Chromatin structure can introduce systematic biases in genome-wide analyses of Plasmodium falciparum.
Background: The maintenance, regulation, and dynamics of heterochromatin in the human malaria parasite, Plasmodium falciparum, has drawn increasing attention due to its regulatory role in mutually exclusive virulence gene expression and the silencing of key developmental regulators. The advent of genome-wide analyses such as chromatin-immunoprecipitation followed by sequencing (ChIP-seq) has been instrumental in understanding chromatin composition; however, even in model organisms, ChIP-seq experiments are susceptible to intrinsic experimental biases arising from underlying chromatin structure. Methods: We performed a control ChIP-seq experiment, re-analyzed previously published ChIP-seq datasets and compared different analysis approaches to characterize biases of genome-wide analyses in P. falciparum. Results: We found that heterochromatic regions in input control samples used for ChIP-seq normalization are systematically underrepresented in regard to sequencing coverage across the P. falciparum genome. This underrepresentation, in combination with a non-specific or inefficient immunoprecipitation, can lead to the identification of false enrichment and peaks across these regions. We observed that such biases can also be seen at background levels in specific and efficient ChIP-seq experiments. We further report on how different read mapping approaches can also skew sequencing coverage within highly similar subtelomeric regions and virulence gene families. To ameliorate these issues, we discuss orthogonal methods that can be used to characterize bona fide chromatin-associated proteins. Conclusions: Our results highlight the impact of chromatin structure on genome-wide analyses in the parasite and the need for caution when characterizing chromatin-associated proteins and features.


Molecular identification and prevalence of tick-borne pathogens in zebu and taurine cattle in North Cameroon.
Public interest for tick-borne pathogens in cattle livestock is rising due to their veterinary and zoonotic importance. Consequently, correct identification of these potential pathogens is crucial to estimate the level of exposition, the risk and the detrimental impact on livestock and the human population. Conventional PCR with generic primers was used to identify groups of tick-borne pathogens in cattle breeds from northern Cameroon. The overall prevalence in 1260 blood samples was 89.1%, with 993 (78.8%) positive for Theileria/Babesia spp., 959 (76.1%) for Anaplasma/Ehrlichia spp., 225 (17.9%) for Borrelia spp., and 180 (14.3%) for Rickettsia spp. Sanger sequencing of a subset of positively-tested samples revealed the presence of Theileria mutans (92.2%, 130/141), T. velifera (16.3%, 23/141), Anaplasma centrale (10.9%, 15/137), A. marginale (30.7%, 42/137), A. platys (51.1%, 70/137), Anaplasma sp. 'Hadesa' (10.9%, 15/137), Ehrlichia ruminantium (0.7%, 1/137), E. canis (0.7%, 1/137), Borrelia theileri (91.3%, 42/46), Rickettsia africae (59.4%, 19/32) and R. felis (12.5%, 4/32). A high level of both intra- and inter-generic co-infections (76.0%) was observed. To the best of our knowledge, B. theileri, T. mutans, T. velifera, A. platys, Anaplasma sp. 'Hadesa', R. felis and E. canis are reported for the first time in cattle from Cameroon, and for R. felis it is the first discovery in the cattle host. Babesia spp. were not detected by sequencing. The highest number of still identifiable species co-infections was up to four pathogens per genus group. Multifactorial analyses revealed a significant association of infection with Borrelia theileri and anemia. Whereas animals of older age had a higher risk of infection, the Gudali cattle had a lower risk compared to the other local breeds. Co-infections of tick-borne pathogens with an overall high prevalence were found in all five study sites, and were more likely to occur than single infections. Fulani, Namchi and Kapsiki were the most infected breed in general; however, with regions as significant risk factor. A better-adapted approach for tick-borne pathogen identification in co-infected samples is a requirement for epidemiological investigations and tailored control measures.


Incremental benefit of myocardial contrast to combined dipyridamole-exercise stress echocardiography for the assessment of coronary artery disease.
Although assessment of myocardial perfusion by myocardial contrast echocardiography (MCE) is feasible, its incremental benefit to stress echocardiography is not well defined. We examined whether the addition of MCE to combined dipyridamole-exercise echocardiography (DExE) provides incremental benefit for evaluation of coronary artery disease (CAD). MCE was combined with DExE in 85 patients, 70 of whom were undergoing quantitative coronary angiography and 15 patients with a low probability of CAD. MCE was acquired by low-mechanical-index imaging in 3 apical views after acquisition of standard resting and poststress images. Wall motion, left ventricular opacification, and MCE components of the study were interpreted sequentially, blinded to other data. Significant (>50%) stenoses were present in 43 patients and involved 69 coronary territories. The addition of qualitative MCE improved sensitivity for the detection of CAD (91% versus 74%, P=0.02) and accurate recognition of disease extent (87% versus 65% of territories, P=0.003), with a nonsignificant reduction in specificity. The addition of low-mechanical-index MCE to standard imaging during DExE improves detection of CAD and enables a more accurate determination of disease extent.


Crystal structure and the paraelectric-to-ferroelectric phase transition of nanoscale BaTiO3.
We have investigated the paraelectric-to-ferroelectric phase transition of various sizes of nanocrystalline barium titanate (BaTiO3) by using temperature-dependent Raman spectroscopy and powder X-ray diffraction (XRD). Synchrotron X-ray scattering has been used to elucidate the room temperature structures of particles of different sizes by using both Rietveld refinement and pair distribution function (PDF) analysis. We observe the ferroelectric tetragonal phase even for the smallest particles at 26 nm. By using temperature-dependent Raman spectroscopy and XRD, we find that the phase transition is diffuse in temperature for the smaller particles, in contrast to the sharp transition that is found for the bulk sample. However, the actual transition temperature is almost unchanged. Rietveld and PDF analyses suggest increased distortions with decreasing particle size, albeit in conjunction with a tendency to a cubic average structure. These results suggest that although structural distortions are robust to changes in particle size, what is affected is the coherency of the distortions, which is decreased in the smaller particles.


[Characteristics of cellular immune reactions in animals vaccinated with smallpox and ADPT vaccines].
Immunization with absorbed DPT vaccine in 1 or 3 subcutaneous injections, as well as the intradermal or intracardiac immunization of guinea pigs with smallpox vaccine resulted in the accumulation of 0-positive lymphoyctes, determined by the cytotoxic test, in the lymph nodes. On the 7th day after the intracardiac injection of smallpox vaccine in massive doses a sharp decrease in the content of 0-positive lymphocytes was observed. The number of small lymphocytes with surface immunoglobulin receptors detected by immunofluorescence remains unchanged in the animals of all groups at any period of the experiment.


Photometric and Colorimetric Assessment of LED Chip Scale Packages by Using a Step-Stress Accelerated Degradation Test (SSADT) Method.
By solving the problem of very long test time on reliability qualification for Light-emitting Diode (LED) products, the accelerated degradation test with a thermal overstress at a proper range is regarded as a promising and effective approach. For a comprehensive survey of the application of step-stress accelerated degradation test (SSADT) in LEDs, the thermal, photometric, and colorimetric properties of two types of LED chip scale packages (CSPs), i.e., 4000 °K and 5000 °K samples each of which was driven by two different levels of currents (i.e., 120 mA and 350 mA, respectively), were investigated under an increasing temperature from 55 °C to 150 °C and a systemic study of driving current effect on the SSADT results were also reported in this paper. During SSADT, junction temperatures of the test samples have a positive relationship with their driving currents. However, the temperature-voltage curve, which represents the thermal resistance property of the test samples, does not show significant variance as long as the driving current is no more than the sample's rated current. But when the test sample is tested under an overdrive current, its temperature-voltage curve is observed as obviously shifted to the left when compared to that before SSADT. Similar overdrive current affected the degradation scenario is also found in the attenuation of Spectral Power Distributions (SPDs) of the test samples. As used in the reliability qualification, SSADT provides explicit scenes on color shift and correlated color temperature (CCT) depreciation of the test samples, but not on lumen maintenance depreciation. It is also proved that the varying rates of the color shift and CCT depreciation failures can be effectively accelerated with an increase of the driving current, for instance, from 120 mA to 350 mA. For these reasons, SSADT is considered as a suitable accelerated test method for qualifying these two failure modes of LED CSPs.


Reference Value Profile for Healthy Individuals From the Aljouf region of Saudi Arabia.
Many factors influence hematological values such as sex, age, ethnic origin, geographic location, season, and genetic disease. The aim of this study was to detect the hematological reference value profile for healthy adults from the Aljouf region of Saudi Arabia. The project was carried out on 2,040 healthy individuals, 1,152 males and 888 females, with ages ranging from 17 to 28 years. A group of participants were recruited from the higher secondary schools, university students and premarital centers of Aljouf cities. Hematological reference value profile, hemoglobin (Hb) concentration, red blood cell (RBC) count, RBC indices, white blood cell (WBC) count, differential WBC and platelet (Plt) count were measured. Moreover, a peripheral blood film was prepared in order to detect abnormalities of RBC and all samples were examined for liver function tests (LFTs) and renal function test (RFT) performed, along with a lipid profile. Hb concentration, hematocrit (Hct) and RBCs were found to be significantly higher in males than in females (P < 0.01). On the contrary, Plt ranges were significantly lower in male as compared to female (P < 0.01). No significant differences in the study population were determined in the other hematological parameters (P > 0.05). Our findings reflect that healthy adults from the Aljouf region have some hematological parameters differing quantitatively from Caucasians. The hematological reference value profile reported here can be used as normal reference values for Saudi people of the Aljouf region to help in diagnosis and consequently treatment of individuals with hematological disorders.


Explaining the long reach of prenatal behaviors and attitudes in unmarried men at birth on father engagement in early and middle childhood and adolescence.
The current study examined three research questions: (a) Are unmarried at birth fathers' prenatal and birth-related behavioral, attitudinal, and identity adjustments directly related to father engagement in child-related activities during early childhood and father-child closeness in middle childhood and adolescence? (b) Do father engagement in child-related activities during early childhood, coresidence, and coparenting at age 5 mediate the association between unmarried fathers' prenatal and birth-related variables and father-child relationship in middle childhood and adolescence? (c) Do father-child closeness, coresidence, and coparenting in middle childhood mediate the association between fathers' prenatal and birth-related variables and father-child relationship during adolescence? Using a subsample of Fragile Families and Child Wellbeing data (N = 2,647), we found support for our hypotheses that fathers' prenatal and birth-related variables significantly predicted father-child engagement during early childhood and father-child closeness during middle childhood and adolescence, although not all prenatal and birth-related variables are related to outcomes during each stage of childhood development. Father involvement and coparenting cooperation significantly mediated the associations among fathers' prenatal and birth-related variables and father-child closeness at ages 9 and 15. Our findings indicate that researchers, practitioners, and policymakers should take advantage of the prenatal period and direct resources to facilitate and strengthen prospective unmarried fathers' early relationships with their partners and children. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Histologic and histomorphometric report of three immediately loaded screw implants retrieved from man after a three-year loading period.
The aim of the present study, in man, was an histologic and histomorphometric analysis of the peri-implant tissues in 3 immediately loaded screw implants retrieved, because of fracture, after a 3 year loading period. Mineralized tissue was present at the interface of all implants. The mean bone to implant contact percentage for the 3 implants was 56.3% +/- 5%. The histologic and histomorphometric data showed that mineralized tissues can be obtained in immediately loaded screw implants and that these mineralized tissues are maintained over a period of 3 years. The response of the osseous tissue was not disturbed by the stresses and strains transmitted at the interface. A high primary stability, obtained by the large threads of the screw-shaped implant, was probably a key factor in the long-term maintenance of a mineralized bone interface. The primary stability helps to resist micromotion, i.e., the relative movements between the implant surface and surrounding bone during functional loading.


Topical treatment of major omphalocoele: Acacia nilotica versus povidone-iodine: A randomised controlled study.
Conservative management for major omphalocoele with topical agents as escharotics therapy is well established in practice. Different agents have been used in the past, including mercurochrome and alcohol, proved later to be unsafe. The aim of this study is to evaluate the efficacy and safety of the application of Acacia nilotica paste compared to povidone-iodine solution as a primary non-surgical treatment of major omphalocoele. A double-blind, randomised study was conducted on 24 cases of major omphalocoele where they were randomly divided into two equal groups; Group A treated with topical application of A. nilotica paste and Group B treated with topical application of povidone-iodine solution. Cases with gastroschisis, ruptured major omphalocoele or minor omphalocoele were excluded from the study. The evaluating parameters were size of the fascial defect in cm, period of mechanical ventilation if needed, time required for full oral feeding tolerance, duration of hospital stay and any short- or long-term complications. There was no statistical significant difference between both groups regarding their gestational or post-natal age, weight and the mean umbilical port defect. Patients from Group A tolerated full oral feeding earlier and had shorter total hospital stay duration than those from Group B, but without a statistical significant difference (P = 0.347 and 0.242, respectively). The overall mortality rate was 33.3% without a statistical significant difference between both groups (P = 0.667). Application of A. nilotica is a safe and effective treatment of major omphalocoele as it was associated with rapid full enteral feeding tolerance, short duration of hospital stay and low mortality rate.


[Urolesan in the treatment of stenocardia patients].
Urolesan was used in the treatment of patients with chronic ischemic heart disease. It was established that urolesan favours improvement of the state of the vascular component of microcirculation, aggregation of erythrocytes, increase of the peripheral circulation, normalization of the blood coagulation system, optimization of external respiration, normalization of the activity of serum aminotransferases. These changes were particularly pronounced in exertion stenocardia.


Lengthening of the humerus with intramedullary lengthening nails-preliminary report.
Distraction osteogenesis of the humerus with fully implantable lengthening is now possible since the diameter of the available nails was reduced to 10 mm and below. We report on the first intramedullary lengthening cases of the humerus with two different lengthening devices (FITBONE and PRECICE). Two different approaches and implantation techniques were used. We retrospectively reviewed clinical and radiographic data and pointed out results, pitfalls and complications of the procedure. Four adult patients with relevant length discrepancy of the humerus were treated with fully implantable systems in two centers between 2012 and 2015. Three patients were treated with FITBONE by an antegrade approach; one patient had lengthening with a PRECICE and a retrograde approach. Average nail lengthening was 55 mm (40-65 mm), and the average duration of lengthening was 70 days (52-95 days). The average distraction index was 0.72 mm/day (range 0.4-1.0 mm/day) or 12.5 days/cm (range 8.0-16.2 days/cm). The average consolidation index was 33.6 days/cm (range 25-45 days/cm). There was an implant failure (arrest) with the PRECICE. After consolidation and exchange with a technically improved implant, the course of treatment was uneventful. In patients with antegrade lengthening shoulder abduction decreased, and in the patient with the retrograde approach it improved but elbow extension decreased marginally. Reduced motion of the adjacent joints can be a major problem in intramedullary lengthening of the humerus. This first case series in the field of a rare indication suggests that lengthening of the humerus by fully implantable lengthening nails might be a valuable alternative to lengthening with external fixation. Main advantage of the PRECICE technology is the possible shortening in-between of lengthening.


Model substrates for an RNA enzyme.
M1 RNA, the catalytic RNA subunit of Escherichia coli ribonuclease P, can cleave novel transfer RNA (tRNA) precursors that lack specific domains of the normal tRNA sequence. The smallest tRNA precursor that was cleaved efficiently retained only the domain of the amino acid acceptor stem and the T stem and loop. The importance of the 3' terminal CCA nucleotide residues in the processing of both novel and normal tRNA precursors implies that the same enzymatic function of M1 RNA is involved.


Dichotic-listening task performance of mildly mentally retarded and nonretarded individuals.
An investigation of hemispheric lateralization of mildly mentally retarded individuals was undertaken employing the dichotic-listening paradigm. Exclusively right-handed, right-eyed, and right-footed equal-CA, equal-MA, and mildly retarded subjects were required to recognize and reject dichotic presentation-probe stimulus combinations of words and tones correctly. The dichotic presentation-probe stimulus technique was employed to minimize the influence of memory and to equate methodologically for the handling of the verbal (spoken digits) and nonverbal (complex tones) stimuli. The findings revealed a pattern of dichotic performance that was similar for the three subject groups. A right-ear advantage was obtained for the verbal stimuli; and a left-ear advantage, for the nonverbal stimuli. Performance accuracy, however, was significantly poorer for the mildly retarded subjects relative to the nonretarded subjects. Structural vs. control process differences in the information-processing system were discussed.


Conformational Stability Adaptation of a Double-Stranded RNA-Binding Domain to Transfer RNA Ligand.
The double-stranded RNA-binding domain (dsRBD) is a broadly distributed domain among RNA-maturing enzymes. Although this domain recognizes dsRNA's structures via a conserved canonical structure adopting an α1-β1β2β3-α2 topology, several dsRBDs can accommodate discrete structural extensions expanding further their functional repertoire. How these structural elements engage cooperative communications with the canonical structure and how they contribute to the dsRBD's overall folding are poorly understood. Here, we addressed these issues using the dsRBD of human dihydrouridine synthase-2 (hDus2) (hDus2-dsRBD) as a model. This dsRBD harbors N- and C-terminal extensions, the former being directly involved in the recognition of tRNA substrate of hDus2. These extensions engage residues that form a long-range hydrophobic network (LHN) outside the RNA-binding interface. We show by coarse-grain Brownian dynamics that the Nt-extension and its residues F359 and Y364 rigidify the major folding nucleus of the canonical structure via an indirect effect. hDus2-dsRBD unfolds following a two-state cooperative model, whereas both F359A and Y364A mutants, designed to destabilize this LHN, unfold irreversibly. Structural and computational analyses show that these mutants are unstable due to an increase in the dynamics of the two extensions favoring solvent exposure of α2-helix and weakening the main folding nucleus rigidity. This LHN appears essential for maintaining a thermodynamic stability of the overall system and eventually a functional conformation for tRNA recognition. Altogether, our findings suggest that functional adaptability of extended dsRBDs is promoted by a cooperative hydrophobic coupling between the extensions acting as effectors and the folding nucleus of the canonical structure.


Income is an independent risk factor for worse asthma outcomes.
Socioeconomic status (SES) is associated with asthma morbidity in observational studies, but the factors underlying this association are uncertain. We investigated whether 3 SES correlates-low income, low education, and high perceived stress-were independent risk factors for treatment failure and asthma exacerbations in the context of a randomized controlled trial. The effect of low SES (household income of <$50,000/y and household educational level of less than a Bachelor's degree) and high perceived stress (defined as a score of >20 on a perceived stress scale) on asthma morbidity was analyzed in 381 participants by using Poisson regression models. The primary outcome was treatment failure (defined in the trial protocol as a significant clinical or airflow deterioration), and the secondary outcome was asthma exacerbations requiring systemic corticosteroids. Fifty-four percent of participants had a low income, 40% had a low educational level, and 17% had high perceived stress levels. Even after adjusting for race and other important confounders, participants with lower income had higher rates of both treatment failures (rate ratio, 1.6; 95% CI, 1.1-2.3; P = .03) and exacerbations (rate ratio, 1.9; 95% CI, 1.1-3.3; P = .02). Adherence with inhaled corticosteroids was similarly high for both income categories. Education and perceived stress were not significantly associated with either outcome. In the context of a randomized controlled trial, participants with lower income were more likely to experience adverse asthma outcomes independent of education, perceived stress, race, and medication adherence.


FoxO family members in cancer.
The PI3K-Akt-FoxO signaling pathway plays a central role in diverse physiological processes including cellular energy storage, growth, and survival, among others. As an important effector of this pathway, FoxO is involved in versatile activities that protect organisms from stress and aging. Recent studies on mammalian FoxO have established a direct role for this transcription factor family in cellular proliferation, oxidative stress response, and tumorigenesis. The review will focus on the recent developments pertaining to the function of FoxO as well as discuss the various contexts in which FoxO exerts distinct biological activity such as drug resistance and autophagy in cancer pathogenesis and therapy.


Analysis of ecstasy in oral fluid by ion mobility spectrometry and infrared spectroscopy after liquid-liquid extraction.
We developed and evaluated two different strategies for determining abuse drugs based on (i) the analysis of saliva by ion mobility spectrometry (IMS) after thermal desorption and (ii) the joint use of IMS and infrared (IR) spectroscopy after liquid-liquid microextraction (LLME) to enable the sensitivity-enhanced detection and double confirmation of ecstasy (MDMA) abuse. Both strategies proved effective for the intended purpose. Analysing saliva by IMS after thermal desorption, which provides a limit of detection (LOD) of 160μgL(-1), requires adding 0.2M acetic acid to the sample and using the truncated negative second derivative of the ion mobility spectrum. The joint use of IMS and IR spectroscopy after LLME provides an LOD of 11μgL(-1) with the former technique and 800μgL(-1) with the latter, in addition to a limit of confirmation (LOC) of 1.5mgL(-1). Using IMS after thermal desorption simplifies the operational procedure, and using it jointly with IR spectroscopy after LLME allows double confirmation of MDMA abuse with two techniques based on different principles (viz., IMS drift times and IR spectra). Also, it affords on-site analyses, albeit at a lower throughput.


Mobilization of intracellular calcium from the sarcoplasmic reticulum of B-lymphocytes using cyclic guanosine 3'5'-monophosphate (cGMP).
Cyclic guanosine-3'5' monophosphate (cGMP) can mobilize intracellular calcium from the microsomal fraction of B-lymphocytes of the mouse spleen as a result of activation of cGMP-dependent microsomal proteinkinases. The existence of such a mechanism makes B-lymphocytes independent of extracellular calcium in response to agents whose effect on B-lymphocytes is mediated by calcium mechanisms.


A transplantable human medullary thyroid carcinoma as a model for RET tyrosine kinase-driven tumorigenesis.
Hereditary medullary thyroid carcinoma (MTC) is caused by germline mutations in the RET proto-oncogene, resulting in constitutive activation of the RET tyrosine kinase. A substantial proportion of sporadic MTCs also have RET mutations, making the RET tyrosine kinase a potential therapeutic target in MTC. We have established a transplantable MTC in nude mice from a sporadic human MTC carrying a RET C634R mutation. Transplanted tumors had an exponential growth rate with an approximate doubling time of about 3 weeks, and expressed a neuroendocrine phenotype characteristic of MTC, e.g., expression of calcitonin, chromogranin A (CgA), synaptophysin, synaptic vesicle protein 2 (SV2), vesicular monoamine transporter-1 and -2, carcinoembryonic antigen, cytokeratin 8/18, epithelial cadherin, and neural cell adhesion molecule. Plasma calcitonin and CgA levels were elevated in tumor-bearing mice and correlated with tumor size. Cytogenetic analysis, including spectral karyotyping, confirmed the human origin of the xenografted tumors and demonstrated an abnormal, near triploid karyotype. Treatment of tumor-bearing nude mice with the tyrosine kinase inhibitor ZD6474, which specifically inhibits RET, epidermal growth factor receptor (EGFR), and vascular endothelium growth factor receptor (VEGFR) tyrosine kinases, resulted in a dose-dependent inhibition of tumor growth. Oral ZD6474 given once daily (250 mg/kg, 5 days/week) reduced tumor volume to 11% when compared with controls after 4 weeks. Our results show that this transplantable MTC, designated GOT2, represents a novel and useful model for studies of MTC and RET tyrosine kinase-dependent tumor growth.


Nucleotide sequence of a DRw10 beta chain cDNA clone. Identity of the third D region with that of the DRw53 allele of the beta 2 locus and as the probable site encoding a polymorphic MHC class II epitope.
The nucleotide and inferred amino acid sequence of a DRw10 beta chain was obtained from cDNA clones isolated from a DR1, DRw10 heterozygous cell line. The sequence of this beta chain gene was distinctive, differing from those of all other defined DR types. The DRw10 beta chain gene was shown by transfection experiments to encode a polymorphic epitope recognized by mAb 109d6 that is also encoded by the DRw53 beta 2 chain gene. Comparison of the nucleotide sequence of both genes revealed that their third D regions (amino acids 67 to 73) were identical. This suggested first that the 109d6 epitope could be encoded by residues of this region, and second, that a putative gene conversion event transferred this sequence along with the information encoding the 109d6 epitope from a donor gene such as DRw53 beta 2. The sequence of the DRw10 beta chain gene was observed to be identical to that of clone pII beta 4 derived from the non-DR3 haplotype in the Raji cell line, which was also demonstrated to express the determinant recognized by antibody 109d6, suggesting that the typing of this cell line is HLA-DR3/DRw10. No evidence was found for the existence of a DR beta 2 chain gene product encoded by the DRw10 haplotype. The DRw10 haplotype was of particular interest because it was present along with a DR1 haplotype in the propositus who had rheumatoid arthritis, and was shared by the DR4-positive son of the propositus, who also had rheumatoid arthritis. This raised the possibility that the DRw10 haplotype, and most probably one or more specific conformations encoded by the DR beta chain, are involved in the definition of the disease susceptibility phenotype.


Systematic investigation of protein-small molecule interactions.
Cell signaling is extensively wired between cellular components to sustain cell proliferation, differentiation, and adaptation. The interaction network is often manifested in how protein function is regulated through interacting with other cellular components including small molecule metabolites. While many biochemical interactions have been established as reactions between protein enzymes and their substrates and products, much less is known at the system level about how small metabolites regulate protein functions through allosteric binding. In the past decade, study of protein-small molecule interactions has been lagging behind other types of interactions. Recent technological advances have explored several high-throughput platforms to reveal many "unexpected" protein-small molecule interactions that could have profound impact on our understanding of cell signaling. These interactions will help bridge gaps in existing regulatory loops of cell signaling and serve as new targets for medical intervention. In this review, we summarize recent advances of systematic investigation of protein-metabolite/small molecule interactions, and discuss the impact of such studies and their potential impact on both biological researches and medicine.


Melanin affinity and its possible role in neurodegeneration.
Certain drugs with melanin affinity are known to have caused pigmentary lesions in the eye and skin. This was the basis for the hypothesis that compounds with melanin affinity may cause damage also in other melanin-bearing tissues such as the substantia nigra. The heterogeneity of compounds that binds to melanin is large. Toxins, drugs, and several other compounds have melanin affinity. Compounds showing the highest affinity are mainly organic amines and metal ions. The binding of toxicants to melanin probably protects the cells initially. However, the binding is normally, slowly reversible and melanin may accumulate the toxicant and gradually release it into the cytosol. Several studies indicate that neuromelanin may play a significant role both in the initiation and in the progression of neurodegeneration. MPTP/MPP(+) that has been causally linked with Parkinsonism has high affinity for neuromelanin, and the induced dopaminergic denervation correlates with the neuromelanin content in the cells. This shows that the toxicological implications of the accumulation of toxicants in pigmented neurons and its possible role in neurodegeneration should not be neglected. Extracellular neuromelanin has been reported to activate dendritic cells and microglia. An initial neuronal damage induced by a neurotoxicant that leaks neuromelanin from the cells may therefore lead to a vicious cycle of neuroinflammation and further neurodegeneration. Although there are many clues to the particular vulnerability of dopaminergic neurons of substantia nigra in Parkinson's disease, the critical factors are not known. Further studies to determine the importance of neuromelanin in neurodegeneration and Parkinson's disease are warranted.


Seasonal Variation in Fungi in Beach Sand in Summertime: Stintino (Italy).
The goal of this study was to monitor the microbial biodiversity in beach sand that is heavily visited by tourists during the summer, and to determinate whether the high presence of bathers (around 5000 per day) can modify sand microbial composition. Between 2016 and 2020, 150 sand samples were collected from nine different points at La Pelosa beach in Sardinia, Italy. Non-culturing methods were used; DNA extraction and meta-barcode sequencing were performed. All samples were analyzed with sequencing methods for 16S and ITS sequences. Fungal genera differ on the three beaches and in the winter/summer zones. The ITS sequence showed the most common presence of Candida during summer and Paradendryphiella in the winter. The greatest diversity was found in the dune during winter, while in other parts of the beach, there are differences between bacteria and fungi, particularly in the wash zone during the winter, with high diversity for 16S sequences but low diversity for ITS sequences. It appears reasonable that the sands, even on non-urban beaches, should be included in health monitoring programs in addition to the waters, and that access to them should be regulated by limiting the number of bathers with the aim of reducing the presence of pathogenic fungal species.


Caffeine consumption among eating disorder patients: epidemiology, motivations, and potential of abuse.
Aim of the study was to investigate caffeine use in different types of eating disorders (ED) patients either using a categorical approach [Diagnostic and Statistical Manual of Mental Disorders - Fourth Edition - Text Revision (DSM-IV-TR) diagnostic criteria] or a dimensional perspective. Fifty-eight ED female patients [anorexia nervosa (AN), restricting and binge-eating/purging type, N=15; bulimia nervosa (BN) purging type/nonpurging type, N=26; binge eating disorder (BED), N=17] referred to an Eating Disorder Unit and 15 non-clinical controls were administered the Eating Disorder Inventory-2 (EDI-2), the Clinical Global Impression (CGI) and the Caffeine Use Test, an interview specifically developed to investigate caffeine intake. Statistical analyses were then repeated clustering patients according to the presence/absence of purging behaviors (purgers, N=22; non-purgers, N=19; BED, N=17). Current and lifetime caffeine use, measured as mg/day, were similar comparing controls and ED patients as a whole. BN patients showed a significantly higher maximum lifetime caffeine intake (817.4+/-528,9 vs 325.0+/-294.6 mg/die, F=3.246, p<0.05); the same for purgers vs controls (p<0.05). Caffeine abuse was significantly more represented among patients vs controls (p<0.01), but similar among different patients' groups. As for diagnoses according to DSM-IV-TR Substance Use modified for caffeine, no significant difference was found among the different groups, for either Dependence, Intoxication or Withdrawal. Most of patients and controls reported pleasure as the main motivation for caffeine use, followed by increased vigilance and attention and appetite suppression in AN and BN patients. Note that a shift in diagnosis in the course of the ED from non-purging to purging type was associated with an increase in caffeine current, lifetime and maximum lifetime intake (F=1.667 p<0.05), except for BED patients. Severity of the ED measured as CGI score or comorbidity did not affect caffeine intake in patients as a whole, but in the purging subgroup current caffeine use was increased in presence of an anxiety disorder (p<0.05), and decreased in presence of a mood disorder (p<0.01). Data from the present study are in agreement with previous evidence in literature that a high percentage of ED patients ordinarily use caffeine with an average intake similar to that of the general population, however with a kind of binge attitude. Among heavy drinkers, daily caffeine intake and alcohol/cigarettes use are associated supporting the link with the dimension of impulse disregulation. The substantial number of subjects from our sample satisfying research criteria for Dependence, together with increasing reports of caffeine intoxication, suggests the growing relevance of these issues that deserve further investigation.


Soy isoflavones and glucose metabolism in menopausal women: A systematic review and meta-analysis of randomized controlled trials.
The aim of this meta-analysis was to investigate whether soy isoflavones, a type of phytoestrogen, would affect glucose homeostasis in menopausal women. Studies concerning about the relationship between soy isoflavone treatment and glucose metabolism were searched on MEDLINE and WEB OF SCIENCE (updated through April 2015) and EMBASE (1990-April 2015). Seventeen randomized controlled trials (RCTs) with a total number of 1529 menopausal women were identified for meta-analysis. Soy isoflavones were found to show great significance for the improvement of glucose metabolism, though marked heterogeneity was found between studies. The overall results showed that the average difference in fasting blood glucose values between women assigned to soy isoflavones and women in placebo groups was -0.22 mmol/L (95% CI: -0.38 to -0.07 mmol/L) under a random-effects model. In addition, the effect of soy isoflavones on insulin was also significant: -0.43 μIU/mL (95% CI: -0.71 to -0.14 μIU/mL), as was the effect on homeostasis model assessment insulin resistance (HOMA-IR): -0.52 (95% CI: -0.76 to -0.28). Although the results displayed a significant tendency in favor of soy isoflavones, it appears that genistein alone played an important role in improving glucose metabolism due to its low heterogeneity.


A Short Series of Laparoscopic Mesenteric Bypasses for Chronic Mesenteric Ischemia.
Laparoscopic aortomesenteric bypass may be performed to treat the chronic mesenteric ischemia patients who are not suitable for endovascular treatment. This study presents an initial experience with a limited series of laparoscopic mesenteric artery revascularization for the treatment of mesenteric ischemia. Chronic mesenteric ischemia (CMI) patients with previous unsuccessful endovascular treatment or with arterial occlusion and extensive calcification precluding safe endovascular treatment were offered laparoscopic mesenteric revascularization. From October 2015 until November 2018, nine patients with CMI underwent laparoscopic revascularization. In addition to demographic data and perioperative results of the treatment, graft patency was assessed with Duplex ultrasound at 1, 3, 6 and 12 months, and annually thereafter. A descriptive analysis of the data was performed. All bypasses were constructed with an 8 mm ring enforced expanded polytetrafluoroethylene graft in a retrograde fashion (from infrarenal aorta or iliac artery) to either superior mesenteric artery or splenic artery (2 cases). Median operation time was 356 mins (range 247-492 mins). Five patients had a history of unsuccessful endovascular treatment. Laparoscopic technical success was 78%, and the primary open conversion rate was 22%. All laparoscopic revascularization procedures remained patent after discharge during a median follow-up time of 26 months (range 18-49 months). The primary graft patency at 30 days was 78%. Primary assisted, and secondary graft patency was 78% and 100%, respectively. Median weight gain was 2 kg (range 2-18 kg), and all patients achieved relief from postprandial pain and nausea. No mortality was observed during the follow-up period. Laparoscopic aortomesenteric revascularization procedures for chronic mesenteric ischemia are feasible but require careful patient selection. These procedures should only be performed at referral centers by vascular surgeons with prior experience in laparoscopic vascular surgery.


Optimal collection and storage conditions for catecholamine measurements in human plasma and urine.
Improvements in methodologies for measuring concentrations of catecholamines (CA) have led to an increasing use of these compounds as markers in the screening of patients and in long-term clinical trials. Because of the associated logistical problems, we have investigated the unresolved question of optimal conditions for sample preparation and for storage of plasma and urine samples. Results show that blood should be centrifuged within 1 h after collection; the use of a refrigerated centrifuge is not necessary. Once plasma is prepared, CA are stable for 1 day at 20 degrees C, 2 days at 4 degrees C, 1 month at -20 degrees C (or 6 months with added glutathione), and up to 1 year at -70 degrees C. CA are stable at 4 degrees C for 1 month in unpreserved urine and for 4 months in urine preserved with EDTA and sodium metabisulfite. In acidified urine, CA were nearly unchanged after 1 year at 4 and -20 degrees C.


Quantum Monte Carlo study of the optical and diffusive properties of the vacancy defect in diamond.
Fixed-node diffusion quantum Monte Carlo (DMC) calculations of the ground and excited state energetics of the neutral vacancy defect in diamond are reported. The multiplet structure of the defect is modeled using guiding wave functions of the Slater-Jastrow type with symmetrized multideterminant Slater parts. For the ground state we obtain the 1E state in agreement with experiment. The calculated energy of the lowest dipole allowed transition is consistent with the experimentally observed GR1 band, which has long been identified with the neutral vacancy in diamond, although no previous first-principles ab initio calculation of this transition exists. The calculated multiplet splitting of over 2 eV indicates the importance of a proper treatment of electron exchange and correlation in this system. DMC calculations of the formation and migration energy of the vacancy defect are presented.


Antioxidant activity of Siamese neem tree (VP1209).
Leaves, fruits, flowers and stem bark extracts from the Siamese neem tree (Azadirachta indica A. Juss var. siamensis Valeton, Meliaceae) were assessed for antioxidant activity in vitro using the 1,1-diphenyl-2-picryl hydrazyl (DPPH) scavenging assay, total antioxidant activity and inhibition of lipid peroxidation in Chago K1 cancer cell culture by the thiobarbituric acid reactive substances (TBARS) method. The results showed that leaf aqueous extract, flower and stem bark ethanol extracts exhibited higher free radical scavenging effect on the DPPH assay with 50% scavenging activity at 26.5, 27.9 and 30.6 microg/ml, respectively. The total antioxidant activity of these extracts was found to be 0.959, 0.988 and 1.064 mM of standard trolox, respectively. At 100 microg/ml, the flower ethanol and leaf aqueous extracts significantly decreased malondialdehyde (MDA) levels (46.0 and 50.6%, respectively) by the TBARS method. The results suggest that extracts from leaf, flower and stem bark of the Siamese neem tree have strong antioxidant potential. This report supports the ethnomedical use of young leaves and flowers of this plant as a vegetable bitter tonic to promote good health.


Antioxidative system parameters and level of IL-18 after surgery in patients with renal cell carcinoma according to gender.
Functions of the antioxidative system and cytokines are important factors that can influence the development of cancer. The aim of this study was to determine gender-related differences in the concentration of inflammatory cytokine IL-18 and alterations of antioxidative system parameters after surgical treatment of renal cell carcinoma patients. The study comprised 40 patients (22 women and 18 men). The level of peroxidation product malonodialdehyde (MDA), the status of the antioxidative system and the concentration of interleukin-18 (IL-18) were analyzed. A significant difference in the changes of antioxidative system reflecting parameters was observed according to the gender. In the male group with higher IL-18 concentration after surgery the level of MDA was lower, while in the women group it did not change and the level of catalase activity was higher; superoxide dismutase activity was higher in both groups. It is important to notice that our previous study (where the data were not analyzed in respect of gender) showed that the level of IL-18 was significantly higher after surgical treatment if MDA content was increased. More detailed studies are required to confirm in which cases association between IL-18 concentration and MDA level is related to cancer progression and in which cases - to the better prognosis depending on the patients' gender.


Encephalopathy associated with Hashimoto thyroiditis: pediatric perspective.
In recent years, neuropsychiatric symptoms associated with Hashimoto thyroiditis have been increasingly recognized in both adult and pediatric patients. This neurologic complication has been termed "Hashimoto encephalopathy," and it can begin abruptly, in the form of seizures or agitation, with or without other neurologic complaints, or it can develop gradually, in a relapsing-remiting manner, including, among others, cognitive deterioration and psychiatric illness. The occurrence of Hashimoto encephalopathy is unrelated to the patient's thyroid function status. Although the pathogenesis of this encephalopathy is not fully understood, evidence for both autoimmune and vasculitic mechanisms exists. Owing to a low index of suspicion, this encephalopathy is very likely underdiagnosed in children. The diagnosis of Hashimoto encephalopathy is based on detecting antithyroid antibodies in these patients, although there is no correlation between antibody levels and the severity of the illness. Cerebrospinal fluid analysis, electroencephalography, and neuroimaging studies do not show consistent findings to support the diagnosis. Physicians' awareness of this complication is of great importance because most patients respond dramatically to corticosteroid therapy. Moreover, early recognition might also avoid an expensive diagnostic work-up in patients with unexplained encephalopathy (J Child Neurol 2006;21:1-5).


Enhanced Vitamin K (Menaquinone-7) Production by Bacillus subtilis natto in Biofilm Reactors by Optimization of Glucose-based Medium.
Benefits of vitamin K have been reported by many studies recently, due to its ability to reduce the risk of cardiovascular diseases and its potential benefits against osteoporosis. Specifically, menaquinone-7 (MK-7), being the most potent form of vitamin K, has definitely received most of the attention. Currently, solid or static liquid fermentation strategies are utilized for industrial production of MK-7 by Bacillus strains. However, these strategies face fundamental operational and scale-up issues as well as intense pellicle and biofilm formations which is problematic in static liquid fermentation, due to heat and mass transfer inefficiencies they create. The purpose of this study was to demonstrate that biofilm reactors will overcome the issues associated with suspended cell reactors when using Bacillus strains to produce MK-7. The expectation is that the use of biofilm reactors will result in a significant increase in the production of MK-7. Vitamin K production by Bacillus subtilis natto when grown in a biofilm reactor was evaluated at various concentrations of the three major nutrients, glucose, yeast extract and casein. The data was analyzed using response surface methodology (RSM). The maximum concentration of MK-7 in the biofilm reactors was 20.5±0.5 mg/L, which was a 344 % increase when compared to the amount produced in suspended-cell reactors containing the same optimum media composition. These results demonstrate the potential of utilizing biofilm reactors for MK-7 production on an industrial scale.


A Phex mutation in a murine model of X-linked hypophosphatemia alters phosphate responsiveness of bone cells.
Mutations in the PHEX gene cause X-linked hypophosphatemia (XLH). Hypophosphatemia in XLH results from increased circulating levels of a phosphaturic hormone, fibroblast growth factor 23 (FGF23), which inhibits renal phosphate reabsorption and 1,25-dihydroxyvitamin D (calcitriol) synthesis. The current standard therapy for XLH--high-dose phosphate and calcitriol--further increases FGF23 concentrations, suggesting that patients with XLH may have an altered response to extracellular phosphate. To test for the presence of abnormal phosphate responsiveness, we compared serum biochemistries and femoral Fgf23 mRNA expression between wild-type mice, murine models of XLH (Phex(K496X)) and hyperphosphatemic tumoral calcinosis (Galnt3(-/-)), and Galnt3/Phex double-mutant mice. Phex mutant mice had not only increased Fgf23 expression but also reduced proteolytic cleavage of intact Fgf23 protein, resulting in markedly elevated intact Fgf23 levels and consequent hypophosphatemia. In contrast, despite markedly increased Fgf23 expression, Galnt3 knockout mice had significantly high proteolytic cleavage of Fgf23 protein, leading to low intact Fgf23 concentrations and hyperphosphatemia. Galnt3/Phex double-mutant mice had an intermediate biochemical phenotype between wild-type and Phex mutant mice, including slightly elevated intact Fgf23 concentrations with milder hypophosphatemia. Despite the hypophosphatemia, double-mutant mice attempted to reduce serum phosphate back to the level of Phex mutant mice by upregulating Fgf23 expression as much as 24-fold higher than Phex mutant mice. These data suggest that Phex mutations alter the responsiveness of bone cells to extracellular phosphate concentrations and may create a lower set point for "normal" phosphate levels.


Biotyping of Malassezia pachydermatis strains using the killer system.
The killer phenomenon has been used as epidemiological marker for Candida albicans, where hundreds of biotypes can be obtained. The objective of this study is to observe the behaviour of 30 strains of Malassezia pachydermatis isolated from dogs with otitis (15) or dermatitis (15) against 9 killer yeasts, which, when grouped in triplets produced a 3 digit code (biotype). The growth inhibition of the 30 strains of M. pachydermatis due to the effect of the killer yeasts used permitted the determination of the following biotypes: 888 (33.3%), 212 (26.7%), 111 (16.7%), 312 (6.7%), 512 (6.7%), 242 (3.3%), 311 (3.3%) and 411 (3.3%). Biotypes 888, 212 and 111 occurred most frequently in both ear canal and skin samples.


Photothermal Responsivity of van der Waals Material-Based Nanomechanical Resonators.
Nanomechanical resonators made from van der Waals materials (vdW NMRs) provide a new tool for sensing absorbed laser power. The photothermal response of vdW NMRs, quantified from the resonant frequency shifts induced by optical absorption, is enhanced when incorporated in a Fabry-Pérot (FP) interferometer. Along with the enhancement comes the dependence of the photothermal response on NMR displacement, which lacks investigation. Here, we address the knowledge gap by studying electromotively driven niobium diselenide drumheads fabricated on highly reflective substrates. We use a FP-mediated absorptive heating model to explain the measured variations of the photothermal response. The model predicts a higher magnitude and tuning range of photothermal responses on few-layer and monolayer NbSe2 drumheads, which outperform other clamped vdW drum-type NMRs at a laser wavelength of 532 nm. Further analysis of the model shows that both the magnitude and tuning range of NbSe2 drumheads scale with thickness, establishing a displacement-based framework for building bolometers using FP-mediated vdW NMRs.


Risk of Melanoma Recurrence After Diagnosis of a High-Risk Primary Tumor.
With emerging new systemic treatments for metastatic melanoma, early detection of disease recurrence is increasingly important. To investigate the risk of melanoma recurrence in patients with a localized melanoma at a high risk of metastasis. A total of 1254 patients with newly diagnosed, histologically confirmed tumor category T1b to T4b melanoma in Queensland, Australia, were recruited prospectively between October 1, 2010, and October 1, 2014, for participation in a cohort study. Data analysis was conducted from February 8, 2018, to February 20, 2019. We used Cox proportional hazards regression analysis to examine associations between patient and tumor factors and melanoma recurrence. Disease-free survival (DFS) by melanoma tumor category defined by the 7th vs 8th editions of the AJCC Cancer Staging Manual (AJCC 7 vs AJCC 8). Melanoma recurrences were self-reported through follow-up questionnaires administered every 6 months and confirmed by histologic or imaging findings. Of 1254 patients recruited, 825 individuals (65.8%) agreed to participate. Thirty-six were found to be ineligible after providing consent and a further 89 patients were excluded after reclassifying tumors using AJCC 8, leaving 700 participants with high-risk primary melanoma (mean [SD] age, 62.2 [13.5] years; 410 [58.6%] men). Independent predictors of recurrence were head or neck site of primary tumor, ulceration, thickness, and mitotic rate greater than 3/mm2 (hazard ratio, 2.36; 95% CI, 1.19-4.71). Ninety-four patients (13.4%) developed a recurrence within 2 years of diagnosis: 66 tumors (70.2%) were locoregional, and 28 tumors (29.8%) developed at distant sites. After surgery for locoregional disease, 37 of 64 patients (57.8%) remained disease free at 2 years, 7 patients (10.9%) developed new locoregional recurrence, and 20 patients (31.3%), developed distant disease. Two-year DFS was similar when comparing AJCC 7 and AJCC 8, for T1b (AJCC 7, 253 [93.3% DFS]; AJCC 8, 242 [93.0% DFS]) and T4b (AJCC 7 and AJCC 8, 50 [68.0% DFS] category tumors in both editions. Patients with T2a to T4a tumors who did not have a sentinel lymph node biopsy (SLNB) at diagnosis had lower DFS than patients with the same tumor category and a negative SLNB (T2a: 136 [91.1%; 95% CI, 86.4-95.9] vs 96 [96.9%; 95 % CI, 93.4-100.0]; T4a: 33 [78.8%; 95% CI, 64.8-92.7] vs 6 [83.3; 95% CI, 53.5-100.0]). These findings suggest that 13.4% of patients with a high-risk primary melanoma will experience disease recurrence within 2 years. Head or neck location of initial tumor, SLNB positivity, and signs of rapid tumor growth may be associated with primary melanoma recurrence.


Terrestrial discharge into the Great Barrier Reef Lagoon: nutrient behavior in coastal waters.
Pollution of coastal regions of the Great Barrier Reef World Heritage Area (GBRWHA) is dominated by river discharge associated with agricultural development of the adjacent catchments. Runoff of sediment, nutrients and pesticides has sharply increased since European settlement. Since 1991 plumes from river discharge entering the GBRWHA have been mapped by aerial mapping of plume edges and concentrations of contaminants in plumes measured. Plume dispersion is governed primarily by wind speed and direction. Most plumes spread in a band up to 50 km from the coast. Particulate material discharged in the plumes is trapped within 10 km of the coast. Dissolved nutrients disperse much further and elevated nutrient concentrations are measurable at distances of hundreds of kilometres from river mouths. This differential transport of particulate versus dissolved nutrients is important for the potential effects of these materials and management of their generation on the Great Barrier Reef catchment.


Finite Element-Based Numerical Simulations to Evaluate the Influence of Wollastonite Microfibers on the Dynamic Compressive Behavior of Cementitious Composites.
This paper investigates the dynamic compressive behavior of wollastonite fiber-reinforced cementitious mortars using multiscale numerical simulations. The rate dependent behavior of the multiphase heterogeneous systems is captured in a multiscale framework that implements continuum damage towards effective property prediction. The influence of wollastonite fiber content (% by mass) as cement replacement on the dynamic compressive strength and energy absorption capacity is thereafter elucidated. An average compressive strength gain of 40% is obtained for mortars with 10% wollastonite fiber content as cement replacement, as compared to the control mortar at a strain rate of 200/s. The rate dependent constitutive responses enable the computation of energy absorption, which serves as a comparative measure for elucidating the material resistance to impact loads. Approximately a 45% increase in the dynamic energy absorption capacity is observed for the mixture containing 10% wollastonite fibers, as compared to the control case. Overall, the study establishes wollastonite fibers as a sustainable and dynamic performance-enhanced alternative for partial cement replacement. Moreover, the multiscale numerical simulation approach for performance prediction can provide an efficient means for the materials designers and engineers to optimize the size and dosage of wollastonite fibers for desired mechanical performance under dynamic loading conditions.


Activation of the human thymidine kinase (TK) promoter by simian virus 40 large T antigen requires both the T antigen pRb family-binding domain and TK promoter sequences resembling E2F-binding sites.
Infection of quiescent cells with the DNA tumor virus simian virus 40 induces expression of the cellular thymidine kinase (TK) gene a minimum of 10- to 20-fold, and this induction depends upon the viral protein large T antigen (T-Ag). To define both human TK promoter elements and T-Ag functional domains required for transcriptional induction, we have established a system in which stable Rat-1 transfectants harboring TK promoter-luciferase hybrid genes are infected with recombinant adenoviruses expressing either wild-type or mutant forms of T-Ag and luciferase expression is measured as an indicator of promoter activity. The results show that (i) a 135-bp TK promoter fragment is activated 10- to 15-fold by viral infection; (ii) this activation is the result of both T-Ag-dependent and -independent mechanisms; (iii) the T-Ag pRb family-binding domain, but not the p53-binding, helicase, or ATPase domain, is required for activation; and (iv) activation is severely diminished with a TK promoter fragment in which E2F-like-binding sites have been removed. These data demonstrate a requirement for both an E2F-related factor and a pRb family member in activation of the TK promoter by T-Ag. This contrasts with the promiscuous activation of many cellular and viral genes by T-Ag, which is independent of its ability to bind pRb.


Associations between genomic stratification of breast cancer and centrally reviewed tumour pathology in the METABRIC cohort.
The integration of genomic and transcriptomic profiles of 2000 breast tumours from the METABRIC [Molecular Taxonomy of Breast Cancer International Consortium] cohort revealed ten subtypes, termed integrative clusters (IntClust/s), characterised by distinct genomic drivers. Central histopathology (N = 1643) review was undertaken to explore the relationship between these ten molecular subtypes and traditional clinicopathological features. IntClust subtypes were significantly associated with histological type, tumour grade, receptor status, and lymphocytic infiltration (p < 0.0001). Lymph node status and Nottingham Prognostic Index [NPI] categories were also significantly associated with IntClust subtype. IntClust 3 was enriched for tubular and lobular carcinomas, the latter largely accounting for the association with CDH1 mutations in this cluster. Mucinous carcinomas were not present in IntClusts 5 or 10, but did not show an association with any of the remaining IntClusts. In contrast, medullary-like cancers were associated with IntClust 10 (15/26). Hormone receptor-positive tumours were scattered across all IntClusts. IntClust 5 was dominated by HER2 positivity (127/151), including both hormone receptor-positive (60/72) and hormone receptor-negative tumours (67/77). Triple-negative tumours comprised the majority of IntClust 10 (132/159) and around a quarter of IntClust 4 (52/217). Whilst the ten IntClust subtypes of breast cancer show characteristic patterns of association with traditional clinicopathological variables, no IntClust can be adequately identified by these variables alone. Hence, the addition of genomic stratification has the potential to enhance the biological relevance of the current clinical evaluation and facilitate genome-guided therapeutic strategies.


Activity of aspartate aminotransferase in the hippocampal formation of the rat during post-natal development and after lesion of the hippocampal Schaffer's collaterals: A quantitative histochemical study.
The activity of histochemically demonstrable aspartate aminotransferase was measured histophotometrically in the hippocampal formation, in dorsal root ganglia, and in superior cervical ganglia during postnatal development of the rat. The enzyme activity showed a marked (up to 15 times) increase in the dendritic layers of the hippocampal formation during the first weeks of life. Just as previous biochemical findings, the present data coincide with parameters of maturation of glutamatergic/aspartatergic structures in the hippocampal formation and other brain regions. For comparison, aspartate aminotransferase activities in both superior cervical ganglia and dorsal root ganglia were remarkably stable in the period studied. A nearly total transection of the CA3 pyramidal cell axon collaterals (Schaffer's collaterals) resulted in a striking reduction in aspartate aminotransferase activity within their target areas of the CA1 region (by 51.4% in stratum oriens, and by 60.2% in stratum radiatum). Our histochemical and histophotometrical data strongly account for the assumption that aspartate aminotransferase, beyond its numerous functions in general amino acid and nitrogen metabolism, is involved in the synthesis and/or catabolism of glutamate and aspartate used as excitatory transmitters.


Ultrasound Shear Wave Elastography: A Novel Method to Evaluate Bladder Pressure.
Children with bladder dysfunction resulting in increased storage pressure are at risk for renal deterioration. The current gold standard for evaluation of bladder pressure is urodynamics, an invasive test requiring catheterization. We evaluated ultrasound shear wave elastography as a novel means of assessing bladder biomechanical properties associated with increased bladder pressure. Concurrent shear wave elastography and urodynamics were performed. Ultrasound shear wave elastography images were obtained of the anterior and posterior wall when empty and at 25%, 50%, 75% and 100% expected bladder capacity, and end fill volume. Regions of interest were confirmed by a pediatric radiologist. Bladder cohorts were defined as compliant (capacity detrusor pressure less than 25 cm H2O) and noncompliant (25 cm H2O or greater). Pearson correlation coefficients and a mixed effects model evaluated the relationship between shear wave speed and detrusor pressure, compliance and normalized compliance. An unpaired t-test was used for between cohort analyses. In all 23 subjects mean shear wave speed of the anterior and posterior bladder walls significantly correlated with detrusor pressure throughout filling. When comparing compliant and noncompliant bladders, mean shear wave speed and detrusor shear wave speed of the anterior wall significantly increased with filling of noncompliant bladders. Shear wave speed remained at baseline levels in compliant bladders. Mean shear wave speed of the anterior wall was significantly correlated with compliance and normalized compliance. Ultrasound shear wave elastography bladder measurements correlate well with bladder storage pressure, and shear wave speed measurements differ between compliant and noncompliant bladders. This is the first known study to demonstrate that shear wave elastography is promising as a bedside modality for the assessment of bladder dysfunction in children.


An unusual outbreak of norovirus associated with a Halloween-themed swimming pool party in England, 2016.
In October 2016, an outbreak of norovirus occurred among attendees of a Halloween-themed party at a public swimming pool in the south-east of England. Norovirus genogroup II was confirmed in 11 cases. In the retrospective cohort study of pool users, 68 individuals (37 female and 31 male), with a median age of 11 years (range: 0-50 years), met the case definition of developing diarrhoea or vomiting between 6 and 72 h after the pool visit. Multivariable analysis showed that increasing age was associated with a reduced risk of illness (odds ratio = 0.91; 95% confidence interval: 0.83-0.99). Pool behaviours (swallowing water) and the timing of visit (attending pool party after automatic dosing system was switched off) were independently associated with increased risk. Environmental investigations revealed that the automatic dosing system was switched off to reduce chlorine levels to an intended range of 0.5-1 parts per million to facilitate the use of a commercial red dye. There was a lack of compliance with the operator's own pool operating procedures, particularly on maintaining effective chlorine levels in pool water, recording of test results and recording of actions undertaken. This outbreak highlights the risks of lowering chlorine levels when using pool water colourants.


[Rehabilitation service in a primary care unit].
to evaluate the impact of rehabilitation service in two primary care units (PCU). a comparative cross-sectional study was performed in two PCU (No. 53 and No. 51). The former has a rehabilitation service and the second one refers to a rehabilitation consultant at a specialized hospital. The variables evaluated were: amount of cases, days of disability, average days/cases, and average cost/case according to specific disease considering the five main medical disease required disability for general or risk work disease. We used descriptive statistics for results and Student's t test to evaluate significance. back pain was the first cause of disablement; the average in opportunity care was seven days. The average number of days disability benefit for hand trauma, ankle sprain, and for back pain was lower in PCU No. 53, as well as for cervical sprain, but slightly higher for knee sprain. Evaluating the global incapacity days average and cost for each case, we found a significant difference to PCU No. 53. recovery time to return work was faster and lower the average cost in patients from to the PCU No. 53 which has its own rehabilitation service.


Smoking behavior on the first day of a quit attempt predicts long-term abstinence.
The nicotine patch has been widely used for smoking cessation, but not all smokers quit using the patch. Knowing which smokers are likely to succeed with the nicotine patch may improve the efficiency of nicotine patch use. To identify predictors of smoking abstinence using baseline characteristics, smoking behavior, and withdrawal symptoms. Using 2 randomized clinical trials of pharmacologic treatment, brief counseling, and quit date formats in the outpatient research clinic setting, predictors of smoking cessation were derived in 1 sample (n = 159), then prospectively validated in an independent sample (n = 48). Subjects smoked 1 pack of cigarettes per day or more and were motivated to quit smoking. Self-report of abstinence at 6 months verified by exhaled carbon monoxide of 8 ppm or less was used. Abstinence at 6 months was 24% in the derivation set and 25% in the validation set. Using logistic regression, a model containing quit date abstinence (odds ratio, 10.6; 95% confidence interval [CI], 2.9-38.7) and baseline nicotine dependence (odds ratio, 0.75; 95% CI, 0.6-1.0 per unit increase in Fagerstrom score) provided the optimal predictive ability and was validated in the independent set. Quit date abstinence improved the likelihood of 6-month abstinence by 4.1 over baseline (95% CI, 2.6-6.4) for low-nicotine-dependent smokers and 1.2 (95% CI, 0.6-2.2) for high-nicotine-dependent smokers. Quit date smoking altered the likelihood of 6-month abstinence by 0.2 (95% CI, 0.0-0.8) for low-dependent smokers and 0.1 for high-dependent smokers (95% CI, 0.0-0.6). Abstinence on the quit date and low-nicotine dependence improve the likelihood of smoking abstinence at 6 months. Smoking on the quit date may be an indication for postponing the cessation attempt or adjusting the therapy for smoking cessation.


[Is calcium indicated in resuscitation? Experimental studies in dogs].
This study compares the effect of epinephrine (11 dogs) with that of the combination of epinephrine with calcium (10 dogs) in CPR after anoxial cardiac arrest. In the epinephrine group resuscitation was successful in all 11 dogs within 4 minutes. In the calcium group only 7 out of 10 dogs could be resuscitated: 3 dogs died during CPR in cause of irreversible fibrillation (2 cases) and "stone heart" (1 case). High CPK-activity reflected severe myocardial cell damage. In the survivors of the calcium group cardiac function was significantly impaired: 1. decreased left ventricular pressure (LVP) and contractility (dp/dt max), 2. increased afterload, 3. decreased cardiac output inspite of increased heart rate, 4. as a consequence, reduced perfusion of the vital organs. - These results foster the conclusion that calcium should not be used henceforth in CPR. Epinephrine was and is still the drug of choice in resuscitation after cardiac arrest.


Combined Medial Column Arthrodesis with Open Reduction Internal Fixation of Central Column for Treatment of Lisfranc Fracture-Dislocation: A Review of Consecutive Cases.
Lisfranc fracture-dislocations can be devastating injuries with significant long-term sequelae with or without surgical intervention. The main goal of treatment is to minimize the common long-term complications including pain, progressive arch collapse, degenerative joint disease, hardware failure, and reoperation. Partial primary fusion involving the first, second, and third tarsometatarsal joints has become a common approach for primarily dislocation injuries, with open reduction and internal fixation (ORIF) favored for Lisfranc injuries involving fracture. ORIF commonly requires revision surgery for hardware removal or delayed fusion. Major revision creates hardship for the patient due to the prolonged recovery required, and even "simple" hardware removal can be traumatic to local nerve, artery, and tendon structures. A common injury pattern includes the findings of primary dislocation and instability of the first tarsometatarsal joint with oftentimes comminuted fracture to the second and third tarsometatarsal joints, which does not fit the standard surgical approach. We report a review of our preferred surgical approach consisting of medial column primary arthrodesis combined with central column ORIF and lateral column temporary pinning. We undertook an institutional review board-approved review of 35 consecutive Lisfranc injuries treated with this hybrid approach. Mean follow-up time was 22.14 ± 22.39 (range 2.5 to 84) months. All but 2 (5.71%) patients had radiographic evidence of union at 10 weeks. Complications included 3 with neuritis, 1 with medial column nonunion that was treated with a bone stimulator, and 1 with revision of second metatarsal nonunion. The present retrospective series highlights our experience with isolated primary fusion of the medial column in both subtle and obvious Lisfranc injuries.


Macrophage complexity in human atherosclerosis: opportunities for treatment?
The pivotal role of macrophages in experimental atherosclerosis is firmly established, but their contribution to human disease is less well defined. In this review we have outlined the current insights on macrophage phenotypes and their presumed precursors, monocytes, in clinical atherosclerosis, and their association with disease progression. Moreover, we will assess major clinical modifiers of macrophage-mediated plaque inflammation and define the outstanding questions for further study. Our survey indicates that macrophage accumulation and status in human plaques are linked with lesion progression and destabilization as well as with symptomatic coronary artery disease. Likewise, levels of their precursors, circulating monocytes were repeatedly seen to associate with atherosclerosis and to predict clinical outcome. Furthermore, the presence and phenotype of both macrophages and monocytes appears to be responsive to the traditional risk factors of atherosclerosis, including hypercholesterolemia, hypertension, and type 2 diabetes, and to treatment thereof, with clear repercussions on disease development. Although plaque macrophages and their precursor cells do represent attractive targets for treating cardiovascular diseases, this therapeutic avenue requires much deeper understanding of the complexity of macrophage biology in human atherosclerosis than available at present.


The Sooner, the Better? The Importance of Optimal Timing of Cholecystectomy in Acute Cholecystitis: Data from the National Swedish Registry for Gallstone Surgery, GallRiks.
Up-front cholecystectomy is the recommended therapy for acute cholecystitis (AC). However, the scientific basis for the definition of the optimal timing for surgery is scarce. The aim of this study was to analyze how the timing of surgery, after the admission to hospital for AC, affects the intra- and postoperative outcomes. Within the national Swedish Registry for Gallstone Surgery and Endoscopic Retrograde Cholangiopancreatography (GallRiks), all patients undergoing cholecystectomy for acute cholecystitis between January 2006 and December 2014 were identified. Data regarding patient characteristics, intra- and postoperative adverse events (AEs), bile duct injuries, and 30- and 90-day mortality risk were captured, and the correlation between the surgical timing and these parameters was analyzed. In total, data on 87,108 cholecystectomies were analyzed of which 15,760 (18.1 %) were performed due to AC. Bile duct injury, 30- and 90-day mortality risk, and intra- and postoperative AEs were significantly higher if the time from admission to surgery exceeded 4 days. The time course between surgery and complication risks seemed to be optimal if surgery was done within 2 days after hospital admission. Although AC patients operated on the day of hospital admission had a slightly increased AE rate as well as 30- and 90-day mortality rates than those operated during the interval of 1-2 days after admission, the bile duct injury and conversion rates were, in fact, significantly lower. The optimal timing of cholecystectomy for patients with AC seems to be within 2 days after admission. However, the somewhat higher frequency of AE on admission day may emphasize the importance of optimizing the patient before surgery as well as ensuring that adequate surgical resources are available.


The impact of associated tenotomies on the outcome of incomplete phalangeal osteotomies for lesser toe deformities.
Partial or incomplete osteotomy (IO) of the phalanx is recently described in the literature. However, the clinical outcome and the rate of complications when applied to lesser toe deformities (LTD) have been never addressed. This study aims to find out if the association of tenotomies to incomplete or partial phalanx osteotomies has a significant impact on the clinical outcomes, the occurrence of complications, and the recovery time after surgery. A retrospective review of two cohorts of cases operated in our institution for hallux abductus valgus (HAV) and associated LTD from 2008 to 2014 was carried out. The surgical correction of both HAV and the associated LTD was always performed by minimally invasive techniques. The study included a total of 223 patients (723 IO in 556 toes). In 129 cases, the IO for LTD correction was performed without tenotomies, and in 94, the procedure was combined with flexor and/or extensor tenotomies. Patients were assessed with the American Orthopaedic Foot and Ankle Society (AOFAS) questionnaire before surgery and at 6- and 12-month follow-up. The mean preoperative AOFAS score before surgery was similar in both cohorts. At 12-month follow-up, the cohort without tenotomies showed better recovery (95.7 ± 2.8 versus 92.5 ± 6.8; p < 0.01). AOFAS scores decreased as the number of associated LTD increased (r = - 0.814; p < 0.001). Cases operated on by PO + tenotomy showed a high rate of complications such as delayed union of the osteotomy (p < 0.01), hypertrophic callus (p < 0.01), phalangeal fracture at the osteotomy site (p < 0.01), and lack of correction (p < 0.05). The overall occurrence of adverse events was 38.6% in cases operated by PO + tenotomy and 13.9% in cases receiving PO alone (p < 0.0001). Cases operated on without tenotomy showed a shorter time to complete recovery for daily life activities (37.4 ± 2.3 versus 43.0 ± 1.7 days; p < 0.01). The performance of associated tenotomies to incomplete phalanx osteotomies provides worse clinical outcomes, higher complication rates, and longer recovery time as compared to similar forefoot surgeries without tenotomies. The study was based on retrospectively registered data starting on May 24, 2008.


Functional heterogeneity among herpes simplex virus-specific human CD4+ T cells.
One hundred thirteen HSV-specific CD4+ T cell clones were established from the PBL of a healthy person and their functional heterogeneity was investigated. All clones proliferated in response to stimulation with HSV in the presence of autologous APC. Among those, 48 clones showed cytotoxic activity to HSV-infected autologous EBV-transformed lymphoblastoid cell line, but not to HSV-infected autologous fibroblasts, HSV-infected allogeneic cells, or K562 cells (group 1). Five clones showed cytotoxicity against HSV-infected autologous cells as well as HSV-infected allogeneic cells and K562 cells (group 2). The cytotoxicity of these clones was found to be mediated by the direct killing but not by the "innocent bystander" killing of target cells. Sixty clones showed no cytotoxic activity, however, among these, 23 revealed HLA-unrestricted and nonspecific cytotoxicity in the presence of PHA in culture (group 3), and the remaining 37 did not show any cytotoxic activity even in the presence of PHA (group 4). The cytotoxic patterns of these clones did not change in activated and resting phases, suggesting that the difference in cytotoxic ability does not depend on cell cycles. The cytotoxic activity of group 1 was inhibited by addition of anti-HLA-DR or anti-CD3 mAb to the culture, whereas these mAb had no effect on the cytotoxicity of group 2. All four groups of clones had helper activity for anti-HSV antibody production by autologous B cells. Moreover it was found that all groups of clones simultaneously produced IL-2, IL-4, and IFN-gamma after culture with APC followed by HSV Ag stimulation. The surface phenotype of all clones was uniformly CD2+, CD3+, CD4+, CD8-, CD29+, CD45RA-, but expression of Leu 8 was varied. These data therefore indicate that HSV-specific human CD4+ T cells are classified into at least four groups according to the presence and specificity of cytotoxicity, i.e., Th cells with HSV-specific and HLA-class II-restricted cytotoxicity, Th cells with HLA-unrestricted and nonspecific cytotoxicity, Th cells with lectin-dependent cytotoxicity, and Th cells without cytotoxic activity. The present finding of functional heterogeneity among virus-specific human CD4+ T cells might shed light on the pathogenesis of CD4+ T cell immunodeficiency, such as human retrovirus infections.


Postoperative Outcomes of Local Skin Flaps Used in Oncologic Reconstructive Surgery of the Nasal Ala: A Systematic Review.
Despite numerous options for nasal ala reconstruction, advantages and disadvantages of each method are unclear. To summarize reported outcomes of local flaps without the use of grafts for nasal ala oncologic reconstructive surgery. A nasal ala-specific protocol was adapted from a previous head- and neck-specific PROSPERO submission (CRD42017071596). The search was conducted in MEDLINE, EMBASE, and CENTRAL on December 23, 2017 and updated on May 10, 2019. Two reviewers screened 9,313 results from head and neck literature. Study bias was evaluated with the ROBINS-I tool. Subunit-based categorization of included studies identified 12 nasal ala-specific publications. Complications (flap necrosis, hematoma, wound infections, trapdoor deformities, and dehiscence), functional (nasal valve or respiratory dysfunction), and cosmetic (alar rim distortion/asymmetry/notching, secondary/revisionary procedures, and patient satisfaction) outcomes were extracted. Generally favorable outcomes are seen in all flaps. Careful consideration of donor sites for interpolation flaps is needed for optimal cosmetic outcomes. Transposition flaps, including laterally based bilobed and trilobed flaps, created good outcomes, although melolabial transposition flaps may produce poorer outcomes compared with melolabial island pedicle advancement flaps. Caution is needed for rotation flaps to prevent nasal valve/respiratory dysfunction due to alar crease contracture or ridge elevation. Further research is needed.


Insanity: A Legal and Cinematic Diagnosis.
The term "insanity" has been retired from medical nomenclature for about 100 years. Formerly interchangeable with the legal term, implying unsoundness of mind, it persists as a legal determination, mainly in criminal matters. However, the most prevalent uses of "insanity" are in colloquial speech and media. We track "insanity" in medical and legal parlance, reasons for its disappearance from psychiatry, and its persistence in popular culture. During the 19th century, specific types of legal insanity fell out of favor, especially "moral insanity," referring to irresistible impulses. The term persisted, for example, in some civil cases and in criminal cases, both denoting lack of capacity. In America, early 20th century focus on disease classification and nomenclature shifted from catchall terms (such as insanity, dementia, mania, and idiocy) to medical labels (psychosis and neurosis). Psychiatrist William Alanson White led the movement to change nomenclature. In 1921, the American Journal of Insanity became the American Journal of Psychiatry. By the time White was the American Psychiatric Association president in 1925, the medical use of "insanity" had been replaced in textbooks by progressive terminology. However, variations on "insane," suggesting loss of reason without diagnostic specificity, have become a staple among film tropes.


[The therapeutic value of alpha-fetoprotein and cholinesterase in patients with hepatitis B virus related acute onset chronic liver failure].
To evaluate the therapeutic value of alpha-fetoprotein (AFP) and cholinesterase (ChE) in patients with hepatitis B virus related acute onset chronic liver failure (HBV-ACLF). A case-control observation was conducted. Sixty-seven patients with HBV-ACLF admitted to Tianjin Second People's Hospital from January 2009 to October 2015 were enrolled. According to the diagnostic criteria of ACLF, the patients were divided into early, middle, and late groups, and alternatively, according to the outcome, they were divided into survival group and death group. Serum samples were collected after 0, 2, 4, 8 weeks to determine the value of AFP and ChE and analyze the value of AFP and ChE in reflecting the changes during HBV-ACLF progression. The differences in AFP and ChE between the survival group and the death group were compared. The prognostic values of AFP and ChE for HBV-ACLF patients were evaluated. Among 67 patients, there were 24, 24, and 19 patients in the early, middle and late stage, respectively, and there were 0, 9, 18 deaths at 8 week. With the advance of HBV-ACLF, the levels of both AFP and ChE were decreased in the early, middle, and late stage, with the AFP value of 40.205 (14.663, 90.550), 23.445 (8.233, 64.213), 8.990 (6.120, 14.340) μg/L (F = 36.149, P = 0.000) and the ChE value of (4.217±1.408), (3.619±1.200), (2.503±1.248) kU/L, respectively (F = 19.575, P = 0.000). In the death group, the levels of serum AFP at 0, 2, 4, 8 weeks were significantly lower than those in survival group [μg/L: 21.540 (7.670, 50.470) vs. 60.680 (16.383, 146.100), 10.560 (6.170, 20.100) vs. 60.090 (27.662, 100.700), 8.750 (3.045, 10.105) vs. 51.875 (16.778, 88.833), 3.900 (2.120, 7.660) vs. 20.400 (12.950, 50.430), P < 0.05 or P < 0.01]. The levels of serum ChE at 2, 4, 8 weeks in the death group were significantly lower than those in the survival group (kU/L: 3.206±1.480 vs. 4.008±1.227, 2.893±1.478 vs. 4.140±1.236, 2.948±1.355 vs. 4.329±1.390, P < 0.05 or P < 0.01). The levels of AFP in 67 patients were 30.100 (10.100, 90.100) μg/L, and ChE was (3.685±1.382) kU/L at 2 weeks, and they showed no correlation between AFP and ChE according to the linear correlation analysis (r = 0.082, P = 0.508), suggesting that AFP and ChE could be used as two independent prognostic factors for HBV-ACLF patients. It was showed by receiver operating characteristic curve (ROC) analysis that the area under the curve of AFP (AUC) was 0.847 (P = 0.000), while the AUC of ChE was 0.681 (P = 0.012). The highest values of Youden index and the maximum effectiveness of testing were obtained when AFP and ChE reached 20.520 μg/L and 2.924 kU/L, respectively, with the sensitivity and the specificity of AFP being 85.0% and 77.8%, respectively, and of ChE being 77.5% and 59.3%, respectively. When using the value of AFP ≥ 20.520 μg/L combined with the value of ChE ≥ 2.924 kU/L, the sensitivity for predicting HBV-ACLF outcome was 65.9%, and its specificity was 91.0%. Both AFP and ChE were helpful in providing accurate information for the progression and prognosis of HBV-ACLF patients due to the facts that their values were less interfered by the clinical treatment and that they have higher specificity.


Structural characterization of a trapped folding intermediate of pyrrolidone carboxyl peptidase from a hyperthermophile.
The refolding of cysteine-free pyrrolidone carboxyl peptidase (PCP-0SH) from a hyperthermophile is unusually slow. PCP-0SH is trapped in the denatured (D1) state at 4 °C and pH 2.3, which is different from the highly denatured state in the presence of concentrated denaturant. In order to elucidate the mechanism of the unusually slow folding, we investigated the structure of the D1 state using NMR techniques with amino acid selectively labeled PCP-0SH. The HSQC spectrum of the D1 state showed that most of the resonances arising from the 114-208 residues are broadened, indicating that conformations of the 114-208 residues are in intermediate exchange on the microsecond to millisecond time scale. Paramagnetic relaxation enhancement data indicated the lack of long-range interactions between the 1-113 and the 114-208 segments in the D1 state. Furthermore, proline scanning mutagenesis showed that the 114-208 segment in the D1 state forms a loosely packed hydrophobic core composed of α4- and α6-helices. From these findings, we conclude that the 114-208 segment of PCP-0SH folds into a stable compact structure with non-native helix-helix association in the D1 state. Therefore, in the folding process from the D1 state to the native state, the α4- and α6-helices become separated and the central β-sheet is folded between these helices. That is, the non-native interaction between the α4- and α6-helices may be responsible for the unusually slow folding of PCP-0SH.


Placenta and milk as excretory routes of lipophilic pesticides in women.
Age, diet and ethnic-dependent excretion of chlorinated pesticides through placenta and milk was studied in women from the general population around Lucknow in India by using gas-liquid chromatography equipped with an electron-capture detector (3H+). Pesticidal contaminants in both placenta and milk were 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT), its metabolites 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p'-DDE) and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDD), alpha, beta, gamma - isomers of benzene hexachloride (BHC) and aldrin. The study revealed higher excretion of pesticides through the placentas of older and non-vegetarian women than younger and vegetarian women. There was higher excretion of total DDT in rural than in urban women while a reverse trend was found in the case of total BHC in two ethnic groups. Similarly, age and dietary habits influenced the excretion of these pesticides in milk. Women undergoing premature delivery and stillbirths were found to excrete more DDT in their milk than those undergoing full-term normal delivery.


[Essential hypertension as an occupational disease in engine crew members].
The paper raises the question as to whether essential hypertension should be included into a list of occupational diseases (intellectual, sensory, and emotional workloads and working conditions) in engine crew members. In the engine crew members of the South-Eastern Railway, the cases and days lost because of illness and temporary disability are higher than those in the whole railway. 32% of the engine drivers (assistants) are followed up for diagnosed essential hypertension. Hypertension (49.7%) is the main reason for suspending engine crew members from work as evidenced by train pre-trip medical examinations. The working conditions (noise, infrasound, mcroclimate) in train engineer's cabs do not meet the sanitary requirements; in this connection the occupational disease sensorineural hearing loss is also registered.


Acute acrylonitrile exposure inhibits endogenous H2S biosynthesis in rat brain and liver: The role of CBS/3-MPST-H2S pathway in its astrocytic toxicity.
Hydrogen sulfide (H2S) as the third gasotransmitter molecule serves various biological regulatory roles in health and disease. Acrylonitrile (AN) is a common occupational toxicant and environmental pollutant, causing brain and liver damage in mammals. The biotransformation of AN is dependent-upon reduced glutathione (GSH), cysteine and other sulfur-containing compounds. However, the effects of AN on the endogenous H2S biosynthesis pathway have yet to be determined. Herein, we demonstrated that a single exposure to AN (at 25, 50, or 75 mg/kg for 1, 6 or 24 h) decreased the endogenous H2S content and H2S-producing capacity in a dose-dependent manner, both in the cerebral cortex and liver of rats in vivo. In addition, the inhibitory effects of AN (1, 2.5, 5, 10 mM for 12 h) on the H2S content and/or the expression of H2S-producing enzymes were also found both in primary rat astrocytes and rat liver cell line (BRL cells). Impairment in the H2S biosynthesis pathway was also assessed in primary rat astrocytes treated with AN. It was found that inhibition of the cystathionine-β-synthase (CBS)/3-mercaptopyruvate sulfurtransferase (3-MPST)-H2S pathway with the CBS inhibitor or 3-MPST-targeted siRNA significantly increased the AN-induced (5 mM for 12 h) cytotoxicity in astrocytes. In turn, CBS activation or 3-MPST overexpression as well as exogenous NaHS supplementation significantly attenuated AN-induced cytotoxicity. Taken together, endogenous H2S biosynthesis pathway was disrupted in rats acutely exposed to AN, which contributes to acute AN neurotoxicity in primary rat astrocytes.


Cyclophosphamide-refractory scleroderma-associated interstitial lung disease: remarkable clinical and radiological response to a single course of rituximab combined with high-dose corticosteroids.
We would like to report our experience of using rituximab in cyclophosphamide refractory, rapidly progressive interstitial lung disease (ILD) in a patient with limited scleroderma. A 40-year-old man presented with 10-week history of inflammatory polyarthritis, which responded to a short course of oral corticosteroids. However, 3 weeks later, he developed new onset of exertional dyspnoea. High-resolution CT of the thorax was suggestive of early ILD. Surgical lung biopsy showed features of fibrotic non-specific interstitial pneumonia. He was diagnosed with scleroderma on the basis of: presence of anticentromere antibodies, Raynaud's phenomenon, pulmonary fibrosis, digital oedema and hypomotility along with a dilated oesophagus. He was treated aggressively with pulse doses of corticosteroids and cyclophosphamide; however, his ILD continued to deteriorate. At this stage, he received rituximab (two pulses of 1 g each), which led to a gradual clinical improvement. Now, 12 months since his rituximab infusion, he walks 2 miles daily without any exertional dyspnoea.


Nonketotic Hyperglycinemia: Two Case Reports and Review.
Nonketotic hyperglycinemia (NKH) or glycine encephalopathy is an autosomal recessive disorder of glycine metabolism resulting in an excessive accumulation of glycine in all body tissues, including the central nervous system. It is caused by a biochemical defect in the glycine cleavage system and considered as a rare disorder with an estimated prevalence of 1:60,000. The neonatal form presents in the first few days of life with progressive encephalopathy, hypotonia, myoclonic jerks, hiccups, seizures, rapid progression to coma and often death due to central apnea. Surviving infants often have severe developmental delay and refractory seizures. Atypical forms of NKH present with heterogeneous and nonspecific disease course. Classical glycine encephalopathy usually carries a very poor prognosis. We describe two neonates who presented with neonatal encephalopathy, apnea, and progressive lethargy. Increased CSF glycine level along with an elevated CSF to plasma glycine ratio was suggestive of classic NKH. Burst suppression EEG and agenesis of the corpus callosum were supportive findings. Evolution of the EEG patterns and course of the disease are discussed in detail. Transient phases of clinical stabilization and normalized plasma biochemical results may not necessarily reflect the actual encephalopathic process. Serial EEGs are helpful to assess the efficacy of treatment and to modify the therapeutic approach.


Processing of herpes simplex virus type 1 glycoproteins: two-dimensional gel analysis using monoclonal antibodies.
The number of discrete species of glycoprotein induced by herpes simplex virus type 1 (HSV-1, strain 17 syn+) and their processing has been examined by a combination of immunoprecipitation with monoclonal antibodies and analysis of immune precipitates by two-dimensional (2D) gel electrophoresis. Seventeen different monoclonal antibodies directed against glycoproteins A/B, C, D and E were used. Polypeptides intermediate in the synthesis of gA/B, C and D were visualized and two early intermediates of glycoprotein A/B (pgA/B1 and pgA/B2), both mannose-containing, were identified. Comparison on 2D gels of polypeptides synthesized in vitro from HSV-1-infected cell mRNA with those synthesized in pulse-chase experiments in vivo has allowed identification of early precursors of pgA/B and pgD. Their apparent mol. wt. were 105000 and 47000 respectively. The 2D gel analysis of glycoproteins induced in HFL cells infected with 17 syn+ revealed a number of previously unreported glycoprotein species. One had mobility on both gradient and single concentration SDS-polyacrylamide gels similar to that of gC but was resolved from gC on non-equilibrium pH gradient gels. This glycoprotein was produced in relatively large amounts and was not precipitated by any of the monoclonal antibodies used in this study. The data suggest that this glycoprotein either has a polypeptide chain unrelated to those of glycoproteins A/B, C, D or E or alternatively is derived from one of them and modified in such a way as to mask or remove the antigenic sites with which the monoclonal antibodies interact. Sixteen other previously unreported glycoprotein species not obviously related, as judged by their electrophoretic mobility, to gA/B, C, D or E were also identified: two reacted with gA/B-specific monoclonal antibodies and five with glycoprotein C-specific monoclonal antibodies. The origin of the remaining nine new glycoproteins has still to be ascertained.


Interferometric analysis of sub-nanosecond laser-induced optical breakdown dynamics in the bulk of fused-silica glass.
Dynamics of laser-induced optical breakdown in the bulk of fused-silica glass irradiated by a sub-nanosecond laser pulse at a wavelength of 790 nm with a fluence of 522 J/cm2 was studied by the femtosecond time-resolved complex interferometry in Nomarski arrangement utilising a Fresnel bi-prism. Evolution of the plasma channel and the development of the free electron density were in focus of the investigation. The measured ultimate length of the plasma channel was equal to 30 μm and almost doubled the length estimated within the moving breakdown model. The history of the transient electron density distribution in the plasma was reconstructed from the phase shift maps using the inverse Abel transform and it revealed further deviation from this model. The core of the plasma channel exhibited at the last stages of the development a considerable level of the electron density up to 2.4×1020 cm-3. The signature of the pre-breakdown phase has been identified as radiation caused by ionization-released electrons interacting with ions and has been demonstrated in solids for the first time in this way. Origin of the discrepancy between the theoretical prediction of the moving breakdown model and the measured values of the channel length is discussed as well.


Clinical profiles and symptoms of childhood depression.
The identification of depression poses significant clinical challenges to physicians, parents and teachers. Symptoms of depression may differ from those of the adult depression. Clinical depression varies in the nature and intensity of its presenting symptoms. It may present with the cardinal symptoms of major depressive disorder. It often goes unidentified and undiagnosed when it presents as masked depression or depressive equivalents.