Dysregulated Response of Follicular Helper T Cells to Hepatitis B Surface Antigen Promotes HBV Persistence in Mice and Associates With Outcomes of Patients.
Production of neutralizing antibodies against hepatitis B surface antigen (HBsAg) is dysregulated in patients with persistent hepatitis B virus (HBV) infection. We investigated mechanisms by which this immune response to the virus is disrupted and whether it can be restored to promote clearance of HBV. Immune-competent C57BL/6N and C57BL/6J, as well as mice deficient in follicular helper T cells (Tfh-cell-deficient), B cells, or Foxp3+ T-regulatory cells (Treg cell deficient), were given hydrodynamic injections of pAAV/HBV1.2 plasmids. Some mice were given injections of sorted Tfh cells, pan-B cells, Treg cells, or a blocking antibody against CTLA4. Production of antibodies against HBsAg and clearance of HBV were assessed by flow cytometry, enzyme-linked immunosorbent assay, polymerase chain reaction, and immunohistochemical analyses. We obtained blood samples from patients with HBV infection and isolated Treg cells. We measured the ability of Treg cells to suppress production of interleukin 21 (IL21) in CD4+ T cells. Immune-competent C57BL/6N and C57BL/6J mice transfected with the plasmid encoding HBV had features of viral clearance and viral persistence observed in humans. A Tfh-cell response to HBsAg was required for clearance of HBV and was suppressed by Treg cells in mice with persistent HBV infection. Depletion of Treg cells or inhibition of Treg-cell function (with blocking antibody against CTLA4) restored the Tfh-cell response against HBsAg and clearance of HBV in mice. Impaired Tfh-cell response to HBsAg was observed in blood from patients with chronic HBV infection, responsiveness was restored by depletion of Treg cells or blocking antibody against CTLA4. In studies of HBV-infected mice and blood from patients with chronic HBV infection, we found a Tfh-cell response to HBsAg of to be required for HBV clearance, and that this response was blocked by Treg cells. Inhibiting Treg-cell activity using neutralizing antibody against CTLA4 restored the ability of Tfh cells to clear HBV infection; this approach might be developed for treatment of patients with chronic HBV infection.


Primary angiosarcoma of the pulmonary trunk mimicking pulmonary thromboembolism.
We present a case of primary angiosarcoma of the pulmonary trunk that was initially misdiagnosed as a subacute massive pulmonary thromboembolism in a 26-year-old woman. This is an extremely rare disease that is usually indistinguishable from acute or chronic thromboembolic disease of the pulmonary arteries because the clinical and radiologic findings of pulmonary artery angiosarcoma are similar to those of pulmonary thromboembolism. Although the incidence of pulmonary artery angiosarcoma is very low, our case demonstrates that this disease entity should be included in the differential diagnosis of pulmonary thromboembolism, especially in patients who do not respond to anticoagulant therapy or present with no identifiable source of thromboembolic events.


Music Perception in Bone-Anchored Hearing Implant Users.
Music is a highly complex acoustic stimulus in both spectral and temporal contents. Accurate representation and delivery of high-fidelity information are essential for music perception. However, it is unclear how well bone-anchored hearing implants (BAHIs) transmit music. The study objective is to establish music perception performance baselines for BAHI users and normal hearing (NH) listeners and compare outcomes between the cohorts. A case-controlled, cross-sectional study was conducted among 18 BAHI users and 11 NH controls. Music perception was assessed via performance on seven major musical element tasks: pitch discrimination, melodic contour identification, rhythmic clocking, basic tempo discrimination, timbre identification, polyphonic pitch detection, and harmonic chord discrimination. BAHI users performed comparably well on all music perception tasks with their device compared with the unilateral condition with their better-hearing ear. BAHI performance was not statistically significantly different from NH listeners' performance. BAHI users performed just as well, if not better than NH listeners when using their control contralateral ear; there was no significant difference between the two groups except for the rhythmic timing (BAHI non-implanted ear 69% [95% CI: 62%-75%], NH 56% [95% CI: 49%-63%], p = 0.02), and basic tempo tasks (BAHI non-implanted ear 80% [95% CI: 65%-95%]; NH 75% [95% CI: 68%-82%, p = 0.03]). This study represents the first comprehensive study of basic music perception performance in BAHI users. Our results demonstrate that BAHI users perform as well with their implanted ear as with their contralateral better-hearing ear and NH controls in the major elements of music perception. III Laryngoscope, 2023.


Multiple domains in a ribozyme construct confer increased suppressive activity in monkey cells.
An expression vector designed to express a long (approximately 1 kb) RNA containing multiple ribozyme domains was cotransfected into mammalian cells with a plasmid encoding, as target, messenger RNA for chloramphenicol acetyltransferase (CAT). In comparative studies the multimeric ribozyme construct proved to be significantly more effective at suppressing CAT expression than either the corresponding antisense RNA, or a transcript carrying a single ribozyme domain. Suppression of gene activity was apparently specific because expression of an independently expressed gene for human growth hormone was unaffected. The profile of CAT RNA extracted from transfected cells was consistent with RNA cleavage.


Physiological Significance of the Heterogeneous Distribution of Zeaxanthin and Lutein in the Retina of the Human Eye.
Zeaxanthin and lutein are xanthophyll pigments present in the human retina and particularly concentrated in its center referred to as the yellow spot (macula lutea). The fact that zeaxanthin, including its isomer meso-zeaxanthin, is concentrated in the central part of the retina, in contrast to lutein also present in the peripheral regions, raises questions about the possible physiological significance of such a heterogeneous distribution of macular xanthophylls. Here, we attempt to address this problem using resonance Raman spectroscopy and confocal imaging, with different laser lines selected to effectively distinguish the spectral contribution of lutein and zeaxanthin. Additionally, fluorescence lifetime imaging microscopy (FLIM) is used to solve the problem of xanthophyll localization in the axon membranes. The obtained results allow us to conclude that one of the key advantages of a particularly high concentration of zeaxanthin in the central part of the retina is the high efficiency of this pigment in the dynamic filtration of light with excessive intensity, potentially harmful for the photoreceptors.


Characterization of new pancreatic cancer-reactive monoclonal antibodies directed against purified mucin.
Although mucins have been found to be useful in the diagnosis of pancreatic cancer, the carbohydrate and peptide structures of pancreatic mucins are still not well characterized. Monoclonal antibodies were produced using mucins purified from xenografts of a human pancreatic cancer cell line as the immunogen. One of these, Ia3, reacted with almost all pancreatic, gastric, and colorectal carcinomas examined by immunoperoxidase staining, but with few normal tissues. Ia3 antigen was elevated in sera of 50.4% of individuals with gastrointestinal tumors, but its levels did not correlate with those of CA15-3, CA19-9, or DU-PAN-2. Serum Ia3 antigens migrated more slowly in sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the polymorphic epithelial mucins recognized by DF3 or 115D8. Ia3 reacted only with native, and not with partially deglycosylated, pancreatic cancer xenograft mucins. Periodate or neuraminidase treatment destroyed this reactivity, but protease had little effect. The antigen recognized by another antibody, Nd2, was not detected in normal pancreatic, colonic, or gastric tissues but was present in approximately 60% of the pancreatic and gastric carcinomas examined. Nd2 reactivity with native and partially deglycosylated mucin was lost after pretreatment with protease and beta-mercaptoethanol. We conclude that, while Ia3 reacts with carbohydrates, Nd2 reactivity appears to be dependent on the integrity of the mucin protein core. The antigenic determinants of Ia3 and Nd2 are different from those of B72.3, CA19-9, DU-PAN-2, SPan-1, and several breast cancer mucin-directed antibodies. These results suggest that the malignancy-associated structures identified by Ia3 and Nd2 may provide new information on the carbohydrate and peptide structure of pancreatic cancer mucins.


Ni3V2O8 nanoflower mimetic enzyme constructs a dual-signal system for ascorbic acid detection.
Ascorbic acid is a nutritional small molecule essential to human life activities and health, playing a vital role in many physiological processes. Fresh fruits and beverages can provide abundant AA to maintain human metabolic balance. Therefore, it is of great significance to develop a nanomaterial with superior nanozyme activity for rapid and convenient detection of ascorbic acid (AA) in fruits and beverages. Herein, a dual-signal sensing platform based on UV-vis absorption and test strip chromaticity for the quantitative determination of AA is presented. The sensing platform is based on the horseradish peroxidase-like activity of Ni3V2O8 nanoflowers, which catalyzes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by hydrogen peroxide to the blue oxide TMB (ox TMB). The ox TMB produced by the oxidation has a characteristic absorption peak at 650 nm. In the presence of AA, the blue ox TMB is reduced to colorless TMB, and the quantitative detection of AA can be achieved by detecting the decrease in intensity of the absorption peak by UV-Vis spectrophotometry. Under the optimal experimental conditions, the sensing platform exhibited excellent sensitivity and selectivity. A wide linear range of 0.1 μM to 40 μM with a detection limit of 0.032 μM was obtained. The linear equation is ΔA = 0.02513c + 0.1164 with a correlation coefficient of 0.9979. It showed excellent properties in the detection of real samples of fruit juices and beverages, meanwhile, a method for the rapid detection of AA based on chromaticity change of test strips was constructed with high sensitivity and convenience. The linearity range for the ascorbic acid was 1-50 μM with LOD of 0.42 μM. The developed sensing platform has the capability to quickly and accurately detect ascorbic acid (AA) in fresh fruits and beverages. This proposed method offers a new and promising approach for the rapid and cost-effective detection of ascorbic acid, which has a wide range of potential applications.


Terminal phase males stimulate ovarian function and inhibit sex change in the protogynous wrasse Thalassoma duperrey.
Under experimental conditions, the probability of sex change in the protogynous wrasse Thalassoma duperrey is determined largely by an individual's relative size within a social group. Natural populations, however, contain two distinct male phenotypes that may also play a role in regulating sex change. To investigate potential effects of male phenotype, the ability to change sex, ovarian histology and serum estradiol-17 beta levels were examined in females maintained under controlled social settings. Large females housed with smaller or larger terminal phase males had significantly larger gonadosomatic indices than females housed singly, with other females or with smaller initial phase males. Similarly, ovaries of females housed with terminal phase males showed no histological evidence of sex change, whereas large females from other social groupings were in advanced stages of sex change. These results demonstrate terminal phase males inhibit sex change regardless of their size relative to the female. Furthermore, gonadosomatic indices, ovarian histology, and serum estradiol-17 beta levels of females housed with terminal phase males indicate normal ovarian function whereas ovaries of other treatment groups appear quiescent or are undergoing sex change. Consequently, terminal phase males may be required for normal ovarian development which may, in turn, inhibit sex change in T. duperrey.


Signet-ring cell sinus histiocytosis: report of the clinicopathologic characteristics of 4 cases with emphasis on the differential diagnosis of signet-ring cell lesions.
Signet-ring cell sinus histiocytosis (SRCSH) represents a distinctly rare reactive phenomenon predominantly affecting axillary and pelvic lymph nodes (LNs) of individuals with breast or prostatic adenocarcinoma. Reports of SRCSH in the literature are sparse with only 12 previous examples, thus underscoring the rarity of this process. Here, we report 4 additional SRCSH cases affecting 2 women and 2 men (M/F = 1:1; age range: 50-71 years; mean age = 61 years). In the 2 men, pelvic LNs were excised during radical cystoprostatectomy for genitourinary cancer, whereas in one woman, SRCSH was incidentally discovered in axillary LNs during mastectomy for breast adenocarcinoma. The other female patient presented with a history of aortic valve replacement and enlarged supraclavicular LNs. Microscopically, all involved LNs exhibited marked distention with filling of the subcapsular and medullary sinuses by sheets of signet-ring histiocytes containing a singular large, cytoplasmic vacuole and a crescentic nucleus. Overt cytologic atypia, pleomorphism, and mitoses were absent. Erythrophagocytosis and occasional fibrosis were appreciated. None of the LNs with SRCSH showed evidence of metastatic tumor. Immunohistochemically, signet-ring sinus histiocytes were invariably positive for CD68 and CD163 but were negative for pancytokeratins. The histopathologic characteristics of SRCSH, albeit bland, in conjunction with the patient's medical history, may be misinterpreted as metastatic adenocarcinoma with signet-ring cell configuration. Immunohistochemical confirmation of the histiocytic lineage of the lesional cells in SRCSH usually suffices for rendering an accurate diagnosis. The underlying pathogenetic mechanism and possible biologic significance of SRCSH remain currently unknown.


[Targeting of anticancer drug using intelligent polymers].
To fabricate the drug targeting system, it is very effective to combine passive targeting with intelligent polymeric system such as temperature-responsive polymers. We have been studying the passive targeting of anticancer drug bonded with or physically trapped in polymeric micelle derived from poly (ethylene oxide) (PEO) and poly (aspartic acid) block copolymer. On the other hand, Poly(N-isopropylacrylamide) (PIPAAm) is known to be a thermo-responsive polymer which has lower critical solution temperature (LCST, 32 degrees C). PIPAAm shows the reversible phase transition above and below LCST. Attempts to apply thermo-responsive drug carriers were made to enhance the site-specificity of polymeric micelle. Namely, novel type of polymeric micelle were prepared from block copolymers comprising N-isopropylacrylamide (IPAAm)-hydrophobic polymers. This temperature responsive micelles were expected to induce selective accumulation controlled by temperature modulation.


Effect of observing response requirements to sample and comparison stimuli on the establishment of reject control (sample/S- relations).
The establishment of sample/S- relations (or reject control) during conditional discrimination training (AB, BC) affects transitivity (AC), equivalence (CA) and reflexivity (AA, BB, CC) test outcomes. The present study parametrically evaluated the effects of different observing patterns to comparison stimuli on the establishment of reject control during baseline conditional relation training. A matching-to-sample with observing requirements (MTS-OR) procedure was implemented during AB and BC conditional discrimination training. During training, the participants were required to observe the sample and incorrect comparison on every trial before responding. In addition, the participants were divided into three groups that differed regarding the percentage of training trials on which they were prevented from observing the correct comparison stimuli: 25%, 50%, and 75%. Once the mastery criteria were achieved during training, transitivity (AC), symmetry (BA, CB), equivalence (CA), and reflexivity (AA, BB, CC) tests were conducted with all comparison stimuli visible from the beginning. The results suggest that the number of errors during transitivity, equivalence, and reflexivity tests progressively increased as participants were prevented from observing the correct comparison on a greater number of trials during training. Symmetry test results, however, were not affected by the experimental manipulation. Moreover, the number of participants showing reject-control patterns during tests slightly increased and the number of participants showing select-control patterns decreased as a function of the number of trials on which the participants were prevented from observing the correct comparison. Thus, we suggest that observing patterns during training is a relevant variable that affects equivalence test outcomes.


[Chronological mass change of intracranial transplanted omentum in accord with decrease/increase in a patient's body weight--case report].
Moyamoya disease is a chronically progressive and occlusive cerebrovascular disease. Steno-occlusion of the terminal portion of the internal carotid artery occurs bilaterally, and fine vascular network gradually develops at the base of the brain and they act as collaterals to the distal portion of the anterior and middle cerebral arteries. Steno-occlusion of the posterior cerebral artery develops in accord with progression of the disease. Intracranial omental transplantation is one of the indirect bypass surgeries for the treatment of cerebral ischemia, especially in the territories of the anterior/posterior cerebral artery in moyamoya disease. In the present report, one of sixty-seven moyamoya disease patients who had been treated with intracranial omental transplantation, showed chronological change of the mass of the transplanted omentum in accord with decrease/increase in the patient's body weight. In addition, the patient experienced headache which appeared and disappeared in accord with increase and decrease in the patient's body weight, respectively. The transplanted omentum was thought to act in the same way as fat tissue in other portions of the body. Measurement of body weight is thought to be as important as radiological examination using computed tomography/magnetic resonance imaging in the follow-up period after intracranial omental transplantation in moyamoya disease.


Human induced pluripotent stem cell differentiation and direct transdifferentiation into corneal epithelial-like cells.
The corneal epithelium is maintained by a small pool of tissue stem cells located at the limbus. Through certain injuries or diseases this pool of stem cells may get depleted. This leads to visual impairment. Standard treatment options include autologous or allogeneic limbal stem cell (LSC) transplantation, however graft rejection and chronic inflammation lowers the success rate over long time. Induced pluripotent stem (iPS) cells have opened new possibilities for treating various diseases with patient specific cells, eliminating the risk of immune rejection. In recent years, several protocols have been developed, aimed at the differentiation of iPS cells into the corneal epithelial lineage by mimicking the environmental niche of limbal stem cells. However, the risk of teratoma formation associated with the use of iPS cells hinders most applications from lab into clinics. Here we show that the differentiation of iPS cells into corneal epithelial cells results in the expression of corneal epithelial markers showing a successful differentiation, but the process is long and the level of gene expression for the pluripotency markers does not vanish completely. Therefore we set out to determine a direct transdifferentiation approach to circumvent the intermediate state of pluripotency (iPS-stage). The resulting cells, obtained by direct transdifferentiation of fibroblasts into limbal cells, exhibited corneal epithelial cell morphology and expressed corneal epithelial markers. Hence we shows for the first time a direct transdifferentiation of human dermal fibroblasts into the corneal epithelial lineage that may serve as source for corneal epithelial cells for transplantation approaches.


Results of liver dearterialization combined with regional infusion of 5-fluorouracil for liver cancer.
40 patients with primary or metastatic liver malignancies have been treated with hepatic dearterialization, 19 of them in combination with regional infusion with 5-fluoroacil. The survival was longer than expected from untreated materials. This was especially shown to be valid for patients with primary liver cancer and liver metastases from cancer coli-recti, in which the prolongation of survival was associated with an acceptable quality of life. The long term results were correlated with the "aggressiveness" in therapeutic approach. Only studies of prospective randomized series can settle whether the results depend on the specific measures against the liver tumours or on the general treatment.


Social reactions to disclosure of interpersonal violence and psychopathology: A systematic review and meta-analysis.
Public attention has been increasingly paid to how friends, family members, and others can best support survivors of sexual assault and other forms of violence. The broader social support literature posits that perceiving social support positively is more important to mental health than the degree to which social support is actually received, and that negative interactions with social supporters are more harmful than positive interactions are helpful (potentially because negative reactions violate survivors' expectations of their social supporters). This may be especially true after a crisis, such as interpersonal violence. Thus, this systematic review and meta-analysis summarizes the literature on social reactions to interpersonal violence. Meta-regression analyses were performed on 1871 correlations from 51 studies reflecting the degree to which receiving specific reactions more frequently, or perceiving reactions more positively, was associated with psychopathology. Results indicated that negative social reactions to disclosure-especially reactions involving controlling, distracting, and treating survivors differently-were associated with worse psychopathology, whereas positive social reactions did not appear to be protective. Perceiving reactions more positively was associated with less severe psychopathology, but (although causation cannot be concluded) positive perceptions' potential benefit appeared to be smaller than the potential risk conveyed by negative reactions. These findings indicate that interventions which reduce the degree to which survivors receive negative social reactions are needed.


Cell-mediated immune reactions as probes of sensitization to tumour-associated antigens of methylcholanthrene-induced mouse sarcomas.
The evaluation of the Field-Caspary macrophage electrophoretic mobility (MEM) assay as a probe of sensitization to tumour-associated antigens was performed in the genetically defined experimental system of methylcholanthrene-induced sarcomas in C57BL10/ScSnPh mice. Six sarcomas carrying noncross-reacting tumour-associated transplantation antigens were examined. For comparison, antigenic specificity of the six sarcomas was simultaneously assessed by the microcytotoxicity assay with 3H-thymidine. Immunization of mice with syngeneic tumour cells as well as sensitization of lymph node cells (LNC) in vitro lead to the generation of cytotoxic lymphocytes. Tumour bearers' LNC were not cytotoxic; when added to preimmunized LNC, the tumour bearers' LNC suppressed cytotoxicity. In contrast to the results obtained with the microcytotoxicity assay, the MEM test gave positive results with LNC from both tumour-bearing and tumour-immunized mice. Whereas some cross-reactions were observed among the fibrosarcomas in the microcytotoxicity assay (6 out of 28 combinations examined), the results of the Field-Caspary MEM test showed the same individually specific pattern of antigenicity as observed in transplantation tests and revealed no cross-reactivity in the 25 combinations tested.


Preoperative evaluation and management of the pancreatic head mass.
The differential diagnosis of a pancreatic head mass encompasses a wide range of clinical entities that include both solid and cystic lesions. This chapter focuses on our approach to the patient presenting with a newly found pancreatic head mass with the main goals of determining the risk of the lesion being malignant or premalignant, resectability if the patient is appropriate for surgical intervention, assessment of need for multimodality treatment and determination the patient's surgical risk.


A meta-analysis of child physical abuse prevalence in China.
This study estimated the prevalence of child physical abuse in China, compared Chinese prevalence with international and Asian estimates, and ascertained whether some differences in sample characteristics and methodological factors (e.g., time prevalence, definitional or regional difference) help explain variations in Chinese rates. Based on a meta-analysis of 47 studies found in English- and Chinese-language peer-reviewed journals that involved general populations of students or residents reporting child physical abuse prior to age 18, the life time prevalence of any child physical abuse in China was estimated at 36.6% (95% CI: 30.4-42.7), which was significantly higher than either the international or the Asian estimate in Stoltenborgh et al.'s (2013) study. Chinese prevalence was estimated at 43.1% (95% CI: 36.6-52.5) for minor physical abuse, 26.6% (95% CI: 21.4-31.8) for severe physical abuse, and 7.8% (95% CI: 5.0-10.5) for very severe physical abuse. Subgroup analysis found a significant difference between lifetime and 12-month or less prevalence only for minor physical abuse. The prevalence of any and minor child physical abuse in mainland China was significantly higher than that in non-mainland China. The mainland and non-mainland difference was significant even controlling for definitional and methodological factors as well as sample characteristics. The findings suggested the need to develop educational programs to promote non-violent parenting particularly in mainland China.


Eikenella corrodens as a cause of brain abscess.
A case of brain abscess due to Eikenella corrodens in a 67-year-old woman is reported. To our knowledge, this is the first case in which the causative organism has been isolated from the occipital lobe. E. corrodens may be the cause of apparently sterile brain abscesses encountered in neurosurgery and should be considered in the differential etiology of brain abscess.


Congenital lobar emphysema. A case report.
One infant, age 19 days, with congenital lobar emphysema whose main symptoms included dyspnea, cyanosis and severe infant Respiratory Distress Syndrome, was presented in this paper. The most reliable diagnostic tool was chest x-ray films with anteroposterior and lateral views. As additional diagnostic method, thorax computerized tomography was also used. The affected lobe was the left upper lobe. Resection of the affected lobe was performed with good results.


MacroH2A allows ATP-dependent chromatin remodeling by SWI/SNF and ACF complexes but specifically reduces recruitment of SWI/SNF.
The variant histone macroH2A helps maintain X inactivation and gene silencing. Previous work implied that nucleosomes containing macroH2A cannot be remodeled by ISWI and SWI/SNF chromatin remodeling enzymes. Using approaches that prevent misassembly of macroH2A nucleosomes, we find that macroH2A nucleosomes are excellent substrates for both enzyme families. Interestingly, SWI/SNF, which is involved in gene activation, preferentially binds H2A nucleosomes over macroH2A nucleosomes, but ACF, an ISWI complex implicated in gene repression, shows no preference. Thus, macroH2A may help regulate the balance between activating and repressive remodeling complexes.


Cinnamon (Cinnamomum zeylanicum) Oil as a Potential Alternative to Antibiotics in Poultry.
The removal of antibiotic growth promoters (AGPs) as feed additives in poultry nutrition from the market in many countries has compelled researchers to find unconventional and safe alternatives to AGPs. Probiotics, prebiotics, enzymes, organic acids, herbs, immune-stimulants and essential oils (EO) have been investigated as feed additives in poultry production. Cinnamon (Cinnamomum zeylanicum), one of the oldest medicinal plants and widely used around the world, can be used in poultry rations in the form of powder or essential oil. Essential oils produced from aromatic plants have become more interesting owing to their potential effects as hypocholesterolaemic agents, antioxidants, antimicrobials, antifungals and stimulants of digestive enzymes. The potential insecticidal and antimicrobial activities of EO against pathogens that cause spoilage in agriculture crops and human diseases might be attributed mainly to the high content of volatile components (mainly cinnamaldehyde, eugenol and carvacrol) in cinnamon oil. The present review focuses on the effects of cinnamon oil as a feed additive on poultry performance, carcass traits, meat quality, hypocholesterolaemic impact, antioxidant activity, immunity and microbiological aspects.


Pharmacological characterization of the nucleotide receptors that mobilize Ca2+ ions in human parathyroid cells.
We have used the fluorescent probe fura-2 to perform agonist studies of the receptor(s) that mobilizes Ca2+ ions in response to extracellular ATP in human parathyroid cells. Extracellular ATP induced Ca2+ responses in both normal and adenomatous parathyroid cells. Activation resulted in an initial small transient response during which Ca2+ ions were released from intracellular stores, followed by a prominent plateau response during which Ca2+ ions entered the cells from the extracellular fluid. The responses exhibited moderate desensitization upon repeated stimulation with ATP, and the ratio of the plateau to the peak response remained constant for any given group of activated cells. The baseline intracellular calcium concentration was 100 +/- 4.3 nM (mean +/- S.E.M., n = 3). Following maximal activation by extracellular ATP it rose to a peak of 684 +/- 45.7 nM (n = 3) and a plateau level of 415 +/- 9.9 nM (n = 3). We examined the effects of a variety of nucleotide species. The order of potency was: adenosine, AMP < alpha, beta-methylene ATP < ADP < ATP approximately UTP. In the concentration range 1-1000 microM, UTP (the concentration of agonist inducing a half-maximal response, EC50 = 2.4 microM) was slightly more potent than ATP (EC50 = 3.6 microM), and the two nucleotides evoked similar maximal responses. In the concentration range 0.01-1.0 microM, however, there was a clear difference in the behaviour of the two nucleotides. In particular, ATP, but not UTP, evoked responses that suggested the presence of a second receptor of higher potency but markedly lower efficacy.(ABSTRACT TRUNCATED AT 250 WORDS)


Differential features of early childhood motor skill development and working memory processing: evidence from fNIRS.
The study investigated the differential characteristics associated with motor skill development and working memory processing during early childhood, thereby providing insights for understanding motor learning and cognitive development in young children. In total, 101 preschool children (age: 4-6 years) were recruited for this study. The motor skill development level and the working memory capacity of the children were assessed using the MOBAK Motor Development Assessment Scale and a block task paradigm, respectively. Functional near-infrared spectroscopy brain imaging technology was used to monitor hemodynamic signals in the prefrontal cortex (PFC) of the children while they completed different memory tasks. MATLAB software and the Homer2 plugin were used to calculate the oxygenated hemoglobin (Oxy-Hb) concentration in relevant brain regions during the tasks. (1) The low motor skill group exhibited significantly lower accuracy during the three-memory load condition than during the two-memory load condition. Under both two-memory and three-memory load conditions, the high motor skill group exhibited significantly higher accuracy than the low motor skill group. (2) Significant differences in the Oxy-Hb concentration were observed in the left dorsolateral PFC (L-DLPFC), and right and left triangular part of the Broca's area (R-PTBA and L-PTBA, respectively) between the two memory difficulty levels for the high motor skill group. The Oxy-Hb concentration was significantly higher during the three-memory load condition than during the two-memory load condition. Under the two-memory load condition, the high motor skill group exhibited significantly higher Oxy-Hb concentration in the L-DLPFC and L-PTBA regions than in the low motor skill group. Under the three-memory load condition, the high motor skill group exhibited significantly higher Oxy-Hb concentration in the L-DLPFC, R-PTBA, and L-PTBA regions than the low motor skill group. A close association was observed between the motor skill levels and working memory in young children, with higher motor skill levels being associated with more pronounced brain activation patterns during working memory tasks.


Suppression of recurrent genital herpes by single daily dosages of acyclovir.
Patients with frequent recurrences of genital herpes were treated with oral acyclovir tablets, 800 mg, or placebo once daily for two years. Confirmed recurrences for all patients were treated with acyclovir capsules, 200 mg orally five times per day, for five days. Of 46 patients enrolled, 18 of 22 acyclovir recipients and 14 of 24 placebo recipients completed two years of study. Patients receiving acyclovir experienced a mean of 0.184 recurrences/month compared with a mean of 0.977 recurrences/month for patients receiving placebo (p less than 0.0001). A total of 28 percent of acyclovir recipients and no placebo recipients remained free of recurrences for two years. The low rate of recurrences in the acyclovir group remained consistent throughout the study. No serious clinical or laboratory abnormalities associated with acyclovir were observed. The 800-mg acyclovir tablet given daily was well-tolerated and effective for two years in the management of recurrent genital herpes.


[Vascular reconstructive surgery for elderly patients with internal carotid artery stenosis at the cervical portion].
We encounted increasing numbers of elderly patients suffering from stenotic lesions of the cervical internal carotid artery. Most of them have been considered to indicate a need for carotid artery stenting. The purpose of this study was to clarify with regard to the modality of treatment and perioperative complications the effectiveness of vascular reconstruction procedure in elderly patients. Ninety eight lesions in 91 patients with stenosis of the internal carotid artery were treated surgically. Eighty lesions received carotid endarterectomy (CEA), and 18 lesions received percutaneous transluminal angioplasty (PTA) with or without stent. Treatment with PTA-with-or-without-stent has been opted in cases of patient with such conditions as radiation-induced stenosis, re-stenosis after CEA, unfitness for general anesthesia, bilateral lesions both of which need to be reconstructed with in a short interval. We divided the patients into 4 groups according to their age; under 70-year-old group, 70-to-74-year old group, 75-to-79-year old group, and over 80-year-old group. Vasoreconstructive procedures were performed for 41 patients in the under 70-year-old group, for 31 in the 70-to-74-year old group, for 21 in the 75-to-79-year old group, and for 5 in the over 80-year-old group. The overall surgical morbidity rate was 2% (2 of 98 cases) and there was no mortality. Tweleve patients (12.2%) experienced transient neurological deficits. Two patients exhibited perioperative complications in gastrointestinal organs, but none of the patients experienced cardiac complications. Elderly patients tend to experience systemic complications such as gastrointestinal complications as well as transient neurological deficits, which appear as restlessness, possibly due to hyperperfusion syndrome. The perioperative complication rate in elderly patients (putting the patients of the 70-to-74-year old group, the 75-to-79-year old group, and the over 80-year-old group together) was, statistically, significantly higher than those in patients of under the 70-year-old group. However, when two treatment modalities, CEA and PTA-with-or-without stent, were compared, there was no significant difference in the perioperative complication rate. Careful patient selection and prudent perioperative management enabled us to perform vasoreconstructive surgery even for elderly patients with internal carotid artery stenosis in a relatively safe manner with an acceptable complication rate. Decision making in selecting treatment modality, CEA or PTA with or without stent, should not be based solely on aging.


Treatment of cytomegalovirus retinitis with intravitreal injection of liposome encapsulated ganciclovir in a patient with AIDS.
To study its safety and efficacy in treating cytomegalovirus (CMV) retinitis, an AIDS patient received an intravitreal injection of liposome encapsulated ganciclovir in the right eye. The left eye served as a control, receiving intravitreal free ganciclovir. The right eye showed no retinal haemorrhages or detachment; however, vision declined initially, stabilising later. Weekly examination showed neither progression of the CMV retinitis nor new lesions in the right eye. The left eye showed reactivation of old CMV retinitis. Liposome encapsulated ganciclovir reduced the number of intravitreal injections, stabilising CMV retinitis, and warrants further study.


Critical evaluation of platelet aggregation in whole human blood.
Platelet aggregation studies generally are performed in platelet-rich plasma (PRP) by the turbidometric method. The authors compared this technic with the recently introduced impedance aggregometry in PRP and whole blood (WB). In healthy controls there was a good correlation between the two technics when aggregation was induced by ADP or collagen. As compared with PRP, platelets in WB were more sensitive to the aggregating effect of thrombin, ristocetin, and arachidonic acid. Platelet sensitivity to prostacyclin was increased in WB. The anti-platelet effect of a single oral dose of aspirin could be detected for a longer period in WB than in PRP. Platelet aggregation tests in WB from patients with platelet dysfunctions showed the same response pattern to different aggregating agents as in PRP. In contrast to turbidometry, the impedance method in PRP and WB enabled registration of platelet aggregation in a dose-dependent fashion in a sample from a patient with severe hyperlipoproteinemia. It is concluded that platelet aggregation can be studied conveniently with the impedance method in the more physiologic medium of WB. Providing the same information as the well-established turbidometry, the time-sparing impedance method needs less citrated blood. Moreover, our results show an increased sensitivity of the WB system to some aggregating and anti-platelet agents.


The origin and identity of embryonic stem cells.
Embryonic stem (ES) cells are used extensively in biomedical research and as a model with which to study early mammalian development, but their exact origin has been subject to much debate. They are routinely derived from pre-implantation embryos, but it has been suggested that the cells that give rise to ES cells might arise from epiblast cells that are already predisposed to a primordial germ cell (PGC) fate, which then progress to ES cell status via the PGC lineage. Based on recent findings, we propose here that ES cells can be derived directly from early epiblast cells and that ES cells might arise via two different routes that are dictated by their culture conditions.


Code Pseudo Stroke - A Case of Hypokalaemic Periodic Paralysis Mimicking Stroke.
We present a case of thyrotoxic periodic paralysis (TPP) presenting with stroke symptoms as a harbinger of Grave's disease. A 61-year-old female presented with symptoms of abdominal pain and fatigue two weeks prior to admission and reported acute diarrhoea and unintentional weight loss. Investigation revealed thyrotoxicosis with undetectable thyroid stimulating hormone (TSH), elevated free T4 and elevated thyroid stimulating immunoglobulin (TSI). On the third day of admission, while undergoing physical therapy, code stroke was called on account of the onset of right-side predominant acute flaccid paralysis of upper and lower extremities, right-side facial droop, dysarthria and hyporeflexia bilaterally. The patient was alert and fully oriented with stable vitals with no increased labour in breathing at room air. An emergent head and neck CT, angiography, and magnetic resonance imaging (MRI) were negative. Serum potassium was 2.7 mmol/l, requiring prompt replacement. The patient's paralysis and dysarthria improved over the following three days with a complete reversal of symptoms following the correction of serum potassium. Thyrotoxic periodic paralysis can occur in association with any of the causes of hyperthyroidism. It is due to a significant intracellular shift of potassium, subsequently manifesting clinically with hypokalaemia and muscle paralysis. Hypokalaemic periodic paralysis is an acute flaccid paralysis secondary to hypokalaemia, triggered by hyperthyroidism, environmental elements such as a high carbohydrate intake or rest after intense exercise, and an underlying genetic component with mutations in the muscle ion channels.The pursuit of code stroke protocol in the in-patient setting must bear in mind reversible but potentially fatal and disabling alternative diagnoses such as stroke mimics including hypokalaemic periodic paralysis.Stroke mimics defined as acute onset neurological deficits that are not related to vascular cause are a true challenge, multiple scales have been created to distinguish them from an actual stroke, such as the Recognition of Stroke in the Emergency Room (ROSIER) scale.


Crystallization and preliminary X-ray characterization of arylamine N-acetyltransferase C (BanatC) from Bacillus anthracis.
The arylamine N-acetyltransferase (NAT) enzymes are xenobiotic metabolizing enzymes that have been found in a large range of eukaryotes and prokaryotes. These enzymes catalyse the acetylation of arylamine drugs and/or pollutants. Recently, a Bacillus anthracis NAT isoform (BanatC) has been cloned and shown to acetylate the sulfonamide antimicrobial sulfamethoxazole (SMX). Subsequently, it was shown that BanatC contributes to the resistance of this bacterium to SMX. Here, the crystallization and the X-ray characterization of BanatC (Y38F mutant) are reported. The crystals belong to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 53.70, c = 172.40 A, and diffract to 1.95 A resolution on a synchrotron source.


Receptor-mediated endocytosis of an extracellular steroid-binding protein (TeBG) in MCF-7 human breast cancer cells.
Previous studies suggested that an extracellular steroid-binding protein, testosterone-estradiol-binding globulin (TeBG), can enter a variety of cells. Experiments were conducted to determine whether uptake of TeBG occurs by nonspecific fluid phase endocytosis or via a specific receptor-mediated process. In human breast carcinoma cells (MCF-7) maintained on serum-free medium, exposure to radiolabeled TeBG resulted in cellular uptake, which reached a plateau by 6 h and could be inhibited 80% by competition with unlabeled TeBG. Uptake was temperature dependent with cell-associated radioactivity at 37 C being 1.6-fold higher than at 4 C. Subsequent exposure of cells to pronase resulted in release of the cell-associated TeBG by 88% and 44% at 4 C and 37 C, respectively. After transfer to media devoid of TeBG, approximately 35% of cell-associated radioactivity was release into the medium at 37 C; it was not possible to distinguish whether this was released from the cell surface or from inside the cell. Investigation of the localization of TeBG-gold complexes by electron microscopy revealed that TeBG first binds to the plasmalemma. Within 15 min label appears in receptosomes, which fuse to form multivesicular endosomes. By 1 h all label is observed in multivesicular endosomes and lysosomes, most of which are in the Golgi zone. Localization of the internalized radioactivity using classical cell fractionation techniques showed it appears in a symmetrical band exhibiting the same buoyant density as the lysosomal marker acid phosphatase. The observations reported here show that: 1) TeBG binds to MCF-7 cells; 2) some of the bound TeBG is taken up via a mechanism with all the characteristics of receptor-mediated endocytosis; and 3) within these cells TeBG is localized in endosomes and lysosomes.


Evaluation of real-time RT-PCR compared with conventional RT-PCR for detecting human metapneumovirus RNA from clinical specimens.
Human metapneumovirus (hMPV) is an etiologic agent of respiratory tract infections. In this study, we compared the sensitivity and specificity of real-time reverse transcription (RT)-polymerase chain reaction (PCR), conventional RT-PCR, and nested PCR in detecting hMPV genes. A total of 146 clinical specimens from 143 patients who showed acute respiratory tract infection symptoms were tested by real-time RT-PCR, conventional RT-PCR, and nested PCR targeting for the fusion gene. We detected hMPV RNA from 14 (9.6%) clinical specimens (real-time RT-PCR, 8; conventional RT-PCR, 5; and nested PCR, 13). When conventional RT-PCR was the reference standard, the sensitivity and specificity of real-time RT-PCR were 100 and 97.9%, respectively. When nested PCR was the standard, the sensitivity and specificity of real-time RT-PCR were 53.8 and 99.2%, respectively. Therefore, real-time RT-PCR was more sensitive than conventional RT-PCR but less so than nested PCR. Phylogenetic analysis showed that the real-time RT-PCR detected four genetic sublineages of hMPV. These results taken together indicate that real-time RT-PCR is an efficient method for detecting four genetic sublineages of hMPV from clinical specimens.


Pleiotropic effects and cholesterol-lowering therapy.
HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitors, or 'statins', have revolutionized the management of patients with atherosclerotic vascular disease. Following 2 large acute coronary syndrome trials, additional clinical benefit outside their effect of low-density lipoprotein (LDL) lowering was proposed. This concept was introduced following the observation of cardiovascular event rate reduction, only weeks after initiation of treatment and supposedly before the effect of LDL lowering could have influenced atheroma volume burden. Furthermore, there has been a substantial compilation of experimental data demonstrating beneficial effects of statins on inflammation, thrombosis, platelet aggregation, immunomodulation and endothelial function. These are hypothesized to occur via the interruption of the mevalonate pathway. However, the absolute benefit of these non-lipid-lowering effects, often referred to as 'pleiotropic effects', has been challenged by meta-analysis data. Anti-inflammatory actions have also been proposed to occur by the process of LDL lowering alone rather than due to a unique property of statins. Furthermore, some experimental data reports have shown evidence of pleiotropic effects in non-statin lipid-modifying agents. In this review article, we consolidate what is known so far and critically analyse the literature in order to highlight the outstanding issues.


Late morbidity in long-term survivors of childhood brain tumors: a nationwide registry-based study in Finland.
The population of long-term survivors of childhood brain tumors (BTs) is growing. The aim of our study was to evaluate late-appearing morbidity in BT survivors. Patients diagnosed with a BT at the age of 0-15 years between 1970 and 2004, and surviving at least 5 years, were identified from the Finnish Cancer Registry (n = 740). Their late new morbidity ≥ 5 years after cancer diagnosis was assessed using the Hospital Discharge Registry containing hospitalizations and outpatient visits in specialized health care settings. The morbidity of BT survivors was compared with that of the sibling cohort (n = 3615). The 5-year survivors had a significantly increased hazard ratio (HR) for endocrine diseases (HR, 14.7), psychiatric disorders (HR, 1.8), cognitive and developmental disorders (HR, 16.6), neurological diseases (HR, 9.8), disorders of vision and hearing (HR, 10.5), and diseases of the circulatory system (HR, 2.7) compared with the sibling cohort. The HRs for disorders of musculoskeletal system (HR, 1.4) and diseases of the kidney (HR, 2.1) were not significantly increased. Radiation treatment did not explain all of the excess morbidity. Female survivors had a higher risk for disorders of vision and hearing (P = .046). Age at diagnosis did not show an effect on HRs. The HRs for endocrine diseases and disorders of vision or hearing loss were highest for survivors treated in the 1980s or later. Pediatric BT survivors had significant neurocognitive consequences. This, together with the considerable risk for endocrine morbidity, will motivate us to organize systematic follow-up procedures for pediatric BT survivors.


Psychosocial predictors of supervisor-, peer-, subordinate-, and service-provider-targeted aggression.
The authors investigate whether known person predictors (trait anger, trait aggression) and situational predictors (perceived interpersonal mistreatment, perceived organizational sanctions against aggression) of supervisor-targeted aggression also predict employee's aggression toward other workplace targets, namely peers, subordinates, and customers' aggression toward service providers. The authors also investigate the moderating impact of situational factors on the relationship between person factors and aggression. Participants (N = 308) were asked whether they had a conflict with their supervisor, a subordinate, a work peer, and/or a service provider in the past 6 months. Different patterns of main and interaction effects emerged across the 4 targets, suggesting the importance of accounting for the target of aggression in workplace aggression research.


[Cryptorchidism: pathogenesis and histology].
An impairment of the hypothalamopituitarygonadal axis was found to be the main cause of undescended testicles in 80 per cent of patients. Basing himself on personal observation, the author develops the theory that the mechanism of testicular descent is the stimulation by GnRH of the pituitary gland to release gonadotrophins, which in turn stimulate the interstitial cells to secrete testosterone. High local testosterone concentration fosters the development of the differentiating portion of the mesonephric duct into the epididymis. The epididymis induces descent by pushing rather than pulling the testis into the scrotal position. Any interference with this mechanism during the embryonal life leads to cryptorchid testes, which, in the vast majority of cases are localized at the external inguinal ring. The earlier the scrotal position is reached, the greater the chances of subsequent fertility.


Mindfulness and the psychology of presence.
Mindfulness, as 'being in the present on purpose' has become of practical and theoretical interest to clinical psychologists. It is an element in several influential and effective treatment methods. Research concentrates on the effects of practice but neglects the experience of presence. A common requirement that the clinician also practice mindfulness leads to an interest in describing clinical presence. This stance has been of interest in psychoanalysis but not studied in mainstream clinical psychology. The description of that as a state of being and the similarity between mindfulness practice and the phenomenological method provide a link with a 6-year series of seminars given to psychotherapists by the philosopher Martin Heidegger. His demonstrations of the relation between experiential and scientific knowledge offer a way to make links between a reflective clinical psychology, mindfulness, and current developments in the understanding of consciousness.


Residues of pesticide co-formulants in lettuce and parsley: Identification of decline processes using field trials in different cropping systems.
Although co-formulants constitute a substantial portion of the total plant protection product (PPP) mass applied to crops, data on residue formation and the behaviour of these substances on plants are scarce. In an earlier study we demonstrated that co-formulants commonly used in PPPs can form considerable residues, i.e., in the low to medium mg/kg range, but normally decline rapidly within few days. In the field trial reported here, we aimed to identify the major decline processes of co-formulants. Residues of co-formulants were therefore monitored in parsley and lettuce grown in an open field as well as under foil tunnels equipped with either an overhead or a drip irrigation system. Dissipation of three anionic surfactants was markedly faster when crops (parsley and lettuce) were exposed to natural rainfall or irrigation from above compared to drip irrigation. In contrast, the decline of three volatile organic solvents was not affected by rain or irrigation, but was dependent on the crop, with much shorter half-lives in lettuce than in parsley. Furthermore, dilution through plant growth contributed significantly to the reduction of residues over time. In this work we substantiate earlier findings on the magnitude and dissipation of residues of anionic surfactants and solvents representing the most important co-formulant classes. The chosen experimental setup allowed differentiation between decline processes and we confirm that foliar wash-off is a major dissipation process for anionic surfactants. For volatile organic solvents, dissipation appears to depend on the properties not only of the substance but also of the plant (surface). © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Cephalometric craniofacial characteristics in patients with temporomandibular joint ankylosis.
The sequelae of temporomanibular joint (TMJ) ankylosis include limitation of jaw movement, interference of oral function and affects on the craniofacial growth. Analysis of the craniofacial form of TMJ ankylosis offers guidelines for managing this disease. Forty-five patients with intraarticular TMJ ankylosis were collected from the files at the Chang Gung Craniofacial Center. There were 21 male and 24 female patients, aged 3 to 47 years. Thirty-seven patients were unilaterally affected and eight had bilateral involvement. Patients were grouped according to gender and age. Both the medical history and onset of the disease were investigated in all patients. The pretreatment lateral cephalograms were used for analysis. The variables were compared with the Chinese norms with corresponding sex and age groups. The etiology included 48.9% facial trauma history, 17.8% traumatic delivery or birth injury, 15.6% middle ear or dental infection, 2.2% chronic arthritis and 15.6% unknown causes. The onset of mouth opening limitation was under 16 years of age. The average total mandibular length was less than the norm by 30 mm. Each patient presented with a mandible that had backward rotation with chin recession. Accentuated antegonial notch and inferiorly located condyle were observed on the affected side. The maxilla was shorter and the ANB was larger than the norm by 10 degrees but the overbite and overjet were within normal ranges. The facial growth was severely disturbed in terms of dimension, morphology and direction of growth in patients with TMJ ankylosis. Better management of mandibular fractures, good infection control and early treatment intervention are ways to reduce the influence on craniofacial growth.


The role of liver microenvironment in hepatic metastasis.
Metastasis is still poorly understood and thus further research must be conducted to provide insight into the driving factors. Novel research has revealed the significance of the microenvironment in the delegation of metastasis, expanding the field of cancer metastasis to cells and cell environments surrounding the migrated tumor cells. Research on hepatic metastasis is an ever-growing domain of this field, as several primary tumors can metastasize to the liver. The two features within the liver that promote metastasis-cellular and acellular-are found in the current interpretation of liver microenvironment. Novel findings of both are included in this review. Different hypotheses detailing the methods by which metastasis can occur must be included to understand the significance of the microenvironment, as well as a brief overview of the methods that can be used during research. This review aims to highlight the importance of liver microenvironment on the development or potential regression of hepatic metastasis through discussing both acellular and cellular components of liver microenvironment and their interaction with metastasis.


The spontaneous loss of calcitonin and somatostatin in thyroid C cells of a guinea pig.
A guinea pig showing spontaneous abnormalities in both calcitonin and somatostatin syntheses was found. Thyroid C cells revealed an almost complete absence of secretory granules immunoreactive to calcitonin and somatostatin. Instead, vesicular inclusions of various sizes, which exhibited positive immunoreactions for calcitonin and somatostatin, were found in some C cells. There were several C cells with enlarged and eosinophil cytoplasm, showing a hyaline degeneration. The number of C cells per thyroid and their distribution pattern were not different from those of normal controls. Follicular cells did not exhibit any detectable changes, whereas parathyroid glands were somewhat degenerated.


Observations on nonconverting phage, c-n71, obtained from a nontoxigenic strain of Clostridium botulinum type C.
A nontoxigenic mutant (C-N71) obtained from a toxigenic strain of Clostridium botulinum type C, Stockholm, with nitrosoguanidine treatment was found to be lysogenic by the lysis test. Although the filtrate of a passaged lysate of this nontoxigenic but lysogenic strain, C-N71, lysed cells of the nontoxigenic strain C-AO2 equally as well as the converting phage c-st obtained from the strain C-Stockholm, it did not convert C-AO2 to the toxigenic state. The lysis spectrum of this filtrate was the same as that of the c-st phage. The ability of the filtrate to lyse the indicator cells, C-AO2, was destroyed neither by trypsin nor DNase but was inactivated by heat treatment at 80 C for 10 min. This suggested that the agent which caused lysis was not boticin but probably a phage. An electron micrograph of the complete phage, c-n71, which was similar in morphology to that of the c-st phage was obtained from the filtrate of strain C-N71. Anti-c-n71 phage rabbit serum neutralized both the lytic and the converting activities of the c-st phage. These findings strongly suggest that the c-n71 phage is a mutant of the c-st phage which lacks the gene controlling production of botulinum type C toxin.


A Scalable Weight-Free Learning Algorithm for Regulatory Control of Cell Activity in Spiking Neuronal Networks.
Recent developments in neural stimulation and recording technologies are providing scientists with the ability of recording and controlling the activity of individual neurons in vitro or in vivo, with very high spatial and temporal resolution. Tools such as optogenetics, for example, are having a significant impact in the neuroscience field by delivering optical firing control with the precision and spatiotemporal resolution required for investigating information processing and plasticity in biological brains. While a number of training algorithms have been developed to date for spiking neural network (SNN) models of biological neuronal circuits, exiting methods rely on learning rules that adjust the synaptic strengths (or weights) directly, in order to obtain the desired network-level (or functional-level) performance. As such, they are not applicable to modifying plasticity in biological neuronal circuits, in which synaptic strengths only change as a result of pre- and post-synaptic neuron firings or biological mechanisms beyond our control. This paper presents a weight-free training algorithm that relies solely on adjusting the spatiotemporal delivery of neuron firings in order to optimize the network performance. The proposed weight-free algorithm does not require any knowledge of the SNN model or its plasticity mechanisms. As a result, this training approach is potentially realizable in vitro or in vivo via neural stimulation and recording technologies, such as optogenetics and multielectrode arrays, and could be utilized to control plasticity at multiple scales of biological neuronal circuits. The approach is demonstrated by training SNNs with hundreds of units to control a virtual insect navigating in an unknown environment.


[Spontaneous changes of ventricular ectopic beats in ischemic patients and subjects without other signs of cardiopathy].
Spontaneous variability of premature ventricular complexes was evaluated by 72 hour continuous ambulatory electrocardiographic monitoring in 38 patients. Nineteen patients had coronary artery disease and 19 subjects had no documented signs of cardiac disease. Using multifactorial analysis of variance we determined the minimal percent reduction of premature ventricular complexes frequency that could not be ascribed to spontaneous variability alone in both single subject and in groups of 10 and 19 individuals and analyzed the difference between the two groups of ischemic and healthy patients. Our results show that in each subject with or without coronary artery disease the minimal reduction of premature ventricular complexes is similar: 71% and 72% respectively. On the other hand when the two groups were compared the percentage of reduction was 49% in healthy subjects and 40% in patients with coronary artery disease. Spontaneous variability of premature ventricular complexes decreases as the period of monitoring lengthens; however the improvement obtained with longer electrocardiographic monitoring doesn't justify the prolongation of the examination beyond 24 hours.


Safety of disease-modifying drugs for multiple sclerosis in pregnancy: current challenges and future considerations for effective pharmacovigilance.
When contemplating a pregnancy, women treated for multiple sclerosis (MS) with a disease-modifying drug must decide to discontinue their medication before conception or risk exposing their unborn child to potential drug toxicity. Few studies exist as reference for patients and physicians, and of those available, the majority are less than ideal due to real-world constraints, ethical issues and methodological shortcomings. The authors provide a brief summary of existing animal and human data with current recommendations regarding the safety of IFN-β, glatiramer acetate, natalizumab, mitoxantrone, fingolimod and teriflunomide during pregnancy and lactation in women with MS. We also assess the quality, strengths and limitations of the existing studies including challenges with study design. The investigation of outcomes such as spontaneous abortion and congenital anomalies are highlighted with potential methodological improvements for future studies on drug safety in pregnancy suggested. The authors explore the pharmacokinetics and pharmacodynamics of the MS disease-modifying drugs for their possible mechanistic role in fetal harm and discuss the potential role of clinical trials. Future pharmacovigilance studies should continue to pursue multicenter collaboration with an emphasis on appropriate study design.


Proteomic identification of membrane-associated placental protein 4 (MP4) as perlecan and characterization of its placental expression in normal and pathologic pregnancies.
More than 50 human placental proteins were isolated and physico-chemically characterized in the 70-80s by Hans Bohn and co-workers. Many of these proteins turned to have important role in placental functions and diagnostic significance in pregnancy complications. Among these proteins was membrane-associated placental protein 4 (MP4), for which identity or function has not been identified yet. Our aim was to analyze the sequence and placental expression of this protein in normal and complicated pregnancies including miscarriage, preeclampsia and HELLP syndrome. Lyophilized MP4 protein and frozen healthy placental tissue were analyzed using HPLC-MS/MS. Placental tissue samples were obtained from women with elective termination of pregnancy (first trimester controls, n = 31), early pregnancy loss (EPL) (n = 13), early preeclampsia without HELLP syndrome (n = 7) and with HELLP syndrome (n = 8), late preeclampsia (n = 8), third trimester early controls (n = 5) and third trimester late controls (n = 9). Tissue microarrays were constructed from paraffin-embedded placentas (n = 81). Slides were immunostained with monoclonal perlecan antibody and evaluated using light microscopy and virtual microscopy. Perlecan was also analyzed for its expression in placentas from normal pregnancies using microarray data. Mass spectrometry-based proteomics of MP4 resulted in the identification of basement membrane-specific heparan sulfate proteoglycan core protein also known as perlecan. Immunohistochemistry showed cytoplasmic perlecan localization in syncytiotrophoblast and cytotrophoblasts of the villi. Perlecan immunoscore decreased with gestational age in the placenta. Perlecan immunoscores were higher in EPL compared to controls. Perlecan immunoscores were higher in early preeclampsia without and with HELLP syndrome and lower in late preeclampsia than in respective controls. Among patients with preeclampsia, placental perlecan expression positively correlated with maternal vascular malperfusion and negatively correlated with placental weight. Our findings suggest that an increased placental perlecan expression may be associated with hypoxic ischaemic injury of the placenta in miscarriages and in early preeclampsia with or without HELLP syndrome.


External Quality Assessment Scheme for reference laboratories - review of 8 years' experience.
We describe an External Quality Assessment Scheme (EQAS) intended for reference (calibration) laboratories in laboratory medicine and supervised by the Scientific Division of the International Federation of Clinical Chemistry and Laboratory Medicine and the responsible Committee on Traceability in Laboratory Medicine. The official EQAS website, RELA (www.dgkl-rfb.de:81), is open to interested parties. Information on all requirements for participation and results of surveys are published annually. As an additional feature, the identity of every participant in relation to the respective results is disclosed. The results of various groups of measurands (metabolites and substrates, enzymes, electrolytes, glycated hemoglobins, proteins, hormones, thyroid hormones, therapeutic drugs) are discussed in detail. The RELA system supports reference measurement laboratories preparing for accreditation according to ISO 17025 and ISO 15195. Participation in a scheme such as RELA is one of the requirements for listing of the services of a calibration laboratory by the Joint Committee on Traceability in Laboratory Medicine.


Secondary interactions in 4-bromo-N,N-dimethylanilinium bromide.
The crystal packing of the title compound, C(8)H(11)BrN(+).Br(-), involves three types of secondary interaction: a classical N-H.Br(-) hydrogen bond, a 'weak' but short C-H.Br(-) interaction (normalized H.Br distance of 2.66 A) and a cation-anion Br.Br contact of 3.6331 (4) A. The hydrogen bonds connect two cations and two anions to form rings of graph set R(4)(2)(14). The Br.Br contacts link these rings to form layers parallel to the bc plane.


The Shelf Life of a Safety Climate Assessment: How Long Until the Relationship with Safety-Critical Incidents Expires?
This study investigates safety climate as both a leading (climate → incident) and a lagging (incident → climate) indicator of safety-critical incidents. This study examines the "shelf life" of a safety climate assessment and its relationships with incidents, both past and future, by examining series of incident rates in order to determine when these predictive relationships expire. A survey was conducted at a large, multinational chemical manufacturing company, with 7,467 responses at 42 worksites in 12 countries linked to over 14,000 incident records during the 2 years prior and 2 years following the survey period. Regressions revealed that safety climate predicts incidents of varying levels of severity, but it predicts the most severe incidents over the shortest period of time. The same is true for incidents predicting safety climate, with more severe incidents having a shorter predictive window. For the most critical relationship (climate predicting more severe incidents), the ability of a safety climate assessment to predict incidents expires after 3 months. The choice of aggregation period in constructing incident rates is essential in understanding the safety climate-incident relationship. The common yearly count of incidents would make it seem that more severe incidents cannot be predicted by safety climate and also fails to show the strongest predictive effects of less severe incidents. This research is the first to examine assumptions regarding aggregation periods when constructing safety-related incident rates. Our work guides organizations in planning their survey program, recommending more frequent measurement of safety climate.


Site-directed mutagenesis of a plant viral satellite RNA changes its phenotype from ameliorative to necrogenic.
Comparison of the nucleotide sequences of cucumber mosaic virus (CMV) satellite RNAs, which induce necrosis on tomatoes, reveals a highly conserved region within their 3' halves. The sequence of WL1 satellite (WL1-sat) RNA, which attenuates CMV symptoms on tomatoes, differs from all necrogenic satellite RNAs at three nucleotide positions within this conserved region. These nucleotides were progressively mutated to determine what sequence is required for the induction of necrosis in tomatoes. Infectious transcripts from a cDNA clone of WL1-sat RNA, and its mutated derivatives, were assayed on tomato. Three of the four mutant satellite RNAs, in which two of the three nucleotides correspond to those present in necrogenic satellite RNAs, and the rest of the molecule corresponded to WL1-sat RNA, attenuated CMV symptoms on tomatoes, and were phenotypically identical to the parental WL1-sat RNA. One other mutant satellite RNA, in which all three mutated nucleotides corresponded to the sequence of necrogenic satellite RNAs and the rest of the molecule corresponded to WL1-sat RNA, induced a lethal necrosis on tomatoes. Necrosis was also observed when the same mutant satellite RNA was associated with a second CMV strain as helper virus. These results indicate that a single nucleotide change at any of the three nucleotides examined distinguishes necrogenic from nonnecrogenic satellite RNAs. This necrosis was similar to that induced by a naturally necrogenic satellite RNA. The various mutations did not modify the effect of WL1-sat RNA on the symptoms induced by CMV on tobacco or squash.


Suprapubic catheterization: challenges in changing catheters.
Suprapubic catheterization of the bladder is used as a short- or long-term alternative to urethral catheterization. As with any indwelling urinary catheter, correct insertion, care and removal are vitally important to minimize problems. A particular problem that affects suprapubic catheters is 'cuffing', which on its own or combined with encrustation can potentially cause a great deal of difficulty on removal or discomfort for the patient. This article discusses the causes of cuffing, and suggests using catheters with integral balloons to reduce the incidence of the problem.


Abnormal neuronal metabolism and storage in mucopolysaccharidosis type VI (Maroteaux-Lamy) disease.
Mucopolysaccharidosis (MPS) type VI, also known as Maroteaux-Lamy disease, is an inherited disorder of glycosaminoglycan catabolism caused by deficient activity of the lysosomal hydrolase, N-acetylgalactosamine 4-sulphatase (4S). A variety of prominent visceral and skeletal defects are characteristic, but primary neurological involvement has generally been considered absent. We report here that the feline model of MPS VI exhibits abnormal lysosomal storage in occasional neurones and glia distributed throughout the cerebral cortex. Abnormal lysosomal inclusions were pleiomorphic with some resembling zebra bodies and dense core inclusions typical of other MPS diseases or the membranous storage bodies characteristic of the gangliosidoses. Pyramidal neurones were shown to contain abnormal amounts of GM2 and GM3 gangliosides by immunocytochemical staining and unesterified cholesterol by histochemical (filipin) staining. Further, Golgi staining of pyramidal neurones revealed that some possessed ectopic axon hillock neurites and meganeurites similar to those described in Tay-Sachs and other neuronal storage diseases with ganglioside storage. Some animals evaluated in this study also received allogeneic bone marrow transplants, but no significant differences in neuronal storage were noted between treated and untreated individuals. These studies demonstrate that deficiency of 4S activity can lead to metabolic abnormalities in the neurones of central nervous system in cats, and that these changes may not be readily amenable to correction by bone marrow transplantation. Given the close pathological and biochemical similarities between feline and human MPS VI, it is conceivable that children with this disease have similar neuronal involvement.


The Timing of Stimulation and IL-2 Signaling Regulate Secondary CD8 T Cell Responses.
Memory CD8 T cells provide protection to immune hosts by eliminating pathogen-infected cells during re-infection. While parameters influencing the generation of primary (1°) CD8 T cells are well established, the factors controlling the development of secondary (2°) CD8 T cell responses remain largely unknown. Here, we address the mechanisms involved in the generation and development of 2° memory (M) CD8 T cells. We observed that the time at which 1° M CD8 T cells enter into immune response impacts their fate and differentiation into 2° M CD8 T cells. Late-entry of 1° M CD8 T cells into an immune response (relative to the onset of infection) not only facilitated the expression of transcription factors associated with memory formation in 2° effector CD8 T cells, but also influenced the ability of 2° M CD8 T cells to localize within the lymph nodes, produce IL-2, and undergo Ag-driven proliferation. The timing of stimulation of 1° M CD8 T cells also impacted the duration of expression of the high-affinity IL-2 receptor (CD25) on 2° effector CD8 T cells and their sensitivity to IL-2 signaling. Importantly, by blocking or enhancing IL-2 signaling in developing 2° CD8 T cells, we provide direct evidence for the role of IL-2 in controlling the differentiation of Ag-driven 2° CD8 T cell responses. Thus, our data suggest that the process of 1° M to 2° M CD8 T cell differentiation is not fixed and can be manipulated, a notion with relevance for the design of future prime-boost vaccination approaches.


Becoming a Mother During COVID-19 Pandemic: How to Protect Maternal Mental Health Against Stress Factors.
During the COVID-19 pandemic, there were an increasing prevalence of perinatal psychiatric symptoms, such as perinatal anxiety, depression, and post-traumatic stress disorders. This growth could be caused by a range of direct and indirect stress factors related to the virus and changes in health, social and economic organization. In this review, we explore the impact of COVID-19 pandemic on perinatal mental health, and propose a range of hypothesis about their etiological mechanisms. We suggest first that the fear of being infected or infected others (intrauterine transmission, passage of the virus from mother to baby during childbirth, infection through breast milk), and the uncertainty about the effect of the virus on the fetuses and infants may have played a key-role to weakening the mental health of mothers. We also highlight that public health policies such as lockdown, limiting prenatal visits, social distancing measures, and their many associated socio-economic consequences (unemployment, loss of income, and domestic violence) may have been an additional challenge for perinatal mental health. Ground on these hypotheses, we finally purpose some recommendations to protect perinatal mental health during a pandemic, including a range of specific support based on digital technologies (video consultations, phone applications) during pregnancy and the postpartum period.


Brainstem node for loss of consciousness due to GABA(A) receptor-active anesthetics.
The molecular agents that induce loss of consciousness during anesthesia are classically believed to act by binding to cognate transmembrane receptors widely distributed in the CNS and critically suppressing local processing and network connectivity. However, previous work has shown that microinjection of anesthetics into a localized region of the brainstem mesopontine tegmentum (MPTA) rapidly and reversibly induces anesthesia in the absence of global spread. This implies that functional extinction is determined by neural pathways rather than vascular distribution of the anesthetic agent. But does clinical (systemic-induced) anesthesia employ MPTA-linked circuitry? Here we show that cell-selective lesioning of the MPTA in rats does not, in itself, induce anesthesia or coma. However, it increases the systemic dose of pentobarbital required to induce anesthesia, in a manner proportional to the extent of the lesion. Such lesions also affect emergence, extending the duration of anesthesia. Off-target and sham lesions were ineffective. Combined with the prior microinjection data, we conclude that drug delivery to the MPTA is sufficient to induce loss-of-consciousness and that neurons in this locus are necessary for anesthetic induction at clinically relevant doses. Together, the results support an architecture for anesthesia with the MPTA serving as a key node in an endogenous network of dedicated pathways that switch between wake and unconsciousness. As such, the MPTA might also play a role in syncope, concussion and sleep.


Angiogenic effect of platelet-rich plasma combined with gelatin hydrogel granules injected into murine subcutis.
Platelet-rich plasma (PRP), which contains highly concentrated platelets, is produced by centrifuging whole blood. It is a safe and readily available source of a wide range of growth factors necessary for angiogenesis. Gelatin hydrogel granules have been designed and prepared for the controlled release of many growth factors. The angiogenic effect of human PRP was examined in vitro, and the effect of its subcutaneous injection with gelatin hydrogel granules into murine subcutis was evaluated. Human PRP was prepared using a double-spin method. The concentration of growth factors and the platelet count were examined in PRP and in vitro, and the angiogenic activity of human umbilical vein endothelial cells (HUVECs) in co-culture with human dermal fibroblast cells (NHDFs) in the presence and absence of PRP was evaluated. Then, in vivo, PRP, either free or with gelatin hydrogel granules, was injected subcutaneously into tiebacks on mice. Using a microscope and Kurabo angiogenesis image analyser software, the area containing newly formed capillaries was evaluated histologically and the microvascular network score was calculated. PRP was shown to contain high concentrations of PDGF, VEGF and TGFβ and had an angiogenic effect on the co-culture system. PRP with gelatin hydrogel granules significantly enlarged the area containing newly formed capillaries and promoted the microvascular network in murine subcutaneous tissue. PRP encapsulated in gelatin hydrogel microspheres shows promise for enhancing angiogenic effects in murine subcutis and could represent a potential therapeutic combination for the treatment of ischaemic disorders. Copyright © 2015 John Wiley & Sons, Ltd.


Two new species of Fomitiporia (Hymenochaetales, Basidiomycota) from Tibet, southwest China.
Two new species of Fomitiporia growing on Hippophae trees, F. norbulingka and F. subhippophaëicola, are described from southwest China based on morphological characters and phylogenetic analysis. Fomitiporia norbulingka is characterized by pileate basidiomata, mostly angular pores (6-9 per mm), slightly thick-walled generative hyphae, subglobose to globose basidiospores (6.5-7 × 5.5-7 μm), and absence of cystidioles. Fomitiporia subhippophaëicola is diagnostic by effused-reflexed to pileate basidiomata, angular pores (8-10 per mm), thick-walled generative hyphae, subglobose to obovoid basidiospores (6-8 × 5.5-7 μm), and presence of ventricose to fusoid cystidioles. Phylogenetic analysis inferred from combined sequences including the nuc rDNA ITS1-5.8-ITS2 region, 28S rDNA D1-D2 domains, partial sequences of translation elongation factor 1-α, and RNA polymerase II second largest subunits genes indicated that F. norbulingka and F. subhippophaëicola represent two new lineages which group together with F. hippophaëicola.


Root endophytic bacterial and fungal communities in a natural hot desert are differentially regulated in dry and wet seasons by stochastic processes and functional traits.
Dryland ecosystems experience seasonal cycles of severe drought and moderate precipitation. Desert plants may develop symbiotic relationships with root endophytic microbes to survive under the repeated wet and extremely dry conditions. Although community coalescence has been found in many systems, the colonization by functional microbes and its relationship to seasonal transitions in arid regions are not well understood. Here we examined root endophytic microbial taxa, and their traits in relation to their root colonization, during the dry and wet seasons in a hot desert of the southwestern United States. We used high-throughput DNA sequencing of 16S rRNA and internal transcribed spacer gene profiling of five desert shrubs, and analyzed the seasonal change in endophytic microbial lineages. Goodness of fit to the neutral community model in relationship to microbial traits was evaluated. In summer, Actinobacteria and Bacteroidia increased, although this was not genus-specific. For fungi, Glomeraceae selectively increased in summer. In winter, Gram-negative bacterial genera, including those capable of nitrogen fixation and plant growth promotion, increased. Neutral model analysis revealed a strong stochastic influence on endophytic bacteria but a weak effect for fungi, especially in summer. The taxa with higher frequency than that predicted by neutral model shared environmental adaptability and symbiotic traits, whereas the frequency of pathogenic fungi was at or under the predicted value. These results suggest that community assembly of bacteria and fungi is regulated differently. The bacterial community was affected by stochastic and deterministic processes via bacterial response to drought (response trait), beneficial effect on plants (effect trait), and likely stable mutualistic interactions with plants suggested by the frequency of nodule bacteria. For fungi, mycorrhizal fungi were selected by plants in summer. The regulation of beneficial microbes by plants in both dry and wet seasons suggests the presence of plant-soil positive feedback in this natural desert ecosystem.


Litomosoides yutajensis n. sp., first record of this filarial genus in a mormoopid bat.
Twenty-five bats were trapped in Yutaje (Amazonas, Venezuela) and examined for Litomosoides (Filarioidea: Onchocercidae). Of the nine recovered bat species, only Pteronotus parnelli was infected; it is a cave-dwelling species belonging to a family, Mormoopidae, which has not previously been included in the host range of the genus. The new species, L. yutajensis n. sp., has two median cephalic bosses covered with rugosities and differs from the 15 recognized species and subspecies from bats in several characters. Alike L. molossi Esslinger, 1973, L. chandleri Esslinger, 1973 and L. chitwoodi Bain, Guerrero, Rodriguez 2003, the new species has cuticular lateral bosses on the body. Eight of 10 P. parnelli were microfilaraemic, but only three had adult worms, showing that microfilariae survive longer than adults, which could lengthen the period of transmission. No infective larvae were detected in the following macronyssid mites: 58 Ornithonyssus bacoti, Ornithonyssinae, experimentally fed on microfilaraemic bats and dissected 15 days later, and a few Radfordiella sp., Macronyssinae, recovered from P. parnelli.


Single germline VH and V kappa genes encode predominating antibody variable regions elicited in strain A mice by immunization with p-azophenylarsonate.
We have cloned and sequenced the predominant germline V kappa gene segment expressed by B cells of strain A origin that synthesize antibodies with specificity for Ars. In hybridomas synthesizing anti-Ars antibodies, this V kappa gene segment (V kappa IdCR) has been found exclusively associated with the J kappa 1 gene segment without exhibiting junctional sequence variation. Sequence comparisons of the germline V kappa IdCR gene with expressed derivatives reveals that the latter frequently contain somatically introduced amino acid replacements. Taken together with results of previous structural analyses, these results show that the predominant population of IdCR+ V regions elicited in the secondary immune response is encoded by one or two combinations of V gene segments, has little junctional diversity, and is extensively diversified by somatic mutation in both heavy and light chains.


Effect of Nanoalumina on Sex Hormones and Fetuses in Pregnant Rats.
This study aimed at investigating the effect of nanoalumina on sex hormones, and fetuses in pregnant rats. In this study, sixty-four pregnant rats were divided into eight groups. The control and the injection-control group received normal food and water, and 0.5 ml of distilled water, respectively. Treatment groups were treated with 25, 50, 100, 250, 500, and 1000µg/ml concentrations of Nanoalumina from the 7th day until the 18th day of pregnancy. On the 18th day, the rats were investigated in terms of their hormone levels. We evaluated the number of healthy and aborted offspring, as well as fetus size. Nanoalumina caused an increase in progesterone hormones at the concentrations of 250, and 500µg/ml, and a significant reduction in estrogen hormone and aborted fetuses at the concentrations of 250 and 500µg/ml (p<0.05). The largest and smallest size of fetuses were observed in 500µg/ml and 1000µg/ml, respectively. The highest number of aborted fetuses was observed in the group treated with the 500µg/ml concentration. There was no aborted fetuses with 25, 50,100, control, and injection-control groups. Due to nanoalumina toxicity, it must be used with caution.


Discovery of a New Antibiotic Demethoxytetronasin Using a Dual-Sided Agar Plate Assay (DAPA).
For over a century, researchers have cultured microorganisms together on solid support─typically agar─in order to observe growth inhibition via antibiotic production. These simple bioassays have been critical to both academic researchers that study antibiotic production in microorganisms and to the pharmaceutical industry's global effort to discover drugs. Despite the utility of agar assays to researchers around the globe, several limitations have prevented their widespread adoption in advanced high-throughput compound discovery and dereplication campaigns. To address a list of specific shortcomings, we developed the dual-sided agar plate assay (DAPA), which exists in a 96-well plate format, allows microorganisms to compete through opposing sides of a solid support in individual wells, is amenable to high-throughput screening and automation, is reusable, and is low-cost. Herein, we validate the use of DAPA as a tool for drug discovery and show its utility to discover new antibiotic natural products. From the screening of 217 bacterial isolates on multiple nutrient media against 3 pathogens, 55 hits were observed, 9 known antibiotics were dereplicated directly from agar plugs, and a new antibiotic, demethoxytetronasin (1), was isolated from a Streptomyces sp. These results demonstrate that DAPA is an effective, accessible, and low-cost tool to screen, dereplicate, and prioritize bacteria directly from solid support in the front end of antibiotic discovery pipelines.


Regional intelligence and suicide rate: new data for Australia and a synthesis of research.
Previous research has shown for the most part positive correlations between intelligence and suicide prevalence on the national level. However, this study found proxies for regional intelligence in Australia (international average domain scores from the PISA 2000 study) to be significantly negatively correlated with the total, male, and female suicide rates of the different administrative divisions of Australia, and this finding was independent of regional wealth. A research synthesis of the current results and those from similar studies of other countries (positive correlations for Austria, Belarus, The British Isles, Denmark, and The Netherlands; inconclusive findings for France, Germany, and the USA) was conducted. This synthesis of research findings showed that positive ecological correlations of intelligence with suicide rate were more likely observed for nations with higher suicide rates and poorer general living conditions, whereas there was no relation with national IQ.


A Novel PTCH1 Frameshift Mutation Leading to Nevoid Basal Cell Carcinoma Syndrome.
Nevoid basal cell carcinoma syndrome (NBCCS), also known as Gorlin syndrome, is a rare multisystemic autosomal dominant disorder typically presenting with cutaneous basal cell carcinomas, multiple keratocysts, and skeletal anomalies. NBCCS is caused by heterozygous mutations in the PTCH1 gene in chromosome 9q22, in the PTCH2 gene in 1p34, or the SUFU gene in 10q24.32. Here, we report on an 18-month-old boy presenting with medulloblastoma, frontal bossing, and multiple skeletal anomalies and his father who has basal cell carcinomas, palmar pits, macrocephaly, bifid ribs, calcification of falx cerebri, and a history of surgery for odontogenic keratocyst. These clinical findings were compatible with the diagnosis of NBCCS, and a novel mutation, c.1249delC; p.Gln417Lysfs*15, was found in PTCH1 causing a premature stop codon.


Diazepam fails to alter anxiety-like responses but affects motor function in a white-black test paradigm in larval zebrafish (Danio rerio).
The growing popularity of zebrafish in psychopharmacology and behavioral brain research is partly due to the practicality and simplicity of drug administration in this species. Several drugs may be administered to zebrafish by immersing the fish in the drug solution. Furthermore, numerous drugs developed for mammals, including humans, have been found to show a similar effect profile in the zebrafish. Thus, the zebrafish has been suggested as a potentially useful animal screening tool. Despite decades of drug development, anxiety still represents a major unmet medical need, and the search for anxiolytic compounds is continuing. The zebrafish has been proposed for high throughput screens for anxiolytic compounds, and the effects of anxiolytic compounds on the behavior of zebrafish have started to be explored. Diazepam (Valium®) is a frequently prescribed human anxiolytic, a GABAA receptor agonist, has also started to be tested in zebrafish, but with occasional contradicting results. Here, we investigate the effects of diazepam in larval (6-day post-fertilization old) zebrafish in a black-white preference paradigm. We found significant white preference and thigmotaxis (edge preference) in our control fish, anxiety-like responses that habituated over time. However, unexpectedly, we observed no anxiolytic effects of diazepam on these behaviors, and only detected significant motor activity reducing effect of the drug. We discuss the complex interpretation of light/dark tests in zebrafish, and also speculate about the possibility of differential GABAergic mechanisms that diazepam affects in larval vs adult zebrafish.


Ectoparasitic copepod infestation on a wild population of Neotropical catfish Sciades herzbergii Bloch, 1794: Histological evidences of lesions on host.
We analyzed the infestation of the attached copepod species Lepeophtheirus sp on a wild population of Sciades herzbergii. The infestation and occurrence of attached copepods were related to body size, maturity and sex of host and the presence of lesions on fish skin were described. In 61 fish specimens (37 males and 24 females), total of 218 ectoparasitic copepods, including 204 mature and 14 immature stages were found. Copepods were attached to different regions of fish body without any regular pattern. The prevalence of infestation was 80.3% and intensity between 1 and 15 copepods/fish. No significant differences were found between sex or maturity and the presence of attached Lepeophtheirus sp. However, a contingency table including both sex and maturity status, and the total number of attached copepod per combined category showed a significant association. A positive correlation was found between body length of fish and intensity of infestation. Similarly, when comparing the infested fish according to group size, we found more copepods on larger individual. Attached copepods were associated with the presence of lesions visible to the naked eye. Histological analyses showed changes in cell architecture when sections of copepod-free tissues and attached copepods were compared.


Noncovalent Self-Assembly of Single-Layer MoS2 Nanosheets and Zinc Porphyrin into Stable SURMOF Nanohybrids with Multimodal Photocatalytic Properties.
A noncovalent integration of nanosheets of molybdenum disulfide (MoS2) and the zinc porphyrin complex Zn(II) 5,10,15,20-tetrakis(4-carboxyphenyl)porphine (ZnTCPP) through coordination bonding with metal clusters of zinc acetate (Zn[OAc]2) was applied for synthesis of stable hybrid nanomaterial avoiding surface prefunctionalization. The X-ray powder diffraction in combination with the BET nitrogen adsorption method confirms formation of a ZnTCPP-based surface-attached metal-organic framework (SURMOF) with micropores of 1.63 nm on the MoS2 nanosheets. Fluorescence spectroscopy confirmed Forster resonance energy transfer (FRET) between MoS2 and ZnTCPP without contact quenching. Fluorescent trapping with terephthalic acid for hydroxyl radicals and Sensor Green for singlet oxygen was applied for studying the pathways of photodegradation of model organic pollutant 1,5-dihydroxynaphthalene (DHN) in the presence of SURMOF/MoS2. Visible light initiates sensitization through the excitation of ZnTCPP generating singlet oxygen, whereas UV-light promotes either aerobic FRET-mediated "Z scheme" or anaerobic "Type II heterojunction" mechanisms. Owing to its multimodal photochemistry, the SURMOF/MoS2 hybrid showed comparatively high photocatalytic activity in UV-assisted degradation of DHN (keffUV = 4.0 × 10-2 min-1) as well as the antibacterial activity confirmed by E. coli survival test under visible light. Noncovalent self-assembly utilizing coordination bonding in SURMOFs as supramolecular adhesive to avoid surface premodification provides a basis for new types of multicomponent nanosystems with switchable functionalities by combining different 2D materials and chromophores in one hybrid structure.


Engineering phenylalanine ammonia lyase to limit feedback inhibition by cinnamate and enhance biotransformation.
Phenylalanine ammonia-lyase (PAL) is a crucial enzyme for various biotechnology applications, such as producing phenols, antioxidants, and nutraceuticals. However, feedback inhibition from its product, cinnamic acid, limits its forward reaction rate. Therefore, this study aims to address the feedback inhibition in PAL using enzyme engineering strategies. Random and site-directed mutagenesis approaches were utilized to screen mutant enzymes with ameliorated tolerance against cinnamic acid. A thermotolerant and cinnamate-tolerant mutant was rationally identified using a high throughput screening method and subsequent biochemical characterization. We evaluated cinnamate affinity among the seven rationally selected mutations, and the T102E mutation was identified as the most promising mutant. This mutant showed a six-fold reduction in the affinity of PAL for cinnamic acid and a two-fold increase in operational stability compared with native PAL. Furthermore, the enzyme was immobilized on carbon nanotubes to increase its robustness and reusability. The immobilized mutant PAL showed greater efficiency in the deamination of phenylalanine present in protein hydrolysate than its free form. The rationale behind the enhancement of cinnamate tolerance was validated using molecular dynamic simulations. Overall, the knowledge of the sequence-function relationship of PAL was applied to drive enzyme engineering to develop highly tolerant PAL.