The magnetosome proteins MamX, MamZ and MamH are involved in redox control of magnetite biomineralization in Magnetospirillum gryphiswaldense.
Magnetospirillum gryphiswaldense uses intracellular chains of membrane-enveloped magnetite crystals, the magnetosomes, to navigate within magnetic fields. The biomineralization of magnetite nanocrystals requires several magnetosome-associated proteins, whose precise functions so far have remained mostly unknown. Here, we analysed the functions of MamX and the Major Facilitator Superfamily (MFS) proteins MamZ and MamH. Deletion of either the entire mamX gene or elimination of its putative haem c-binding magnetochrome domains, and deletion of either mamZ or its C-terminal ferric reductase-like component resulted in an identical phenotype. All mutants displayed WT-like magnetite crystals, flanked within the magnetosome chains by poorly crystalline flake-like particles partly consisting of haematite. Double deletions of both mamZ and its homologue mamH further impaired magnetite crystallization in an additive manner, indicating that the two MFS proteins have partially redundant functions. Deprivation of ΔmamX and ΔmamZ cells from nitrate, or additional loss of the respiratory nitrate reductase Nap from ΔmamX severely exacerbated the magnetosome defects and entirely inhibited the formation of regular crystals, suggesting that MamXZ and Nap have similar, but independent roles in redox control of biomineralization. We propose a model in which MamX, MamZ and MamH functionally interact to balance the redox state of iron within the magnetosome compartment.


Pancreatic secretion in response to an elemental diet and intravenous hyperalimentation.
Pancreatic exocrine responses to oral and intraduodenal administration of an elemental diet and to an intravenously administered hyperalimentation solution were determined in dogs with fistulas of the stomach and pancreas. Administration of an intraduodenal elemental diet prevented the responses to water and bicarbonate seen with oral administration but did not prevent the protein response. Intravenously administered hyperalimentation solution produced only minor effects on pancreatic secretion when compared with the elemental diet.


EV@LUTIL: An open access database on occupational exposures to asbestos and man-made mineral fibres.
The aim of Evalutil is to document occupational exposure to asbestos and man-made mineral fibers. These databases provide grouped descriptive and metrological data from observed situations of occupational exposure, collected through the analysis of scientific articles and technical reports by industrial hygienists. Over 5,000 measurements were collected. We describe the occupations, economic activities, fiber-containing products, and operations on them that have been documented most often. Graphical measurement syntheses of these data show that the situations presented for asbestos and RCF, except mineral wools, report fiber concentrations mainly above historical occupational exposure limits. Free access to these data in French and in English on the Internet (https://ssl2.isped.u-bordeaux2.fr/eva_003/) helps public health and prevention professionals to identify and characterize occupational exposures to fibers. Extended recently to nanoscale particles, Evalutil continues to contribute to the improvement of knowledge about exposure to inhaled particles and the health risks associated with them.


Relationship between pancreatic lesions and serum glucose values in Mystromys albicaudatus.
Serum glucose levels and pancreatic islet cell changes were studied in combination in 127 Mystromys albicaudatus in an effort to develop better criteria for defining diabetes mellitus in thiss rodent. The study supports a previous report in which 170 mg/100 ml was selected as indicative of hyperglycemia based on statistical analysis of serum glucose distributions. The presence of exceptions to predicted correlations between islet cell changes and serum glucose levels was identified for further study.


Frequency of the Rendement Napole RN- allele in a population of American Hampshire pigs.
A total of 204 purebred Hampshire pigs were obtained from 23 breeders. These animals were the progeny of 41 sires and 123 dams. A sample of purebred Yorkshire (n = 24) pigs were also used in the study. Animals were classified by glycolytic potential determined on a live-animal longissimus muscle biopsy sample. Hampshire pigs (n = 176) with glycolytic potential greater than 185 micromol/g (x = 238.8 micromol/g; SD = 29.54) were classified as heterozygous or homozygous for the dominant RN- allele (RN-rn+ or RN-RN-, respectively), whereas cohorts (n = 28) with glycolytic potential less than 185 micromol/g (x = 141.3 micromol/g; SD = 24.48) were considered as homozygous normal (rn+rn+). All Yorkshire pigs (n = 24) had a mean glycolytic potential level of 146.1 micromol/g (SD = 20.18) and were considered as homozygous normal (rn+rn+). The Hardy-Weinberg equilibrium yielded frequencies of .630 and .370 for the dominant RN- allele and normal rn+ alleles in the Hampshire population, respectively, and genotypic frequencies of .397 (RN-RN-), .466 (RN-rn+), and .137 (rn+rn+). Hampshires with glycolytic potential > or = 185 micromol/g had significantly lower longissimus muscle ultimate pH, intramuscular fat, subjective marbling scores, and percentage of protein (P < .001) and had greater longissimus muscle percentage of moisture (P < .001), drip loss (P < .01), and cooking loss (P < .001) than rn+rn+ Hampshires and Yorkshires. These data suggest the RN- allele exists at a high frequency within the American Hampshire breed. Higher glycolytic potential levels, which accompany the allele, may cause decreased meat quality.


Navigating healthcare systems before and after resettlement: Exploring experiences and recommendations for improvement from the perspectives of a Bhutanese refugee community.
Though refugees often survive in refugee camps for many years, little is known about the impact of their interactions within the healthcare system during that time and how it may affect current concerns with the healthcare system after resettlement. Guiding our analysis was the Community Health Development model, which emphasizes the importance of understanding the impact of historical experiences on a community's health to identify specific current health needs, and plan solutions alongside the community to address and improve health concerns. To better understand the healthcare system related experiences of Bhutanese refugees before and after resettlement, and describe potential solutions based on their perspectives to improve their health status. This study used an explorative qualitative research design. Four focus group discussions were conducted with 40 female participants to examine their experiences within the healthcare system in Nepal (e.g. before resettlement) and the US (after resettlement). Focus group data were audio-recorded, translated, coded, and reported based on qualitative thematic analysis. Findings revealed that Bhutanese refugees were mistreated in the Nepalese healthcare system, often neglected from healthcare access and services because of their refugee status. Upon arrival to the United States after resettlement, study participants also reported experiencing challenges within the US health care system including cultural and linguistic barriers when interacting with medical interpreters during visits with their providers, as well as having inadequate time during the visit to fully express their concerns. Respondents' recommendations to improve their overall health centered on their experiences with the US health care system including initiatives developing leadership skills for building community capacity towards advocating for the refuges, while increasing access to external resources. The result of this study outlines an account of Bhutanese refugees' experiences and recommendations for improving their community's health based on such past experiences and their current needs. These findings provide a starting point for future research with underserved refugee migrant groups and indicate a need for health programs to be historically and culturally sensitive in order to be more effective. Further, the understanding of refugees' collective history should inform the development of collaborative interventions with community members in order to be effective.


Increased resistance to acetaminophen-induced hepatotoxicity in retrovirus-infected mice.
In this research we examined the influence of chronic retrovirus infection on the hepatic metabolism of a model substrate, acetaminophen (APAP), and its induced liver injury in mice inoculated with LP-BM5 murine leukemia viruses. Female C57BL/6 mice at 15-17 wks after LP-BM5 retrovirus inoculation and age-matched control animals were used in the studies. APAP treatment (300 mg kg-1, p.o.) resulted in moderate to severe centrilobular necrosis in control animals, with the necrotic area accounting for 40-60% of the total area. In contrast, the APAP-treated (300 mg kg-1, p.o.) infected animals exhibited mild zonal necrosis with the necrotic area accounting for 1-6% of the total area. In the same study, a statistically significant higher percentage of APAP glucuronide and a lower percentage of unchanged APAP were recovered from the urine of the LP-BM5-inoculated animals than from that of controls. No statistically significant differences between infected and uninfected animals in the urinary recovery of APAP sulfate, APAP cysteine, or APAP mercapturate were observed. The formation of APAP metabolites and APAP-associated biochemical changes were also determined from liver slice preparation to avoid in vivo complicating factors. Consistently more significant depletion of the intracellular glutathione levels and K+ content were observed in slices from the uninfected animals at high concentrations of APAP (1 and 2 mM) than in slices from the retrovirus-infected animals. The differences in APAP-associated biochemical changes were accompanied by a 1.4-1.5-fold increase in the formation of APAP glucuronide, sulfate, and glutathione metabolites in slices prepared from animals inoculated with LP-BM5. We concluded that, based on histological examination and hepatic biochemical measurements, the retrovirus-infected animals were more resistant to APAP-induced liver injury. This could be due, in part, to alterations in the detoxification and activation metabolic pathways of APAP.


Model-based clustering of microarray expression data via latent Gaussian mixture models.
In recent years, work has been carried out on clustering gene expression microarray data. Some approaches are developed from an algorithmic viewpoint whereas others are developed via the application of mixture models. In this article, a family of eight mixture models which utilizes the factor analysis covariance structure is extended to 12 models and applied to gene expression microarray data. This modelling approach builds on previous work by introducing a modified factor analysis covariance structure, leading to a family of 12 mixture models, including parsimonious models. This family of models allows for the modelling of the correlation between gene expression levels even when the number of samples is small. Parameter estimation is carried out using a variant of the expectation-maximization algorithm and model selection is achieved using the Bayesian information criterion. This expanded family of Gaussian mixture models, known as the expanded parsimonious Gaussian mixture model (EPGMM) family, is then applied to two well-known gene expression data sets. The performance of the EPGMM family of models is quantified using the adjusted Rand index. This family of models gives very good performance, relative to existing popular clustering techniques, when applied to real gene expression microarray data. The reduced, preprocessed data that were analysed are available at www.paulmcnicholas.info


A Dioxygenase Catalyzes Steroid 16α-Hydroxylation in Steroidal Glycoalkaloid Biosynthesis.
Steroidal glycoalkaloids (SGAs) are toxic specialized metabolites that are found in the Solanaceae. Potato (Solanum tuberosum) contains the SGAs α-solanine and α-chaconine, while tomato (Solanum lycopersicum) contains α-tomatine, all of which are biosynthesized from cholesterol. However, although two cytochrome P450 monooxygenases that catalyze the 22- and 26-hydroxylation of cholesterol have been identified, the 16-hydroxylase remains unknown. Feeding with deuterium-labeled cholesterol indicated that the 16α- and 16β-hydrogen atoms of cholesterol were eliminated to form α-solanine and α-chaconine in potato, while only the 16α-hydrogen atom was eliminated in α-tomatine biosynthesis, suggesting that a single oxidation at C-16 takes place during tomato SGA biosynthesis while a two-step oxidation occurs in potato. Here, we show that a 2-oxoglutarate-dependent dioxygenase, designated as 16DOX, is involved in SGA biosynthesis. We found that the transcript of potato 16DOX (St16DOX) was expressed at high levels in the tuber sprouts, where large amounts of SGAs are accumulated. Biochemical analysis of the recombinant St16DOX protein revealed that St16DOX catalyzes the 16α-hydroxylation of hydroxycholesterols and that (22S)-22,26-dihydroxycholesterol was the best substrate among the nine compounds tested. St16DOX-silenced potato plants contained significantly lower levels of SGAs, and a detailed metabolite analysis revealed that they accumulated the glycosides of (22S)-22,26-dihydroxycholesterol. Analysis of the tomato 16DOX (Sl16DOX) gene gave essentially the same results. These findings clearly indicate that 16DOX is a steroid 16α-hydroxylase that functions in the SGA biosynthetic pathway. Furthermore, St16DOX silencing did not affect potato tuber yield, indicating that 16DOX may be a suitable target for controlling toxic SGA levels in potato.


[Decompensated sinusitis complicated by cancer of the nasal cavity and the maxillary sinus].
This paper was designed to illustrate the probability of the development of an oncological disease of the nasal cavity and the paranasal sinuses in young subjects presenting with long-standing decompensated chronic sinusitis; also, it demonstrates the possibility of long-term remission after surgical elimination of the inflammatory process. A 28 year-old patient is described having decompensated chronic pansinusitis complicated by sepsis and secondary suppurative dacryocystitis. Multiple surgical interventions on several paranasal sinuses for the treatment of this pathology proved inefficient and were followed by sanation rhinosinusotomy with the simultaneous removal of the malignant tumour. The long-term monitoring of paranasal sinus condition after radical pansinusotomy and ablation of the tumour followed by a course of radiotherapy gave evidence of the malignant process with simultaneous elimination of inflammatory changes in paranasal sinuses.


Skin Care Products: Age-Appropriate Cosmetics.
In order to maintain skin in "good condition" one can use cosmetic products. Importantly, those skin care products should fulfil specific requirements for specific life phases and specific skin conditions. In this review, we focused on 2 different age groups - namely, infants and the elderly - as well as on 2 specific skin conditions occurring in both age groups - very dry skin (Xerosis) and hyperhydrated skin (diaper rash). The goal in both conditions should be to maintain skin surface in its physiological acidic state, which is in turn crucial for the permeability barrier function, stratum corneum integrity/cohesion and antimicrobial defense. Skin care products formulated with an effective buffer system at a more acidic pH, for example 4, may be the best option to improve the acid mantle and skin barrier function and thus keep the skin in "good condition."


Sexual activity and vaginal topography in women with symptomatic pelvic floor disorders.
Pelvic floor disorders affect vaginal anatomy and may affect sexual function. The aims of this study were to explore the relationship between vaginal anatomy and sexual activity in women with symptomatic pelvic floor disorders and to assess whether vaginal measurements (topography) correlate with sexual function. This is a retrospective cohort study comparing sexually active and nonsexually active women planning urogynecologic surgery. Our primary outcome was the difference in vaginal topography based on Pelvic Organ Prolapse Quantification (POP-Q) exam between cohorts. Correlations between POP-Q measurements and Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire short form (PISQ-12) scores were assessed in sexually active women. The POP-Q is a quantitative and standardized examination for prolapse. The PISQ-12 is a condition-specific sexual function questionnaire validated in sexually active women with pelvic floor disorders. Of 535 women, 208 (39%) were sexually active and 327 (61%) were not. Median genital hiatus (GH) and perineal body (PB) measurements and a PB:GH ratio were not significantly different between the two cohorts. Total vaginal length (TVL) was longer in sexually active women (median 9 vs. 8 cm, P<0.001). In a linear regression analysis controlling for potential confounders, sexually active women still had a longer TVL by 0.4 cm (95% confidence interval 0.07, 0.6 cm) compared with those who were not sexually active. Of the 327 nonsexually active women, 28% indicated they avoided sexual activity because of pelvic floor symptoms. There was poor correlation between TVL, GH, PB, and PB : GH ratio with PISQ-12 scores (r=0.10, -0.05, -0.09, -0.03, respectively). In women with pelvic floor disorders, sexual activity is associated with a longer vaginal length. One-quarter of women indicated they avoided sexual activity because of pelvic floor symptoms. Vaginal topography does not correlate with sexual function based on PISQ-12 scores.


Nucleosome Positioning by an Evolutionarily Conserved Chromatin Remodeler Prevents Aberrant DNA Methylation in Neurospora.
In the filamentous fungus Neurospora crassa, constitutive heterochromatin is marked by tri-methylation of histone H3 lysine 9 (H3K9me3) and DNA methylation. We identified mutations in the Neurospora defective in methylation-1 (dim-1) gene that cause defects in cytosine methylation and implicate a putative AAA-ATPase chromatin remodeler. Although it was well-established that chromatin remodelers can affect transcription by influencing DNA accessibility with nucleosomes, little was known about the role of remodelers on chromatin that is normally not transcribed, including regions of constitutive heterochromatin. We found that dim-1 mutants display both reduced DNA methylation in heterochromatic regions as well as increased DNA methylation and H3K9me3 in some intergenic regions associated with highly expressed genes. Deletion of dim-1 leads to atypically spaced nucleosomes throughout the genome and numerous changes in gene expression. DIM-1 localizes to both heterochromatin and intergenic regions that become hyper-methylated in dim-1 strains. Our findings indicate that DIM-1 normally positions nucleosomes in both heterochromatin and euchromatin and that the standard arrangement and density of nucleosomes is required for the proper function of heterochromatin machinery.


[Clinical electro-ophthalmology at the Max Planck Institute of the Frankfurt University Ophthalmology Clinic 1970-1991].
The survey shows the frequency and distribution of diseases evaluated by electroophthalmological methods. Patients with retinal diseases (51.2%) and those with diseases of the optic nerve (21.8%) were examined most frequently. In a high percentage these investigations lead to a clinically useful assessment: described as confirmation or exclusion of a clinical diagnosis, as establishing a possible differential diagnosis or clearing up formerly unknown aspects of a disease. In cases of hereditary retinal disorders only 11% remained unclear, with presumed optic neuritis only 6%. The importance of electroophthalmological investigations is there ability to assess functional deficits in the visual system especially in somehow more rare retinal and centrally located disorders, functional deficits of unknown origins or in general diseases including the visual system.


Transcriptional control of high molecular weight keratin gene expression in multistage mouse skin carcinogenesis.
Monospecific antikeratin antisera and specific complementary DNA probes were used to analyze expression of keratin genes in newborn mouse skin and skin papillomas and carcinomas by indirect immunofluorescence, immunoblotting, and in situ hybridization. Tumors were induced by initiation with 7,12-dimethylbenz[a]anthracene and promotion with 12-O-tetradecanoylphorbol-13-acetate. Type I epidermal keratin K14 protein (Mr 55,000) is found in all living layers of the newborn skin but is most abundant in the lower strata. K1 (Mr 67,000) and K10 (Mr 59,000) proteins are predominantly suprabasal and K1 is processed in the stratum corneum. Transcripts for K14 were confined largely to the basal cell layer by in situ hybridization. Transcripts for K1 and K10 were highly expressed in suprabasal cells including the granular cell layer. In benign tumors, distribution of K14 protein is similar to that in newborn skin, while the abundance of K1 and K10 appears to be somewhat reduced although the tissue distribution remains suprabasal. Transcription of K14 is aberrant in benign tumors and transcripts persist throughout much of the suprabasal cell layers. Transcripts of K1 and K10 are normally distributed in papillomas but grain density is less intense than in newborn epidermis. Keratin expression in carcinomas is highly disturbed. K14 protein and transcripts are highly expressed in all strata in carcinomas while protein and transcripts for K1 and K10 are essentially absent. These results suggest that papilloma cells fail to respond to or generate signals to regulate K14 expression in the differentiating suprabasal cell layers and may not fully express their suprabasal cell keratins. Carcinomas fail to express suprabasal cell keratins and this is regulated at the transcriptional level. The loss of suprabasal keratin expression may provide a marker for malignant conversion in the mouse skin carcinogenesis model.


The oligosaccharide component of alpha 1-adrenergic receptors from BC3H1 and DDT1 muscle cells. Studies with glycosidases and photoaffinity labelling of intact cells.
In this study, we clarify the structural aspects of the oligosaccharides associated with the alpha 1-adrenergic receptor in two muscle cell lines. Photoaffinity labelling of intact BC3H1 or DDT1 muscle cells with 2-[4-(4-azido-3-[125I]iodobenzoyl)piperazin-1-yl]-4-amino-6, 7-dimethoxyquinazoline ([125I]azidoprazosin) followed by SDS/polyacrylamide-gel electrophoresis (PAGE) and autoradiography revealed specifically labelled proteins of molecular mass = 87,000 and 81,000, respectively. Treatment of photoaffinity-labelled receptors in DDT1 cells with 33 u. of endoglycosidase F/ml for 24 h resulted in the loss of the 81 kDa receptor and the appearance of a 52.5 kDa protein. When lower concentrations of glycosidase or shorter incubation times were used, the 81 kDa receptor was converted to a 66 kDa protein. Treatment of the photoaffinity-labelled BC3H1 receptor with endoglycosidase F resulted in the appearance of a 50.5 kDa protein. Neither alpha-mannosidase nor endoglycosidase H had an effect on the photoaffinity labelling patterns of the receptor from the two cell types. alpha 1-Adrenergic receptors, solubilized from membranes prepared from BC3H1 and DDT1 cells, bound to wheat germ agglutinin-Sepharose and were displaced by N-acetylglucosamine. Taken together, these results indicate that alpha 1-adrenergic receptors in BC3H1 and DDT1 cells contain complex, but not high, mannose oligosaccharide chains; differences in the composition or number of chains partially accounts for the different molecular mass of the receptor in the two cell lines. The results further indicate that the oligosaccharide chains contribute substantially to the apparent molecular mass of alpha 1-adrenergic receptors, as detected by SDS/PAGE, and that the protein backbone of these receptors is likely to be approximately 50 kDa.


Cerebrovascular effects of carbon monoxide.
This review examines the influence of endogenous and exogenous carbon monoxide (CO) on the cerebral circulation. Although CO generated from neuronal heme oxygenase can modulate neurotransmission, evidence supporting its role in cerebral vasodilation is limited. In newborn piglets, heme oxygenase is enriched in microvessels and contributes to hypoxic vasodilation. Low CO concentrations dilate piglet arterioles by opening calcium-activated potassium channels. With inhalation of CO and formation of carboxyhemoglobin, cerebral vasodilation can be greater than that occurring with hypoxic hypoxia at equivalent reductions of arterial oxygen content. This additional vasodilation is probably attributable to additional release of hypoxic vasodilators secondary to increased oxyhemoglobin affinity, although direct effects of CO on cerebral arterioles may also occur. When CO exposure is prolonged, cerebral endothelium undergoes oxidant stress as evident by nitrotyrosine formation. As CO levels increase, modest decreases in oxygen consumption are detectable, which may reflect CO or nitric oxide interactions with cytochrome oxidase in regions with very low oxygen availability. If subsequent CO concentration increases sufficiently to depress cardiac function and limit cerebral perfusion, cerebral oxygen consumption becomes further reduced, and oxidant stress becomes amplified by leukocyte sequestration and xanthine oxidase activity with consequent lipid peroxidation. Specific regions of the brain, such as central white matter, globus pallidus, and hippocampus, are selectively vulnerable to CO toxicity, but whether the mechanisms involved in selective injury differ from other forms of hypoxia-ischemia needs to be clarified.


The legal framework for mental health nursing.
This article sets out basic legal principles underlying the care and treatment of mentally ill clients. Legal considerations are important for nurses, because it is now felt that the care of the mentally ill is as much a legal matter as a medical one. The frequent ordering of compulsory care, detention and administration of medication takes place in a climate of ever-increasing emphasis on autonomy, and has resulted in the need for nurses to ensure that they do not violate clients' human and legal rights. Legislation is concerned not primarily with the finer points of what mental illness is--that is left to the medical profession--but with criteria which warrant the detention and treatment of a person against their will. The article points to international documents which set out principles for the care of the mentally ill, as well as both Commonwealth, state and territory legislation. It summarises the main provisions of these in relation to involuntary detention and treatment, as well as electroconvulsive therapy and psychosurgery. Nurses who care for mentally ill clients need, therefore, to understand in some detail the legislation that applies to them in their professional capacity, as well as the human rights it seeks to protect.


Potential dissemination of antibiotic resistance genes from transgenic plants to microorganisms.
Evidence that genes were transferred during evolution from plants to bacteria was obtained from nucleotide and protein sequence analyses. However, the extent of such transfers among phylogenetically distant organisms is limited by various factors, including those related to complexity of the environment and those endogenous to the bacteria, designed to prevent a drift of the genome integrity. The goal of this article is to give an overview of the potentials and limits of natural interkingdom gene transfers, with a particular focus on prokaryote originating sequences fitting the nuclear genome of transgenic plants.


Development and validation of the maternal blues scale through bonding attachments in predicting postpartum blues.
Postpartum blues in Indonesia has a high prevalence at 37% to 67%. Postpartum blues syndrome has been described as varying changes in the affective domain, such as feelings, behavior, or thoughts, that can be influenced by the roles and tasks of women, along with their social, cultural, and economic support. Instruments that measure maternal blues through bonding attachment behavior have never before been developed in Indonesia. This study aimed to develop a maternal blues scale through bonding attachments to predict postpartum blues. The research design consisted of three stages: 1) phenomenology design and focus group discussion; 2) development and construction of the maternal blues scale, and 3) a cross-sectional study to measure validation of the scales. Respondents were postpartum mothers in the first week after birth. The sample comprised 501 participants. Sampling was done by consecutive sampling at the Public Health Center (PUSKESMAS) in the South Jakarta area. Data analysis used exploratory factor analysis (EFA) and confirmatory factor analysis (CFA), correlation, and a diagnostic testing . Item analysis produced 32 items consisting of 24 items regarding the mother's role and duties as internal factors and eight factors involving social, cultural, and economic support as external factors. Both factors were valid and reliable in predicting postpartum blues with indicators (t loading factors ≥ 1.96, standardized loading factor (SLF) ≥.50, internal factors: construct reliability (CR) ≥ .70 and extraction variants (VE) ≥ .50 and external factors: CR ≥ .74 to .83 VE ≥ .50 to .63). The relationship with Kennerley's maternity blues as a gold standard was significant. Internal factors had a score of 53, with a sensitivity of 60.2%. The external factors score was 19, with a sensitivity of 77.3%. The new scale for postpartum blues prediction developed displayed internal consistency and validity of each indicator (internal and external factors) that was good (CR ≥ .70; VE ≥ .50). This scale provides a feasible tool to predict postpartum blues.


Investigation of glyceraldehyde-3-phosphate dehydrogenase from human sperms.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDs) was purified from human sperms and properties of the enzyme were investigated. After sonication of sperms, the most part of GAPDs is associated with the insoluble cell fraction. Trypsin treatment results in the cleavage of part of the N-terminal domain of the enzyme yielding a soluble fragment that was purified by hydrophobic chromatography on Phenyl-Sepharose. The isolated fragment was shown to be a tetramer with molecular weight of approximately 150 kD (according to Blue Native PAGE) and composed of subunits of 40 kD (according to SDS-PAGE). The specific activity of the isolated fragment reached 374 U/mg. It is supposed that GAPDs exists in sperms as the tetrameric molecule bound to the fibrous sheath of the flagellum through the N-terminus of one or two subunits. Comparative study of the amino acid sequences of mammalian GAPDs revealed conservative cysteine residues (C21, C94, and C150) that are specific for the sperm isoenzyme and absent in the somatic isoenzyme. Residue C21 can be involved in the formation of the disulfide bond between the N-terminal domain of GAPDs and fibrous sheath proteins.


Anxiolytic-like effects of Moringa oleifera in Swiss mice.
Moringa oleifera is evident to act against many neurological diseases, including muscle spasm, epilepsy, nervousness, fatigue, memory impairment, convulsion, and epilepsy. Anxiety represents the most common and disabling psychiatric condition, being often associated with depressive symptoms. This study investigated the anxiolytic-like effects of crude organic fractions of M. oleifera leaves in different behavioral paradigms that evaluate anxiety in mice. To this end, mice were administered with crude extracts (500 mg/kg, p.o.) and/or diazepam (2 mg/kg, p.o.), and submitted to behavioral tests. In the open-field test, the number of square field cross, grooming and rearing were calculated, while in light-dark and swing test were, respectively, the time spent in dark portion and number of swings. Each test was performed for 3 min. M. oleifera leaf methanol and n-hexane extracts elicited an anxiolytic-like effect observed by increased total time in the center and decreased number of rearings and groomings responses in the open field and swing tests, and residence in the dark portion in the light-dark box, similar to the diazepam group. A moderate anxiolytic effect was observed in the aqueous fraction group, while insignificant effects were recorded in the ethyl acetate fraction group in all test paradigms. In addition, both extracts potentiate the calming effects of diazepam in experimental animals. Preliminary phytochemical reports suggest that M. oleifera contains alkaloids, flavonoids, phenols, steroids, glycosides, saponins, tannin, terpenes, and gums. Of note, the results expand the understanding of M. oleifera effects in central nervous system and suggest that plant metabolites may be helpful for anxiety-related disorders management.


Gene expression of transforming growth factor-beta 3 and tissue inhibitor of metalloproteinase type 1 during membranous bone healing in rats.
A number of growth factors have been implicated in fracture repair. Transforming growth factor-beta 3 (TGF-beta 3) is believed to be involved in osteoblast proliferation, chemotaxis, and collagen synthesis. The collagens act as the scaffolding for new bone matrix formation, whereas tissue inhibitors of metalloproteinases (TIMPs) may help regulate matrix remodeling in bone repair. Despite their hypothesized integral role in fracture repair, the temporal expression of these molecules in membranous bone fracture healing remains unknown. The objective of this study was to assess the temporal pattern of TGF-beta 3 and TIMP type 1 (TIMP-1) expression in rat mandibular fracture healing. Twenty-eight adult male Sprague-Dawley rats underwent a mandibular osteotomy, and the healing regenerate was harvested on postoperative days 3, 5, 7, 9, 23, and 37. Total cellular ribonucleic acid was isolated, and Northern analysis was performed. TGF-beta 3 expression was downregulated dramatically 3 days after the osteotomy and remained less than 20% of control levels throughout repair. In marked contrast, TIMP-1 gene expression, low during early repair, increased more than twofold over control at later time points. Understanding the temporal pattern of gene expression during membranous fracture healing has important clinical implications because elucidating these mechanisms may lead to appropriate biomolecular approaches to augment membranous bone fracture healing.


[Therapy of the Steinbrinck-Chediak-Higashi-Syndrom (author's transl)].
The Steinbrinck-Chediak-Higashi Syndrome is characterized by morphological and functional alterations of polymorphonuclear leukocytes. The impaired function in the polymorphonuclear leukocytes may be related to abnormal microtubular assembly. The ingestion of 200 mg/d of ascorbic acid potentiated chemotaxis and bactericidal activity of polymorphonuclear leukocytes from a patient with this disease. Furthermore, the treatment with ascorbic acid reduced the spontaneous cap formation by these leukocytes, demonstrating the improvement of microtubular assembly. The life long treatment with ascorbic acid may affect the clinical course of these patients favourably.


Homeostatic role of B-1 cells in tissue immunity.
To date, studies of tissue-resident immunity have mainly focused on innate immune cells and T cells, with limited data on B cells. B-1 B cells are a unique subset of B cells with innate-like properties, enriched in murine pleural and peritoneal cavities and distinct from conventional B-2 cells in their ontogeny, phenotype and function. Here we discuss how B-1 cells represent exemplar tissue-resident immune cells, summarizing the evidence for their long-term persistence & self-renewal within tissues, differential transcriptional programming shaped by organ-specific environmental cues, as well as their tissue-homeostatic functions. Finally, we review the emerging data supporting the presence and homeostatic role of B-1 cells across non-lymphoid organs (NLOs) both in mouse and human.


The dopamine D Receptor (DRD4) gene exon III polymorphism, problematic alcohol use and novelty seeking: direct and mediated genetic effects.
The present study sought to integrate convergent lines of research on the associations among the dopamine D(4) receptor (DRD4) gene, novelty seeking and drinking behaviors with the overall goal of elucidating genetic influences on problematic drinking in young adulthood. Specifically, this study tested a model in which novelty seeking mediated the relationship between DRD4 variable number of tandem repeats (VNTR) genotype and problematic alcohol use. Participants (n = 90, 40 females) were heavy-drinking college students. Analyses using a structural equation modeling framework suggested that the significant direct path between DRD4 VNTR genotype and problematic alcohol use was reduced to a trend level in the context of a model that included novelty seeking as a mediator, thereby suggesting that the effects of DRD4 VNTR genotype on problematic alcohol use among heavy-drinking young adults were partially mediated by novelty seeking. Cross-group comparisons indicated that the relationships among the model variables were not significantly different in models for men versus women. These results extend recent findings of the association between this polymorphism of the DRD4 receptor gene, problematic alcohol use and novelty seeking. These findings may also help elucidate the specific pathways of risk associated with genetic influences on alcohol use and abuse phenotypes.


Resistance to Marek's disease at hatching in chickens vaccinated as embryos with the turkey herpesvirus.
Chickens vaccinated with herpesvirus of turkey (HVT) as 18-day embryos or at hatching were challenged as neonates with pathogenic Marek's disease (MD) virus (MDV). Embryonally vaccinated chickens had much greater resistant to challenge than chickens vaccinated post-hatch. Embryos became readily infected with HVT regardless of whether the vaccine was deposited into the body of the embryo or extraembryonally, such as in the amniotic sac. Embryonally vaccinated chickens were viremic with HVT at hatching and remained persistently viremic through the duration of the experiment. The titer of recoverable virus was higher in the embryonally vaccinated chickens than in the chickens vaccinated post-hatch. Embryonal vaccination did not affect hatchability. Vaccination at any stage of embryonation tested protected better against neonatal challenge than did vaccination at hatching. Protection against an early challenge was greatest when the embryos were 17 or 18 days old at the time of vaccination. Lower protection in chickens vaccinated as 11-day embryos was not due to humoral immunologic tolerance. Chickens vaccinated at the 11th day of embryonation were poorly protected against MDV challenge at three or eight days of age but were well protected if the challenge was delayed until the 14th day of age.


The Brazilian-Portuguese version of the Upper Extremity Functional Index (UEFI): Translation, cross-cultural adaptation and measurement properties for Brazilian adults after a burn injury.
To translate, cross-culturally adapt, validate, verify the reliability and estimate the minimal detectable change (MDC) of the UEFI to Brazilian Portuguese (UEFI-Br) for burns. The study was carried out with 131 Brazilian burn cases at two time points, at discharge and at the first outpatient follow-up (15-21 days after discharge) in a Burn Treatment Center. The adaptation process of the UEFI was based on international recommendations. The Burn Specific Health Scale-Brief-Br (BSHS-B-Br) was also applied in order to analyze construct validity of the UEFI-Br and distribution-based methods were used to estimate its MDC. Intra- and inter-rater reliability were good with ICC of 0.986 (CI 95 %: 0.98-0.99) and 0.969 (CI 95 %: 0.955-0.979), respectively, at discharge and 0.997 (CI 95 %: 0.996-0.998) and 0.987 (CI 95 %: 0.981-0.991), respectively, at the first outpatient follow-up appointment. We found good internal consistency with Cronbach's α values of 0.987 and 0.996, respectively, at the two times. The SEM was 4.42 and 2.31 at the first and second time points, respectively. The UEFI-Br scores demonstrated strong correlation with the Burn Specific Health Scale-Brief-Br (BSHS-B-Br) function domain scores (r = 0.87-0.90). No significant correlation was found between UEFI-Br scores and participants' characteristics. The MDC of the UEFI-Br lies between 11 and 13 points. The Brazilian version of the UEFI-Br, a useful tool to assess upper limb function and disability, is a valid and reliable tool for use with Brazilian burn survivors. The MDC for the instrument was determined to be 11-13 points.


Barriers to the use of clinical practice guidelines: a qualitative study of Danish physiotherapists and chiropractors.
Low back pain (LBP) is the leading cause of disability worldwide. Providing evidence-based practice (EBP) for patients with LBP is more cost-effective compared with non-EBP. To help health care professionals provide EBP, several clinical practice guidelines have been published. However, a relatively poor uptake of the guidelines has been identified across various countries. To enhance future implementation of EBP, the aim of this study was to explore barriers to using LBP guidelines in clinical practice. A qualitative constructivist grounded theory design was employed in order to gain an in-depth understanding of the barriers. Semi-structured interviews (+/- observations) of nine physiotherapists and nine chiropractors from primary care in the Central Denmark Region were conducted. Two key barriers were found to using guidelines in practice: (1) a scepticism due to doubts about validity and applicability of the guidelines, which emerged particularly among physiotherapists; and (2) a deep biomechanical professional identity, due to perceived role, interest, lack of skills, and patient preferences, which emerged particularly among chiropractors. For guidelines to be better implemented in practice, these key barriers must be addressed in a tailored strategy. Furthermore, this study showed a difference in barriers between the two professions.


[Refeeding after ingestion of diets containing 10% soybean protein associated with various concentrations of alginate or sodium carrageenan. Effects on growth and lipid parameters in the rat].
Male 21-d-old Wistar rats were fed over 3 experimental periods. During the first period of 4 wk, diets contained 10% casein or defatted soy flour proteins, with or without 0.5, 1, 2 or 3% sodium alginate or sodium carrageenan, and were heated. During the second period, they were fed a standard diet for 16 wk with 17% proteins, and during the third period, they received the same diet as in the first period, but with 20% proteins. Rat body weights were measured throughout the study period; plasma lipid levels were then determined after fasting. Presence of sodium alginate in the diet had no effect on growth, but rats fed carrageenan presented growth retardation at the end of the experimental period, which was not altered by refeeding the standard diet. Sodium alginate did not modify rat triglyceridemia, except at the 1% level. Carrageenan had a hypotriglyceridemic effect. Alginate and carrageenan had no effect on blood cholesterol. Compared to soybean protein, casein intake did not increase plasma cholesterol levels as generally described. The effect of carrageenans on growth and plasma triglyceride levels could be a result of their physico-chemical properties.


GALA, a database for genomic sequence alignments and annotations.
We have developed a relational database to contain whole genome sequence alignments between human and mouse with extensive annotations of the human sequence. Complex queries are supported on recorded features, both directly and on proximity among them. Searches can reveal a wide variety of relationships, such as finding all genes expressed in a designated tissue that have a highly conserved noncoding sequence 5' to the start site. Other examples are finding single nucleotide polymorphisms that occur in conserved noncoding regions upstream of genes and identifying CpG islands that overlap the 5' ends of divergently transcribed genes. The database is available online at http://globin.cse.psu.edu/ and http://bio.cse.psu.edu/.


A Bayesian evidence synthesis for estimating campylobacteriosis prevalence.
Stakeholders making decisions in public health and world trade need improved estimations of the burden-of-illness of foodborne infectious diseases. In this article, we propose a Bayesian meta-analysis or more precisely a Bayesian evidence synthesis to assess the burden-of-illness of campylobacteriosis in France. Using this case study, we investigate campylobacteriosis prevalence, as well as the probabilities of different events that guide the disease pathway, by (i) employing a Bayesian approach on French and foreign human studies (from active surveillance systems, laboratory surveys, physician surveys, epidemiological surveys, and so on) through the chain of events that occur during an episode of illness and (ii) including expert knowledge about this chain of events. We split the target population using an exhaustive and exclusive partition based on health status and the level of disease investigation. We assume an approximate multinomial model over this population partition. Thereby, each observed data set related to the partition brings information on the parameters of the multinomial model, improving burden-of-illness parameter estimates that can be deduced from the parameters of the basic multinomial model. This multinomial model serves as a core model to perform a Bayesian evidence synthesis. Expert knowledge is introduced by way of pseudo-data. The result is a global estimation of the burden-of-illness parameters with their accompanying uncertainty.


Congenital and developmental anomalies of the shoulder girdle.
An understanding of congenital and developmental anomalies of the shoulder girdle is facilitated by a knowledge of embryology, physeal appearance and closure, and phylogenetic changes. Scapular, clavicular, and proximal humeral anomalies are classified and discussed with reference to diagnosis and management. Finally anomalies of the shoulder girdle musculature are delineated.


Electroretinogram in response to x-ray stimulation.
The retina of the grass frog, Rana pipiens, responds to flashes of highintensity x-rays and produces an electroretinogram indistinguishable in form from the electroretinogram produced in response to light stimulation at low and intermediate intensities. At higher intensities the form changes and, for maximal responses, the electroretinogram in response to x-rays shows a lower amplitude and a longer latent period than that in response to light. The prolonged latent period indicates additional intermediate reactions for the x-rays response.


Effects of oral narcotics on sleep-disordered breathing in healthy adults.
Alcohol and benzodiazepines may increase sleep-disordered breathing by decreasing activity of pharyngeal dilating muscles, favoring the development of obstructive apneas and hypopneas. Narcotics cause greater depression of wakeful respiration than the previously mentioned drugs; however, the influence of narcotics on the upper airway and breathing during sleep has not been studied. We, therefore, examined, in 12 healthy adults, the effects of oral hydromorphone hydrochloride (2 and 4 mg) on breathing during sleep and on a variety of awake respiratory variables (minute ventilation, gas exchange, and chemoresponsiveness). In addition, awake pharyngeal inspiratory airflow resistance was determined before and after narcotic administration to assess the drug's influence on patency of the upper airway. Following both doses, minute ventilation decreased, and carbon dioxide pressure increased. The 4-mg dose of hydromorphone hydrochloride also produced a significant decrement in the hypoxic ventilatory response, whereas hypercapnic responsiveness and pharyngeal resistance did not change following either dose of the drug. Despite the respiratory depression during wakefulness described previously, no significant change was observed in any measure of sleep-disordered breathing after either dose of narcotic. We conclude that in healthy individuals without suspected sleep apnea, oral hydromorphone in standard dosages does not significantly increase sleep-disordered breathing. This result may be due to a lack of selective depression of upper-airway muscular function by the doses of narcotic used.


The necrophagous fly anthrax transmission pathway: empirical and genetic evidence from wildlife epizootics.
Early studies confirmed Bacillus anthracis in emesis and feces of flies under laboratory conditions, but there is little empirical field evidence supporting the roles of flies in anthrax transmission. We collected samples during outbreaks of anthrax affecting livestock and native and exotic wildlife on two ranches in West Texas (2009-2010). Sampling included animal carcasses, maggots, adult flies feeding on or within several meters of carcasses, and leaves from surrounding vegetation. Microbiology and PCR were used to detect B. anthracis in the samples. Viable B. anthracis and/or PCR-positive results were obtained from all represented sample types. Genetic analysis of B. anthracis samples using multilocus variable number tandem repeat analysis (MLVA) confirmed that each ranch represented a distinct genetic lineage. Within each ranch, we detected the same genotype of B. anthracis from carcasses, maggots, and adult flies. The results of this study provide evidence supporting a transmission cycle in which blowflies contaminate vegetation near carcasses that may then infect additional browsing animals during anthrax outbreaks in the shrubland environment of West Texas.


2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) modulates epidermal growth factor (EGF) binding to basal cells from a human keratinocyte cell line.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) modulates the proliferation, differentiation, or both, of epidermal keratinocytes in vivo and in culture. The growth of epidermal cells in culture is regulated by several biochemical mediators including epidermal growth factor (EGF). In this report the actions of TCDD on EGF binding in a basal cell population from a human keratinocyte cell line were examined. TCDD decreased the specific binding of 125I-EGF to basal cells by 40% within 96 hr. This reduction in EGF binding could not be attributed to changes in the state of differentiation as assessed by cell size and morphology, and cornified envelope competence, a marker of terminal differentiation. Modulation of EGF binding by TCDD was concentration-dependent (EC50 = 1 nM) and stereospecific, suggesting involvement of the Ah receptor. Scatchard analysis of EGF binding to the basal cells indicated a single class of high-affinity sites in both control (Kd = 0.14 nM) and treated (Kd = 0.11 nM) cultures and confirmed a decrease in the number of these sites in response to TCDD. The reduction in EGF binding correlated with a decrease in EGF-stimulated DNA synthesis and cell proliferation. Comparison of differentiation-competent squamous cell carcinoma (SCC) lines treated with TCDD supported an association between modulation of EGF binding and enhanced differentiation. The data indicate that basal cells are a target for TCDD. We propose that the modulation of EGF binding in basal keratinocytes by TCDD is one of the critical regulatory events resulting in enhanced differentiation.


Hybrid plasmids containing the pyruvate dehydrogenase complex genes and gene-DNA relationships in the 2 to 3 minute region of the Escherichia coli chromosome.
A sample of colonies from the Clarke-Carbon ColE1-Escherichia coli DNA plasmid gene bank was screened by conjugation for complementation of the lipoamide dehydrogenase lesion of a deletion strain lacking all components of the pyruvate dehydrogenase complex, delta (aroP aceE aceF lpd). Two ColE1-lpd+ hybrid plasmids were identified: pGS2 (ColE1-ace lpd+; 24 kb) and pGS5 (ColE1-lpd+; 14 kb). Enzymological studies confirmed that pGS2 expressed all the activities of the pyruvate dehydrogenase complex, whereas pGS5 expressed the lipoamide dehydrogenase and acetyltransferase activities (the latter from a ColE1 promoter). These and other plasmids were used to construct a 47-site (15 enzymes) restriction map for a 24.2 kb segment of bacterial DNA in the nadC-lpd region. A further 13 sites (six enzymes) were defined in a 5.4 kb sub-segment containing the lpd gene. lambda phage derivatives containing specific fragments were constructed and used in transduction studies which located the ace and lpd genes in a 7.78 kb sub-segment flanked by AccI and NruI sites.


Chemotherapy dosing part I: scientific basis for current practice and use of body surface area.
Cytotoxic chemotherapy is characterised by a low therapeutic index and significant variability in therapeutic and toxic effects. In an attempt to reduce this variability, most chemotherapy doses are individualised according to patient body surface area (BSA). This practice, which was introduced almost 50 years ago, clearly has practical and economic implications for the healthcare system. Furthermore, the clinical value of this approach has, in recent years, been questioned. Despite established practice, chemotherapy dose selection remains complicated, partly because treatment effects are difficult to measure, partly because drugs are used in combination with other treatment modalities, and also because the patient's condition may change with disease progression. Various patient-related factors can affect drug pharmacokinetics (PK) and pharmacodynamics (PD), for example organ function, expression and activity of metabolising enzymes, drug resistance, body size, gender, age, concomitant disease and co-administration of other drugs. These factors may be of clinical significance in chemotherapy dose determination and measures of PK, PD or both feature in attempts to devise more rigorous methods for chemotherapy dosing. Part I of this series of two reviews describes the history and clinical impact of BSA-based chemotherapy, and examines the scientific evidence to support BSA dosing. It evaluates the factors affecting PK and PD for specific drugs that could inform and refine dose determination.


Diet complexity and l-threonine supplementation: effects on nutrient digestibility, nitrogen and energy balance, and body composition in nursery pigs.
This study was conducted to investigate the effects of dietary complexity and l-Thr supplementation on energy and nutrient utilization in nursery pigs. Thirty-two nursery pigs (7.23 ± 0.48 kg) were randomly assigned to a 2 × 2 factorial treatment arrangement based on diet complexity (complex vs. simple) with different levels of l-Thr supplementation. The complex diet contained animal protein sources (e.g., fish meal and plasma) and a dairy product (e.g., dried whey) to mimic a conventional nursery diet. The simple diet was formulated with corn, wheat, and soybean meal. Both diets were supplemented with l-Thr to contain either 100% or 115% (SUP Thr) of the estimated standardized ileal digestible Thr requirement for 9 kg body weight pigs (NRC, 2012). The pigs were individually housed in metabolism crates and fed an experimental diet ad libitum for a 7-d adaptation period and 5 d of total but separate urine and fecal collection. On day 14, all pigs were euthanized to determine body composition. The diet complexity, l-Thr supplementation, and their interactions were considered main effects. Pigs fed the complex diet tended to exhibit greater (P < 0.10) apparent total tract digestibility (ATTD) of ash and urinary energy output than those fed the simple diet. The complex diet had greater (P < 0.05) digestible energy and net energy contents than the simple diet. Furthermore, the complex diet-fed pigs had lower (P < 0.05) plasma urea nitrogen concentration on day 14 than simple diet-fed pigs. The SUP Thr decreased (P < 0.05) ATTD of acid detergent fiber but trended (P < 0.10) toward a decrease in urinary nitrogen (N) output and an increase in N retention and body N mass. In conclusion, the simple diet for nursery pigs had lower digestible and net energy contents than a complex diet. The SUP Thr can improve N utilization and body protein deposition, irrespective of diet complexity.


Metastatic carcinoma of the oral region: An analysis of 21 cases.
Metastatic carcinoma to the jaws and oral region are very rare, representing less than 1% of all oral tumors. Unfortunately, oral metastasis is usually manifestation of an advanced stage of primary cancer, and indicates widespread disease and poor prognosis. In this retrospective study, a total of 2039 patients with history of oral malignant tumor between 1980 and 2012 at Seoul National University Dental Hospital were evaluated. We analyzed the dental and medical records, and histopathological database of 2039 patients to assess the prevalence of oral metastasis of carcinoma in terms of sex and age, as well as, the most common origin of primary cancer, and prevalent site and histopathological type of metastatic carcinoma. Among 2039 patients, 21 (1.03%) were finally diagnosed with metastatic carcinoma of the jaws and oral region. Among the 21 patients, only 11 had a working diagnosis as oral metastasis upon clinical evaluation before performing a biopsy. The mean age at the time of diagnosis with a metastatic carcinoma was 56.86, and there was a male preponderance. Metastatic carcinoma was more frequent in the jaws than in the soft tissue, especially in the mandible compared to the maxilla. The most frequent primary site was the lungs, followed by the liver and breasts. The predominant histopathological types were hepatocellular carcinoma and adenocarcinoma. Patient outcomes indicated a poor prognosis with the time from the appearance of the metastasis to death was only 12 months. According to these cases, oral metastases of carcinoma were exceedingly rare in Koreans. It can allow the clinicians take into account the possible presence of metastases and lead to early diagnosis.


Enhanced sensitivity of self-assembled-monolayer-based SPR immunosensor for detection of benzaldehyde using a single-step multi-sandwich immunoassay.
This paper describes the fabrication and sensing characteristics of a self-assembled monolayer (SAM)-based surface plasmon resonance (SPR) immunosensor for detection of benzaldehyde (BZ). The functional sensing surface was fabricated by the immobilization of a benzaldehyde-ovalbumin conjugate (BZ-OVA) on Au-thiolate SAMs containing carboxyl end groups. Covalent binding of BZ-OVA on SAM was found to be dependent on the composition of the base SAM, and it is improved very much with the use of a mixed monolayer strategy. Based on SPR angle measurements, the functional sensor surface is established as a compact monolayer of BZ-OVA bound on the mixed SAM. The BZ-OVA-bound sensor surface undergoes immunoaffinity binding with anti-benzaldehyde antibody (BZ-Ab) selectively. An indirect inhibition immunoassay principle has been applied, in which analyte benzaldehyde solution was incubated with an optimal concentration of BZ-Ab for 5 min and injected over the sensor chip. Analyte benzaldehyde undergoes immunoreaction with BZ-Ab and makes it inactive for binding to BZ-OVA on the sensor chip. As a result, the SPR angle response decreases with an increase in the concentration of benzaldehyde. The fabricated immunosensor demonstrates a low detection limit (LDL) of 50 ppt (pg mL(-1)) with a response time of 5 min. Antibodies bound to the sensor chip during an immunoassay could be detached by a brief exposure to acidic pepsin. With this surface regeneration, reusability of the same sensor chip for as many as 30 determination cycles has been established. Sensitivity has been enhanced further with the application of an additional single-step multi-sandwich immunoassay step, in which the BZ-Ab bound to the sensor chip was treated with a mixture of biotin-labeled secondary antibody, streptavidin and biotin-bovine serum albumin (Bio-BSA) conjugate. With this approach, the SPR sensor signal increased by ca. 12 times and the low detection limit improved to 5 ppt with a total response time of no more than ca. 10 min. Figure A single-step multi-sandwich immunoassay step increases SPR sensor signal by ca. 12 times affording a low detection limit for benzaldehyde of 5 ppt.


Increased game frequency period crossing Ramadan intermittent fasting decreases fat mass, sleep duration, and recovery in male professional basketball players.
Increased basketball game frequency may affect athlete performances, especially during Ramadan intermittent fasting (RIF). The objective of the present investigation was to assess the impacts of increased game frequency periods crossing the RIF on body composition, sleep habits, indices of well-being, recovery state, and dietary intake in professional male basketball players. Twenty-eight professional basketball players participated in this study and were divided into increased-games-frequency (INCR) or normal-games-frequency (NORM) groups. INCR trained four times and completed two games per week, whereas NORM completed only one game per week. During the first and fourth weeks of RIF, the following variables were assessed: internal load (weekly session rating of perceived exertion (s-RPE), heartrate (HR)), dietary intake, body composition, sleep quality (PSQI survey), well-being indices questionnaire (sleep, fatigue, stress, delayed onset of muscle soreness (DOMS)), and recovery state with the Total Quality Recovery (TQR) questionnaire. The internal load significantly increased after 4 weeks of RIF in INCR compared to NORM (p < 0.001). Significant decrease of TQR, sleep duration, and a significant increase of DOMS only for INCR (26.93%, p < 0.001, ES = 0.48, small; 33.83%, p < 0.001, ES = 0.40, small; 161.17%, p < 0.001, ES = 0.32, small; respectively). Significant group × time interaction was observed for body mass (p = 0.006, ES = 0.46, small) and body fat percentage (p = 0.025, ES = 0.33, small), with INCR having a greater decrease in all these values. Increased game frequency period crossing RIF decreases fat mass, sleep duration, and recovery in professional basketball players, which may consequently affect performance and health.


Completed Suicide Linked to the COVID-19 Pandemic by Using the Psychological Autopsy Method in Sibiu County, Romania: Case Series and Literature Review.
The COVID-19 pandemic is associated with suicide, as some data suggests. Our study aims to investigate the emergence of eleven completed suicide cases suspected to be linked to the COVID-19 pandemic during the restrictive measures imposed by the Romanian government, and to identify the consequences of mental health, suicidal motivation, and behavioral changes. To this end, we analyzed the deceased's medical records and applied the psychological autopsy method to the relatives/caregivers of the deceased for a suicidal investigation history, within conducted free-flow discussions. To highlight behavioral changes that occurred in the distant antecedents as well as immediately before the suicidal act, we used two sets of closed questions comprised of fifteen alarm signs, including depressive and/or anxiety symptoms. Our results showed that a deterioration of the mental status, especially concerning depressive and anxiety symptoms, was evident in people without or with pre-existing psychiatric pathology. The suicidal motivation proved to be complex including, in addition to the SARS-CoV-2 infection, social and economic consequences of the COVID-19 pandemic. We noted an intensification of the investigated alarm signs and even the emergence of new warning signs in the recent antecedents. Based on our findings, we reaffirmed the important role of the psychological autopsy method in suicide investigation, proving that it can detect the specific impact of the COVID-19 pandemic on people prone to suicide. This impact can be psycho-emotional, social, and/or economical, and thus we can state that the COVID-19 pandemic and its consequences can be, at least, a triggering factor that enhances completed suicide risk. Further studies are needed in this particular area because correlations between the COVID-19 pandemic and completed suicide do not appear to be accidental.


Enzymatic approaches and bisulfite sequencing cannot distinguish between 5-methylcytosine and 5-hydroxymethylcytosine in DNA.
DNA cytosine methylation (5mC) is highly abundant in mammalian cells and is associated with transcriptional repression. Recently, hydroxymethylcytosine (hmC) has been detected at high levels in certain human cell types; however, its roles are unknown. Due to the structural similarity between 5mC and hmC, it is unclear whether 5mC analyses can discriminate between these nucleotides. Here we show that 5mC and hmC are experimentally indistinguishable using established 5mC mapping methods, thereby implying that existing 5mC data sets will require careful re-evaluation in the context of the possible presence of hmC. Potential differential enrichment of 5mC and hmC DNA sequences may be facilitated using a 5mC monoclonal antibody.


Comparison of IMRT treatment plans between linac and helical tomotherapy based on integral dose and inhomogeneity index.
Intensity modulated radiotherapy (IMRT) is an advanced treatment technology for radiation therapy. There are several treatment planning systems (TPS) that can generate IMRT plans. These plans may show different inhomogeneity indices to the planning target volume (PTV) and integral dose to organs at risk (OAR). In this study, we compared clinical cases covering different anatomical treatment sites, including head and neck, brain, lung, prostate, pelvis, and cranio-spinal axis. Two treatment plans were developed for each case using Pinnacle(3) and helical tomotherapy (HT) TPS. The inhomogeneity index of the PTV and the non-tumor integral dose (NTID) were calculated and compared for each case. Despite the difference in the number of effective beams, in several cases, NTID did not increase from HT as compared to the step-and-shoot delivery method. Six helical tomotherapy treatment plans for different treatment sites have been analyzed and compared against corresponding step-and-shoot plans generated with the Pinnacle(3) planning system. Results show that HT may produce plans with smaller integral doses to healthy organs, and fairly homogeneous doses to the target as compared to linac-based step-and-shoot IMRT planning in special treatment site such as cranio-spinal.


miRNA analysis in pancreatic cancer: the Dartmouth experience.
Pancreatic cancer is considered one of the most lethal cancers being the fourth leading cause of cancer deaths in adults in the United States because of the lack of early signs and symptoms and the lack of early detection. Pancreatic ductal adenocarcinoma (PDAC) is the most common histological type among pancreatic cancers, representing 80%-90% of all solid tumors of the pancreas. The majority of PDAC develops from three precursor lesions: pancreatic intraepithelial neoplasia, intraductual papillary mucinous neoplasm and mucinous cystic neoplasm. Although histologic tissue evaluation remains the gold standard for diagnosis, endoscopic ultrasound-guided fine needle aspiration has become the preferred modality for obtaining pathologic confirmation. At Dartmouth-Hitchcock Medical Center (DHMC),we have developed and validated a microRNA (miRNA) panel for patients with pancreatic diseases that can be used in association with the gold standard method for diagnosis. miRNAs have an important role in biological processes, such as apoptosis, metabolism, cell growth and differentiation. In cancer, miRNAs can be classified as either oncogenic or tumor suppressor according to their function in the carcinogenic process. In this study, we describe the expression of many miRNA in benign and malignant pancreatic tissues as well as their clinical significance. For this reason, miRNAs have been considered potential biomarkers of pancreatic diseases that could potentially contribute to an early diagnosis, predict disease progression, accurately monitor disease, contribute to better treatment strategies and reduce mortality by improving disease management.


[Effect of acceleration on the sugar content of rabbit cerebrospinal fluid].
The sugar content in the liquor of rabbits exposed to accelerations ranging from several minutes to several hours was studied. The animals were accelerated at 3 g in a centrifuge with a radius of 55 cm and a velocity of 70 rpm, in the Khilov's swing and in a rotating device simulating angular accelerations. The liquor was sampled by the suboccipital puncture and the sugar content was measured according to Hagedorn-Jensen. A statistically significant increase in the sugar content was noted after every exposure to accelerations of alternating signs than with accelerations of a constant value. The experiments on labyrinth-ectomized animals showed a certain effect of the vestibular analyzer on the carbohydrate metabolism. However, when the equilibrium function reached compensation, the sugar content in the liquor returned to the pretest level. Therefore, this reaction involves not only the vestibular system but also the optic, proprio- and interoceptive afferent systems. A comparison of the intracysternal and intramuscular injections of epinephrine points to the central (in contrast to the hematogenic) nature of an increase in the sugar content during the first minutes of acceleration. The electric stimulation of the anterior and posterior hypothalamus by means of implanted electrodes produced the same elevation of the liquor sugar as accelerations. It can be postulated that afferent impulses from the analyzer systems reach the hypothalamic centers, thus exciting the autonomic system and, first of all, the sympathic system.


Posterior corneal elevation after LASIK with three flap techniques as measured by Pentacam.
To evaluate and compare posterior corneal changes using elevation data obtained from Pentacam (Oculus Optikgeräte GmbH) Scheimpflug imaging in eyes undergoing LASIK with three different modes of flap creation: IntraLase femtosecond laser FS60 (Abbott Medical Optics) (femtosecond group), Amadeus (Ziemer Group AG) mechanical microkeratome (keratome group), or flap formation using 20% alcohol laser epithelial keratomileusis (LASEK) (LASEK group). Ninety myopic patients (90 eyes) undergoing refractive surgery were recruited. The change in posterior corneal elevation at 21 predetermined points in the central 5-mm area was measured using exported elevation data from the Pentacam before LASIK and 18 months postoperative and was compared among and within three modes of flap creation. Mean change in posterior elevation in the central 5-mm area was 5.13±4.16 μm for the femtosecond group, 5.78±4.42 μm for the keratome group, and 6.68±4.72 μm for the LASEK group and was similar among groups (P=.59). Change in posterior elevation before and after LASIK was not significant within any group (P=.342, P=.232, and P=.321 for the femtosecond, keratome, and LASEK groups, respectively). Preoperative spherical equivalent, central corneal thickness, ablation depth, and estimated residual bed thickness did not correlate with change in posterior corneal elevation for the femtosecond, keratome, or LASEK groups (P>.05). Using Pentacam elevation data, there were no significant changes in posterior corneal elevation following LASIK among or within the three methods of flap creation. At 18 months after LASIK, the posterior corneal surface is not displaced anteriorly significantly and is equally stable using these three surgical techniques.


Anterior instability of the glenohumeral joint with humeral avulsion of the glenohumeral ligament. A review of 41 cases.
We studied retrospectively a consecutive series of 547 shoulders in 529 patients undergoing operation for instability. In 41, the cause of instability was considered to be lateral avulsion of the capsule, including the inferior glenohumeral ligament, from the neck of the humerus, the HAGL lesion. In 35, the lesion was found at first exploration, whereas in six it was noted at revision of a previous failed procedure. In both groups, the patients were older on average than those with instability from other causes. Of the primary cases, in 33 (94.3%) the cause of the first dislocation was a violent injury; six (17.4%) had evidence of damage to the rotator cuff and/or the subscapularis. Only four (11.4%) had a Bankart lesion. In patients undergoing a primary operation in whom the cause of the first dislocation was a violent injury, who did not have a Bankart lesion and had no suggestion of multidirectional laxity, the incidence of HAGL was 39%.


Ultrastructural differences during embryonic cell death in normal and peripherally deprived ciliary ganglia.
Normally occurring neuron death and that brought about by prior removal of the peripheral target organ was studied ultrastructurally in embryonic chick ciliary ganglion in order to better understand the mechanism of cell death in this system. Before the period of cell death, all neurons in the normal ganglion developed a well-organized rough endoplasmic reticulum (RER) which coincided with peripheral synapse formation. None of the peripherally deprived neurons underwent this change, suggesting that some interaction with the periphery, possibly synapse formation, triggered them into the secretory state. Cell death in peripherally deprived neurons was signalled by nuclear changes followed by freeing of ribosomes from polysomes and RER and presumably cessation of protein synthesis. In contrast, normal cell death was brought about by dilation of the RER with eventual cytoplasmic disruption, nuclear changes appearing only secondarily. It is suggested that failure to form or maintain peripheral synapses could result in the accumulation of transmission-related proteins with consequent cisternal dilation, and eventual cell death.


Machine learning for radiomics-based multimodality and multiparametric modeling.
Due to the recent developments of both hardware and software technologies, multimodality medical imaging techniques have been increasingly applied in clinical practice and research studies. Previously, the application of multimodality imaging in oncology has been mainly related to combining anatomical and functional imaging to improve diagnostic specificity and/or target definition, such as positron emission tomography/computed tomography (PET/CT) and single-photon emission CT (SPECT)/CT. More recently, the fusion of various images, such as multiparametric magnetic resonance imaging (MRI) sequences, different PET tracer images, PET/MRI, has become more prevalent, which has enabled more comprehensive characterization of the tumor phenotype. In order to take advantage of these valuable multimodal data for clinical decision making using radiomics, we present two ways to implement the multimodal image analysis, namely radiomic (handcrafted feature) based and deep learning (machine learned feature) based methods. Applying advanced machine (deep) learning algorithms across multimodality images have shown better results compared with single modality modeling for prognostic and/or prediction of clinical outcomes. This holds great potentials for providing more personalized treatment for patients and achieve better outcomes.


Effect of steric encumbrance of tris(3-phenylpyrazolyl)borate on the structure and properties of ternary copper(II) complexes having N,N-donor heterocyclic bases.
Complexes of formulation [Cu(Tp(Ph))(L)](ClO(4)) (1-4), where Tp(Ph) is anionic tris(3-phenylpyrazolyl)borate and L is N,N-donor heterocyclic base, viz. 2,2'-bipyridine (bpy, 1), 1,10-phenanthroline (phen, 2), dipyridoquinoxaline (dpq, 3), and dipyridophenazine (dppz, 4), are prepared from a reaction of copper(II) acetate.hydrate with KTp(Ph) and L in CH(2)Cl(2) and isolated as perchlorate salts. The complexes are characterized by analytical, structural, and spectral methods. The crystal structures of complexes 1-4 show the presence of discrete cationic complexes having the metal, Tp(Ph), and L in a 1:1:1 ratio and a noncoordinating perchlorate anion. The complexes have a square-pyramidal 4 + 1 coordination geometry in which two nitrogens of L and two nitrogens of the Tp(Ph) ligand occupy the basal plane and one nitrogen of Tp(Ph) binds at the axial site. Complexes 3 and 4 display distortion from the square-pyramidal geometry. The Cu-N distances for the equatorial and axial positions are approximately 2.0 and 2.2 A, respectively. The phenyl groups of Tp(Ph) form a bowl-shaped structure that encloses the [CuL] moiety. The steric encumbrance is greater for the bpy and phen ligands compared to that for dpq and dppz. The one-electron paramagnetic complexes (mu approximately equal to 1.8 mu(B)) exhibit axial EPR spectra in CH(2)Cl(2) glass at 77 K giving g(parallel) and g(perpendicular) values of approximately 2.18 (A(parallel) = 128 G) and approximately 2.07. The data suggest a [d(x(2)-y(2))](1) ground state. The complexes are redox-active and display a quasireversible cyclic voltammetric response for the Cu(II)/Cu(I) couple near 0.0 V versus SCE with an i(pc)/i(pa) ratio of unity in CH(2)Cl(2) or DMF-0.1 M TBAP. The E(1/2) values of the couple vary in the order 4 > 3 > 2 > 1. A profound effect of steric encumbrance caused by the Tp(Ph) ligand is observed in the reactivity of 1-4 with the calf thymus (CT) and supercoiled (SC) DNA. Complexes 2-4 show similar binding to CT DNA. The propensity for the SC DNA cleavage varies as 4 > 3 > 2. The bpy complex does not show any significant binding or cleavage of DNA. Mechanistic investigations using distamycin reveal minor groove binding for 2 and 3 and a major groove binding for 4. The scission reactions that are found to be inhibited by hydroxyl radical scavenger DMSO are likely to proceed through sugar hydrogen abstraction pathways.


Alternative treatments for chronic spontaneous urticaria beyond the guideline algorithm.
The international EAACI/GALEN/EDF/WAO guideline suggests a stepwise approach for the therapeutic management of chronic spontaneous urticaria (CSU), outlined in an algorithm. The aim of this article is to summarize and review the evidence available on alternative treatment options for CSU outside of this algorithm. Although CSU is a common disease, there are a limited number of high-quality studies, and only antihistamines and omalizumab are licensed for its treatment. Most studies regarding alternative therapies for CSU show methodological limitations and a high risk of bias. For many therapies, only case reports and uncontrolled studies exist. Recent publications on alternative treatments for chronic urticaria/CSU include reports on the use of adalimumab, rituximab, vitamin D, probiotics, histaglobulin, injection of autologous whole blood or serum, and phototherapy. Numerous treatments beyond the guideline algorithm have been evaluated in patients with refractory CSU. The global level of evidence to support their efficacy in CSU is low or very low. Further research is needed to assess the efficacy and safety of alternative therapies of CSU to manage adequately those patients who do not respond to the treatments included in the algorithm.


Audio-Conversion of Biomedical Signals - A Possible Approach to Improve Remote Monitoring of Elderly and Visually Impaired People.
Long-term remote patients monitoring implies minimal discomfort and reliability throughout the study period. These requirements are fulfilled by portable (wearable) patient devices, with low consumption, which transmit data wirelessly, at a short distance, to a mobile communication device (GSM) and through it, to a remote end recipient - doctor, medical center or a hospital server. The data transfer technology requires the monitored person to perform a sequence of actions, such as: selecting the appropriate application on the mobile phone, establishing a connection between the patient module and the phone, recording the data in the phone's memory, starting the data transfer from the phone to the final receiver. Practice shows that often this sequence of activities is difficult for elderly people and especially for visually impaired people, which as a result compromises the remote monitoring process. In this paper are presented an approach and conceptual implementation of a system for remote monitoring of cardiac activity, using the most popular way of remote connectivity - voice (sound) communication. In addition to the ease of use, this type of communication does not require special data protection, due to the lack of RF interfaces for short-distance data transmission. The presented results of laboratory studies, as well as conducted tests under medical supervision of patients in a cardiology clinic, confirm the workability of the proposed approach for remote monitoring of patients by audio conversion of the ECG signal.


Relationship Between Mitochondrial Structure and Bioenergetics in Pseudoxanthoma elasticum Dermal Fibroblasts.
Pseudoxanthoma elasticum (PXE) is a genetic disease considered as a paradigm of ectopic mineralization disorders, being characterized by multisystem clinical manifestations due to progressive calcification of skin, eyes, and the cardiovascular system, resembling an age-related phenotype. Although fibroblasts do not express the pathogenic ABCC6 gene, nevertheless these cells are still under investigation because they regulate connective tissue homeostasis, generating the "arena" where cells and extracellular matrix components can promote pathologic calcification and where activation of pro-osteogenic factors can be associated to pathways involving mitochondrial metabolism. The aim of the present study was to integrate structural and bioenergenetic features to deeply investigate mitochondria from control and from PXE fibroblasts cultured in standard conditions and to explore the role of mitochondria in the development of the PXE fibroblasts' pathologic phenotype. Proteomic, biochemical, and morphological data provide new evidence that in basal culture conditions (1) the protein profile of PXE mitochondria reveals a number of differentially expressed proteins, suggesting changes in redox balance, oxidative phosphorylation, and calcium homeostasis in addition to modified structure and organization, (2) measure of oxygen consumption indicates that the PXE mitochondria have a low ability to cope with a sudden increased need for ATP via oxidative phosphorylation, (3) mitochondrial membranes are highly polarized in PXE fibroblasts, and this condition contributes to increased reactive oxygen species levels, (4) ultrastructural alterations in PXE mitochondria are associated with functional changes, and (5) PXE fibroblasts exhibit a more abundant, branched, and interconnected mitochondrial network compared to control cells, indicating that fusion prevail over fission events. In summary, the present study demonstrates that mitochondria are modified in PXE fibroblasts. Since mitochondria are key players in the development of the aging process, fibroblasts cultured from aged individuals or aged in vitro are more prone to calcify, and in PXE, calcified tissues remind features of premature aging syndromes; it can be hypothesized that mitochondria represent a common link contributing to the development of ectopic calcification in aging and in diseases. Therefore, ameliorating mitochondrial functions and cell metabolism could open new strategies to positively regulate a number of signaling pathways associated to pathologic calcification.


Early-stage diagnosis of bladder cancer using surface-enhanced Raman spectroscopy combined with machine learning algorithms in a rat model.
Early diagnosis and accurate assessment of tumor development facilitate early bladder cancer resection and initiation of drug therapy. This study enabled an early, accurate, label-free, noninvasive diagnosis of bladder tumors by analyzing nano-biomarkers in a single drop of urine through surface-enhanced Raman spectroscopy (SERS). In a standard N-butyl-N-4-hydroxybutyl nitrosamine-induced rat model of bladder cancer, cancer stage and polyp tumor development were monitored using a small endoscope with a diameter of 1.2 mm in a minimally invasive manner without the need to kill the rats. Samples were divided into cancer-free, early-stage, and polyp-form cancer. Training data were classified according to micro-cystoscopic 5-aminolevulinic acid fluorescence diagnosis, and specimens were postmortem verified through histopathological analysis. A drop of urine from each sample group was placed on an Au-coated zinc oxide nanoporous chip to filter nano-biomaterials and selectively enhance the Raman signals of nanoscale analytes via SERS. Principal component analysis was used to reduce the dimensionality of the collected Raman spectra, and partial least squares discriminant analysis was used to find diagnostic clusters based on the labeled samples. The combination of SERS and machine learning achieved an accuracy ≥99.6% in diagnosing both early- and polyp-stage bladder tumors. With an area under the receiver operating characteristic curve greater than 0.996, the accuracy of the diagnosis in the rat model suggests that SERS-based diagnostic methods are promising when coupled with machine learning. Low-cost, label-free, and noninvasive surface-enhanced Raman spectra are ideal for developing clinically relevant point-of-care diagnostic techniques.


[Mortality from traffic accidents in the Tunja-San Gil road area, Colombia, 2001].
To identify the characteristics of mortality related to motor vehicle accidents in the Tunja-San Gil area (Colombia) in the year 2001. A descriptive study was performed on the deaths from road accidents. Autopsy reports, body removal reports, death certificates and blood concentration reports, available at the National Institute for Forensic Sciences and at the local hospitals, were reviewed. Forty five fatalities were analyzed. The mean age of the victims was 34.7 years; males were more affected than females, with a 3:1 ratio. 37% were pedestrians, 26% motorcyclists, 20% drivers, 8.6% cyclists and 8.6% passengers. 65.1% of the road accidents occurred during weekends. The most common road accidents were either collisions or by running over. The places with the highest rates of death from motor vehicle related accidents were the urban areas of Tunja and San Gil and the Vado Real-Oiba road. The mean Injury Severity Score was 42.8. 53% of the deaths at the scene received no medical care; 73% of the victims died within two hours after the motor-vehicle accident. 17.8% of deaths were definitely preventable and 46.7% were possibly preventable, for a combined preventable death rate of 64.5%. It is necessary to develop and implement a road-accident related-trauma care system.


Zeffiro User Interface for Electromagnetic Brain Imaging: a GPU Accelerated FEM Tool for Forward and Inverse Computations in Matlab.
This article introduces the Zeffiro interface (ZI) version 2.2 for brain imaging. ZI aims to provide a simple, accessible and multimodal open source platform for finite element method (FEM) based and graphics processing unit (GPU) accelerated forward and inverse computations in the Matlab environment. It allows one to (1) generate a given multi-compartment head model, (2) to evaluate a lead field matrix as well as (3) to invert and analyze a given set of measurements. GPU acceleration is applied in each of the processing stages (1)-(3). In its current configuration, ZI includes forward solvers for electro-/magnetoencephalography (EEG) and linearized electrical impedance tomography (EIT) as well as a set of inverse solvers based on the hierarchical Bayesian model (HBM). We report the results of EEG and EIT inversion tests performed with real and synthetic data, respectively, and demonstrate numerically how the inversion parameters affect the EEG inversion outcome in HBM. The GPU acceleration was found to be essential in the generation of the FE mesh and the LF matrix in order to achieve a reasonable computing time. The code package can be extended in the future based on the directions given in this article.


Astrobiology as a framework for investigating antibiotic susceptibility: a study of Halomonas hydrothermalis.
Physical and chemical boundaries for microbial multiplication on Earth are strongly influenced by interactions between environmental extremes. However, little is known about how interactions between multiple stress parameters affect the sensitivity of microorganisms to antibiotics. Here, we assessed how 12 distinct permutations of salinity, availability of an essential nutrient (iron) and atmospheric composition (aerobic or microaerobic) affect the susceptibility of a polyextremotolerant bacterium, Halomonas hydrothermalis, to ampicillin, kanamycin and ofloxacin. While salinity had a significant impact on sensitivity to all three antibiotics (as shown by turbidimetric analyses), the nature of this impact was modified by iron availability and the ambient gas composition, with differing effects observed for each compound. These two parameters were found to be of particular importance when considered in combination and, in the case of ampicillin, had a stronger combined influence on antibiotic tolerance than salinity. Our data show how investigating microbial responses to multiple extremes, which are more representative of natural habitats than single extremes, can improve our understanding of the effects of antimicrobial compounds and suggest how studies of habitability, motivated by the desire to map the limits of life, can be used to systematically assess the effectiveness of antibiotics.


Effects of graded levels of dietary pomegranate peel on methane and nitrogen losses, and metabolic and health indicators in dairy cows.
This study aimed to quantify the effects of dietary inclusion of tannin-rich pomegranate peel (PP) on intake, methane and nitrogen (N) losses, and metabolic and health indicators in dairy cows. Four multiparous, late-lactating Brown Swiss dairy cows (796 kg body weight; 29 kg/d of energy corrected milk yield) were randomly allocated to 3 treatments in a randomized cyclic change-over design with 3 periods, each comprising 14 d of adaptation, 7 d of milk, urine, and feces collection, and 2 d of methane measurements. Treatments were formulated using PP that replaced on a dry matter (DM) basis 0% (control), 5%, and 10% of the basal mixed ration (BMR) consisting of corn and grass silage, alfalfa, and concentrate. Gaseous exchange of the cows was determined in open-circuit respiration chambers. Blood samples were collected on d 15 of each period. Individual feed intake as well as feces and urine excretion were quantified, and representative samples were collected for analyses of nutrients and phenol composition. Milk was analyzed for concentrations of fat, protein, lactose, milk urea N, and fatty acids. Total phenols and antioxidant capacity in milk and plasma were determined. In serum, the concentrations of urea and bilirubin as well as the activities of alanine aminotransferase (ALT), aspartate aminotransferase, glutamate dehydrogenase, alkaline phosphatase, and γ-glutamyl transferase were measured. The data was subjected to ANOVA with the Mixed procedure of SAS, with treatment and period as fixed and animal as random effects. The PP and BMR contained 218 and 3.5 g total extractable tannins per kg DM, respectively, and thereof 203 and 3.3 g hydrolyzable tannins. Total DM intake, energy corrected milk, and methane emission (total, yield, and intensity) were not affected by PP supplementation. The proportions of C18:2 n-6 and C18:3 n-3 in milk increased linearly as the amount of PP increased in the diet. Milk urea N, blood urea N, and urinary N excretion decreased linearly with the increase in dietary PP content. Total phenols and antioxidant capacity in milk and plasma were not affected by the inclusion of PP. The activity of ALT increased in a linear manner with the inclusion of PP. In conclusion, replacing up to 10% of BMR with PP improved milk fatty acid composition and alleviated metabolic and environmental N load. However, the elevated serum ALT activity indicates an onset of liver stress even at 5% PP, requiring the development of adaptation protocols for safe inclusion of PP in ruminant diets.


The Effects of Lysophosphatidic Acid on The Incidence of Cell Death in Cultured Vitrified and Non-Vitrified Mouse Ovarian Tissue: Separation of Necrosis and Apoptosis Border.
The aim of the present study was to examine whether lysophosphatidic acid (LPA) could decrease cell death and improve in vitro culture (IVC) conditions in cultured vitrified mouse ovarian tissue. In this experimental study, we collected and randomly divided 7-day-old mouse ovarian tissues into vitrified and non-vitrified groups. The ovaries were cultured in the presence and absence of LPA for one week. Morphology and follicular development were evaluated by hematoxylin and eosin (H&E) and Masson's trichrome (MTC) staining. The incidence of cell death was assessed by flow cytometry using annexin V/propidium iodide (PI) and a caspase-3/7 assay in all studied groups. The vitrified groups had a significantly decreased follicle developmental rate compared to the non-vitrified groups (P<0.05). Overall, qualitative and quantitative results showed prominent follicular degeneration in the vitrified groups compared with the respective non-vitrified groups. Both LPA treated groups had a significantly higher proportion of preantral follicles compared to the non-LPA treated groups (P<0.05). Flow cytometry analysis results showed significantly greater early and late apoptotic cells in all groups (17.83 ± 8.80%) compared to necrotic cells (7.97 ± 0.92%, P<0.05). The percentage of these cells significantly increased in the vitrified groups compared with non-vitrified groups. LPA treated groups had a lower percentage of these cells compared to non-LPA treated groups (P<0.05). The lower enzyme activity was observed in non-vitrified (especially in the LPA+ groups) cultured ovaries compared to the vitrified group (P<0.05). Both vitrification and IVC adversely affected cell survival and caused cell death. We postulated that LPA supplementation of culture medium could improve the developmental rate of follicles and act as an anti-cell death factor in non-vitrified and vitrified ovarian tissues. It could be used for in vitro maturation of ovarian tissue.


Substrate Specificity of the FurE Transporter Is Determined by Cytoplasmic Terminal Domain Interactions.
FurE, a member of the Nucleobase Cation Symporter 1 transporter family in Aspergillus nidulans, is specific for allantoin, uric acid (UA), uracil, and related analogs. Herein, we show that C- or N-terminally-truncated FurE transporters (FurE-ΔC or FurE-ΔΝ) present increased protein stability, but also an inability for UA transport. To better understand the role of cytoplasmic terminal regions, we characterized genetic suppressors that restore FurE-ΔC-mediated UA transport. Suppressors map in the periphery of the substrate-binding site [Thr133 in transmembrane segment (TMS)3 and Val343 in TMS8], an outward-facing gate (Ser296 in TMS7, Ile371 in TMS9, and Tyr392 and Leu394 in TMS10), or in flexible loops (Asp26 in LN, Gly222 in L5, and Asn308 in L7). Selected suppressors were also shown to restore the wild-type specificity of FurE-ΔΝ, suggesting that both C- and/or N-terminal domains are involved in intramolecular dynamics critical for substrate selection. A direct, substrate-sensitive interaction of C- and/or N-terminal domains was supported by bimolecular fluorescence complementation assays. To our knowledge, this is the first case where not only the function, but also the specificity, of a eukaryotic transporter is regulated by its terminal cytoplasmic regions.


Gramicidin S: a peptide model for protein glycation and reversal of glycation using nucleophilic amines.
Nonenzymatic glycation of proteins has been implicated in various diabetic complications and age-related disorders. Proteins undergo glycation at the N-terminus or at the epsilon-amino group of lysine residues. Glycation of proteins proceeds through the stages of Schiff base formation, conversion to ketoamine product and advanced glycation end products. Gramicidin S, which has two ornithine residues, was used as a model system to study the various stages of glycation of proteins using electrospray ionization mass spectrometry. The proximity of two ornithine residues in the peptide favors the glycation reaction. Formation of advanced glycation end products and diglycation on ornithine residues in gramicidin S were observed. The formation of Schiff base adduct is reversible, whereas the Amadori rearrangement to the ketoamine product is irreversible. Nucleophilic amines and hydrazines can deglycate the Schiff base adduct of glucose with peptides and proteins. Hydroxylamine, isonicotinic acid hydrazide and aminoguanidine effectively removed glucose from the Schiff base adduct of gramicidin S. Hydroxylamine is more effective in deglycating the adduct compared with isonicotinic acid hydrazide and aminoguanidine. The observation that the hydrazines are effective in deglycating the Schiff base adduct even in the presence of high concentrations of glucose, may have a possible therapeutic application in preventing complications of diabetes mellitus. Hydrazines may be used to distinguish between the Schiff base and the ketoamine products formed at the initial stages of glycation.


[Expression of aminoglycoside phosphotransferase gene is under the control of recA promoter].
A novel expression vector using the 300 bp promotor-operator fragment of the recA gene of Escherichia coli has been constructed. The strength of the recA promotor was examined by assaying aminoglycoside phosphotransferase (APT) activity expressed from APT gene placed downstream of the promotor. We have observed, that some plasmids, containing N-portion of recA gene caused a large increase in radiosensitivity of host bacteria cells.