradiol in association with interferon beta-1b therapy. CONCLUSION: It is important for patients and providers to be aware of the association between postmenopausal bleeding with the use of interferon beta-1b therapy which could be due to elevated serum estradiol levels.

Formation of trinuclear rhodium-hydride complexes [{Rh(PP*)H}3-(µ2-H)3(µ3-H)][anion]2--during asymmetric hydrogenation?

Recently described and fully characterized trinuclear rhodium-hydride complexes [{Rh(PP*)H}3(µ2-H)3(µ3-H)][anion]2 have been investigated with respect to their formation and role under the conditions of asymmetric hydrogenation. Catalyst-substrate complexes with mac (methyl (Z)-N-acetylaminocinnamate) ([Rh(tBu-BisP*)(mac)]BF4, [Rh(Tangphos)(mac)]BF4, [Rh(Me-BPE)(mac)]BF4, [Rh(DCPE)(mac)]BF4, [Rh(DCPB)(mac)]BF4), as well as rhodium-hydride species, both mono-([Rh(Tangphos)-H2(MeOH)2]BF4, [Rh(Me-BPE)H2(MeOH)2]BF4), and dinuclear ([{Rh(DCPE)H}2(µ2-H)3]BF4, [{Rh(DCPB)H}2(µ2-H)3]BF4), are described. A plausible reaction sequence for the formation of the trinuclear rhodium-hydride complexes is discussed. Evidence is provided that the presence of multinuclear rhodium-hydride complexes should be taken into account when discussing the mechanism of rhodium-promoted asymmetric hydrogenation.

A Holstein cow-calf model for the transfer of ciprofloxacin through milk after a long-term intravenous infusion.

This study is part of an ongoing effort to develop animal models that provide milk and sufficient infant (offspring) plasma samples to fully describe a drug's pharmacokinetics to quantitate the risk to the nursing infant. Ciprofloxacin was administered to six healthy Holstein cows as a constant rate intravenous infusion (flow rate was weight adjusted) to achieve a steady-state concentration of approximately 300 ng/mL for 7 days. Plasma and milk samples were collected from the cow at regular intervals over the course of the 7 days. The plasma and milk samples were analyzed for ciprofloxacin by high-performance liquid chromatography. The milk was fed to calves, and calf plasma samples were analyzed to study the lactational transfer of ciprofloxacin from dam to nursing neonate. Remarkably, concentrations of ciprofloxacin in milk were 45 times higher than plasma drug concentrations in the dam. Approximately 6% of the administered dose was transferred to the milk, resulting in an average oral dose of 0.5 mg/kg to the calves with every feeding. The drug did not accumulate in the calves, and plasma concentrations were between one-tenth and one-fifth the plasma concentrations of the dam.

Tissue/fluid correlation study for the depletion of sulfadimethoxine in bovine kidney, liver, plasma, urine, and oral fluid.

Sulfonamides are among the oldest, but still effective, antimicrobial veterinary medicines. In steers and dairy cows, the sulfonamides are effective in the treatment of respiratory disease and general infections. Sulfadimethoxine (SDM) has been approved by US Food and Drug Administration (FDA) for use in steers and dairy cows with a tolerance of 100 ng/g (ppb) in edible tissues and 10 ppb in milk. The detection of SDM residue above tolerance in the animal slaughtered for food process will result in the whole carcass being discarded. This report describes a comprehensive depletion study of SDM (and its main metabolite) in plasma, urine, oral fluid, kidney, and liver. In this study, nine steers were injected intravenously with the approved dose of SDM; the loading dose was 55 mg/kg, followed by 27.5 mg/kg dose at 24 h and again at 48 h. Fluids (blood, urine, and saliva) and tissue (liver and kidney) samples were collected at intervals after the last dose of SMD. The combination of laparoscopic serial sampling technique with the liquid chromatography/mass spectrometry method provided the data to establish the tissue/fluid correlation in the depletion of SMD. A strong correlation and linearity of the log-scale concentration over time in the depletion stage has been confirmed for kidney, liver, and plasma.

Saliva in the Diagnosis of COVID-19: A Review and New Research Directions.

This review presents literature that highlights saliva's utility as a biofluid in the diagnosis and monitoring of COVID-19. A systematic search was performed in 5 electronic databases (PubMed, Embase, LILACS, Scopus, and Web of Science). Studies were eligible for inclusion if they assessed the potential diagnostic value and/or other discriminatory properties of biological markers in the saliva of patients with COVID-19. As of July 22, 2020, a total of 28 studies have investigated the presence of SARS-CoV-2 RNA in saliva. Several of those studies confirmed reliable detection of SARS-CoV-2 in the saliva of patients with COVID-19. Saliva offered sensitivity and specificity for SARS-CoV-2 detection comparable to that of the current standard of nasopharyngeal and throat swabs. However, the utility of saliva in diagnosing COVID-19 infection remains understudied. Clinical studies with larger patient populations that measure recordings at different stages during the disease are still necessary to confirm the accuracy of COVID-19 diagnosis with saliva. Nevertheless, the utility of saliva as a diagnostic tool opens the possibility of using rapid and less invasive diagnostic strategies by targeting bioanalytes rather than the pathogen.

Chorion laeve accreta - Another manifestation of morbid adherence.

INTRODUCTION: Smooth muscle in the decidua of fetal membranes (membrane myofibers, MMF) is not mentioned in standard textbooks. METHODS: The current report presents collected observations on 52 patients with MMF at 2 institutions between 2004 and 2017 - including placentas, postpartum curettages, and hysterectomies. RESULTS: Clinical presentations include observation of adherent membranes during delivery, disrupted and incomplete membranes in placentas submitted for examination, postpartum bleeding associated with retained fetal membranes, association with membrane hematomas and membrane hemosiderin, morbidly adherent fetal membranes in hysterectomies; and association with grossly adherent pieces of tissue or nodules in fetal membranes. DISCUSSION: Although MMF can be an incidental microscopic observation in a routine placenta, the suggested diagnostic terminology when there are clinical and/or gross presentations is Chorion Laeve Accreta (ChLA). Further study is needed but MMF appears to be the fetal membrane counterpart of BPMF(basal plate myofibers), possibly due to damage of subjacent myometrium by trophoblastic proteases, so that shear stress during delivery causes myofibers to come out attached to the decidua of fetal membranes. Neither the prevalence of MMF, nor its reliability as a marker for placenta accreta is addressed in this collection. Association of MMF with BPMF, and recurrence of MMF, are documented; but the true frequency of these phenomena remains to be established.

Synthesis of Rh(iii) thiophosphinito pincer hydrido complexes by base-free C-H bond activation at room temperature.

Rhodium(iii) thiophosphinito pincer hydrido complexes were synthesised by C-H activation under exceptionally mild conditions at room temperature without additional base or irradiation and fully characterised by multinuclear NMR spectroscopy and X-ray crystallography. C-H activation under these mild conditions contrasts with the reactivity of related systems with POCOP ligands.

Chronic hypoxia in vivo reduces placental oxidative stress.

Decreased placental oxygenation and increased oxidative stress are implicated in the development of preeclampsia. Oxidative stress arises from imbalance between pro-versus anti-oxidants and can lead to biological oxidation and apoptosis. Because pregnant women living at high altitude (3100 m, HA) have lowered arterial PO2 and an increased incidence of preeclampsia, we hypothesized that HA placentas would have decreased anti-oxidant enzyme activity, increased oxidative stress (lipid peroxidation, protein oxidation and nitration) and greater trophoblast apoptosis than low-altitude (LA) placentas. We measured enzymatic activities, lipid and protein oxidation and co-factor concentrations by spectrophotometric techniques and ELISA in 12 LA and 18 HA placentas. Immunohistochemistry (IHC) was used to evaluate nitrated proteins and specific markers of apoptosis (activated caspase 3 and M30). Superoxide dismutase activity was marginally lower (p=0.05), while glutathione peroxidase activity (p<0.05), thioredoxin concentrations (p<0.005) and thioredoxin reductase activity p<0.01 were all reduced in HA placentas. Decreased anti-oxidant activity was not associated with increased oxidative stress: lipid peroxide content and protein carbonyl formation were lower at HA (p<0.01). We found greater nitrotyrosine residues in the syncytiotrophoblast at 3100 m (p<0.05), but apoptosis did not differ between altitudes. Our data suggest that hypoxia does not increase placental oxidative stress in vivo. Nitrative stress may be a consequence of hypoxia but does not appear to contribute to increased apoptosis. Lowered placental concentrations of anti-oxidants may contribute to the susceptibility of women living at HA to the development of preeclampsia, but are unlikely to be etiological.

A highly conserved mouse gene with a propensity to form pseudogenes in mammals.

A mouse cDNA clone corresponding to an abundantly transcribed poly(A)+ mRNA was found to be represented by 200 copies in mammalian genomes. To understand the origin and nature of this sequence family, we studied two genomic members and two cDNA clones from mouse liver. The DNA sequence of the coding strand of a full-length cDNA clone was shown to have an open reading frame capable of encoding a 25-kilodalton polypeptide that has not been previously described. In vitro transcription-translation experiments verified the presence of an open reading frame encoding a protein of the predicted size. Restriction analysis of genomic DNA and DNA sequence analysis of genomic clones indicated that many of the 200 members of this family represent processed pseudogenes, with one or a small number of active structural genes. The vast majority of the genomic copies are heterogeneous in length, truncated at their 5' ends with respect to the mRNA, and do not appear to have intervening sequences. Two distinct genomic members of this family were sequenced and found to represent incomplete copies of the mRNA. Both are 5' truncated at slightly different points with respect to the mRNA. Both pseudogenes have multiple base changes, insertions, and deletions relative to the mRNA, and one of them encodes the poly(A) tail of the mRNA. The expression of this gene family is highest in rapidly dividing cells such as early mouse embryos and testis, but was seen in all tissues tested. This gene shows extremely high sequence conservation, extending to chicken, amphibian, and nematode genomes. Surprisingly, the gene appears to exist in only one copy in these organisms.

Saliva and Serum Protein Exchange at the Tooth Enamel Surface.

The acquired enamel pellicle is an oral, fluid-derived protein layer that forms on the tooth surface. It is a biologically and clinically important integument that protects teeth against enamel demineralization, and abrasion. Tooth surfaces are exposed to different proteinaceous microenvironments depending on the enamel location. For instance, tooth surfaces close to the gingival sulcus contact serum proteins that emanate via this sulcus, which may impact pellicle composition locally. The aims of this study were to define the major salivary and serum components that adsorb to hydroxyapatite, to study competition among them, and to obtain preliminary evidence in an in vivo saliva/serum pellicle model. Hydroxyapatite powder was incubated with saliva and serum, and the proteins that adsorbed were identified by mass spectrometry. To study competition, saliva and serum proteins were labeled with CyDyes, mixed in various proportions, and incubated with hydroxyapatite. In vivo competition was assessed using a split-mouth design, with half the buccal tooth surfaces coated with serum and the other half with saliva. After exposure to the oral environment for 0 min, 30 min and 2 h, the pellicles were analyzed by SDS-PAGE. In pure saliva- or serum-derived pellicles, 82 and 84 proteins were identified, respectively. When present concomitantly, salivary protein adsorbers effectively competed with serum protein adsorbers for the hydroxyapatite surface. Specifically, acidic proline-rich protein, cystatin, statherin and protein S100-A9 proteins competed off apolipoproteins, complement C4-A, haptoglobin, transthyretin and serotransferrin. In vivo evidence further supported the replacement of serum proteins by salivary proteins. In conclusion, although significant numbers of serum proteins emanate from the gingival sulcus, their ability to participate in dental pellicle formation is likely reduced in the presence of strong salivary protein adsorbers. The functional properties of the acquired enamel pellicle will therefore be mostly dictated by the salivary component.

The future of lasers in onychomycosis.

INTRODUCTION: Laser therapy is one of the most promising device-based therapies for onychomycosis. To date, reported clinical efficacies, as well as anecdotal clinical results, have varied greatly, and the specific mechanism of action has not been well-elucidated. METHODS: Here, we provide an overview of the mechanisms of action and detailed analysis of the technical parameters involved in creating a laser that will be successfully fungicidal in onychomycosis. RESULTS: This review provides important insight into why the efficacies of laser studies reported to date vary so greatly and what is critical in order to obtain high efficacy in the clinical treatment of onychomycosis. CONCLUSIONS: There is substantial opportunity to improve the targeting and anti-targeting properties of lasers to address the specific considerations required to treat onychomycosis and, more generally, other dermal pathogens.

The proto-oncogene c-ros codes for a transmembrane tyrosine protein kinase sharing sequence and structural homology with sevenless protein of Drosophila melanogaster.

Our earlier study predicted that proto-oncogene c-ros codes for a receptor-like tyrosine protein kinase (TPK). To further understand its protein structure and physiological function, we have analysed its expression in various tissues of chicken and have isolated and sequenced cDNA clones containing the entire coding region of the gene. Confirming our earlier study, we found that kidney is the organ that expresses the highest level of c-ros mRNA, in addition, we found a lower level of expression in gonad, thymus, bursa and brain. A distinctive 8.3 kb c-ros mRNA is present in kidney. No detectable amount of c-ros mRNA was found in the rest of tissues examined. Nucleotide sequence of the c-ros cDNA predicts that it codes for a transmembrane (TM) TPK molecule of 2311 amino acids (aa). The extracellular domain consists of 1873 amino acids which share 20 to 43% homology with that of the Drosophila sevenless protein and TPK domains of the two genes have 58 to 74% homology. The extracellular domain containing 37 potential N-linked glycosylation sites is preceded by a 5' hydrophobic sequence resembling a typical signal peptide. An internal hydrophobic domain of 26 amino acids, the presumed transmembrane domain, is followed by a spacer sequence of 58 amino acids, a TPK domain of 270 amino acids and a carboxyl tail of 84 amino acids. Overall, our result indicates that c-ros codes for a glycosylated transmembrane TPK molecule which shares a remarkable sequence and structural homology with that of Drosophila sevenless protein.

Diminution of bone mass in childhood diabetes.

Photon absorption measurements of forearm bone density in 196 insulin-dependent patients, age 6--26 years, were compared with findings in 124 controls. Expected density, gm. Ca/cm.2 bone width (M/W), was calculated from regressions of M/W on ulnar length for white and black male and female controls. There were no significant correlations between M/W differences from expected and serum Ca, Mg, P, or alkaline phosphatase levels, estimated physical activity level, insulin dosage, or the presence of joint contracture. White females averaged 8.2 per cent (+/- 1 S.E.M.) loss of M/W, as against white male average loss of 4.7 per cent +/- 1 and black female loss of 2 per cent +/- 2 (p less than 0.001); the black male population was too small for separate analysis. M/W loss greater than 10 per cent was seen in 29 per cent of white males, 19 per cent of blacks, and 48 per cent of white females (p less than 0.02). When the groups were further divided into those with duration of diabetes less than or equal to five years and those with duration greater than five years, significant reduction in M/W average loss over time was seen with white females (10.6 per cent +/- 1.2 to 3.7 per cent+/- 1.5, p less than 0.0001). Expression of this defect in bone mineralization is controlled by race and sex acting independently of each other.

Organization and expression of non-Alu family interspersed repetitive DNA sequences in the mouse genome.

The mouse genome is complex with regard to DNA sequence organization and transcriptional activity. To more fully understand the role of interspersed repetitive DNA sequences we have isolated and characterized five different mouse non-Alu DNA sequence families. We have found that: (1) the distribution of repetitive sequences is non-random in the genome; (2) two of the five families (Bam5 and R) were previously described by Fanning (1982) and Gebhard et al. (1982), respectively. We found that these two families are linked to each other and are found adjacent to seven of seven studied structural genes but in randomly selected DNA fragments showed much less significant linkage. (3) The position of the Bam5 and R family repeat units relative to beta-globin and relative to a housekeeping gene has been evolutionarily conserved in mice and humans. (4) Three previously undescribed families representing from 200 to 40,000 copies per genome have been characterized and shown to have equivalent human sequences. (5) All five families studied are represented in RNA polymerase II transcripts. Little RNA polymerase III transcription homologous to these three families could be detected. The structural and functional features of these five families defined in this paper provide a basis for studies on the functional role of interspersed repetitive DNA in the mouse.

Placental diagnostic criteria and clinical correlation--a workshop report.

Placental pathology is a valuable link explaining how underlying pregnancy risk factors result in adverse pregnancy outcome. This potentially useful information is currently underutilized due to a general lack of rigor in placental diagnosis. This lack of rigor is in large part due to a failure to appreciate the importance of the information provided and hence to demand accurate and clinically responsive reporting. This workshop reviewed the results of a recent initiative by the Perinatal Section of the Society for Pediatric Pathology to systematize and validate diagnostic schema for the description of lesions related to three important pathologic processes: amniotic fluid infection, maternal vascular underperfusion, and fetal vascular obstruction. This was followed by presentation of three studies correlating these pathologic processes with the following clinical outcomes: complications of prematurity, fetal growth restriction, and neurodisability following term delivery.

Levels and role of cytokines in bovine leukemia virus (BLV) infection.

Bovine leukemia virus belongs to a small subfamily of exogenous retroviruses that includes the human retroviruses HTLV-1, HTLV-II and the simian virus, STLV-1. Like other retroviruses, infection with BLV results in deregulation of the host immune system at both humoral and cellular levels. An approach which might help in the elucidation of some immune impairment phenomena is the investigation of the role that cytokines play in the pathogenesis and immune response of BLV infected animals. Here we describe our findings on IL-6 and TNF. We have found that the levels of IL-6 in the sera of BLV infected cows which show persistent lymphocytosis (BLV+ PL+) were significantly higher than those of BLV infected with no lymphocytosis (BLV+ PL-) or BLV negative cows (BLV-). The same results were obtained by measuring the spontaneous production of IL-6 in peripheral blood mononuclear cells (PBMC). Furthermore, PBMC derived from BLV+PL+ cows secrete higher levels of IL-6 and TNF alpha than those derived from BLV+PL- and BLV- ones following in vitro exposure to the BLV gp51 antigen, bacterial endotoxins (LPS) and ConA. Similar results were obtained when supernatants from stimulated adherent (monocytes, macrophages) and non-adherent cells (B and T lymphocytes) were tested. When exogenous IL-6 and TNF alpha were added to BLV infected cells in vitro, the expression of viral antigens was strongly suppressed. Thus, the possibility exists that the elevated production of IL-6 and even more than that of TNF alpha play a role as contributing factors to the latency of the clinical expression in BLV infection.

Child patients in a field hospital during the 2003 Gulf conflict.

During the 2003 conflict in the Gulf, 202 Field Hospital provided the initial Role 3 medical support when hostilities began on 20th March. The first child casualty arrived two days later and over the next four weeks there were 24 admissions of patients aged 15 years or less, amounting to 2% of the total admissions but constituting 19% of ITU admissions. There were 49 operations on children out of a total of 352 theatre cases. The small number of children treated had a disproportionate impact on the functioning of the field hospital. Planned provision for such patients could reduce this impact and should include suitable equipment and specialist staff.