Rapid identification of Campylobacter species using oligonucleotide probes to 16S ribosomal RNA.
A comparison of the 16S ribosomal RNA sequences from various Campylobacter species was used to identify unique sequences which distinguish pathogenically significant campylobacters from other members of the genus. Oligonucleotides complementary to these sequences were synthesized, and under stringent conditions hybridization reactions using these probes and total RNA as the target nucleic acid displayed the desired species specificity. A simple, rapid lysis procedure was developed that allowed the specific detection of C. jejuni or C. coli from as few as 10(6) bacterial cells in less than eight hours. This method provides immediate advantages for the unequivocal identification of Campylobacter species in the microbiology laboratory, and demonstrates the potential for the use of these probes in a rapid diagnostic test of clinical samples for Campylobacter infection.


Young adult's own and parental social characteristics predict injury morbidity: a register-based follow-up of 135,000 men and women.
Sociodemographic differences in injury mortality are well-established, but population-level studies on social patterns of injury morbidity remain few in numbers, particularly among young adults. Yet injuries are the leading cause of mortality, morbidity and disability among young people. Studies among children have shown steep social gradients in severe injuries, but less is known on the social patterning of injuries in late adolescence and early adulthood, when young people are in the process of becoming independent adults. This study examines how young adults' current living arrangements, education, main economic activity, and parental social background are associated with hospital-treated injuries in late adolescence and early adulthood. The study uses prospective, individual-level data gathered from several administrative sources. From a representative 11% sample of the total Finnish population, we included young people between ages 17-29 years during the follow-up (N = 134,938). We used incidence rates and Cox proportional hazards models to study hospital-treated injuries and poisonings in 1998-2008. Higher rates of injury were found among young adults living alone, single mothers, the lower educated and the non-employed, as well as those with lower parental social background, experience of childhood family changes or living with a single parent, and those who had left the parental home at a young age. Injury risks were consistently higher among young adults with lower education, but current living arrangements and main economic activity showed some age-related nuances in the associations: both earlier and later than average transitions in education, employment, and family formation associated with increased injury risks. The social differentials were strongest in poisonings, intentional self-harm, and assaults, but social patterns were also found in falls, traffic-related injuries and other unintentional injuries, underlining the existence of multiple distinct mechanisms and pathways behind the differentials. The transition to adulthood is a life period of heightened risk of injury, during which both parental social background and the young people's own social position are important determinants of serious injuries that require inpatient care.


Food insecurity is associated with glycemic markers, and socioeconomic status and low-cost diets are associated with lipid metabolism in Mexican mothers.
The association between socioeconomic status (SES) and chronic disease has recently become more evident in middle- and low-income countries. We hypothesized that poor socioeconomic conditions, such as food insecurity, low educational level, or low SES, may restrict access to a healthy diet and may be associated with cardiometabolic risk independently of body fat. This study examined the relation between socioeconomic indicators, body fat, and cardiometabolic disease risk markers in a random sample of mothers living in Queretaro, Mexico. Young and middle-aged mothers (n = 321) answered validated questionnaires to determine SES, food insecurity, and educational level and a semiquantitative food frequency questionnaire to determine dietary patterns and the cost of individual diet. Clinical measurements included anthropometry, blood pressure, lipids profile, glucose, and insulin. Obesity was present in 29% of the participants. Women with moderate food insecurity had higher waist circumference, glucose, insulin, and homeostasis model assessment of insulin resistance than women with food security. High triglyceride concentration and lower levels of high-density lipoprotein and low-density lipoprotein cholesterol were associated with lower SES and lower educational level. Women who consumed a lower carbohydrate diet had higher SES, higher education, and better cardiovascular risk markers. The higher carbohydrate diet profile was the least expensive diet. There was an inverse association between the cost and energy-density of foods. In conclusion, food insecurity was associated with glycemic control markers, and lower SES and education were related to a low-cost, higher carbohydrate diet and to a greater cardiovascular risk. The influence of the social environment on obesity and cardiovascular diseases needs to be further explored.


Enantiomeric ratio of shikonin derivatives as a possible key for the determination of the origin of lithospermi radix.
An HPLC method was developed to resolve the enantiomers of shikonin derivatives of the Lithospermi Radix. The optimum mobile phase on a Chiracel AD column was 5% isopropanol in n-hexane with flow rate of 1 ml/min. Establishment of this method made possible to determine the ratios of shikonin/acetylshikonin or alkanin/acetylalkanin in the same root. The correlation of the ratios of these substance pairs appeared characteristic for the country where they were originated from. All of the Korean species showed significantly higher ratios of shikonin/acetylshikonin and alkanin/acetylalkanin than the Chinese ones. This method would be useful to determine the origin of Lithospermi Radix.


Autologous and allogeneic stem cell transplantation in multiple myeloma.
Multiple myeloma is still an incurable disease. The standard conventional chemotherapy comprises melphalan and prednisone (MP). Combination chemotherapy regimens could not improve the median survival of 36 months observed with MP. In the French IFM90 study, HD therapy with TBI plus melphalan 140 mg/m2 was shown to prolong overall survival and progression-free survival compared to conventional treatment. Nonetheless, most patients eventually succumb due to disease progression. Allogeneic transplantation may induce long-term remissions and even cure, but is hampered by a high transplantation-related mortality (TRM). Currently, efforts are made to reduce this TRM and to evaluate the graft-versus-myeloma effect.


The association of ultraviolet radiation-B (305 nm), season of diagnosis, and latitude on the survival outcome of prostate cancer in the high UV environment of Australia.
Positive associations between sun exposure and cancer survival have been observed in regions of high latitudes, where ambient solar ultraviolet (SUV) radiation is generally low. We examined the effects of ambient ultraviolet-B radiation (UVB) at time of diagnosis, season of diagnosis and latitude of residence on survival outcome from prostate cancer. Regression models for relative survival were used to estimate relative excess risks (RER) of death after diagnosis of prostate cancer from cancer registries in Eastern Australia (Queensland, New South Wales, Victoria and Tasmania). Relative excess risks was increased with diagnosis in summer (RER = 1.20; 95 % CI 1.14-1.26) relative to winter, high ambient UVB at the time of diagnosis (>60 mW/m(2); RER = 1.10; 95 % CI 1.05-1.15) relative to low SUV (<30 mW/m(2)), and with residence in high latitudes (35°S-43°S; RER = 1.20; 95 % CI 1.14-1.26) relative to low latitudes (9°S-29.9°S). RER was highest for summer diagnosis in all three latitude bands, after adjusting for age, follow-up period, and socioeconomic status. The contradictory outcome from season and latitude suggests that their use as surrogates for UV warrants validation. Our data suggest that high ambient solar ultraviolet radiation at the time of diagnosis of prostate cancer increases the risk of dying from this cancer.


dsRNA binding properties of RDE-4 and TRBP reflect their distinct roles in RNAi.
Double-stranded RNA (dsRNA)-binding proteins facilitate Dicer functions in RNA interference. Caenorhabditis elegans RDE-4 facilitates cleavage of long dsRNA to small interfering RNA (siRNA), while human trans-activation response RNA-binding protein (TRBP) functions downstream to pass siRNA to the RNA-induced silencing complex. We show that these distinct in vivo roles are reflected in in vitro binding properties. RDE-4 preferentially binds long dsRNA, while TRBP binds siRNA with an affinity that is independent of dsRNA length. These properties are mechanistically based on the fact that RDE-4 binds cooperatively, via contributions from multiple domains, while TRBP binds noncooperatively. Our studies offer a paradigm for how dsRNA-binding proteins, which are not sequence specific, discern dsRNA length. Additionally, analyses of the ability of RDE-4 deletion constructs and RDE-4/TRBP chimeras to reconstitute Dicer activity suggest RDE-4 promotes activity using its dsRNA-binding motif 2 to bind dsRNA, its linker region to interact with Dicer, and its C-terminus for Dicer activation.


Development of an in vitro coculture of primary sensitive pig neurons and keratinocytes for the study of cutaneous neurogenic inflammation.
Cutaneous neurogenic inflammation (CNI) is often associated with skin disorders. Activated sensory neurons secrete neuropeptides, such as substance P (SP), which initiate or aggravate inflammation in the skin. The discovery of new molecules acting on these neurons is hampered by the difficulty of reproducing the interactions between nerve endings and skin in vitro. We developed an in vitro model based on the coculture of porcine primary keratinocytes and sensory neurons, which mimics skin innervation. To test the relevance of this model, we compared the effects of different substances on CNI by measuring SP secretion in vitro using a sensitive enzyme immunoassay. Collectively, our results indicate that the use of porcine cells could be very useful to perform an in vitro model of CNI. By adding capsaicin, which induces the secretion of SP by neurons, to the culture, we show that our model mimics CNI in vitro, allowing us to screen for molecules that inhibit this inflammatory response. Such a model can be used to test the effects of different substances on CNI and may be useful for dermatological or cosmetic applications. Based on our screen, we found that extracts of Laminaria digitata and Vernonia sublutea inhibit CNI.


[Radiofrequency treatment of an osteoid osteoma of the wrist: report of 2 cases].
Two cases of osteoid osteomas of the lunate and ulnar styloid process are described, treated by thermocoagulation. Follow-up was done clinically and by computed tomography (scanner) at one year, and clinically at three years. A quick and complete resolution of pain occurred in both cases, with bony sclerosis around the treated lesion on computed tomography, with no evidence of recurrence. A mild reduction in grip strength without functional impairment was noted in both cases, possibly due to a stronger controlateral dominant hand, and to a previous failed surgical excision in one case. No damage to neurovascular, musculotendinous or cutaneous structures occurred. Thermocoagulation of osteoid osteomas seems to be an efficient and safe technique even in critical and superficial anatomical regions such as the wrist.


Sequencing batch airlift reactors (SBAR): a suitable technology for treatment and valorization of mineral oil wastewaters towards lipids production.
Produced water (PW) and spent oil-based wastewaters are some of the largest mineral oil wastewaters produced. Due to the high toxicity of hydrocarbons, several countries set stringent discharge limits and its treatment is compulsory before discharge. In this work, biological treatment of mineral oil wastewaters coupled with the production of bacterial lipids is demonstrated in sequential batch airlift reactors (SBAR). Two SBAR (2 L working volume) were used for treatment of PW and lubricant-based wastewater (LW), inoculated with Alcanivorax borkumensis SK2 (SBARAb+PW) and Rhodococcus opacus B4 (SBARR.o+LW), respectively. A total petroleum hydrocarbon removal (TPH) efficiency up to 96% and 80% were achieved for SBARAb+PW and SBARR.o+LW, respectively. Intracellular lipids production in SBARAb+PW increased when lower TPH/N ratios and higher feast stage duration were applied (up to 0.74 g g-1 cell dry weight (CDW)), whereas in SBARR.o+LW higher lipids production was observed for higher TPH/N ratios (0.94 g g-1 in CDW). Triacylglycerols (TAG) were the main intracellular lipid accumulated in both SBARAb+PW and SBARR.o+LW operations, while wax ester (WE) production was only observed extracellularly in the SBARAb+PW.


Blood glucose regulation using an open-loop insulin delivery system in pancreatectomized dogs given glucose infusions. I. Portal square waves.
This study characterizes the glycaemic and insulin responses of a group of 5 anaesthetized dogs to a portal glucose infusion of 10 mg/kg/min before and after pancreatectomy. Insulin was administered intraportally to the pancreatectomized dogs according to a simple preprogrammed waveform composed of a constant basal rate of 0.35 +/- 0.02 mU/kg/min which was increased to 2.00 mU/kg/min at the time of the 60 minute glucose challenge. When this square waveform was applied the glycaemic response was similar to that seen in the normal controls in the baseline and challenge periods. Blood glucose concentration differed significantly (p less than 0.05) only from 20 to 100 minutes after the end of the challenge when it was higher by 20 +/- 1 mg/dl. Insulin levels were not significantly different from controls. It may be concluded that normoglycaemia and normoinsulinaemia can be maintained by a simple constant rate of portal insulin delivery while the blood glucose response to a glucose infusion can be ostensibly normalized without hyperinsulinaemia simply by enhancing insulin delivery during the challenge. The feasibility of this approach implies that with further development of the preprogrammed waveforms and with a greater understanding of their characteristics portable insulin delivery systems may be realized which accomodate more physiological challenges. The portal route for insulin delivery may however be necessary if peripheral hyperinsulinism is inappropriate.


In Vivo MR Imaging of Dual MRI Reporter Genes and Deltex-1 Gene-modified Human Mesenchymal Stem Cells in the Treatment of Closed Penile Fracture.
The purpose of this study was to investigate the feasibility of dual magnetic resonance imaging (MRI) reporter genes, including ferritin heavy subunit (Fth) and transferrin receptor (TfR), which provide sufficient MRI contrast for in vivo MRI tracking, and the Deltex-1 (DTX1) gene, which promotes human mesenchymal stem cell (hMSC) differentiation to smooth muscle cells (SMCs), to treat closed penile fracture (CPF). Multi-gene co-expressing hMSCs were generated. The expression of mRNA and proteins was assessed, and the original biological properties of hMSCs were determined and compared. The intracellular uptake of iron was evaluated, and the ability to differentiate into SMCs was detected. Fifty rabbits with CPF were randomly transplanted with PBS, hMSCs, Fth-TfR-hMSCs, DTX1-hMSCs, and Fth-TfR-DTX1-hMSCs. In vivo MRI was performed to detect the distribution and migration of the grafted cells and healing progress of CPF, and the results were correlated with histology. The mRNA and proteins of the multi-gene were highly expressed. The transgenes could not influence the original biological properties of hMSCs. The dual MRI reporter genes increased the iron accumulation capacity, and the DTX1 gene promoted hMSC differentiation into SMCs. The distribution and migration of the dual MRI reporter gene-modified hMSCs, and the healing state of CPF could be obviously detected by MRI and confirmed by histology. The dual MRI reporter genes could provide sufficient MRI contrast, and the distribution and migration of MSCs could be detected in vivo. The DTX1 gene can promote MSC differentiation into SMCs for the treatment of CPF and effectively inhibit granulation tissue formation.


The zone of social abandonment in cultural geography: on the street in the United States, inside the family in India.
This essay examines the spaces across societies in which persons with severe mental illness lose meaningful social roles and are reduced to "bare life." Comparing ethnographic and interview data from the United States and India, we suggest that these processes of exclusion take place differently: on the street in the United States, and in the family household in India. We argue that cultural, historical, and economic factors determine which spaces become zones of social abandonment across societies. We compare strategies for managing and treating persons with psychosis across the United States and India, and demonstrate that the relative efficiency of state surveillance of populations and availability of public social and psychiatric services, the relative importance of family honor, the extent to which a culture of psychopharmaceutical use has penetrated social life, and other historical features, contribute to circumstances in which disordered Indian persons are more likely to be forcefully "hidden" in domestic space, whereas mentally ill persons in the United States are more likely to be expelled to the street. However, in all locations, social marginalization takes place by stripping away the subject's efficacy in social communication. That is, the socially "dead" lose communicative efficacy, a predicament, following Agamben, we describe as "bare voice."


Sphingosine 1-phosphate receptors are dysregulated in endometriosis: possible implication in transforming growth factor β-induced fibrosis.
To study the molecular mechanisms involved in the appearance of the fibrotic trait in endometriosis by investigating whether the signaling pathway of the bioactive sphingolipid sphingosine 1-phosphate (S1P) was altered in endometriotic lesions. Case-control laboratory study. University research institute and university hospital. A total of 75 women, with and without endometriosis, were included in the study. Endometrial samples were obtained from women affected (n = 15 endometrioma [OMA]; n = 30 deep infiltrating endometriosis [DIE]) and not (n = 30) by endometriosis by means of laparoscopic surgery, followed by clinical and imaging investigation and checking for the expression of fibrosis markers and genes implicated in S1P metabolism and signaling by means of real-time polymerase chain reaction. The role of the S1P signaling axis in endometriosis-associated fibrosis was studied in vitro, where RNA interference approaches were used to investigate if S1P synthesis by sphingosine kinases (SKs) and specific S1P receptors (S1PRs) are implicated in the profibrotic effect of the cytokine transforming growth factor (TGF) β1. mRNA expression analysis of S1PR demonstrated a deep dysregulation of S1P signaling in endometriosis, characterized by increased expression of fibrosis markers: S1P1 was transcriptionally more expressed in OMA, and S1P3 and S1P5 mRNA levels were significantly augmented in both OMA and DIE. SK1 and its activating protein calcium- and integrin-binding protein 1 (CIB1) were significantly up-regulated in OMA and DIE. A crucial role for the SK/S1PR axis in the profibrotic effect elicited by TGFβ1 was highlighted in vitro. The S1P signaling axis may represent a useful biomarker or innovative pharmacologic target for endometriosis.


Could natural products modulate early inflammatory responses, preventing acute respiratory distress syndrome in COVID-19-confirmed patients?
The ARDS (Acute Respiratory Distress Syndrome) is a severe respiratory syndrome that was recently associated as the main death cause in the COVID-19 pandemic outbreak. Hence, in order to prevent ARDS, the pulmonary function maintenance has been the target of several pharmacological approaches. However, there is a lack of reports regarding the use of effective pharmaceutical active natural products (PANPs) for early treatment and prevention of COVID-19-related ARDS. Therefore, the aim of this work was to conduct a systematic review regarding the PANPs that could be further studied as alternatives to prevent ARDS. Consequently, this work can pave the way to spread the use of PANPs on the prevention of ARDS in COVID-19-confirmed or -suspected patients. The search strategy included scientific studies published in English from 2015 to 2020 that promoted the elucidation of anti-inflammatory pathways targeting ARDS by in vitro and/or in vivo experiments using PANPs. Then, 74 studies regarding PANPs, able to maintain or improve the pulmonary function, were reported. The PANPs may present different pulmonary anti-inflammatory pathways, wherein (i) reduction/attenuation of pro-inflammatory cytokines, (ii) increase of the anti-inflammatory mediators' levels, (iii) pulmonary edema inhibition and (iv) attenuation of lung injury were the most observed biological effects of such products in in vitro experiments or in clinical studies. Finally, this work highlighted the PANPs with promising potential to be used on respiratory syndromes, allowing their possible use as alternative treatment at the prevention of ARDS in COVID-19-infected or -suspected patients.


Memory rescue and learning in synaptic impaired neuronal circuits.
Neuronal impairment is a characteristic of Alzheimer's disease (AD), but its effect on neural activity dynamics underlying memory deficits is unclear. Here, we studied the effects of synaptic impairment on neural activities associated with memory recall, memory rescue, and learning a new memory, in an integrate-and-fire neuronal network. Our results showed that reducing connectivity decreases the neuronal synchronization of memory neurons and impairs memory recall performance. Although, slow-gamma stimulation rescued memory recall and slow-gamma oscillations, the rescue caused a side effect of activating mixed memories. During the learning of a new memory, reducing connectivity caused impairment in storing the new memory, but did not affect previously stored memories. We also explored the effects of other types of impairments including neuronal loss and excitation-inhibition imbalance and the rescue by general increase of excitability. Our results reveal potential computational mechanisms underlying the memory deficits caused by impairment in AD.


Metal binding and interdomain thermodynamics of mammalian metallothionein-3: enthalpically favoured Cu+ supplants entropically favoured Zn2+ to form Cu4 + clusters under physiological conditions.
Metallothioneins (MTs) are a ubiquitous class of small metal-binding proteins involved in metal homeostasis and detoxification. While known for their high affinity for d10 metal ions, there is a surprising dearth of thermodynamic data on metals binding to MTs. In this study, Zn2+ and Cu+ binding to mammalian metallothionein-3 (MT-3) were quantified at pH 7.4 by isothermal titration calorimetry (ITC). Zn2+ binding was measured by chelation titrations of Zn7MT-3, while Cu+ binding was measured by Zn2+ displacement from Zn7MT-3 with competition from glutathione (GSH). Titrations in multiple buffers enabled a detailed analysis that yielded condition-independent values for the association constant (K) and the change in enthalpy (ΔH) and entropy (ΔS) for these metal ions binding to MT-3. Zn2+ was also chelated from the individual α and β domains of MT-3 to quantify the thermodynamics of inter-domain interactions in metal binding. Comparative titrations of Zn7MT-2 with Cu+ revealed that both MT isoforms have similar Cu+ affinities and binding thermodynamics, indicating that ΔH and ΔS are determined primarily by the conserved Cys residues. Inductively coupled plasma mass spectrometry (ICP-MS) analysis and low temperature luminescence measurements of Cu-replete samples showed that both proteins form two Cu4 +-thiolate clusters when Cu+ displaces Zn2+ under physiological conditions. Comparison of the Zn2+ and Cu+ binding thermodynamics reveal that enthalpically-favoured Cu+, which forms Cu4 +-thiolate clusters, displaces the entropically-favoured Zn2+. These results provide a detailed thermodynamic analysis of d10 metal binding to these thiolate-rich proteins and quantitative support for, as well as molecular insight into, the role that MT-3 plays in the neuronal chemistry of copper.


Substance P-induced release of Met5-enkephalin from striatal and periaqueductal gray slices.
Substance P(SP), the heptapeptide SP and the stable analogue (p-Glu5-MePhe8-MeGly9) SP (DiMe-C7) induce a Ca2+-dependent release of Met5-enkephalin (MET) from slices of periaqueductal gray matter (PAG) and striatum of rats. The MET release from striatal slices is greater than that from PAG slices because of the higher MET content of striatum. Intraventricular injection of SP and of the two related peptides induce analgesia in the rat, and their analgesic potency is in line with their capacity to release MET. Other neuropeptides which possess antinociceptive activity such as bombesin, neurotensin, vasopressin and somatostatin fail to release MET from PAG slices.


Coordinate regulation of NAD(P)H:quinone oxidoreductase and glutathione-S-transferases in primary cultures of rat neurons and glia: role of the antioxidant/electrophile responsive element.
NAD(P)H:quinone oxidoreductase (QR) and glutathione-S-transferases (GSTs) are among the enzymes believed to protect an organism against oxidative stress. To test if redox-cycling compounds regulate the expression of these enzymes in cells of neural origin, primary cultures of rat cerebellar neurons and glia were treated with tert-butylhydroquinone (tBHQ) and hydroquinone (HQ). Basal levels of endogenous QR and GST activity were significantly greater in glia than neurons; and QR, GSTP1, and A3 were increased in glial but not neuronal cultures by treatment with tBHQ and HQ. A possible role for protein kinase C (PKC) in the tBHQ-mediated increase in QR and GST was evaluated by activating PKC with phorbol 12-myristate 13-acetate or inhibiting PKC with bisindolylmaleimide I. PKC was not involved in maintaining basal expression or mediating the increased expression of GST or QR by tBHQ. Transcriptional activation of QR and rGSTP1 by tBHQ could be mediated through a common responsive element present in the 5'-flanking region of both genes, the antioxidant/electrophile responsive element (ARE/EpRE). Transient transfection of the glial cultures with rGSTP1- or rQR1-ARE/EpRE-luciferase reporter constructs demonstrated that tBHQ transcriptionally activates the ARE/EpRE. Thus, the increased expression of genes regulated by the ARE/EpRE in cells of the central nervous system may provide protection against oxidative stress.


Relative binding affinities of bisphosphonates for human bone and relationship to antiresorptive efficacy.
Potent bisphosphonates (BPs) preferentially bind bone at sites of active osteoclastic bone resorption, where they are taken up by the osteoclast and inhibit resorption. We tested the hypothesis that BP affinity to human bone affects antiresorptive potency. [(1)(4)C]-Alendronate binding to human bone was saturable and reversible with an apparent Kd of 72 microM by Scatchard analysis. In competition binding assays, unlabeled alendronate (Ki: 61 microM) was slightly more potent than pyrophosphate (Ki = 156 microM) in blocking [(1)(4)C]-alendronate binding. Likewise, most tested BPs, including etidronate (Ki: 91 microM), ibandronate (116 microM), pamidronate (83 microM), risedronate (85 microM) and zoledronate (81 microM), showed comparable affinities. Interestingly, tiludronate (173 microM; P < 0.05 vs. all other BPs) and especially clodronate (806 microM; P > 0.0001 vs. all other BPs) displayed significantly weaker affinity for bone. The weak affinity of clodronate translated into a requirement for 10-fold higher dosing in in vitro bone resorption assays when bone was pretreated with BP and subsequently washed prior to adding osteoclasts. In stark contrast, neither alendronate nor risedronate lost any efficacy after washing the bone surface. These findings suggest that most clinically tested BPs may have similar affinities for human bone. For those with reduced affinity, this may translate into lower potency that necessitates higher dosing.


Distribution and transfer of antibiotic resistance genes in different soil-plant systems.
The extensive application of farm manure that is contaminated with pharmaceutical antibiotics not only causes substantial soil pollution but additionally leads to the input of antibiotic resistance genes (ARGs) into the soil. These ARGs would proliferate and affect human health via the food chain. The effects of cultivated crops and wild plants on ARGs in rhizosphere soil are unclear. Therefore, we chose potted plants of cultivated crops (pakchoi, lettuce, corn) and wild plants (barnyard grass, crabgrass, dog tail), and set up test groups, i.e., treatment group, antibiotic-contaminated soil; control group, no antibiotic-contaminated soil; and a blank group without plants. The aim was to explore differences in the distribution and transfer of ARGs in the soil-plant system between cultivated crops and wild plants and at the same time to explore the influence of bacterial community evolution on ARGs in the rhizosphere soil of cultivated crops and wild plants. We concluded that under the pressure of antibiotic selection, ARGs can be transferred to the root endophytes of plants through the soil and further to the phyllosphere of plants, and cultivated crops such as pakchoi and wild plants barnyard grass have a strong ability to transport ARGs. Regardless of cultivated crops or wild plants, the abundance of ARGs in rhizosphere soil can be substantially reduced by 66.53 ~ 85.35%. Redundancy analysis and network analysis indicated that bacterial community succession is the main mechanism affecting changes of ARGs in rhizosphere soil. The reduction of Firmicutes due to the plant was the main factor responsible for the reduction of the abundance of ARGs in rhizosphere soil. The tetA, tetG, tetX, sul2, and qnrS genes are highly related to some potential pathogens, and the health risks they bring are a red flag that deserves attention.


[Application of melanocytes and bone marrow mesenchymal stem cells in tissue engineered skin construction].
To explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells (BMSCs) in vivo. Melanocytes were isolated from human foreskin. BMSCs were isolated from human bone marrow. Both of them were co-cultured at a ratio of 1:10, and then were implanted into the collagen membrane to construct the tissue-engineered skin, which was applied for wound repair in nude mice. The effectiveness of wound repair and the distribution of melanocytes were evaluated by morphological observation, in vivo 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI) fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy. The wounds were satisfactorily repaired among the nude mice. The melanocytes were distributed in the skin with normal structure, as confirmed by DAPI fluorescent staining tracing, HE staining, S-100 immunohistochemistry, and transmission electron microscopy. Melanocytes and BMSCs, after proper in vitro culture at an appropriate ratio, can construct the tissue-engineered skin with I type collagen membrane.


Noise analysis and relaxation experiments of transport of hydrophobic anions across lipid membranes at equilibrium and nonequilibrium.
Under equilibrium and nonequilibrium steady-state conditions, the spectral intensity of current noise SJ(f) generated by the transport of hydrophobic anions across lipid bilayer membranes was investigated. The experimental results were compared with different reaction models. SJ(f) showed a characteristic increase proportional to f2 between frequency-independent tails at low and high frequencies. This gradient was found to be independent of applied voltage which indicates the contribution of a single voltage-dependent reaction step of ion translocation across the membrane. From the shape of SJ(f) at low frequencies the rate constant of ion desorption from the membrane into the aqueous phase could be estimated. Unambiguous evidence for the application of a general model, which includes the coupling of slow ion diffusion in the aqueous phase to ion adsorption/desorption at the membrane interface, could not be obtained from the low-frequency shape of SJ(f). The shot noise of this ion transport determines the amplitude of SJ(f) at high frequencies which decreases with increasing voltage applied. Analysis of voltage-jump current-relaxation experiments and of current noise carried out on one membrane yielded significant differences of the derived ion partition coefficient. This deviation is qualitatively described on the basis of incomplete reaction steps.


Microwave assisted facile synthesis of {1/1,3-bis/1,3,5-tris-[(2-nitroxyethylnitramino)-2,4,6-trinitrobenzene]} using bismuth nitrate pentahydrate as an eco-friendly nitrating agent.
1-(2-Nitroxyethylnitramino)-2,4,6-trinitrobenzene (3a), 1,3-bis(2-nitroxyethyl nitramino)-2,4,6-trinitrobenzene (3b) and 1,3,5-tris(2-nitroxyethylnitramino)-2,4,6-trinitrobenzene (3c) were prepared by the nitration of 1-(2-hydroxyethylamino)-2,4,6-trinitrobenzene (2a) 1,3-bis(2-hydroxyethylamino)-2,4,6-trinitrobenzene (2b) and 1,3,5-tris(2-hydroxyethylamino)-2,4,6-trinitrobenzene (2c) using bismuth nitrate pentahydrate (eco-friendly nitrating agent) in tetrahydrofuran adsorbed on silica gel under microwave irradiation, respectively. Key intermediate compounds viz., 2a, 2b and 2c were synthesized by condensing picryl chloride, styphnyl chloride and 1,3,5-trichloro-2,4,6-trinitrobenzene with ethanol amine, respectively, based on the lines of the reported method. The synthesized compounds were characterized based on their physical constant, infrared (IR) spectroscopy and (1)H nuclear magnetic resonance (NMR) spectroscopy. The spectroscopic data obtained indicated the formation of nitrate esters (3a-3c). The nitration methodology adopted in the present study is of relevance in the context of green chemistry. The target compounds (3a-3c) synthesized using eco-friendly approach are of interest from the point of high energy materials (HEMs).


2017: beginning of a new era for Chlamydia research in China and the rest of the world.
The First Chinese Chlamydia Research Meeting was held in Lanzhou, China in May 2017, 60 years after the disclosure of reproducible isolation of Chlamydia trachomatis by (Fei-fan Tang). We report current state of the Chlamydia research community in China, and briefly review recent progress in Chlamydia vaccinology. The meeting represents a new milestone for Chlamydia research in the country. The Chinese Chlamydia Research Society (CCRS) was formed during the meeting. Future meetings will be held biennially and should facilitate collaboration of Chinese researchers with their domestic and international colleagues.


Sharpey-Schafer lecture: gas channels.
The traditional dogma has been that all gases diffuse through all membranes simply by dissolving in the lipid phase of the membrane. Although this mechanism may explain how most gases move through most membranes, it is now clear that some membranes have no demonstrable gas permeability, and that at least two families of membrane proteins, the aquaporins (AQPs) and the Rhesus (Rh) proteins, can each serve as pathways for the diffusion of both CO₂ and NH₃. The knockout of RhCG in the renal collecting duct leads to the predicted consequences in acid-base physiology, providing a clear-cut role for at least one gas channel in the normal physiology of mammals. In our laboratory, we have found that surface-pH (pH(S)) transients provide a sensitive approach for detecting CO₂ and NH₃ movement across the cell membranes of Xenopus oocytes. Using this approach, we have found that each tested AQP and Rh protein has its own characteristic CO₂/NH₃ permeability ratio, which provides the first demonstration of gas selectivity by a channel. Our preliminary AQP1 data suggest that all the NH₃ and less than half of the CO₂ move along with H₂O through the four monomeric aquapores. The majority of CO₂ takes an alternative route through AQP1, possibly the central pore at the four-fold axis of symmetry. Preliminary data with two Rh proteins, bacterial AmtB and human erythroid RhAG, suggest a similar story, with all the NH₃ moving through the three monomeric NH₃ pores and the CO₂ taking a separate route, perhaps the central pore at the three-fold axis of symmetry. The movement of different gases via different pathways is likely to underlie the gas selectivity that these channels exhibit.


Vitamin C mediated protection on cisplatin induced mutagenicity in mice.
In present studies the development of chromosomal aberrations, micronuclei in bone marrow cells and sperm head abnormalities were used as mutagenic bioassay in Swiss albino mice treated with cisplatin alone or ascorbic plus cisplatin. It was noted that in the combined treated hosts the frequency of all the mutagenic parameters were always significantly less than that treated with cisplatin alone. These findings suggest a protective role of ascorbic acid against cisplatin induced mutagenic potentials. Interestingly, in combined treated hosts glutathione (GSH) level in bone marrow cells increased significantly which may suggest a possible mechanism of ascorbic acid mediated protection against cisplatin induced mutagenic potentials in the hosts.


Effectiveness of polydeoxyribonucleotide injection versus normal saline injection for treatment of chronic plantar fasciitis: a prospective randomised clinical trial.
Polydeoxyribonucleotide (PDRN) has been used for the treatment of chronic tendinosis. This prospective randomised study was conducted to evaluate the efficacy and complications of PDRN injection for treatment of plantar fasciitis. Forty patients with a clinical diagnosis of plantar fasciitis were randomly allocated to PDRN injection (PDRN group, n = 20) or normal saline injection (placebo group, n = 20). Injections were performed weekly for three weeks. Clinical evaluations were done at baseline and four and 12 weeks after treatment began using the visual analogue scale (VAS) for foot pain and Manchester-Oxford Foot Questionnaire (MOXFQ). We also monitored the complications in both groups at one, two, four and 12 weeks after initial treatment. The PDRN group achieved a significant improvement in VAS and MOXFQ scores at four weeks after treatment, and this improvement continued until 12 weeks after treatment. On the other hand, the placebo group did not achieve a significant improvement in the VAS or MOXFQ scores at four or 12 weeks. The initial VAS and MOXFQ scores of the PDRN group were not significantly different from those of the placebo group. At four weeks after treatment, the VAS and MOXFQ scores of the PDRN group were better than those of the placebo group, but the difference was not statistically significant. At 12 weeks after treatment, the VAS and MOXFQ scores of the PDRN group were significantly better than those of the placebo group. We noticed no injection-related complications, such as itching, urticaria, redness or infection signs around the injection site in either group. PDRN injection is an effective and safe treatment option and may be considered for plantar fasciitis.


Three-pathway combination for glutathione biosynthesis in Saccharomyces cerevisiae.
Glutathione (GSH), a pivotal non-protein thiol, can be biosynthesized through three pathways in different organisms: (1) two consecutive enzymatic reactions catalyzed by γ-glutamylcysteine synthetase (Gsh1 or GshA) and glutathione synthetase (Gsh2 or GshB); (2) a bifunctional γ-glutamylcysteine synthetase/glutathione synthetase (GshF); (3) an alternative condensation of γ-glutamyl phosphate synthesized by γ-glutamyl kinase (Pro1 or ProB) with cysteine to form γ-glutamylcysteine which was further conjugated to glycine by glutathione synthetase. The Gsh1 and Gsh2 of conventional GSH biosynthetic pathway or the bifunctional GshF reported previously have been independently modulated for GSH production. This study developed a novel three-pathway combination method to improve GSH production in Saccharomyces cerevisiae. A bifunctional enzyme GshF of Actinobacillus pleuropneumoniae was functionally expressed in S. cerevisiae and Pro1 in proline biosynthetic pathway was exploited for improving GSH yield. Moreover, two fusion proteins Gsh2-Gsh1 and Pro1-GshB were constructed to increase the two-step coupling efficiency of GSH synthesis by mimicking the native domain fusion of GshF. The engineered strain W303-1b/FGP with three biosynthetic pathways presented the highest GSH concentration (216.50 mg/L) and GSH production of W303-1b/FGP was further improved by 61.37 % when amino acid precursors (5 mM glutamic acid, 5 mM cysteine and 5 mM glycine) were fed in shake flask cultures. In batch culture process, the recombinant strain W303-1b/FGP also kept high efficiency in GSH production and reached an intracellular GSH content of 2.27 % after 24-h fermentation. The engineered strains harbouring three GSH pathways displayed higher GSH producing capacity than those with individually modulated pathways. Three-pathway combinatorial biosynthesis of GSH promises more effective industrial production of GSH using S. cerevisiae.


Sodium-calcium exchange influences the response to endothelin-1 in lens epithelium.
Studies were conducted to examine the possible involvement of Na+-Ca2+ exchanger in determining the magnitude of the endothelin-1 (ET-1)-receptor-mediated calcium signal in porcine lens epithelial cells. Cytoplasmic calcium concentration was measured in primary cultured cells loaded with Fura-2. ET-1 (100 nM) caused cytoplasmic calcium to increase transiently to approximately 250 nM from a baseline of approximately 65 nM. The calcium increase decayed to a sustained plateau 35-45 nM above the baseline. Both the peak and plateau component of the ET-1 calcium response were abolished by PD145065, an ET receptor antagonist, and by cyclopiazonic acid (CPA) (10 microM). In calcium-free bathing solution, only the plateau was abolished. In the presence of ouabain, low-sodium bathing solution or bepridil, a sodium-calcium exchange inhibitor, peak height more than doubled. Bepridil also increased the peak height of the calcium response to ATP. The half-time for decay of the ET-1 and ATP calcium peak was increased several folds by bepridil, ouabain and low-sodium conditions. Measurements of ionomycin-releasable calcium suggested calcium store size was not increased in bepridil-treated cells. Taken together findings suggest inhibition of sodium-calcium exchange increases the magnitude of the receptor-initiated store-release phase of the ET-1 calcium signaling response as the result of impaired calcium clearance from the cytoplasm.


Monocyte-related cytokines in acute myocardial infarction.
To elucidate the role of monocytes in the cytokine system in acute myocardial infarction (AMI), we examined the time courses of plasma concentrations and generation capacities of monocyte-related cytokines in 17 consecutive patients with uncomplicated AMI (from day 1 to 28) and in 10 control subjects. The concentrations of monocyte-related cytokines were measured by enzyme immunoassay with horseradish peroxidase. Cytokine generation capacity was evaluated by cytokine concentrations in the culture solution 24 hours after incubation of 0.5 ml whole blood with 5 micrograms lipopolysaccharide. Two distinct patterns of increases in the cytokine concentrations were noted: transient (plasma interleukin [IL]-6) and sustained (plasma macrophage colony-stimulating factor and generation capacities of IL-1 alpha, IL-6, granulocyte colony-stimulating factor, and tumor necrosis factor alpha). There was no significant increase in the concentrations of other cytokines. These results indicate that the concentrations of the monocyte-related cytokines dynamically change during the course of AMI, suggesting that they may contribute to the inflammatory and subsequent proliferative responses in AMI.


The spermatozoon of Myzostomum cirriferum (Annelida, Myzostomida).
The Myzostomum spermatozoon has several unique features: The chromatin material is condensed into a variable number of dense balls (usually 42); there is an incomplete myelin-like cover rather than a regular nuclear envelope; a palisade of microtubules surround two mitochondria which extend along the 30-microns-long cell body; an axoneme of the 9+0 formula originates at one end of the cell body, runs intracellularly for a short distance (less than 1 micron), then continues outside the cell body although attached to it by means of a "flagellum-cell body junction," which consists of two sets of strands connecting the membrane of the flagellum to that of the cell body. Two "dorsal rods" in the microtubular palisade evidently are connected to these strands. The flagellum also has a free portion (around 70 microns) which terminates in a pointed spine, called the "sting." No typical centriole and no acrosome have been found. The spermatozoon usually swims with flagellum foremost but may reverse the direction. The cell body also undulates but with an independent wavelength. Presumably some of these peculiarities are specializations for locomotion within body tissues rather than in water; Myzostomum has "dermal copulation."


The pace of modern life II: from rates of contemporary microevolution to pattern and process.
We compiled a database of microevolution on contemporary time scales in nature (47 source articles; 30 animal species), comprising 2649 evolutionary rates in darwins (proportional change per million years) and 2151 evolutionary rates in haldanes (standard deviations per generation). Here we demonstrate how quantitative rate measures can provide general insights into patterns and processes of evolution. The frequency distribution of evolutionary rates was approximately log-normal, with many slow rates and few fast rates. Net selection intensities estimated from haldanes were on average lower than selection intensities commonly measured directly in natural populations. This difference suggests that natural selection could easily accomplish observed microevolution but that the intensities of selection typically measured in nature are rarely maintained for long (otherwise observed evolutionary rates would be higher). Traits closely associated with fitness (life history traits) appear to evolve at least as fast as traits less closely tied to fitness (morphology). The magnitude of evolutionary difference increased with the length of the time interval, particularly when maximum rates from a given study were considered. This pattern suggests a general underlying tendency toward increasing evolutionary diversification with time. However, evolutionary rates also tended to decrease with time, perhaps because longer time intervals average increasingly disparate rates over time, or because evolution slows when populations approach new optima or as genetic variation is depleted. In combination, our results suggest that macroevolutionary transitions may ultimately arise through microevolution occasionally 'writ large' but are perhaps temporally characterized by microevolution 'writ in fits and starts'.


Archaeosome adjuvants: immunological capabilities and mechanism(s) of action.
Archaeosomes (liposomes comprised of glycerolipids of Archaea) constitute potent adjuvants for the induction of Th1, Th2 and CD8(+) T cell responses to the entrapped soluble antigen. Archaeal lipids are uniquely constituted of ether-linked isoprenoid phytanyl cores conferring stability to the membranes. Additionally, varied head groups displayed on the glycerol-lipid cores facilitate unique immunostimulating interactions with mammalian antigen-presenting cells (APCs). The polar lipid from the archaeon, Methanobrevibacter smithii has been well characterized for its adjuvant potential, and is abundant in archaetidyl serine, promoting interaction with a phosphatidylserine receptor on APCs. These archaeosomes mediate MHC class I cross-priming via the phagosome-to-cytosol TAP-dependent classical processing pathway, and also upregulate costimulation by APCs without overt inflammatory cytokine production. Furthermore, they facilitate potent CD8(+) T cell memory to co-delivered antigen, comparable in magnitude and quality to live bacterial vaccine vectors. Archaeosome vaccines provide profound protection in murine models of infection and cancer. This technology is being developed for clinical application and offers a novel prospect for rational design and development of safe and potent subunit vaccines capable of eliciting T cell immunity against intracellular infections and cancers.


Cytoreductive surgery combined with intraoperative chemo-hyperthermia and postoperative radiotherapy in the management of advanced pancreatic adenocarcinoma: feasibility aspects and efficacy.
The aim of our study was to evaluate the feasibility and the efficacy of cytoreductive surgery (CS) with intraoperative chemo-hyperthermia in the management of advanced stage IVA (T4N0M0) pancreatic cancer. From August 1995 through March 1996, seven patients with unresectable adenocarcinoma of the pancreas underwent CS, with preoperative chemotherapy (5-fluorouracil [FU] for 96 h), plus 45-Gy external beam postoperative irradiation with a 6-MeV linear accelerator (1.8 Gy per fraction, 5 days per week). A single session of intraoperative hyperthermia was performed with a waveguide-type applicator operating at 433 MHz, and temperature was measured by inserting a flexiguide needle catheter carrying a thermometry probe with three measuring points into the tumor. The tumor region was heated to 43 degrees C-45 degrees C for up to 60 min, while 5-FU 500 mg was injected simultaneously through the gastroduodenal artery into the splenic artery (intraoperative regional chemotherapy). Postoperative recovery was uneventful for all patients. After the combined treatment, there was a significant decrease in the values of both serum carcinoembryonic antigen (CEA; P = 0.017, Wilcoxon test) and carbohydrate antigen (CA)19-9 ( P = 0.016; Wilcoxon test), from 7.6 +/- 1.5 ng/ml CEA and 869.6 +/- 126.9 U/ml CA to 3.5 +/- 0.8 ng/ml CEA and 104.7 +/- 35.4 U/ml CA19-9. Moreover, there was a significant improvement ( P = 0.016; Wilcoxon test) in Eastern Cooperative Oncology Group performance status, pain score, and body mass index. The median overall survival was 18.5 (SE, 1.8) months. Our preliminary clinical results suggest the tolerability and the considerable potential advantage of using cytoreductive resection with preoperative chemotherapy, intraoperative chemo-hyperthermia, and external beam postoperative radiotherapy for the management of advanced adenocarcinoma of the pancreas.


Two TNF receptors.
In the past few years, considerable progress has been made on the identification and characterization of tumor necrosis factor (TNF) receptors. The relative roles played by the two receptor types in signaling the diverse functions of TNF are less clear. Here, Louis Tartaglia and David Goeddel summarize progress to date and propose a model of TNF receptor triggering that reconciles the seemingly conflicting data.


Postmortem Attraction of Sarcosaprophagous Diptera to Tramadol-Treated Rats and Morphometric Aspects of the Developed Larvae.
The presence of some specific drugs in animal tissues may affect the time of minimal postmortem intervals estimated during forensic entomological investigations. To test the effects of a specific drug on decomposition, a field study was conducted at Fayoum University campus, Egypt, from March to May 2013, using tramadol, a synthetic analgesic opioid used to treat moderate to severe pain in humans. Albino rats were used as the animal model during this study. The duration of the fresh stage of tramadol treated rat (Ttr) carcasses was significantly shorter (2.4 ± 0.27 days) compared to tramadol free rat (Tfr) carcasses (6.4 ± 0.49 days). The dry carcass stage of Ttr lasted longer (10.3 ± 0.99 days) as compared to (7.4 ± 0.18 days) the Tfr carcass. The decomposition process of the (Ttr) carcass was not significantly faster (24.9 ± 1.58 days) as compared to (Tfr) carcasses (29.5 ± 1.69). Lucilia cuprina (Wiedemann), Chrysomya albiceps (Wiedemann), and Musca domestica L. were less attracted to Ttr carcass-baited traps than traps with Tfr carcasses. However, females of Sarcophaga spp. showed a greater attraction to Ttr carcasses. Females of another sarcophagid fly, Wohlfahrtia spp. exhibited similar attraction tendencies to both types of trap baits. Larvae of S. argyrostoma (Robineau-Desvoidy) collected from Ttr carcasses developed to a significantly longer total body length (10.4 ± 0.04 mm) as compared to the average length of the larvae collected from Tfr carcasses (8.9 ± 0.34 mm). During days 9-13 after rat death, the relative lengths of larvae from Ttr carcasses were not significantly different from Tfr carcasses. Larvae fed on Ttr carcasses pupated 2 days later than the control larvae.


Facile access to 3-sulfonylquinolines via Knoevenagel condensation/aza-Wittig reaction cascade involving ortho-azidobenzaldehydes and β-ketosulfonamides and sulfones.
Quinoline-based sulfonyl derivatives, and especially sulfonamides, are relevant and promising structures for drug design. We have developed a new convenient protocol for the synthesis of 3-sulfonyl-substituted quinolines (sulfonamides and sulfones). The approach is based on a Knoevenagel condensation/aza-Wittig reaction cascade involving o-azidobenzaldehydes and ketosulfonamides or ketosulfones as key building blocks. The protocol is appropriate for both ketosulfonyl reagents and α-sulfonyl-substituted alkyl acetates providing the target quinoline derivatives in good to excellent yields.


The sales learning curve.
When a company launches a new product into a new market, the temptation is to immediately ramp up sales force capacity to gain customers as quickly as possible. But hiring a full sales force too early just causes the firm to burn through cash and fail to meet revenue expectations. Before it can sell an innovative product efficiently, the entire organization needs to learn how customers will acquire and use it, a process the authors call the sales learning curve. The concept of a learning curve is well understood in manufacturing. Employees transfer knowledge and experience back and forth between the production line and purchasing, manufacturing, engineering, planning, and operations. The sales learning curve unfolds similarly through the give-and-take between the company--marketing, sales, product support, and product development--and its customers. As customers adopt the product, the firm modifies both the offering and the processes associated with making and selling it. Progress along the manufacturing curve is measured by tracking cost per unit: The more a firm learns about the manufacturing process, the more efficient it becomes, and the lower the unit cost goes. Progress along the sales learning curve is measured in an analogous way: The more a company learns about the sales process, the more efficient it becomes at selling, and the higher the sales yield. As the sales yield increases, the sales learning process unfolds in three distinct phases--initiation, transition, and execution. Each phase requires a different size--and kind--of sales force and represents a different stage in a company's production, marketing, and sales strategies. Adjusting those strategies as the firm progresses along the sales learning curve allows managers to plan resource allocation more accurately, set appropriate expectations, avoid disastrous cash shortfalls, and reduce both the time and money required to turn a profit.


MicroRNA Profiling from Tears as a Potential Non-invasive Method for Early Detection of Diabetic Retinopathy.
Diabetic retinopathy (DR) is a vascular complication of diabetes that can lead to partial or complete loss of vision. Early detection and treatment of DR can prevent blindness. Regular clinical examination is recommended for DR diagnosis; however, it is not always possible or feasible due to limited resources, expertise, time, and infrastructure. Several clinical and molecular biomarkers are proposed for the prediction of DR including microRNAs. MicroRNAs are a class of small non-coding RNAs that are found in biofluids and can be measured using reliable and sensitive methods. The most commonly used biofluid for microRNA profiling is plasma or serum; however, tear fluid (tears) is also demonstrated to contain microRNAs. MicroRNAs isolated from tears present a non-invasive source for DR detection. Different methods of microRNA profiling are available including digital PCR-based methods that can detect up to a single copy of microRNA in the biofluids. Here, we describe microRNA isolation from tears using manual method as well as using a high-throughput automated platform followed by microRNA profiling using digital PCR system.


The cellular basis of hybrid dysgenesis and Stellate regulation in Drosophila.
During normal tissue development, the accumulation of unrepaired cellular and genomic damage can impair growth and ultimately leads to death. To preserve cellular integrity, cells employ a number of defense mechanisms including molecular checkpoints, during which development is halted while dedicated pathways attempt repair. This process is most critical in germline tissues where cellular damage directly threatens an organism's reproductive capacity and offspring viability. In the fruit fly, Drosophila melanogaster, germline development has been extensively studied for over a century and the breadth of our knowledge has flourished in the genomics age. Intriguingly, several peculiar phenomena that trigger catastrophic germline damage described decades ago, still endure only a partial understanding of the underlying molecular causes. A deeper reexamination using new molecular and genetic tools may greatly benefit our understanding of host system biology. Among these, and the focus of this concise review, are hybrid dysgenesis and an intragenomic conflict that pits the X and Y sex chromosomes against each other.


Left ventricular endocardial ecchinococcosis associated with multiple intracranial hydatid cysts.
Cardiac ecchinococcosis is a rare disease. Its incidence varies from 0.02-2%. Commonly seen in the left ventricle arising from the myocardium in the subepicardial region.We report a 15-year-old boy presented with a rare combination of a left ventricular subendocardial hydatid cyst associated with multiple cysts in the left cerebral hemisphere and right posterior occipital lobe. The patient underwent successful surgical excision of the left ventricular hydatid cyst using cardiopulmonary bypass.


Bilateral intralobar pulmonary sequestration: therapeutic implications.
Bilateral intralobar pulmonary sequestration within the lower lobes was suggested by computed tomography and confirmed by arteriography in a 27-year-old woman. Although one side was asymptomatic, both sequestrated areas were resected using staged bilateral thoracotomies. Histopathological examination showed massive inflammation in the symptomatic as well as the asymptomatic sequestrum. To our knowledge, this is the sixth reported case of bilateral intralobar pulmonary sequestration and the third for which bilateral resection has been performed.


Low Accuracy of FIB-4 and NAFLD Fibrosis Scores for Screening for Liver Fibrosis in the Population.
Fibrosis-4 (FIB-4) and the nonalcoholic fatty liver disease fibrosis score (NFS) are the 2 most popular noninvasive blood-based serum tests proposed for widespread fibrosis screening. We therefore aimed to describe the accuracy of FIB-4 and NFS to detect elevated liver stiffness as an indicator of hepatic fibrosis in low-prevalence populations. This study included a total of 5129 patients with concomitant measurement of FIB-4, NFS, and liver stiffness measurement (LSM) by Fibroscan (Echosens, France) from 5 independent population-based cohorts from Spain, Hong Kong, Denmark, England, and France; 3979 participants from the general population and 1150 from at-risk cohorts due to alcohol, diabetes, or obesity. We correlated LSM with FIB-4 and NFS, and calculated pre- and post-test predictive values of FIB-4 and NFS to detect elevated LSM at 8 kPa and 12 kPa cutoffs. The mean age was 53 ± 12 years, the mean body mass index was 27 ± 5 kg/m2, and 2439 (57%) were women. One in 10 patients (552; 11%) had liver stiffness ≥8 kPa, but 239 of those (43%) had a normal FIB-4, and 171 (31%) had normal NFS. The proportion of false-negatives was higher in at-risk patients than the general population. FIB-4 was false-negative in 11% of diabetic subjects, compared with 2.5% false-negatives with NFS. Waist circumference outperformed FIB-4 and NFS for detecting LSM ≥8 kPa in the general population. Almost one-third (28%-29%) of elevated FIB-4/NFS were false-positive in both the general population and at-risk cohorts. FIB-4 and NFS are suboptimal for screening purposes due to a high risk of overdiagnosis and a non-negligible percentage of false-negatives, especially in patients with risk factors for chronic liver disease. Waist circumference emerged as a potential first step to identify patients at risk for liver fibrosis in the general population.


Occurrence of phthalate esters and preliminary data on microplastics in fish from the Tyrrhenian sea (Italy) and impact on human health.
Phthalic acid esters (PAEs) are chemical pollutants widely distributed in the marine environment. They can accumulate in biota, posing a risk to the marine ecosystem and humans. The aim of this study was to measure the content of PAEs in the gills and muscles of three fish species (Mugil cephalus, Diplodus annularis, and Mullus barbatus) caught along the coast of Campania (Italy), as well as to ascertain the dietary exposure to PAEs through the consumption of fish. Secondly, a preliminary insight into microplastics (MPs) pollution in this area was provided through the analysis of Mugil cephalus organs. Solid-phase extraction (SPE) and gas chromatography-mass spectrometry (GC-MS) were used for the PAEs analysis, while an Fourier-transform infrared (FTIR) microscope was used to detect MPs after a pre-digestion of the samples. Risk assessment was based on estimated daily intake (EDI) and lifetime cancer risk (LTCR). The results showed higher bioaccumulation of PAEs in Mullus barbatus than in the other two species and higher concentration in gills than in muscles. MPs (polyamide, polypropylene, and high-density polyethylene) were detected in half of the gill samples, but no particle was detected in the muscle samples of Mugil cephalus. A low carcinogenic and non-carcinogenic risk from the consumption of fish emerged, although a potential risk for the development of cancer was found in the worst-case, especially in toddlers. In conclusion, this study provides insight into PAEs pollution in the Tyrrhenian Sea (Italy), their distribution in fish with different behaviors, and the potential risk to the consumer. Moreover, the data on pollution by MPs in this area could form the basis for future studies.


Sri Lankan FRAX model and country-specific intervention thresholds.
There is a wide variation in fracture probabilities estimated by Asian FRAX models, although the outputs of South Asian models are concordant. Clinicians can choose either fixed or age-specific intervention thresholds when making treatment decisions in postmenopausal women. Cost-effectiveness of such approach, however, needs to be addressed. This study examined suitable fracture probability intervention thresholds (ITs) for Sri Lanka, based on the Sri Lankan FRAX model. Fracture probabilities were estimated using all Asian FRAX models for a postmenopausal woman of BMI 25 kg/m² and has no clinical risk factors apart from a fragility fracture, and they were compared. Age-specific ITs were estimated based on the Sri Lankan FRAX model using the method followed by the National Osteoporosis Guideline Group in the UK. Using the age-specific ITs as the reference standard, suitable fixed ITs were also estimated. Fracture probabilities estimated by different Asian FRAX models varied widely. Japanese and Taiwan models showed higher fracture probabilities while Chinese, Philippine, and Indonesian models gave lower fracture probabilities. Output of remaining FRAX models were generally similar. Age-specific ITs of major osteoporotic fracture probabilities (MOFP) based on the Sri Lankan FRAX model varied from 2.6 to 18% between 50 and 90 years. ITs of hip fracture probabilities (HFP) varied from 0.4 to 6.5% between 50 and 90 years. In finding fixed ITs, MOFP of 11% and HFP of 3.5% gave the lowest misclassification and highest agreement. Sri Lankan FRAX model behaves similar to other Asian FRAX models such as Indian, Singapore-Indian, Thai, and South Korean. Clinicians may use either the fixed or age-specific ITs in making therapeutic decisions in postmenopausal women. The economical aspects of such decisions, however, need to be considered.


Comparative analysis of fresh chondrocytes, cultured chondrocytes and chondroprogenitors derived from human articular cartilage.
Interest in chondroprogenitors arose due to their inherent stem cell like properties, and their initial characterization was based on identification of a small percentage of CD49e positive cells in cultured chondrocytes (CC). It was further noted that when fresh chondrocytes (FC; reported to express low CD49e) were subjected to fibronectin adhesion assay, an isolate of chondroprogenitors was obtained, which was highly positive for CD49e, thus making it a distinguishing marker for this cell population. However, this notion was challenged when reports demonstrated high CD49e expression in CC as well. Therefore, our aim was to compare CD49e expression in FC, CC and chondroprogenitors. Chondrocytes and chondroprogenitors were isolated from articular cartilage of osteoarthritic joints from three patients. Assessment of classic fibronectin receptor (CD49e, CD29), positive (CD105, CD73, CD90) and negative (CD45, CD34) mesenchymal stem cell marker expression in all groups was performed, as chondroprogenitors fulfill the minimal criteria laid down by International Society for Cellular Therapy. Following this, adipogenic, osteogenic and chondrogenic differentiation was assessed by Oil red O, Alizarin Red and Alcian Blue staining respectively. Our observations indicate that FC show significantly low surface marker expression as compared to CC and chondroprogenitors, whereas no significant difference was seen in values when CC and chondroprogenitors were compared. Moreover, comparable results were exhibited when trilineage differentiation potential was compared across groups. Since CC and chondroprogenitors show similar characteristics, there is a pressing need for a specific differentiating marker to isolate a pure population of chondroprogenitors.


Effects of uncertainty on melodic information processing.
In three experiments, musically trained and untrained adults listened to three repetitions of a 5-note melodic sequence followed by a final melody with either the same tune as those preceding it or differing in one position by one semitone. In Experiment 1, ability to recognize the final sequence was examined as a function of redundancy at the levels of musical structure in a sequence, contour complexity of transpositions in a trial, and trial context in a session. Within a sequence, tones were related as the major or augmented triad; within a trial, the four sequences began on successively higher notes (simple macrocontour) or on randomly selected notes (complex macrocontour); and within a session, trials were either blocked (all major or all augmented) or mixed (major and augmented randomly selected). Performance was superior for major melodies, for systematic transpositions within a trial (simple macrocontours), for blocked trials, and for musically trained listeners. In Experiment 2, we examined further the effect of macrocontour. Performance on simple macrocontours exceeded that on complex, and excluded the possibility that repetition of the 20-note sequences provided the entire benefit of systematic transposition in Experiment 1. The effect of musical structure (major/augmented) was also replicated. In Experiment 3, listeners provided structure ratings of ascending 20-note sequences from Experiment 2. Ratings on same trials were higher than those on corresponding different trials, in contrast to performance scores for augmented same and different trials in previous experiments. The concept of functional uncertainty was proposed to account for recognition difficulties on augmented same trials. The significant effects of redundancy on all the levels examined confirm the utility of the information-processing framework for the study of melodic sequence perception.


Pediatric Head and Neck Malignancies.
Head and neck malignancies are rare in pediatric patients, and represent 12% of all pediatric malignancies. The incidence for these head and neck tumors is 1.49 cases per 1,000,000 person-years. Among the most common pediatric head and neck malignancies are lymphomas (27%), neural tumors including primitive neurectodermal tumors (23%), thyroid malignancies (21%), soft tissue sarcomas including rhabdomyosarcoma (12%), nasopharyngeal carcinoma, skeletal and odontogenic malignancies including osteosarcoma, Ewing sarcoma, and ameloblastic carcinoma. This article presents an overview of pediatric head and neck malignancies with emphasis on diagnosis and management.


Stanniocalcin in the euryhaline flounder (Platichthys flesus): primary structure, tissue distribution, and response to altered salinity.
Stanniocalcin (STC) is a homodimeric glycoprotein hormone that was first discovered in fish, where it is largely produced by a unique endocrine gland, the corpuscles of Stannius (CS). In bony fish, it is thought to be an important regulator of calcium and phosphate uptake from the aquatic environment. This report describes the molecular cloning of STC from euryhaline flounder (Platichthys flesus) CS cDNA and genomic DNA. The flounder STC encodes a prehormone of 251 amino acids (aa) with a signal peptide of 17 aa, followed by another 15 aa sequence before the mature protein of 219 aa. The deduced aa sequence of flounder STC shows 62.9-89.0% similarity and 50.4-83.1% identity with other known fish STC sequences, but only 42.3% identity with mouse STC1, 24.4% identity with fugu and zebrafish STC2, and 22.3% identity with mouse STC2. Primary structural analysis demonstrated that flounder STC gene contains five exons in contrast to the four exons present in mammalian STC gene. RT-PCR revealed the expression of flounder STC mRNA to be widely spread in many tissues and organs, similar to the situation in mammals and other fish. Quantitative PCR (Q-PCR) was conducted to measure relative STC expression levels in the CS, which showed STC mRNA expression levels in seawater-adapted fish CS were about 3-fold higher than in freshwater-adapted fish CS.


Synapsins in the vertebrate retina: absence from ribbon synapses and heterogeneous distribution among conventional synapses.
The vertebrate retina contains two ultrastructurally distinct types of vesicle-containing synapses: conventional synapses, made predominantly by amacrine cells, and ribbon synapses, formed by photoreceptor and bipolar cells. To identify molecular differences between these synapse types, we have compared the distribution of the synapsins, a family of nerve terminal phosphoproteins, with that of synaptophysin (p38) and SV2, two intrinsic membrane proteins of synaptic vesicles. We report an absence of synapsin I and II immunoreactivity from all ribbon-containing nerve terminals. These include terminals of rod cells in developing and adult rat retina, rod and cone cells in monkey and salamander retinas, and rat bipolar cells. Furthermore, we show that synapsins I and II are differentially distributed among conventional synapses of amacrine cells. The absence of the synapsins from ribbon synapses suggests that vesicle clustering and mobilization in these terminals differ from that in conventional synapses.


Echocardiographic identification of the mitral valve prolapse syndrome.
The data presented herein would indicate that mitral valve prolapse may be represented by a variable spectrum of abnormalities on the echocardiogram. Utilizing all the abnormalities described, echographic evidence of mitral valve prolapse may be detected in most patients with this disorder. However, evidence of mitral valve prolapse may also be observed in a large contingent of asymptomatic patients without other clinical manifestations of cardiac disease. The biologic significance of these echographic abnormalities remains uncertain and constitutes an important area for future investigation.


Juvenile exposure to methylphenidate reduces cocaine reward and alters netrin-1 receptor expression in adulthood.
The guidance cue netrin-1 acting on mesocorticolimbic dopamine (DA) neurons through its receptor DCC (deleted in colorectal cancer) has been implicated in the neuronal plasticity induced by psychostimulant drugs. We examined in C57/BL6 mice the effects of repeated juvenile methylphenidate (MPH) exposure on cocaine-reward sensitivity in adulthood and determined whether early MPH treatment alters adult expression of DCC in mesocorticolimbic DA regions. By using place conditioning, we show that adult mice exposed to MPH during the juvenile period are less sensitive to cocaine-reward compared to saline-controls. Furthermore, by means of immunoblotting, we demonstrate that early MPH treatment attenuates adult DCC expression in the ventral tegmental area (VTA) selectively. These results support previous evidence that developmental MPH treatment diminishes cocaine-reward in adulthood and are the first to suggest that DCC in the VTA may participate in this enduring effect.


Whole body protein synthesis in relation to basal energy expenditure in healthy children and in children recovering from burn injury.
Dynamic aspects of whole body nitrogen metabolism in children recovering from burn injury have been examined in relation to basal metabolic rate (BMR). A continuous administration of [15N]glycine was used to estimate the rates of whole body protein synthesis (S) and breakdown (C) in five acutely burned children (ages 5-16 years) and in nine healthy subjects (ages 9-18 years). S (grams of protein per kg body wt per day) and BMR were significantly correlated (r = +0.73; P less than 0.01). There was no significant correlation of C with BMR. The ratio of S (grams of protein per day) to BMR (kilocalories per day) was the same in burned and healthy children; the mean value for all children was 0.10 +/- 0.03 g protein synthesis/basal kcal. Calorie intake and S were significantly correlated (r = +0.70; P less than 0.01). There were significant correlations between BMR and percentage of total body surface area burned (r = +0.66; P less than 0.01), and BMR and age (r = -0.57; P less than 0.05). Age did not correlate significantly with percent of total body surface burned.


Anti-Gm and anti-Inv antibodies in children during the course of clinical infectious mononucleosis syndrome.
Forty-eight children with the symptoms of infectious mononucleosis were examined. In six of then anti-Gm(1) antibodies were discovered, in two anti-Gm (21), in one child anti-Inv (1).


E2F1 in gliomas: a paradigm of oncogene addiction.
Cancer arises as a result of a stepwise accumulation of genetic changes. One of these changes, deregulation of the Rb/E2F1 pathway resulting from alterations in members of the pathway, is a hallmark of all human cancers. These mutations promote tumor development by deregulating the E2F family of transcription factors, which results in uncontrolled cell cycle progression. The E2F1 protein functions as a transcription factor that enhances cell proliferation by binding to the promoter region of several genes, including those that are involved in cell cycle regulatory activities and DNA replication. It is now becoming clear that the role of E2F1 in regulating transcription and cell growth is also highly dependent on the cellular context. This complexity is also evident from analyses of perturbations in E2F-modulated tumor development. For example, deregulated E2F1 expression can either promote or inhibit tumorigenesis depending on the nature of the other oncogenic mutations that are present. This explains the ability of E2F1 to behave as both an oncogene and tumor suppressor gene. Here we focus on reviewing the most recent evidence supporting the "addiction" of gliomas to this versatile transcription factor. We also consider the clinical relevance of this by examining the role of E2F1 as a prognosis factor and as a target for the development of novel strategies.


Aroclor 1254 alters the binding of 125I-labeled nerve growth factor in PC12 cells.
The effects of Aroclor 1254, a mixture of polychlorinated biphenyls (PCBs), on nerve growth factor (NGF) receptors and neurite outgrowth in PC12 cells were examined. Aroclor 1254 enhanced the NGF-stimulated neurite elongation and decreased the Kd value for binding of 125I-labeled NGF to the high-affinity NGF receptors. The NGF dose-response curve for neurite outgrowth was also shifted to the left in cells pretreated with Aroclor 1254, suggesting that NGF was more potent in the presence of PCBs. Thus, one mechanism by which PCBs may enhance NGF-stimulated neurotrophic effects is in increasing the affinity of binding of NGF to the high-affinity NGF receptors, which are believed to mediate the neurotrophic effects of NGF. The data suggest that PCBs have the potential to influence the NGF neurotrophic system.


Arterial hypertension related to the injection of onabotulinumtoxinA in the detrusor.
We want to report on a first case reported of a 50-year-old female with repetitive and clinical significant hypertension after each injection of onabotulinumtoxinA. This is a retrospective chart review and prospective evaluation of the natriuresis and blood pressure at baseline and after injection therapy. The aim was to explore the mechanism of action of this apparent onabotulinumtoxinA related hypertension. Retrospectively hypertension appeared after 7 days and vanished after 4-5 months following injection of 300 units of onabotulinumtoxinA in the detrusor, bladder symptoms disappeared after 2 weeks and reoccurred after 5 months. Urological, nephrological, cardiological and endocrinological evaluations were normal. In the prospective evaluation a 3-day bladder diary at baseline revealed a bladder capacity of 131 ± 57 ml and at 1 month when full effect was experienced 173 ± 50 ml. At 1 month there were no leakages with six episodes of intermittent catheterization per day. The 24-hour blood pressure registration demonstrated the onset of hypertension at day 7 together with a reversal of the urinary sodium/creatinine ratios on the renal function profile. The increasing natriuresis coinciding with the hypertension is a normal compensatory mechanism suggesting that the hypertension has a central cause rather than it is caused by haematogenous spreading.


Small RNAs from Drosophila KC-H cells. Characterization of nuclear and cytoplasmic 7S subspecies.
The metabolic characteristics and nuclear and cytoplasmic localization of Drosophila 7 S small RNAs were investigated. It was found that although most Drosophila small RNAs show electrophoretic mobilities similar to those of mammalian cells, distinct metabolic behaviour can be detected in three 7 S subspecies (d1, d3 and d4) which were found not to be class III RNAs as reported for mammalian 7 S RNAs. Drosophila d3 and d4 were found to be localized in the nucleus and to be very rapidly labelled minor subspecies turning over with a half-life of less than 6 h. Although 7 S-d1 migrates close to mammalian 7 S-K, it is a major subspecies in Drosophila, is localized mostly in the nucleus and turns over with a half-life of less than 10 h in contrast with mammalian 7 S-K which has been observed to be quite stable.


Magnetic enrichment of antibody-secreting cells.
AMino acid polymer antigens have been covalently coupled to sheep erythrocytes (SRBC) and these have been used in the high gradient magnetic separation (HGMS) technique to enrich murine lymph node cells which had specific receptors for antigen. The method was rapid and provided excellent cell recovery including the fraction retained on the HGMS column. The system could be operated to enrich preformed rosettes, or as an affinity column when preloaded with antigen-coupled SRBC. A 10-fold enrichment of plaque-forming cells was observed in either mode. The HGMS system employed a conventional electromagnet and the fractionation procedure required about 30 min for a sample of up to 10(8) cells.


Silent ischemic heart disease and pericardial fat volume in HIV-infected patients: a case-control myocardial perfusion scintigraphy study.
to determine the prevalence of asymptomatic ischemic heart disease (IHD) in HIV patients by myocardial perfusion scintigraphy (MPS) and to determine the value of coronary artery calcium score (CACS), carotid intima-media thickness (cIMT) and pericardial fat volume as screening tools for detection of IHD in subjects with HIV. Patients with HIV seem prone to early development of IHD. 105 consecutive HIV patients (mean age 47.4 years; mean duration of HIV 12.3 years; mean CD4+ cell count 636×10⁶/L; all receiving antiretroviral therapy) and 105 controls matched for age, gender and smoking status, without history of IHD were recruited. MPS, CACS, cIMT, pericardial fat volume, and cardiovascular risk scores were measured. HIV patients demonstrated higher prevalence of perfusion defects than controls (18% vs. 0%; p<0.001) despite similar risk scores. Of HIV patients with perfusion defects, 42% had a CACS = 0. CACS and cIMT were similar in HIV patients and controls. HIV patients on average had 35% increased pericardial fat volume and increased concentration of biomarkers of atherosclerosis in the blood. HIV patients with myocardial perfusion defects had increased pericardial fat volume compared with HIV patients without perfusion defects (314±43 vs. 189±12 mL; p<0.001). HIV patients had an increased prevalence of silent IHD compared to controls as demonstrated by MPS. The finding was strongly associated with pericardial fat volume, whereas cardiovascular risk scores, cIMT and CACS seem less useful as screening tools for detection of myocardial perfusion defects in HIV patients.


Systolic left ventricular apical bulging after biventricular pacing mimicking takotsubo cardiomyopathy.
Resolution of the issue of nonresponsiveness to cardiac resynchronization therapy (CRT) remains crucial to the successful treatment of conduction disturbances in heart failure. In this study, a patient with refractory heart failure including left bundle branch block was treated via surgical CRT. The epicardial left ventricular (LV) lead, implanted using thoracoscopic guidance, was unexpectedly located on the apical side. Echocardiographic findings of the LV motion mimicked takotsubo cardiomyopathy. The LV lead was successfully re-implanted along a lateral branch of the cardiac vein using an endovascular approach, resulting in restored contractility and reversal of the LV remodeling.


Impact of cervical intraepithelial neoplasia diagnosis and treatment on self-esteem and body image.
Most women commonly take an abnormal Pap smear to be indicative of invasive cancer. This fear can permeate their lifestyle to the degree that it may influence receptivity to medical procedures as well as promote cervical intraepithelial neoplasia (CIN) to invasive squamous cell carcinoma of the cervix. This longitudinal study was undertaken to provide baseline data on self-esteem and body image as the patient progresses from the initial visit through the postsurgery visit. Twenty patients ranging in age from 15 to 40 years were followed through diagnosis and treatment with colposcopic biopsy showing 25% CIN I, 45% CIN II, and 30% CIN III. Concern for cancer overrode all other concerns except during the postsurgery visit, at which time loss of attractiveness was paramount. Loss of sexual functioning ranked high in all visits. Self-esteem was lowest and anxiety highest during the initial and postsurgery visits. Positive body image was greatest when the results of the biopsy were received and during the postsurgery visit. Patients and sexual partners exhibited similar patterns of anxiety about the medical condition. Psychological repercussions from CIN occurred even prior to diagnosis.


Vaccination of wildlife against rabies: successful use of a vectored vaccine obtained by recombinant technology.
The impact of recombinant technology in veterinary and human medicine can only be hypothesized at this time. The development of vaccines and other biological products that go beyond the abilities of conventional products demonstrates the benefits of this new technology. Raboral V-RG was developed as an alternative rabies vaccine with the novel attribute of being effective by the oral route. Within 10 years after its first application as an experimental vaccine in European, red foxes it developed into a useful tool and is being used to curtail rabies epizootics in three wildlife species in the United States. The use of this vaccine can be considered as monumental in contributing to the control of rabies in species that were at one time considered to be incapable of vaccination in large-scale campaigns.


Developmental toxicity of orally administered technical dimethoate in rats.
Dimethoate (O,O-dimethyl-S-(N-methylcarbamoyl-methyl) phosphorodithioate), an organophosphate insecticide, was examined for its potential to produce developmental toxicity in rats after oral administration. Pregnant Fischer 344 rats were given sublethal doses of 0 (corn oil), 7, 15, and 28 mg/kg/day dimethoate by gavage on gestation days (GD) 6-15. Maternal effects in 15 and 28 mg/kg/day dose groups included cholinergic signs such as tremors, diarrhea, weakness, and salivation, and depression in the maternal and fetal brain acetylcholinesterase (AChE) activities. Other maternal toxicity that included reduction in body weight and feed consumption was observed only in the treated group of 28 mg/kg/day. No maternal toxicity was apparent in the 7 mg/kg/day dose group. Maternal exposure to dimethoate during organogenesis significantly affected the number of live fetuses, early resorption, and mean fetal weight in the 28 mg/kg/day dose group. No external, visceral, and skeletal abnormalities were observed in any of the treated groups compared to the control. On the basis of the present results dimethoate can produce clinical signs of toxicity and significant inhibition of the maternal and fetal AChE activities in dose groups of 15 and 28 mg/kg/day and showed fetotoxicity without teratogenic effects at 28 mg/kg/day.


Johne's disease in Irish dairy herds: considerations for an effective national control programme.
The Irish dairy industry has established a reputation for the production of safe and healthy dairy products and is seeking to further expand its export market for high value dairy products. To support its reputation, stakeholders aim to control Johne's disease. To assist decision-makers determine the most appropriate design for an Irish programme, a narrative review of the scientific literature on the epidemiology of Johne's disease, and selected control programmes throughout the world was undertaken. Two modelling studies specifically commissioned by Animal Health Ireland to assess testing methods used to demonstrate confidence of freedom in herds and to evaluate a range of possible surveillance strategies provided additional information. The majority of control programmes tend to be voluntary, because of the unique epidemiology of Johne's disease and limited support for traditional regulatory approaches. While acknowledging that test performance and sub-clinical sero-negative shedders contributes to the spread of infection, a range of socio-political issues also exist that influence programme activities. The paper provides a rationale for the inclusion of a Veterinary Risk Assessment and Management Plan (VRAMP), including voluntary whole herd testing to identify infected herds and to support assurance-based trading through repeated rounds of negative testing, national surveillance for herd-level case-detection, and improved understanding of biosecurity management practices. Identification and promotion of drivers for industry and producer engagement in Ireland is likely to guide the future evolution of the Irish Johne's Control Programme (IJCP) and further enhance its success. The provision of training, education and extension activities may encourage farmers to adopt relevant farm management practices and help them recognize that they are ultimately responsible for their herd's health and biosecurity.


Cushing syndrome secondary to ectopic adrenocorticotropic hormone secretion from a Meckel diverticulum neuroendocrine tumor: case report.
Ectopic production of adrenocorticotropic hormone (ACTH) by neuroendocrine tumours (NET) is a rare condition, occult presentations often hampering the diagnosis. Although NET are relatively frequent in the ileon and Meckel diverticulum, we describe the first Cushing's syndrome due to ectopic adrenocorticotropic syndrome (CS-EAS) arising from a Meckel diverticulum. A 44-year-old man was admitted with recent onset of diabetes, myopathy, edema and hypokalemic metabolic alkalosis consistent with Cushing's syndrome. Both basal and dynamic laboratory evaluation suggested CS-EAS. Laboratory testing also showed high serum levels of chromogranin A (CgA) and urinary 5-hydroxyindoleacetic acid (5HIAA). Pituitary and neck/thorax/abdomen/pelvis imaging proved to be normal, while somatostatin analogue ((99m)Tc-HYNIC-TOC) scintigraphy revealed increased focalized ileum uptake on the right iliac fossa. Pre-operative ketoconazole and sandostatin treatment controlled the hypercortisolism within a month. Pathological analysis of the resected submucosal 1.8 cm tumour of the Meckel diverticulum and a metastatic local lymph node confirmed a well differentiated neuroendocrine tumour (grade I), whereas immunohistochemistry was positive for ACTH, chromogranin A and synaptophysin. Post-operative clinical and biochemical resolution of Cushing's syndrome was followed by normalization of both CgA and 5HIAA, which were maintained at the 6 month follow-up. The identification, characterization and follow-up of this rare cause of ectopic ACTH secretion is important in order to assess the long-term prognostic and management.


Insight into mycoviruses infecting Fusarium species.
Most of the major fungal families including plant-pathogenic fungi, yeasts, and mushrooms are infected by mycoviruses, and many double-stranded RNA (dsRNA) mycoviruses have been recently identified from diverse plant-pathogenic Fusarium species. The frequency of occurrence of dsRNAs is high in Fusarium poae but low in other Fusarium species. Most Fusarium mycoviruses do not cause any morphological changes in the host but some mycoviruses like Fusarium graminearum virus 1 (FgV1) cause hypovirulence. Available genomic data for seven of the dsRNA mycoviruses infecting Fusarium species indicate that these mycoviruses exist as complexes of one to five dsRNAs. According to phylogenetic analysis, the Fusarium mycoviruses identified to date belong to four families: Chrysoviridae, Hypoviridae, Partitiviridae, and Totiviridae. Proteome and transcriptome analysis have revealed that FgV1 infection of Fusarium causes changes in host transcriptional and translational machineries. Successful transmission of FgV1 via protoplast fusion suggests the possibility that, as biological control agents, mycoviruses could be introduced into diverse species of fungal plant pathogens. Research is now needed on the molecular biology of mycovirus life cycles and mycovirus-host interactions. This research will be facilitated by the further development of omics technologies.


Series of sums of products of higher-order Bernoulli functions.
It is shown in a previous work that Faber-Pandharipande-Zagier's and Miki's identities can be derived from a polynomial identity, which in turn follows from the Fourier series expansion of sums of products of Bernoulli functions. Motivated by and generalizing this, we consider three types of functions given by sums of products of higher-order Bernoulli functions and derive their Fourier series expansions. Moreover, we express each of them in terms of Bernoulli functions.


A quantile regression estimator for interval-censored data.
We describe an estimating equation that can be used to fit quantile regression models to interval-censored data. The proposed estimator presents important advantages over the existing methods, and can be applied when the data are a mixture of interval-censored, left-censored, and right-censored observations. We describe estimation and inference, report simulation results, and apply the proposed method to analyze the Signal Tandmobiel® data. The necessary R code has been incorporated in the existing R package c t q r .


Analysis of B-cell subpopulations. I. Relationships among splenic xid, Ly 1+, and Lyb 5+ B cells.
The phenotypic and functional relationships among the various B-cell subsets is of importance for better understanding studies of the immune system. In this report, the realm of B cells encompassed by Lyb 5, Ly 1, and xid has been examined through the use of the alloantiserum anti-Lyb 5, the unique functional properties of Ly 1+ B cells, and the spontaneous autoantibody producing congenic xid mice, NZB.xid. By functional and phenotypic analysis, we have shown that (i) Lyb 5- B cells and B cells of xid mice are largely, but not completely, overlapping; (ii) xid spleen cells contain a population which is Lyb 5+; (iii) Ly 1+ B cells fall largely in the Lyb 5+ compartment; and (iv) the autoantibody-producing Ly 1+ B cells are predominantly Lyb 5+.


[Molecular genetic investigation of sugar beet (Beta vulgaris L.)].
Molecular genetic studies of sugar beet (Beta vulgaris L.) are reviewed as a basis for the development of genomics of this species. The methods used to study structural and functional genomics are considered. The results and their application to increase the efficiency of sugar beet breeding are discussed.


Assessment of the modulating effects of fructo-oligosaccharides on fecal microbiota using human flora-associated piglets.
We first used human flora-associated (HFA) piglets, a significantly improved model for research on human gut microbiota, to study the effects of short-chain fructo-oligosaccharides (scFOS) on the gut bacterial populations. Ten neonatal HFA piglets were assigned to receive basal diets alone or supplemented with scFOS (0.5 g/kg body weight/day) from 1 to 37 days after birth (DAB). The impact of scFOS on the fecal bacterial populations of the piglets before (12 DAB), during (17 DAB), and after (25 and 37 DAB) weaning were monitored by PCR-denaturing gradient gel electrophoresis and real-time quantitative PCR. The Bifidobacterium genus was stimulated consistently, except during weaning, confirming the bifidogenic property of scFOS. At 12 DAB, the Clostridium leptum subgroup was decreased and two unknown Bacteroides-related species were increased; at 25 DAB, the C. leptum subgroup and Subdoligranulum variabile-like species were elevated, whereas one unknown Faecalibacterium-related species was suppressed; and at 37 DAB, the Bacteroides genus was decreased. The results showed that effects of scFOS on non-bifidobacteria varied at different developmental stages of the animals, warranting further investigation into the host-development-related effects of prebiotics on the gut microbiota and the host physiology using the HFA piglets as a model for humans.


Mismatch Discrimination and Efficient Photomodulation with Split 10-23 DNAzymes.
DNA enzymes (DNAzymes) that catalyze the degradation of complementary RNA molecules have been investigated for many biochemical and sensing applications. Here, we investigated a 10-23 DNAzyme that has been shown previously to possess cellular activity. We determined that it has very low Mg(2+) ion dependence, with DNAzyme activity observed at [Mg(2+)] = 0.01 mM. This metal ion dependence is much lower than is typical for DNAzymes studied to date, and suggests that DNAzymes may be engineered for many additional biological applications. Recently, we demonstrated that this 10-23 DNAzyme can be divided into two parts, which assemble into an active oligonucleotide complex. We investigated in more detail the functionality of the split 10-23 DNAzyme and found that dividing the 15-nucleotide catalytic loop after the 7(th) or 8(th) base maximized its activity. The split DNAzymes required higher metal ion concentrations ([Mg(2+)] = 5 mM), and as we anticipated due to their lower hybridization activity, the split enzymes had the advantage of being more sensitive to single base mismatches in the DNAzyme-RNA duplex. Finally, we demonstrated facile photomodulation of split DNAzyme activity by incorporating a photocleavable biotin moiety bound to streptavidin.


Formic acid poisoning: case report and in vitro study of the hemolytic activity.
A case of fatal oral poisoning with formic acid resulting in shock, metabolic acidosis, and hemolysis is reported. The formic acid concentration on admission was 348 micrograms/mL, which, together with an increase in lactic acid, contributed to the metabolic acidosis. Because it has been suggested in the literature that formic acid might induce hemolysis via a direct cytotoxic action on the RBCs, an in vitro study was performed using human RBCs in saline, phosphate buffered saline, and plasma in order to define the mechanism of the hemolysis. These experiments indicate that the hemolysis is not a cytotoxic effect of formic acid but is related to the degree of acidity in itself.