Reactivity of neonicotinoid insecticides with carbonate radicals.
The reaction of three chloronicotinoid insecticides, namely Imidacloprid (IMD), Thiacloprid (THIA) and Acetamiprid (ACT), with carbonate radicals (CO·₃⁻) was investigated. The second order rate constants (4 ± 1) × 10⁶, (2.8 ± 0.5) × 10⁵, and (1.5 ± 1) × 10⁵ M⁻¹ s⁻¹ were determined for IMD, THIA and ACT, respectively. The absorption spectra of the organic intermediates formed after CO·₃⁻ attack to IMD is in line with those reported for α-aminoalkyl radicals. A reaction mechanism involving an initial charge transfer from the amidine nitrogen of the insecticides to CO·₃⁻ is proposed and further supported by the identified reaction products. The pyridine moiety of the insecticides remains unaffected until nicotinic acid is formed. CO·₃⁻ radical reactivity towards IMD, ACT, and THIA is low compared to that of HO• radicals, excited triplet states, and ¹O₂, and is therefore little effective in depleting neonicotinoid insecticides.


Severe early childhood caries and social determinants in three-year-old children from Northern Thailand: a birth cohort study.
The purpose of this study was to investigate the prevalence and social risk factors of severe early childhood caries in three-year-old children in Northern Thailand, using a birth-cohort study The data utilized in this study were from the prospective cohort study of Thai children (PCTC) from the 28 to 38 weeks gestational age until the children reached the age of 36 months (N = 597) in Mueang Nan district, Northern Thailand. Questionnaires were administered at different time points and dental examination was conducted at the age of 3 years of the child. 44.1% of the 3 year old children had S-ECC. In multivariate logistic regression analysis, environmental factors (the use of rain or well water as drinking water, no schooling of mother of child, being male), and risk behaviour (sleeping with a bottle at 30 months) were associated with S-ECC. Further, in bivariate analysis, psychological distress in the mother, lack of spousal relationship support, suckle to sleep when going to bed, introduction of soft drinks at 12 months, having had more frequently sweet food, and less than daily tooth brushing before 30 months were associated with S-ECC. A very high rate of S-ECC was observed, and oral health may be influenced by social factors.


Identification and characterization of microbial proteins using peptide mass fingerprinting strategies.
Peptide mass fingerprinting is a simple, quick, cheap, and relatively effective method of identifying proteins from mass spectrometry data. Proteins extracted from the complex mixture comprising the proteome of a sample are individually digested with a proteolytic enzyme into a series of peptide fragments. The set of masses of these peptides, determined by mass spectrometry, form a peptide mass fingerprint of the protein. Comparison of this experimental fingerprint with the theoretical fingerprints of all known protein sequences for this organism, derived computationally from a protein sequence database, allows the identification of the particular protein. In this chapter, I discuss the technique including preparation for the peptide mass fingerprinting analysis, the appropriate selection of computational search parameters, and the analysis and interpretation of search results in the context of identifying proteins from microbial samples.


Dose-dependent cysteine-mediated protection of insulin-producing cells from damage by hydrogen peroxide.
Oxidative damage is believed to play a key role in the process of pancreatic beta cell destruction leading to type 1 diabetes. The beta cells are sensitive to oxidative stress because their intracellular anti-oxidative defence mechanisms are weak. The defence mechanisms depend heavily on glutathione, the synthesis of which is dependent on the availability of cysteine. We investigated whether an increased amount of cysteine available could protect beta cells from oxidative damage. Rat insulinoma cells (RINm5F) were exposed to 50 or 100 microM hydrogen peroxide in the presence of three different cysteine concentrations (0.1, 1 and 5mM). Cell viability was analyzed by vital staining and the cellular metabolic status by C,N-diphenyl-N'-4,5-dimethyl thiazol-2-yl tetrazolium bromide (MTT) analysis. Intracellular insulin, DNA and glutathione contents were measured. The mechanism of death was further clarified by gel electrophoretic DNA fragmentation analysis. Hydrogen peroxide decreased cell viability and induced functional impairment. Vital staining indicated that 1mM cysteine effectively protected the cells. The protective effect was confirmed by the MTT assay showing preserved metabolic integrity, and by measurements of intact intracellular insulin and DNA content. Cysteine increased intracellular glutathione. Gel electrophoretic analysis of DNA revealed hydrogen peroxide-induced apoptotic fragmentation. This was also abolished by 1mM cysteine. The therapeutic window of cysteine was narrow: 0.1mM cysteine provided inadequate protection, and 5mM cysteine was already toxic in this setting. A proper dose of cysteine could provide a safe and effective means to protect beta cells from oxidative damage.


Activation of lymphocytes of normal and tumor bearing mice by mangiferin, a naturally occurring glucosylxanthone.
Mangiferin, a naturally occurring glucosylxanthone, was assessed for its immunomodulatory potential. The phytochemical induced extensive in vitro proliferation of murine splenocytes and thymocytes at the doses of 5-40 micrograms ml-1. Suppression of the proliferative response of the cells was observed with higher doses of mangiferin. Mangiferin also activated the splenocytes of tumor hosts at early and late stages of tumor growth. The Phytohemagglutinin (PHA) and Con A unresponsive splenocytes of advanced tumor bearer proliferated extensively in response to mangiferin. Mangiferin when used with Con A produced additive stimulatory effect and induced heightened DNA synthesis of normal and advanced tumor bearers' splenocytes.


Utilization of cytogenetic biomarkers as a tool for assessment of radiation injury and evaluation of radiomodulatory effects of various medicinal plants - a review.
Systematic biological measurement of "cytogenetic endpoints" has helped phenomenally in assessment of risks associated with radiation exposure. There has been a surge in recent times for the usage of radioactive materials in health care, agriculture, industrial, and nuclear power sectors. The likelihood of radiation exposure from accidental or occupational means is always higher in an overburdened ecosystem that is continuously challenged to meet the population demands. Risks associated with radiation exposure in this era of modern industrial growth are minimal as international regulations for maintaining the safety standards are stringent and strictly adhered to, however, a recent disaster like "Fukushima" impels us to think beyond. The major objective of radiobiology is the development of an orally effective radio-modifier that provides protection from radiation exposure. Once available for mass usage, these compounds will not only be useful for providing selective protection against accidental and occupational radiation exposure but also help to permit use of higher doses of radiation during treatment of various malignancies curtailing unwarranted adverse effects imposed on normal tissues. Bio-active compounds isolated from natural sources enriched with antioxidants possess unique immune-modulating properties, thus providing a double edged benefit over synthetic radioprotectors. We aim to provide here a comprehensive overview of the various agents originating from plant sources that portrayed promising radioprotection in various experimental models with special emphasis on studies that used cytogenetic biomarkers. The agents will include crude extracts of various medicinal plants, purified fractions, and herbal preparations.


Morphological changes in dendritic spines of Purkinje cells associated with motor learning.
Experience-dependent changes of spine structure and number may contribute to long-term memory storage. Although several studies demonstrated structural spine plasticity following associative learning, there is limited evidence associating motor learning with alteration of spine morphology. Here, we investigated this issue in the cerebellar Purkinje cells using high voltage electron microscopy (HVEM). Adult rats were trained in an obstacle course, demanding significant motor coordination to complete. Control animals either traversed an obstacle-free runway or remained sedentary. Quantitative analysis of spine morphology showed that the density and length of dendritic spines along the distal dendrites of Purkinje cells were significantly increased in the rats that learned complex motor skills compared to active or inactive controls. Classification of spines into shape categories indicated that the increased spine density and length after motor learning was mainly attributable to an increase in thin spines. These findings suggest that motor learning induces structural spine plasticity in the cerebellar Purkinje neurons, which may play a crucial role in acquiring complex motor skills.


Septic arthritis in childhood.
We studied retrospectively the pattern of septic arthritis in childhood at a major municipal hospital during a ten-year period. Hemophilus influenzae was the most common organism in septic arthritis in patients less than two years old and was associated with upper respiratory tract infections in nine of 12 patients (75%). Staphylococcus aureus was seen in seven of eight (87.5%) children above the age of five and was associated with history of trauma. All patients were black. Despite the high incidence of sickle cell disease in our hospital population, not one patient had sickle cell disease.


Head movement: a novel serotonin-sensitive behavioral endpoint for tail suspension test analysis.
The tail suspension test (TST) as an antidepressant and depression-related behavior screen, has many advantages over the forced swim test (FST) in terms of procedural simplicity and consistent SSRI response. However, the FST has traditionally offered more specific neuromodulatory information by differentiating between serotonin (5-HT) and norepinephrine sensitive behavior categories. Head movement is a newly characterized behavior endpoint in the FST and TST with a selective 5-HT sensitivity. In this investigation, we show that the baseline and drug response profile of head movement previously found in the 129S6 strain of mice (Lockridge et al., 2010) is reproducible in the C57 strain. Head movement is inversely correlated to FST swimming and elevated in the TST by SSRI administration. The use of a weighted bin sample analysis method differentiates TST behaviors into fluoxetine-responsive head movement and desipramine-responsive struggling. The use of 5-HT subtype receptor agonists, after depleting endogenous 5-HT with pCPA, shows the head movement suppressing effect of 5-HT2A and 5-HT2C postsynaptic receptor activation. 5-HT1A and 5-HT1B agonists were ineffective. We propose that a head movement focused analysis can add sensitive and reliable 5-HT detection capability to mouse TST testing with minimal effort but significant reward.


The transverse vaginal septum: a clinicopathologic evaluation.
Twelve cases of transverse vaginal septum were clinically and pathologically evaluated. Seven cases were treated with electrosurgical excision of the septum with encouraging results. Histologic findings in the patients in this study support the view that the epithelium of the vagina and the transverse septum are of mesonephroid origin.


Specific responses to two unrelated antigens in mice made orally tolerant to one of them.
BDF1 and B10 mice, either normal or exclusively fed egg white diluted in water, were immunized with ovalbumin (OVA), sheep erythrocytes (SRBC) or OVA + SRBC. BDF1, but not B10, mice became tolerant to OVA and presented a 100% increase in the total number of immunoglobulin-secreting cells in the spleen. Immunization with OVA + SRBC markedly increased the response to OVA, except in tolerant BDF1 mice, and had no effect on the anti-SRBC PFC response, except in normal (non-tolerant) BDF1 mice.


Kinetic behavior and reversible inhibition of monoamine oxidases--enzymes that many want dead.
Monoamine oxidase (MAO) inhibitors have proven to be valuable tools in pharmacology and therapeutics. This account concerns the behavior of the different types of reversible inhibitor and how an understanding of the kinetic mechanisms of MAO may help in their design.


Nesfatin-1 ameliorates type-2 diabetes-associated reproductive dysfunction in male mice.
The present study was aimed to demonstrate the recuperative effect of nesfatin-1 on testicular dysfunction in the high-fat diet (HFD)/streptozotocin (STZ)-induced type-2 diabetes mellitus (T2DM) mice. Three experimental groups were formed: (1) vehicle control (VC), (2) T2DM mice, (3) T2DM + nesf-1. The mice with blood glucose level higher than 300 mg/dL following HFD and a single dose of STZ were used for the experiment. The T2DM mice showed increases in body mass, blood glucose and insulin levels, reductions in spermatogenesis and steroidogenesis, production of antioxidative enzymes, and disturbed lipid profile. These alterations were all ameliorated by administration of nesfatin-1 at 20 μg/Kg BW for 15 days. Nesfatin-1 treatment also increased the production of testosterone (T), improved insulin sensitivity, and effectively ameliorated the testicular aberrations, and increased spermatogenesis and steroidogenesis. In addition, nesfatin-1 treatment upregulated the PCNA and Bcl2 expression and inhibited the caspase-3 and prohibitin expression in T2DM mice. Nesfatin-1 increased insulin receptor (IR) and GLUT8 expressions, and lactate production, the changes that further substantiate the increase of energy influx to the testis. Altogether, the results suggest the ameliorative effect of nesfatin-1 against T2DM-associated testicular dysfunctions and improved insulin sensitivity along with promoting T production and fertility in T2DM mice.


Mesencephalic astrocyte-derived neurotrophic factor alleviates non-alcoholic steatohepatitis induced by Western diet in mice.
Excessive lipid accumulation, inflammation, and fibrosis in the liver are the major characteristics of non-alcoholic steatohepatitis (NASH). Mesencephalic astrocyte-derived neurotrophic factor (MANF) plays an important role in metabolic homeostasis, raising the possibility that it is involved in NASH. Here, we reduced and increased MANF levels in mice in order to explore its influence on hepatic triglyceride homeostasis, inflammation, and fibrosis during NASH progression. The MANF expression was decreased in Western diet-induced NASH mice. In vivo, liver-specific MANF knockout exacerbated hepatic lipid accumulation, inflammation, and fibrosis of mice induced by Western diet, while liver-specific MANF overexpression mitigated these NASH pathogenic features. In vitro, knocking down MANF in primary hepatocyte cultures aggravated hepatic steatosis and inflammation, which MANF overexpression markedly attenuated. Studies in vitro and in vivo suggested that MANF regulated hepatic lipid synthesis by modulating SREBP1 expression. Inhibiting SREBP1 in primary hepatocytes blocked lipid accumulation after MANF knockdown. MANF overexpression reversed LXRs agonist GW3965 induced SREBP1 and LIPIN1 expression. MANF decreased the expression of pro-inflammatory cytokines by inhibiting NF-κB phosphorylation. These results suggest that MANF can protect against NASH by regulating SREBP1 expression and NF-κB signaling.


A comparison of fast and slow depolarizations evoked by 5-HT in guinea-pig coeliac ganglion cells in vitro.
1. 5-Hydroxytryptamine (5-HT) was applied by pressure ejection to coeliac ganglion cells of the guinea-pig maintained in vitro and responses measured intracellularly. 2. Cells responded in one of three ways to 5-HT: by (a) a fast, transient depolarization (43%), (b) a fast transient followed by a slow depolarization (biphasic response, 30%) or (c) a slow sustained depolarization (25%). 3. Fast depolarizations (response (a) above] were graded according to the duration of the ejection pulse. Maximal responses had a mean amplitude of 12 +/- 0.8 mV, a duration of 6.4 +/- 1.0 s, a latency of 0.4 +/- 0.1 s, were associated with a fall in membrane input resistance, increased in amplitude by hyperpolarization and probably mediated by an increased conductance to Na and K. The estimated reversal potential was -22.8 +/- 2.4 mV (n = 14). The maximal fast response seen in biphasically-responding cells (b) appeared similar to fast response (a). 4. Fast depolarizations (a) showed marked tachyphylaxis and were abolished by superfusion of the ganglion with 5-HT (100 microM). They were reduced in amplitude by tubocurarine (10-100 microM, pIC50 4.4), MDL 72222 (1-5 microM, pIC50 5.8), quipazine (1 microM reduced responses by 65 +/- 15%, n = 3), ICS 205-930 (1 microM reduced responses by 64 +/- 14%, n = 7) and metoclopramide (10 microM reduced responses by about 45%), but were unafected by methysergide (up to 1 microM) or hexamethonium (up to 1 mM). 5. Slow depolarizations (c) varied in amplitude with the duration of the ejection pulse. Maximal responses had a mean amplitude of 6.4 +/- 0.7 mV, a duration of 62 +/- 6 s, a latency of 3.5 +/- 0.8 s and were reduced in amplitude by methysergide (0.1-1 microM, pIC50 6.5) but not by MDL 72222 (1 microM). The maximal slow component in biphasically-responding cells (b) was similar in amplitude and duration to slow response (c), was partially blocked by methysergide (1-5 microM) in 4 of 6 cells and was enhanced by tubocurarine (50 microM) which reduced the fast component. 6. Slow depolarizations (b,c) were associated with either a small reduction or no change in membrane input resistance depending on the cell studied. Hyperpolarization had variable effects on slow depolarization amplitude. 7. It was concluded that the fast, phasic depolarization is mediated by an ionic mechanism and by receptors both of which are distinct from those involved in the slow depolarization. The receptor mediating the fast depolarization is a 5-HT3 receptor while that mediating the slow depolarization has yet to be identified.


A patient with jejunostomy liberated from home intravenous therapy after 14 years; contribution of balance studies.
A patient with a jejunostomy 100 cm from the duodeno-jejunal flexure, following surgery for Crohn's disease, had needed parenteral fluids at home for 14 years because of a negative intestinal balance of sodium. Measurements were made of her oral intake and intestinal output during study periods each of 2 days. Loperamide 4 mg QDS, codeine phosphate 60 mg QDS and both together put her into positive intestinal fluid balance but sodium balance remained negative. Both drugs used together were more effective than either used alone. Ranitidine 300 mg BD made no significant difference to her intestinal output. 1 litre of a glucose-electrolyte solution (120 mmol sodium) sipped during the day resulted in sodium balance, but only with the addition of loperamide and codeine phosphate was positive sodium balance achieved (mean 44 mmol/day). This therapy allowed her to dispense with parenteral fluids which have been stopped for the last year.


Bowel function after transanal endorectal pull-through for Hirschsprung disease - does outcome improve over time?
The reported functional outcome in patients operated with transanal endorectal pull-through (ERPT) for Hirschsprung disease (HD) varies greatly. Some studies suggest better functional outcome in older than in younger HD patients, but there are almost no longitudinal studies that have demonstrated such improvement. Therefore, we aimed to compare functional outcome in a cohort of HD patients over time to assess whether bowel function improves with increasing age. Functional outcome in HD patients operated with ERPT from 1998 to 2007 was recorded by standardized interviews by an independent investigator during 2008-2011 and again in 2017/2018. Bowel function was assessed using the Krickenbeck questionnaire. Clinical data were collected prospectively. 50 of the original 62 patients responded for a second interview. Median age at the two interviews was 8,1 (3,4-16,6) and 15,4 (9,9-25) years respectively. There was no difference in the rate of soiling at first (52%) and second (52%) follow-up. Constipation was reported in 20% of the patients at first, and in 24% at second follow-up. Bowel management was used by 30% and 32% at first and second interview respectively. Soiling and constipation are common in HD patients several years after surgery, and no improvement of bowel function with increasing age could be demonstrated. Level II.


Human soluble TRAIL/Apo2L induces apoptosis in a subpopulation of chemotherapy refractory nodal diffuse large B-cell lymphomas, determined by a highly sensitive in vitro apoptosis assay.
Resistance to chemotherapy in therapy-refractory diffuse large B-cell lymphomas (DLBCL) is related to inhibition of the intrinsic apoptosis pathway. Human soluble tumour necrosis factor (TNF)-related apoptosis-inducing ligand (hsTRAIL/Apo2L) induces apoptosis via the alternative, death-receptor mediated apoptosis pathway and might be an effective alternative form of therapy for these lymphomas. This study investigated whether hsTRAIL/Apo2L could actually induce apoptosis in isolated lymphoma cells of DLBCL biopsies of patients with chemotherapy-refractory DLBCL. Twelve out of a total of 22 DLBCL samples were sensitive to hsTRAIL/Apo2L. These sensitive lymphomas included seven clinically chemotherapy-refractory lymphomas. Furthermore, hsTRAIL/Apo2L induced apoptosis in DLBCL cells and in B-cell lines that showed high expression levels of inhibitors of the intrinsic apoptosis pathway: Bcl-2 and/or X-linked inhibitor of apoptosis (XIAP). hsTRAIL/Apo2L-sensitive lymphoma cells showed expression of the TRAIL receptors R1 and/or R2 and absence of R3 and R4. We conclude that hsTRAIL/Apo2L induced apoptosis in a subpopulation of chemotherapy-refractory nodal DLBCL and that disruption of the intrinsic apoptosis-mediated pathway and expression of Bcl-2 and XIAP did not confer resistance to hsTRAIL/Apo2L-induced apoptosis in DLBCL. Thus, based on our results, further exploration of hsTRAIL/Apo2L as an alternative treatment for patients with chemotherapy-refractory DLBCL should be considered.


Pulmonary response following intratracheal instillation of potroom dust from an aluminum reduction plant into rat lung.
Rats were examined for lung changes 7 days and 22 weeks after a single intratracheal instillation of potroom dust from an aluminum reduction plant. In the acute study, 0.5 mg potroom dust induced changes at the pulmonary surface, and a 5.0-mg dose caused marked changes both at the pulmonary surface and in lung tissue. By contrast, 5.0 mg virginal alumina caused only a mild increase in the number of polymorphonuclear leukocytes in the lung. In the long-term study, potroom dust, virginal alumina, and DQ12 alpha-quartz were instilled at a dose of 5.0 mg. In the cases of potroom dust and virginal alumina, all parameters measured had returned to control levels after 22 weeks, whereas DQ12 induced a massive inflammation and an accumulation of pulmonary surfactant. It is concluded that this air sample of potroom dust causes marked acute irritation and inflammation in rat lung and that this type of dust may therefore be active in human lung and thus contribute to the acute respiratory symptoms experienced by some potroom workers. However, despite the marked acute toxicity of the comparatively high 5.0-mg dose, rat lung appears to return to normal within 22 weeks with no signs of chronic toxicity from this single instillation.


Interactive regulation of grass yield and chemical properties by defoliation, a salivary chemical, and inorganic nutrition.
Clones of 2 C4 grass species, Sprobolus ioclados and S. pyramidalis, were obtained from more and less heavily grazed grasslands, respectively, in Tanzania's Serengeti National Park. Plants were grown in a factorial experiment to determine the effects of severe defoliation, nutrient limitation, and a salivary chemical (thiamine) on plant growth, nitrogen content, and non-structural carbohydrate content. The experimental design included: (1) species; (2) clipping, with plants either unclipped or clipped weekly to a height of 5 cm; (3) 0.2 ml of distilled water of 0.2 ml of 10 ppb aqueous thiamine sprayed on plants from an atomizer after clipping and identical treatments at the same time to unclipped plants; (4) phosphorus (P) at 0.2 or 1 mM; (5) nitrogen (N) at 3 or 15 mM. Clipping was the major variable affecting plant growth. Total and litter yields were reduced to half and residual plant yield was reduced to 30% of the values for unclipped plants. Clipping interacted strongly with other variables since they commonly had minor effects on clipped plants and major effects on unclipped plants. Exceptions to this generalization were generally due to better performance by S. ioclados under clipping. Compared to lower treatment levels, higher treatment levels promoted total yield of unclipped plants by 52% for N, 43% for thiamine, and 33% for P. In general, thiamine had greater effects than P but lesser effects than N. Thiamine promoted yield and modified the chemical balance of plants by promoting carbohydrate (CHO) concentrations and reducing N concentrations. N and P deficiencies promoted CHO accumulation. Clipping promoted the N of leaves and crowns and reduced the N levels in roots. Leaf blade water and N contents were positively correlated with very little scatter. The slope of the line was different for S. ioclados and S. pyramidalis. Leaf blade water and CHO contents were negatively related but there was more scatter and the species could not be distinguished. The species from more heavily grazed grasslands was conspicuously more sensitive to thiamine application. The results indicated that leaf treatment with thiamine, the only likely source of which in natural grasslands is saliva deposited by feeding herbivores, can have major effects on plant yield and metabolic balances at very low application levels. But under defoliation levels as severe as those imposed in this experiment, which reduced above ground plant biomass to a fourth of the level produced by unclipped plants, growth was so strongly limited by defoliation that neither thiamine nor inorganic nutrients affected plant yield residual from clipping. Therefore, whether chemicals such as thiamine that may be introduced onto grass foliage by grazing ungulates and other herbivores will influence the growth of grazed plants will depend upon the grazing intensity associated with the transfer.


Always on alert: How relatives of family members with dementia experience the transition from home to permanent nursing home placement.
Our aim was to gain insight into how the relatives of family members with dementia have experienced their family members' transition from home to permanent nursing home placement. Our research was a qualitative case study. Inspired by Kvale and Brinkmann's phenomenological-hermeneutic approach, we performed the thematic analysis of semi-structured interviews describing how the relatives of family members with dementia have experienced the dementia trajectory. Twelve relatives of family members with dementia living in nursing homes agreed to participate. The descriptions of relatives' experiences indicated four primary themes: mismatches between healthcare services and the family members' needs, witnessing the family members' cognitive decline, the effects of the experiences on the relatives' health and well-being and what matters to the relatives during their family members' transition to nursing home. Mismatches between available resources in the municipalities and the family members' healthcare needs were the most distressing experiences, along with the family members' gradual cognitive decline. In response, the relatives stressed individualized support, continuity in follow-up care and regular information-sharing between them and staff members caring for their family members. They also highlighted the importance of facilitating seamless transitions from home to nursing homes.


Persistence of antibody to hepatitis B and protection from disease among Alaska natives immunized at birth.
Alaska Native (AN) children were at high risk of acquiring hepatitis B virus (HBV) infection before vaccination began in 1983. We evaluated the long-term protection from hepatitis B (HB) vaccination among AN children immunized when infants. During 1984-1995, we recruited a convenience sample of AN children who had received a three dose series of HB vaccine starting at birth and had serum antibody to hepatitis B (anti-HBs) concentrations of >/= 10 mIU/mL at 7-26 months of age. We evaluated anti-HBs concentrations and the presence of anti-HBc in participants' sera every other year up to age 16 years. Anti-HB core antigen (anti-HBc)-positive specimens were tested for hepatitis B surface antigen and for HBV DNA. We followed 334 children for 3151 person-years (median, 10 years per child) with 1610 specimens collected. Anti-HBs concentrations dropped rapidly among all participants. Among children 2, 5 and 10 years of age, 37 of 79 (47%), 33 of 176 (19%) and 8 of 95 (8%), respectively, had anti-HBs concentrations of >/= 10 mIU/mL. Receipt of recombinant vaccine was significantly associated with a more rapid antibody decline (P < 0.001). Six (1.8%) children acquired anti-HBc, 3 of whom had definite breakthrough infections (at least 2 consecutive anti-HBc-positive specimens or at least 1 anti-HBc-positive specimen and HBV DNA detection by PCR). None of these children had detectable hepatitis B surface antigen, and none had symptoms of hepatitis. Anti-HBs concentrations declined over time among AN infants successfully immunized with HB vaccine starting at birth. Transient anti-HBc appeared in a small percentage of children; however, none developed clinical signs of hepatitis or chronic HBV infection.


Cloning, sequencing, and expression of the nhaB gene, encoding a Na+/H+ antiporter in Escherichia coli.
In Escherichia coli, expulsion of sodium ions is driven by proton flux via at least two distinct Na+/H+ antiporters, NhaA and NhaB. When the nhaA gene is deleted from the chromosome, the cell becomes sensitive to high salinity and alkaline pH (Padan, E., Maisler, N., Taglicht, D., Karpel, R., and Schuldiner, S. (1989) J. Biol. Chem. 264, 20297-20302). In the current work we cloned the nhaB gene by complementation of the delta nhaA strain. The gene codes for a membrane protein 504 amino acids long. Hydropathic analysis of the sequence indicates the presence of 12 putative transmembrane helices. NhaB has been specifically labeled with [35S]methionine; it is a membrane protein and displays an apparent M(r) of 47,000, slightly lower than that predicted from its amino acid sequence. Membranes from cells containing multiple dose of nhaB display enhanced Na+/H+ antiporter activity, as measured by the ability of Na+ to collapse a preformed pH gradient or by direct measurement of 22Na+ fluxes. In contrast to NhaA, whose activity increases with pH, NhaB is practically insensitive to pH. Limited homologies with Na+ transporters have been identified.


Lysophospholipase activity of Mycobacterium smegmatis.
Lysophospholipase (LPL) activity found in Mycobacterium smegmatis was characterized. LPL activity was found to exist in high levels in two-to-10-day-old cultures. The enzyme assay was run using 1.4 X 10(9) viable bacteria at an incubation time of 24 h. Enzyme activity was detected between 1 degree C and 80 degrees C with maximal activity between 40 degrees C and 60 degrees C. LPL activity was maximal at pH 9.0, with activity ranging between pH 3.0 and 10.0.


Outcome of incisional hernia repair in patients 80 years and older: results from the Herniamed-Registry.
More and more often complex abdominal surgeries are performed in the elderly. Together with the ageing population these patients are at risk for incisional hernias. We aimed on assessing outcomes following incisional hernia surgery in patients 80 years and older. Using the Herniamed-Registry, a prospective multi-institutional database, data on patients undergoing surgery for incisional hernias were retrospectively assessed. 46,040 patients were included and divided by age. Intraoperative-, general-, and postoperative complications as well as 1-year follow-up outcomes were assessed and compared between patients 80 years and older vs younger than 80 years. Intra- (2.3% vs 1.5%; p < 0.001) and postoperative (8.6% vs 7.2%; p = 0.001) complications, general complications (5.5% vs 3.0%; p < 0.001), as well as reoperations (3.8% vs 3.0%; p = 0.007) were more likely to occur in elderly patients. By contrast, recurrences (3.6% vs 4.5%; p = 0.007), pain at rest (7.3% vs 10.1%; p < 0.001) and on exertion (11.3% vs 18.3%; p < 0.001), as well as pain requiring treatment (5.4% vs 7.7%; p < 0.001) was less likely in the group of patients aged ≥ 80 years. Incisional hernia repair in patients 80 years and older is associated with a slightly higher complication risk but is quite acceptable and also have improved pain scores. The recurrence difference is also clinically unimportant.


Role of tumor angiogenesis in gallbladder carcinoma: with special reference to thymidine phosphorylase.
Angiogenesis plays an important role in tumor growth and metastasis. Thymidine phosphorylase (TP), as well as vascular endothelial growth factor (VEGF), is known to be an angiogenic factor. The aim of this study was to clarify whether TP regulates angiogenesis and to determine the clinicopathological role of TP expression in gallbladder cancer. Thirty-seven patients with gallbladder cancer who underwent curative surgical resection were included in this study, and we evaluated results in 21 of these patients in whom TP gene expression could be measured. Intratumoral TP gene expression was evaluated using the Danenberg tumor profile method. TP gene expression was classified into two groups according to median values: high and low. Tissue TP expression was classified into two groups (positive and negative) by a pathologist. Clinicopathological variables were compared between groups with high and low TP gene expression (TP high and TP low groups). No correlation was observed between TP gene expression (high and low groups) and any clinicopathological variables except survival rate. However, the survival rate was significantly lower in the TP high group (P<0.05). TP gene expression was associated with tissue TP expression evaluated by immunohistochemical staining (P<0.05). In addition, tumor microvessel density was significantly higher in the TP high group (P<0.05). Expression of the TP gene in gallbladder carcinoma is associated with angiogenesis and may be a new independent prognostic parameter.


Comparative study on health care utilization and hospital outcomes of severe acute exacerbation of chronic obstructive pulmonary disease managed by pulmonologists vs internists.
Care for many chronic health conditions is delivered by both specialists and generalists. Differences in patients' quality of care and management between generalists and specialists have been well documented for asthma, whereas a few studies for COPD reported no differences. The objective of this study is to compare consistency with Global initiative for chronic Obstructive Lung Disease guidelines, as well as rate, health care utilization, and hospital outcomes of severe acute exacerbation (AE) of COPD patients managed by pulmonologists and internists. This is a 12-month prospective, comparative observational study among 208 COPD patients who were regularly managed by pulmonologists (Group A) and internists (Group B). Clinical data, health care utilization, and hospital outcomes of the two groups were statistically compared. Out of 208 enrolled patients, 137 (Group A) and 71 (Group B) were managed by pulmonologists and internists, respectively. Pharmacological treatment corresponding to disease severity stages between the two groups was not statistically different. Group A received care consistent with guidelines in terms of annual influenza vaccination (31.4% vs 9.9%, P<0.001) and pulmonary rehabilitation (24.1% vs 0%, P<0.001) greater than Group B. Group A had reduced rates (12.4% vs 23.9%, P=0.033) and numbers of severe AE (0.20±0.63 person-years vs 0.41±0.80 person-years, P=0.029). Among patients with severe AE requiring mechanical ventilation, Group A had reduced mechanical ventilator duration (1.5 [1-7] days vs 5 [3-29] days, P=0.005), hospital length of stay (3.5 [1-20] days vs 16 [6-29] days, P=0.012), and total hospital cost ($863 [247-2,496] vs $2,095 [763-6,792], P=0.049) as compared with Group B. This study demonstrated that pulmonologists followed national COPD guidelines more closely than internists. The rates and frequencies of severe AE were significantly lower in patients managed by pulmonologists, and length of hospital stay and cost were significantly lower among the patients with severe AE who required mechanical ventilation.


A pillar-layered chalcogenide framework assembled by [Mn5S12N12]n layers and [Sb2S5] inorganic pillars.
Reported here is an attractive pillar-layered metal chalcogenide open framework, in which [Sb2S5] building units act as pillars between [Mn5S12(N2H4)6]n layers. The obtained compound exhibits high stability in both acid and base media and good performance in the electrocatalytic oxygen reduction reaction (ORR).


Diagnosing the decline in pharmaceutical R&D efficiency.
The past 60 years have seen huge advances in many of the scientific, technological and managerial factors that should tend to raise the efficiency of commercial drug research and development (RD). Yet the number of new drugs approved per billion US dollars spent on RD has halved roughly every 9 years since 1950, falling around 80-fold in inflation-adjusted terms. There have been many proposed solutions to the problem of declining RD efficiency. However, their apparent lack of impact so far and the contrast between improving inputs and declining output in terms of the number of new drugs make it sensible to ask whether the underlying problems have been correctly diagnosed. Here, we discuss four factors that we consider to be primary causes, which we call the 'better than the Beatles' problem; the 'cautious regulator' problem; the 'throw money at it' tendency; and the 'basic research-brute force' bias. Our aim is to provoke a more systematic analysis of the causes of the decline in RD efficiency.


RNA-FISH and Immunofluorescence of Mouse Preimplantation and Postimplantation Embryos.
There are two modes of X chromosome inactivation (XCI) in the mouse. One mode is imprinted XCI: it is initiated at around the four-cell stage in favor of the paternal X chromosome, and is maintained in the extraembryonic tissues. The other mode is random XCI, which takes place in the epiblast lineage at the periimplantation stage. X-linked noncoding Xist RNA, which becomes upregulated on the X chromosome to be inactivated at the onset of XCI and plays a critical role in both imprinted and random XCI, and its accumulation in the nucleus have been referred to as one of the hallmarks of the presence of the inactivated X chromosome. RNA-FISH has therefore been an invaluable method for the study of XCI. As XCI status changes dynamically during periimplantation development in the mouse, analysis using samples from these developmental stages is absolutely necessary for elucidation of the molecular basis of XCI mechanisms. However, dissection of the embryos at around the periimplantation stages is not easy, and this impedes in vivo analysis of the kinetics of XCI. Here, we describe our methods for dissecting the periimplantation stage embryo and subsequent procedures for RNA-FISH and immunostaining.


The yeast plasmid 2mu circle encodes components required for its high copy propagation.
The yeast plasmid 2mu and certain hybrid plasmids constructed from it are maintained stably and at high copy number in yeast cells. By examining various mutant hybrid 2mu plasmids, we show that these properties require the integrity of four plasmid loci. Two of these, designated REP1 and REP2, are active in trans and correspond to two open coding regions of 2mu. The other two loci are active only in cis and correspond to the origin of replication and to a region, designated REP3, located several hundred bp away from the origin and consisting of direct repeats of a 62 bp sequence. We propose that the REP loci constitute a copy control system that overrides normal cellular restriction on plasmid replication and amplifies the plasmid when copy number is low.


The glycopeptide CSF114(Glc) detects serum antibodies in multiple sclerosis.
Synthetic glycopeptides have the potential to detect antibodies in multiple sclerosis (MS). In the present study, we analyzed the antibodies (IgM class, IgG class and IgG subclasses) to the synthetic glycopeptide CSF114(Glc) in the serum of 186 MS patients, 166 blood donors (BDs), 25 patients affected by meningitis/encephalitis, 41 affected by systemic lupus erythematosus (SLE) and 49 affected by rheumatoid arthritis (RA). The IgM antibody level to CSF114(Glc) was significantly increased in MS patients versus BDs (p<0.001) or versus other autoimmune diseases (SLE or RA, p<0.001). The IgG response was restricted to the subclass IgG2. IgM antibodies to CSF114(Glc) were found in 30% of relapsing/remitting MS patients and, at lower levels, in subjects affected by meningitis/encephalitis. The study of antibodies to CSF114(Glc) is a new, potential immunological marker of MS.


Serial echocardiography for decision making in rapid two-stage arterial switch operation.
Rapid two-stage arterial switch operation is advocated in infants with simple transposition presenting late. Accurate assessment of left ventricular preparation is crucial to successful outcome. The role of echocardiography alone in surgical decision making remains unclear. Seventeen patients with simple transposition (mean age, 4 months) underwent pulmonary artery banding and modified Blalock-Taussig shunt (first stage) to prepare the left ventricle for the arterial switch operation (second stage). Serial echocardiography was performed in the interval phase to assess left ventricular growth. Sixteen patients underwent arterial switch operation after a mean interval of 10.4 +/- 4 days, with 14 successful conversions. There was one mortality (5.9%) and two conversions to a Senning repair. In all patients a mean increase in left ventricular mass (40.8 +/- 17.8 g/m2 to 81.4 +/- 25.4 g/m2) and posterior wall thickness (3.37 +/- 0.47 mm to 4.63 +/- 0.58 mm) was recorded. Left ventricular end-diastolic internal diameter increased in all except the two switch failures. In all the successful cases the left ventricle had assumed a circular shape on cross-section with the interventricular septum contracting in synergy with the left ventricular mass. In the two failures, however, the interventricular septum had remained flat. Echocardiography can be used reliably in surgical decision making in rapid two-stage arterial switch operation. Increase in left ventricular mass, left ventricular posterior wall thickness, and left ventricular end-diastolic internal diameter toward normal combined with an acquisition of circular left ventricular configuration with the interventricular septum contracting in synergy with the left ventricular mass appear to best predict successful outcome.


Complete heart block in a fetus associated with maternal Sjögren's syndrome.
This report illustrates the case of a fetus who was diagnosed by M-mode echocardiography, at the twentieth week of gestation, as having a complete heart block. The mother had a positive antinuclear antibody test result with antibodies directed against the Ro (SSA) and La (SSB) antigen system consistent with the diagnosis of Sjögren's syndrome. Management and outcome of the pregnancy are discussed.


l-Proline improves the cytoplasmic maturation of mouse oocyte by regulating glutathione-related redox homeostasis.
Recent studies have shown that l-proline (proline) is an antioxidant to protect cells from oxidative stress in vivo and in vitro. Glutathione (GSH) is a major cellular redox regulator involved in controlling redox balance and is regarded as one of the key indices to predict the cytoplasmic maturation of oocytes. The objectives of this study are to investigate the effect of proline on the developmental potential of mouse oocytes and to determine the role of gap junctional communication (GJC) on intraoocyte GSH concentration during in vitro maturation (IVM). Compared with control (0 mmol/L), 0.5 mmol/L proline supplementation enhanced rates of activated oocytes, 2-cell and 4-cell embryos, and blastocysts. Furthermore, 0.25 and 0.5 mmol/L proline supplementation markedly upregulated mRNA expression of glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate-cysteine ligase modifier subunit (GCLM) in oocytes and cumulus cells, enhanced GSH concentration in oocytes, and reduced reactive oxygen species (ROS) level in oocytes. Interestingly, carbenoxolone disodium salt (CBX) treatment reduced GSH concentration in oocytes and the rate of early embryo development without proline incubation. Notably, CBX-triggered reduction in the rates of the number of 2-cell and 4-cell embryos and blastocysts were rescued by 0.5 mmol/L proline supplementation. Collectively, these results indicate a novel functional role of proline in oocyte cytoplasmic maturation and regulation of glutathione-related redox homeostasis.


Serum acute phase response induced by different vaccination protocols against circovirus type 2 and Mycoplasma hyopneumoniae in piglets.
The purpose of this study was to compare the acute phase reaction (APR) induced by different vaccination protocols used against Porcine Circovirus (PCV) type-2 and Mycoplasma hyopneumoniae (M.hyo), studying two acute phase proteins (APPs) and changes in rectal temperature (RT). In addition, the possible influence of the time of vaccination and breed were analysed. In the first experiment, 40 commercial crossbred piglets were vaccinated, on the day of weaning, with FLEXcombo® (group A, n=20) or Porcilis PCV® and Stellamune® One (group B, n=20). The second experiment was performed on two farms, on which 40 commercial crossbred piglets or 40 Iberian piglets were vaccinated, 7days post-weaning. On each farm one group (A, n=20) was vaccinated with FLEXcombo® and another group (B, n=20) with Porcilis® PCV-M.hyo. Blood samples were taken before, 24h and 48h after vaccination, and RT were recorded before and 8h after vaccination. Significantly higher increases in group B in RT (P<0.01) and APPs concentrations (P<0.01) were recorded at several sampling times after vaccination compared with group A. The vaccines that produced greater increases in RT also produced higher APPs increases but no influence of the day of vaccination or of the breed were found. Therefore, serum APPs concentrations differed according to the vaccine used, which may be useful, along with RT, for choosing the vaccine or protocol that produces APR of lower magnitude.


Predictors of Mortality among COVID-19 Patients Admitted to Intensive Care Units: A Single-Center Study in Tehran, Iran.
Iran was one of the first countries to become an epicenter of the coronavirus disease 2019 (COVID-19) epidemic. However, there is a dearth of data on the outcomes of COVID-19 and predictors of death in intensive care units (ICUs) in Iran. We collected extensive data from patients admitted to the ICUs of the one of the tertiary referral hospitals in Tehran, Iran, to investigate the predictors of ICU mortality. The study population included 290 COVID-19 patients who were consecutively admitted to the ICUs of the Sina hospital from May 5, 2021, to December 6, 2021, a period that included the peak of the epidemic of the delta (δ) variant. Demographic data, history of prior chronic diseases, laboratory data (including markers of inflammation), radiologic data, and medication data were collected. Of the 290 patients admitted to the ICUs, 187 (64.5%) died and 103 (35.5%) survived. One hundred forty-one (141, 48.6%) were men, and the median age (10th percentile, 90th percentile) was 60 (41, 80). Using logistic regression models, older age, history of hypertension, high levels of inflammatory markers, low oxygen saturation, substantial lung involvement in computed tomography (CT) scans, and gravity of the disease as indicated by the WHO 8-point ordinal scale were primary predictors of mortality at ICU. The use of remdesivir and imatinib was associated with a statistically non-significant reduction in mortality. The use of tocilizumab had almost no effect on mortality. The findings are consistent with and add to the currently existing international literature. The findings may be used to predict risk of mortality from COVID-19 and provide some guidance on potential treatments.


[Modification of the cytogenetic effect of dioxidine in experimental alloxan diabetes in rats].
A dependence was established between the cytogenetic effect of dioxydinum antibacterial drug and the disturbance of carbohydrate metabolism in rats with alloxan diabetes.


The relationship between tear severity, fatty infiltration, and muscle atrophy in the supraspinatus.
Fatty infiltration and muscle atrophy have been described as interrelated characteristic changes that occur within the muscles of the rotator cuff after cuff tears, and both are independently associated with poor outcomes after surgical repair. We hypothesize that fatty infiltration and muscle atrophy are two distinct processes independently associated with supraspinatus tears. A retrospective review of 377 patients who underwent shoulder magnetic resonance imaging at one institution was performed. Multivariate analysis was performed based on parameters including age, sex, rotator cuff tear severity, fatty infiltration grade, and muscle atrophy. A total of 116 patients (30.8%) had full-thickness tears of the supraspinatus, 153 (40.6%) had partial thickness tears, and 108 (28.7%) had no evidence of tear. With increasing tear severity, the prevalence of substantial fatty infiltration (grade ≥2) increased: 6.5% of patients with no tears vs 41.4% for complete tears (P < .001). Similarly, the prevalence of supraspinatus atrophy increased with worsening tear severity: 36.1% of no tears vs 77.6% of complete tears (P < .001). Multivariate analysis demonstrated a significant independent association between fatty infiltration and muscle atrophy when taking into account sex, age, and tear severity. Fatty infiltration and muscle atrophy are independently associated processes. Fatty infiltration is also related to increasing age, muscle tear severity, and sex, whereas muscle atrophy is related to increasing age but not tear severity. In patients without rotator cuff tears, fatty infiltration and atrophy prevalence increased independently with increasing age.


Virologic, immunologic, and clinical observations on a patient during the incubation, acute, and convalescent phases of infectious mononucleosis.
One patient with infectious mononucleosis (IM) was studied from the probable time of Epstein-Barr virus (EBV) infection (38 days before the onset of clinically overt disease), during the incubation and acute phases, until 6 months after clinical remission. Analysis of spontaneous outgrowth of EBV-carrying lymphoblastoid cells, by limiting dilution on feeder layer cultures, showed that virus containing B lymphocytes are already present early during the incubation period. Also low interferon serum levels were detected early after infection, and only before the onset of clinical disease. All other studied clinical laboratory and virus-associated variables were within normal range during the incubation phase, but changed to a pattern characteristic of IM in parallel to the clinical symptoms. During the acute disease EBV-associated nuclear antigen (EBNA)-positive cells could be directly detected among the lymphocytes, and antibodies to EBV antigens appeared. Lymphocytes stained by monoclonal antibodies, detecting Ia-like determinants (activated cells) and suppressor cells, increased dramatically, in parallel to a strong increase of functional suppressor cell activity, measured by inhibition of blastogenesis and PWM-induced immunoglobulin production. During the acute phase there was also a decrease of spontaneous cytotoxicity against the NK-sensitive cell line K562, while cytotoxicity (spontaneous) against an autologous EBV-positive lymphoblastoid cell line (LCL) was detected only during this phase. These reactions correlated to the presence of blasts, and the autologous reaction was exerted mainly by Fc-receptor-negative cells. Lymphokine production in response to EBV antigens was also initiated during the acute phase. During the convalescence period the serological and cellular immune parameters adjusted to the pattern of a normal EBV-seropositive person.


Patient-centered outcome evaluation of the Rural Breast Cancer Survivors Intervention.
Patient-centered evaluation is a critical but often overlooked component of intervention research. The Rural Breast Cancer Survivors (RBCS) Intervention is a survivorship education and support intervention designed for rural breast cancer survivors. Here, we describe evaluation of the RBCS Intervention from the participants' experience. Three hundred sixty-four breast cancer survivors participating in the RBCS Intervention were asked to complete the RBCS Evaluation Survey that consisted of 6 quantitative statements and 12 qualitative questions via mail. Two hundred twenty-one participants (61 %) returned the RBCS Evaluation Survey. All 221 completed the qualitative component, and 91 (25 %) completed the quantitative. Overall, participants indicated that the education materials were easy to understand and addressed their quality of life concerns. Majority (94 %) indicated that the survivorship information was helpful in making health decisions and communicating concerns to family (87 % strongly agree/agree) and oncology team (88 % strongly agree/agree). Only 66 % indicated that the survivorship educational materials were provided at an appropriate time after completion of primary breast cancer treatment. Qualitative data suggested that participants had positive perceptions and experiences and found the RBCS Intervention helpful. One-on-one interaction with the intervention nurses was the single most highly valued aspect. Overall, the RBCS Intervention was helpful. Education materials addressed quality of life concerns. Breast cancer survivors developed skills in communicating with their family and oncology team. Intervention nurses were identified as the most valuable aspect of the program. Suggestions include earlier timing of survivorship education and support, and adaptation using social media access to serve other rural survivors. Patient-centered evaluations, using both quantitative and qualitative data, provide enriched understanding of evidence-based survivorship interventions and should be considered a standard for future work.


Curare has a voltage-dependent blocking action on the glutamate synapse.
The skeletal muscle of members of Orthoptera and Diptera receives an innervation which is probably glutaminergic. Recent study of the ventral muscle fibres in the larvae of the beetle, Tenebrio molitor, has revealed that the transmitter action can be mimicked by the iontophoretic application of L-glutamate to the junctional sites at which the extracellular excitatory postsynaptic potentials (e.p.s.ps) could be recorded (D.Y. and H.W., unpublished observation). Contrary to the evidence favouring glutamate as a transmitter of junctional excitation in insects, some investigators have found that curare (+)tubocrarine, TC), a classic acetylcholine (ACh) antagonist, suppresses the neurally evoked muscle potentials in the fly Sacophaga, and Tenebrio. Here, we have analysed the action of curare on the neuromuscular junction of Tenebrio larvae and found that curare blocked the glutaminergic transmission by antagonising the transmitter at the postsynaptic site.


Absence of human placental lactogen in an otherwise uneventful pregnancy.
Human placental lactogen (hPL) is commonly used in surveying the placental function during normal and pathologic pregnancies. This report describes a pregnancy where hPL could not be found in maternal serum or placental tissue. The pregnancy was in all other respects completely normal, ending with the birth of a normal baby. Some possible reasons and consequences of this unique event are discussed.


Effects of chitosan-glucose complex coating on postharvest quality and shelf life of table grapes.
Surface coatings and films are prospective alternatives for extending the postharvest life of fresh fruits and vegetables. In this study, freshly harvested grapes were treated with chitosan, glucose, chitosan-glucose complex (CGC), or water (control) for up to 60 days at 0 °C in 95% relative humidity followed by 3 days in the air at 20 °C. The results showed that coated samples were effective in terms of senescence inhibition and postharvest diseases prevention. Chitosan-glucose complex showed better effects on delaying the declines of total soluble solids, ascorbic acid and titratable acidity, decreasing decay and weight loss, suppressing respiration rate, inducing POD and SOD activities, in comparison with pure chitosan or glucose. In addition, CGC coating treatment ensured better berry texture and higher sensory scores, compared with those treated with chitosan or glucose alone. The results indicated that CGC may be a promising strategy for improving the postharvest quality and extending the shelf life of table grapes.


FISH analysis of a complex chromosome rearrangement involving nine breakpoints on chromosomes 6, 12, 14 and 16.
We report the prenatal diagnosis of an apparently balanced de novo complex chromosome rearrangement (CCR) which involved nine breakpoints on four different chromosomes. Fluorescence in situ hybridization (FISH) and spectral karyotyping (SKY) were performed as an adjunct to G-banding for characterization of the abnormal chromosomes. The 22-week female fetus showed minor dysmorphic features including dolichocephaly, broad fingernails, tibial bowing, clubfoot, thoracolumbar scoliosis and hypoplastic toenails. Autopsy revealed gall-bladder hypoplasia and an atrial septal defect. Chromosome analysis of fetal tissue confirmed the presence of the complex rearrangement.


Effect of dietary supplementation with vitamin E and stocking density on macrophage recruitment and giant cell formation in the teleost fish, Piaractus mesopotamicus.
The effect of dietary supplementation with 0, 100 and 450 mg of vitamin E (DL-alpha tocopheryl acetate)/kg of a dry diet on the kinetics of macrophage recruitment and giant cell formation in the pacu, maintained at different stocking densities (5 kg/m(3) and 20 kg/m(3)), was investigated by insertion of round glass coverslips into the subcutaneous connective tissue. After a feeding period of 18 weeks, the coverslips were implanted and later removed for examination at 2, 7 and 15 days post-implantation. Fish fed diets supplemented with 450 mg of vitamin E showed an increase (P<0.05) in the accumulation of macrophages, foreign body giant cells and Langhans type cells. The kinetics of macrophage recruitment and giant cell formation on the glass coverslips appeared to be strongly influenced by vitamin E supplementation, since fish fed a basal diet and held at high stocking densities showed low numbers of adhering cells on the coverslips, and high concentrations of plasma corticosteroids. On the other hand, fish given a diet supplemented with 450 mg of vitamin E did not show a similar difference in plasma cortisol concentrations related to stocking density. The effect of cortisol concentrations on carbohydrate metabolism, analysed by assessment of plasma glycaemia, was not clear. Blood glucose concentrations did not vary substantially with the different treatments examined. These results suggest that vitamin E may contribute to the efficiency of the fish's inflammatory response by increasing macrophage recruitment and giant cell formation in the foreign body granulomatous reaction. Vitamin E appeared to act on the stress response of pacus by preventing a stress-related immunosuppression.


Intravenous immunoglobulin treatment of four patients with juvenile polyarticular arthritis associated with persistent parvovirus B19 infection and antiphospholipid antibodies.
Children with rheumatic oligoarthritis and polyarthritis frequently establish persistent parvovirus B19 infections that may be associated with the production of antiphospholipid antibodies (anti-PL IgG). In this study we analysed the influence of high-dose intravenous immunoglobulin (IVIG) therapy on virus load, on the level of anti-PL IgG and its potential capacity to improve the patients' clinical status. Four juvenile patients with long-lasting polyarticular rheumatic diseases and persistent parvovirus B19 infection, associated in three cases with the presence of antibodies against beta2-glycoprotein I (anti-beta2GPI IgG), were treated with two cycles of IVIG on five successive days (0.4 g/kg per day). Clinical parameters including scores of disease activity, virus load and anti-PL IgG levels were determined before, during and after treatment. Two patients showed a complete remission that has lasted 15 months. During that period they showed neither clinical nor laboratory signs of inflammation. Viral DNA was not detectable in serum, and a decrease in anti-beta2GPI IgG was observed. As assessed by the Childhood Health Assessment Questionnaire and the Health-related Quality of Life Questionnaire for Children, both patients were no longer restricted in their activities of daily living and no impact on the health-related quality of life was observed. In one patient the therapy failed: there was no improvement of symptoms and no decrease in virus load or inflammatory parameters. In the fourth patient, clinical and laboratory parameters did not improve despite a decrease in both viral load and anti-PL IgG. Our results show that the use of IVIG to treat parvovirus B19-triggered polyarticular rheumatic disease of childhood might offer an opportunity to improve this disabling condition.


Recurrent headache and visual symptoms in a young man: a rare neuronal intranuclear inclusion disease case report.
Neuronal intranuclear inclusion disease (NIID) is a progressive neurodegenerative disease. Patients with NIID may present with heterogeneous clinical symptoms, including episodic encephalopathy, dementia, limb weakness, cerebellar ataxia, and autonomic dysfunction. Among the NIID cases reported in China, patients often have complicated and severe manifestations. Therefore, many clinicians do not consider the disease when the patient presents with relatively minor complaints. We present the case of a 39-year-old man showing migraine-aura-like symptoms for the past 3 years. Brain magnetic resonance imaging (MRI) revealed hyperintense signals in the splenium of the corpus callosum and corticomedullary junction on diffusion-weighted imaging (DWI) over time. In addition, brain atrophy that was not concomitant with the patient's age was detected while retrospectively reviewing the patient's imaging results. Genetic analysis and skin biopsy confirmed a diagnosis of NIID. The patient was treated with sibelium, and the symptoms did not recur. Migraine-aura-like symptoms may be the predominant clinical presentation in young patients with NIID. Persistent high-intensity signals on DWI in the brain and early-onset brain atrophy might be clues for the diagnosis of NIID.


Development of tumor cell resistance to tumor necrosis factor cytolysis results in reduced fibronectin binding and altered ganglioside expression.
U937A cells are highly susceptible to tumor necrosis factor (TNF) cytolysis. They are also motile and incorporate fibronectin into the extracellular matrix (ECM). This takes the form of a dense fibrillar network in confluent cultures, but in sparse cultures appears as a "snail trail" of insolubilized fibronectin behind the moving cell. In contrast, U937A/R cells selected for resistance to TNF cytolysis are poorly motile and, although they synthesize fibronectin, fail to incorporate it into the ECM. Compared to U937A/R, U937A cells spread more rapidly and extensively on fibronectin-coated plastic and also bound 125I-fibronectin more effectively. Inhibition of U937A spreading on fibronectin required higher doses of GRGDSPK peptide, indicating greater expression on U937A of integrin-type, fibronectin receptors. Gangliosides are non-integrin structures which can bind fibronectin, and there were also qualitative and quantitative differences in ganglioside expression with U937A having two to five times more than U937A/R. Therefore the development of TNF resistance by U937A/R cells is accompanied by a reduced ability to interact with fibronectin, and this probably accounts for the reduced motility and inability to deposit fibronectin in the ECM.


Predictors of Testicular Cancer Mortality in Brazil: A 20-Year Ecological Study.
Testicular cancer is common in young men, and early detection and multimodality treatment can lead to successful outcomes. This study aims to identify sociodemographic and risk factors associated with higher testicular cancer mortality and poorer survival rates, while examining the impact of diagnostic and treatment procedures on reducing mortality. The retrospective ecological study analyzed mortality data from testicular cancer in Brazil from 2001 to 2020. Sociodemographic variables such as marital status, age, birth period, year of death (cohort), race, and geographic region were assessed. Risk factors included cryptorchidism and pesticide exposure. Data were subjected to statistical analysis, which revealed an increasing trend in mortality after 2011 among persons born after 1976 in the 15-40 age group. Individuals in the South Region, whites, and singles had higher age-standardized mortality rates (ASMRs), while singles had lower survival rates. The Northeast region had a higher survival rate. Fungicides and insecticides increase ASMR in Brazil. Herbicides increase ASMR in the Northeast and Midwest regions and insecticides increase ASMR in the Northeast, Southeast, and Midwest regions. High rates of implementation of diagnostic procedures in the Midwest were not sufficient to reduce ASMR. No treatment procedure was associated with mortality at the national or regional level.


Alternatively spliced lipin isoforms exhibit distinct expression pattern, subcellular localization, and role in adipogenesis.
We recently identified mutations in the Lpin1 (lipin) gene to be responsible for lipodystrophy in the fatty liver dystrophy (fld) mouse strain. Previous studies revealed that lipin plays a critical role in adipogenesis, explaining the adipose-deficient phenotype of the fld mouse. In the current study, we demonstrate that alternative mRNA splicing generates two lipin isoforms, lipin-alpha and lipin-beta, which are differentially expressed during adipocyte differentiation. Lipin-alpha expression peaks at day 2 of 3T3-L1 cell differentiation, after which its levels gradually decrease. In contrast, lipin-beta expression is transiently elevated at 10 h, followed by a drop to background levels at 20 h and a gradual increase between days 2 and 6 of differentiation. The two lipin isoforms also exhibit differences in subcellular localization. Lipin-alpha is predominantly nuclear, whereas lipin-beta is primarily located in the cytoplasm of 3T3-L1 adipocytes, suggesting distinct cellular functions. Using primary mouse embryonic fibroblasts expressing either lipin-alpha or lipin-beta, we demonstrate functional differences between the two isoforms. Whereas lipin-alpha is required for adipocyte differentiation, the predominant effect of lipin-beta expression is the induction of lipogenic genes. In vivo, overexpression of lipin-beta specifically in mature adipocytes leads to elevated expression of lipogenic genes and adipocyte hypertrophy, confirming a role of lipin-beta in the regulation of lipogenesis. In conclusion, our data suggest that the two lipin isoforms have distinct, but complementary, functions in adipogenesis, with lipin-alpha playing a primary role in differentiation and lipin-beta being predominantly involved in lipogenesis.


Expression of a human endogenous retrovirus, HERV-K, in the blood cells of leukemia patients.
Human endogenous retroviral sequences (HERVs) are believed to be possible pathogenic agents in carcinogenesis. HERV-K is the most biologically active form, since members of this family have intact open reading frames for the gag, pol or env genes. Antibody response against HERV-K peptides has been reported in leukemia patients, suggesting a possible overexpression of this sequence in leukemic cells. Using real-time quantitative RT-PCR (TaqMan), we found that in six of the eight leukemia samples we collected, transcriptional activity of HERV-K10-like gag gene was 5- to 10-fold higher than in normal peripheral blood mononuclear cells (PBMCs) or mononuclear cells from cord blood. The overexpression was marked enough to be detected by Northern blot. In addition, there was no significant variation of HERV-K expression in normal PBMCs after exposure to different factors (PHA, gamma irradiation, 5-azacytidine) that potentially modulate HERV expression. This suggests that HERV-K relative overexpression in leukemia samples might be specifically associated with tumor development. The origin of these transcriptional variations is therefore worth being investigated further.


Morphological study of 5-HT neurons and astroglial cells on brain of adult rats perinatal or chronically exposed to 2,4-dichlorophenoxyacetic acid.
2,4-D is a chlorophenoxyherbicide used worldwide. We have studied the morphological alterations of 5-HT neurons and glial cells in the mesencephalic nuclei of adult rats exposed to 2,4-D both perinatally (during pregnancy and lactation) and chronically (during pregnancy, lactation and after weaning) with quantitative methods. Pregnant rats were daily exposed to 70 mg/kg of 2,4-D from gestation day (GD) 16 to post-natal day (PND) 23 through diet. After weaning, pups were assigned to one of two sub-groups: T1 (fed with untreated diet until PND 90) and T2 (maintained with 2,4-D diet until PND 90). Brain sections were immunocytochemically stained using polyclonal anti-5-HT, anti-GFAP and anti-S-100 protein antibodies as cells markers. 2,4-D exposure during pregnancy and lactancy (T1 group) produced an increase in 5-HT neuronal area and immunoreactivity (IR) in the mesencephalic nuclei studied. However, with the chronical 2,4-D exposure (T2 group) only the 5-HT neuronal area from the dorsal raphe nucleus (DRN) was increased, suggesting an adaptable response of 5-HT neurons in median raphe nucleus (MRN). The presence of reactive astrocytes in mesencephalic nuclei and in hippocampus were also different for the two 2,4-D exposure designs, showing the existence of a correspondence between neuronal changes and astrogliosis. Results support evidences that 2,4-D alters the serotoninergic system and that 5-HT neurons of each mesencephalic nuclei show different responses to the 2,4-D exposure designs which are parallel to astrogliosis.


Development of pediatric simulation-based education - a systematic review.
This systematic literature review explored the general characteristics, validation, and reliability of pediatric simulation-based education (P-SBE). A literature search was conducted between May 23 and 28 using the PRISMA guidelines, which covered databases such as MEDLINE, EMBASE, CINAHL, and Cochrane Library. In the third selection process, the original texts of 142 studies were selected, and 98 documents were included in the final content analysis. A total of 109 papers have been published in the ten years since 2011. Most of the study designs were experimental studies, including RCT with 76 articles. Among the typologies of simulation, advanced patient simulation was the most common (92), and high-fidelity simulation was the second most common (75). There were 29 compatibility levels and professional levels, with 59 scenarios related to emergency interventions and 19 scenarios related to communication feasibility and decision making. Regarding the effect variable, 65 studies confirmed that skills were the most common. However, validity of the scenarios and effect variables was not verified in 56.1% and 67.3% of studies, respectively. Based on these findings, simulation based-education (SBE) is an effective educational method that can improve the proficiency and competence of medical professionals dealing with child. Learning through simulation provides an immersive environment in which learners interact with the presented patient scenario and make decisions, actively learning the attitudes, knowledge, and skills necessary for medical providers. In the future, it is expected that such research on SBE will be actively followed up and verified for its validity and reliability.


Subjective symptoms and breathing pattern at rest and following hyperventilation in anxiety and somatoform disorders.
The purpose of the present study was to investigate the diagnostic specificity of bodily symptoms and respiratory behavior at rest and after a hyperventilation provocation test (HVPT) in patients that were either grouped according to the DSM classification or diagnosed as suffering from hyperventilation syndrome. Nine hundred three anxiety and somatoform patients, showing symptoms supposedly caused by psychogenic hyperventilation, and 170 healthy subjects, were studied. Breathing pattern and end-tidal CO2 concentration were recorded during breathing at rest and following a HVPT. Subjective symptoms in daily life and after HVPT were measured. A principal-components analysis was performed on both the symptoms and breathing variables and their specificity levels were compared in the two classifications of patients. Some symptoms in daily life were grouped together with the same symptoms after the HVPT, other symptoms were not. This suggests that the HVPT elicited partly specific symptoms, and partly reproduced the symptoms experienced in daily life. Similar findings were observed with respect to the breathing variables. Patients with panic differed from other patients with anxiety disorders by an increased level of symptoms and a FETCO2 decline at rest. The HVPT may be informative for diagnosis because it provokes some of the typical somatic and psychological symptoms, and it identifies the breathing instability that is characteristic of both patients with HVS and with anxiety. The same symptoms and breathing variables characterized the patients, whatever their classification. Overall, the specificity of breathing variables is rather low.


Neurophysiological correlates of Avolition-apathy in schizophrenia: A resting-EEG microstates study.
The "Avolition-apathy" domain of the negative symptoms was found to include different symptoms by factor analytic studies on ratings derived by different scales. In particular, the relationship of anhedonia with this domain is controversial. Recently introduced negative symptom rating scales provide a better assessment of anhedonia, allowing the distinction of anticipatory and consummatory aspects, which might be related to different psychopathological dimensions. The study of associations with external validators, such as electrophysiological, brain imaging or cognitive indices, might shed further light on the status of anhedonia within the Avolition-apathy domain. We used brain electrical microstates (MSs), which represent subsecond periods of quasi-stable scalp electrical field, associated with resting-state neural networks (and thus with global patterns of functional connectivity), to test whether the component symptoms of Avolition-apathy share the same correlates. We analyzed multichannel resting EEGs in 142 individuals with schizophrenia (SCZ) and in 64 healthy controls (HC), recruited within the add-on EEG study of the Italian Network for Research on Psychoses. Relative time contribution, duration and occurrence of four MS classes (MS-A/-B/-C/-D) were calculated. Group differences on MS parameters (contribution and duration) and their associations with negative symptom domains (assessed using the Brief Negative Symptoms Scale) were investigated. SCZ, in comparison to HC, showed increased contribution and duration of MS-C. The contribution of MS-A positively correlated with Avolition-apathy, but not with Expressive deficit. Within the Avolition-apathy domain, anticipatory anhedonia, avolition and asociality, but not consummatory anhedonia, showed the same correlations with MS-A contribution. Our findings support the existence of distinct electrophysiological correlates of Avolition-apathy with respect to Expressive deficit, and lend support to the hypothesis that only the anticipatory component of anhedonia shares the same pathophysiological underpinnings of the Avolition-apathy domain.


Long-term follow-up of patients with candiduria.
Candiduria is commonly encountered in hospitalized patients, particularly those with indwelling urinary catheters. While risk factors and therapy are well described in previous studies, little is known about long-term outcomes and recurrence rates of candiduria. We studied 188 patients with candiduria in a retrospective chart review at a single institution from January 1999 to December 2000. Data were collected regarding risk factors and underlying disease, therapy, follow-up cultures until December 2003, and mortality. Ninety-one patients with at least one follow-up culture >1 month after the initial culture (range 2-48) were available for further study. In this group, patients receiving antifungal therapy for asymptomatic candiduria were paradoxically more likely to have subsequent positive urine cultures than patients who never received antifungal therapy. Six patients developed candidemia during follow-up, although in none was this considered to represent a consequence of candiduria. Mortality rate at the end of the follow-up period (mean of 18 months) was 43%, including one death attributed to candidemia. Therapy for candiduria does not appear to reduce candiduria recurrence rates through 48 months of follow-up and little evidence of treatment benefit was identified.


Complete exon 2 sequence of the HLA-DPA1*03:01 allele by sequence-based typing.
Completion of the first 20 nucleotides of exon 2 of DPA1*03:01 allele.


Predictors of premature termination and completion of randomized controlled trials.
Randomized clinical trials (RCTs) have a key role in progressing biomedical research and guiding clinical decision making, but premature termination remains high (up to 30%), raising concerns regarding funding expenditure and resource allocation. This brief report sought to identify variables associated with RCTs' premature termination and completion.


Regulatory elements that control the lineage-specific expression of myoD.
The molecular basis of skeletal muscle lineage determination was investigated by analyzing DNA control elements that regulate the myogenic determination gene myoD. A distal enhancer was identified that positively regulates expression of the human myoD gene. The myoD enhancer and promoter were active in myogenic and several nonmyogenic cell lines. In transgenic mouse embryos, however, the myoD enhancer and promoter together directed expression of a lacZ transgene specifically to the skeletal muscle lineage. These data suggest that during development myoD is regulated by mechanisms that restrict accessibility of myoD control elements to positive trans-acting factors.


Prenatal diagnosis of recurrence of cystic hygroma with normal chromosomes.
Cyst-like masses in the neck and back region, simulating an encephalocele, were detected by midtrimester ultrasound examinations in two sib fetuses. After termination, the fetuses were found to have large cystic hygromas. The present observation strongly suggests the existence of an autosomal recessive type of cystic hygroma.


Wound care antiseptics - performance differences against Staphylococcus aureus in biofilm.
Staphylococcus aureus is commonly isolated from infected wounds both in animals and humans. It is known to be an excellent biofilm former and biofilms are present in as many as 60% of chronic wounds. Despite that the presence of biofilms in infections are common, antiseptics are usually qualified for in vivo testing according to their effect on planktonic cells. As it is well known that bacteria in biofilms are more tolerant to antiseptics than planktonic bacteria, biofilm infections can be difficult to treat. The aim of the study was to compare three different categories of antiseptics, biguanide (chlorhexidine), quaternary ammonium compound (QAC; Pyrisept) and iodine/iodophores (2% iodine liniment), with regards to efficacy in killing S. aureus in biofilm. If there was observed a difference in efficacy between these antiseptics, a second aim was to find the most effective of the three antiseptics. Large differences in the bactericidal effect of the different antiseptics against S. aureus in biofilm were observed in the present study. Iodine treatment was found to be the most effective followed by Pyrisept and chlorhexidine. The bactericidal effect of the different antiseptics used in the present study was found to vary significantly against S. aureus in biofilm. The present study gives valuable knowledge with regards to selecting the antiseptics that are most likely to be successful in treating biofilm infected wounds. This study also contributes to focus attention on the importance of qualifying antiseptics based on results using biofilm bacteria rather than planktonic bacteria.


The Impaired Surgeon.
Impaired physicians are unable to manage professional activities safely. Impairment can be the result of physical or mental disorders, but impaired physicians commonly refers to those suffering from substance use disorders. These disorders are at least as common in physicians as they are in the general population, and physician health programs have been established in each state to provide expeditious and detailed evaluation, referral to treatment facilities, and post-treatment monitoring to ensure compliance. These programs protect participants against sanctions and safeguard patients. The prognosis for participants is excellent, with more than 70% able to practice medicine at 5 years.


Quantification of the infectivity of Cryptosporidium parvum by monitoring the oocyst discharge from SCID mice.
Doses of 1-10(5) oocysts of Cryptosporidium parvum HNJ-1 were inoculated into severe combined immunodeficient (SCID) mice, and the discharge of oocysts was monitored for 30 days post inoculation. None of the mice discharged any oocysts after oral inoculation of one oocyst. Only one of five SCID mice discharged oocysts after oral inoculation of 10 oocysts, and the prepatent period was 17 days. The other four mice did not discharge any oocysts. All the SCID mice discharged oocysts after oral inoculation of 10(2)-10(5) oocysts. The prepatent periods were 13-17, 8-10, 8, and 10 days in SCID mice inoculated with 10(2), 10(3), 10(4), and 10(5) oocysts, respectively. A proportional correlation was observed between inoculation doses of oocysts ranging from 10 to 10(4) oocysts and the corresponding prepatent periods. The prepatent period can be used to evaluate the infectivity of C. parvum oocysts.


Design, synthesis and binding mode of interaction of novel small molecule o-hydroxy benzamides as HDAC3-selective inhibitors with promising antitumor effects in 4T1-Luc breast cancer xenograft model.
Histone deacetylase 3 (HDAC3) is one of the most promising targets to develop anticancer therapeutics. In continuation of our quest for selective HDAC3 inhibitors, a series of small molecules having o-hydroxy benzamide as the novel zinc binding group (ZBG) has been introduced for the first time that can be able to produce good HDAC3-selectivity over other HDACs. The most promising HDAC3 inhibitors, 11a and 12b, displayed promising in vitro anticancer activities with less toxicity to normal kidney cells. These compounds significantly upregulate histone acetylation and induce apoptosis with a G2/M phase arrest in B16F10 cells. Compound 11a exhibited potent antitumor efficacy in 4T1-Luc breast cancer xenograft mouse model in female Balb/c mice. It also showed significant tumor growth suppression with no general toxicity and extended survival rates post-tumor resection. It significantly induced higher ROS generation, leading to apoptosis. No considerable toxicity was noticed in major organs isolated from the compound 11a-treated mice. Compound 11a also induced the upregulation of acH3K9, acH4K12, caspase-3 and caspase-7 as analyzed by immunoblotting with treated tumor tissue. Overall, HDAC3 selective inhibitor 11a might be a potential lead for the clinical translation as an emerging drug candidate.


Morphological, physiological, and molecular scion traits are determinant for salt-stress tolerance of grafted citrus plants.
Citrus productivity has been decreasing in the last decade in the Mediterranean basin as a consequence of climate change and the high levels of salinity found in the aquifers. Citrus varieties are cultivated grafted onto a rootstock, which has been reported as responsible for plant tolerance to adverse situations. However, other important factors for stress tolerance relying in the scion have been less studied. The aim of this study was to evaluate the effect of the grafted scion on citrus tolerance to salt stress. Four different citrus rootstock/scion combinations were subjected to salt stress for 30 days, using Carrizo citrange (CC) or Citrus macrophylla (CM) as rootstocks, and Navelina orange (NA) or Oronules mandarin (OR) as scions. CM-OR was the most tolerant combination, whereas CC-NA was the most sensitive one. Our results support the idea that the rootstock plays an important role in salt stress tolerance, but scion is also crucial. Thus, photosynthesis and transpiration, processes regulated by abscisic acid and jasmonic acid, are determinant of plant performance. These photosynthetic parameters were not affected in plants of the salt-tolerant combination CM-OR, probably due to the lower intoxication with Cl- ions, allowing a better performance of the photosynthetic machinery under stress conditions. The different stomatal density of the two citrus scions used in this work (higher in the sensitive NA in comparison to the tolerant OR) also contributes to the different tolerance of the grafted plants to this adverse condition. Additionally, CsDTX35.1 and CsDTX35.2, genes codifying for Cl- tonoplast transporters, were exclusively overexpressed in plants of the salt-tolerant combination CM-OR, suggesting that these transporters involved in Cl- compartmentalization could be crucial for salt stress tolerance. It is concluded that to improve citrus tolerance to high salinity, it is important that scions have a versatile photosynthetic system, an adequate stomatal density, and a proper modulation of genes coding for Cl- transporters in the tonoplast.


Effects of Inpatient Rehabilitation in Leg Lymphedema: A Naturalistic Prospective Cohort Study With Intra-individual Control of Effects.
To quantify therapy-attributable effects of a comprehensive inpatient rehabilitation program for lower limb lymphedema (LLL) and to compare the levels of health-related quality of life (HRQL) to population-based norms. Naturalistic prospective cohort study with intra-individual control of effects. Rehabilitation hospital. Patients with LLL (N=67; 46 women). Comprehensive, multidisciplinary inpatient rehabilitation with 45-60 hours of therapy. Short Form 36 (SF-36) for HRQL, lymphedema-specific Freiburg Quality of Life Assessment for lymphatic disorders, Short Version (FLQA-lk), knee-specific Knee Outcome Survey Activities of Daily Living Scale (KOS-ADL), and Symptom Checklist-90Standard (SCL-90S). Observed pre/post rehabilitation effects were individually corrected by subtracting the home waiting-time effects and expressed as standardized effect sizes (ESs) and standardized response means (SRMs). Score differences to norms were quantified by standardized mean differences (SMDs). Participants were on average aged 60.5 years, not yet obese, and had 3 comorbidities (n=67). The greatest improvement was in HRQL on the FLQA-lk with ES=0.767/SRM=0.718, followed by improvements in pain and function with ES/SRM=0.430-0.495 on the SF-36, FLQA-lk, and KOS-ADL (all P<.001). Vitality, mental health, emotional well-being, and interpersonal sensitivity improved most by ES/SRM=0.341-0.456 on all 4 measures (all P≤.003). Post rehabilitation scores were significantly higher than population norms on SF-36 bodily pain (SMD=1.140), vitality (SMD=0.886), mental health (SMD=0.815), and general health (SMD=0.444) (all P<.001), and comparable on the other scales. Those affected by LLL stages II and III benefited substantially from the intervention, attaining equal or higher levels of HRQL than expected compared with the general population norms. Multidisciplinary, inpatient rehabilitation should be recommended for LLL management.


Inborn errors of brain myelin formation.
Inborn errors of brain myelin formation or hypomyelinating leukodystrophies (HLD) represent a heterogeneous group of white matter diseases related to a primitive impairment of oligodendrocytes to produce myelin in the central nervous system (CNS). Cerebral magnetic resonance imaging (MRI) allows an assessment of the myelination pattern. The clinical presentation is related to the degree of hypomyelination and its consequences on axonal functions. When the gene defect interferes with the active infantile phase of myelination, the consequences might be severe, with delayed and loss of psychomotor development, absence of myelin signal on cerebral MRI and of identifiable waves on cerebral evoked potentials, as described by Pelizaeus and Merzbacher (PMD). When the pathophysiological mechanism is less severe, myelin production is maintained, although signs of progressive axonopathy are observed, related to progressive spastic paraplegia (SPG) associated with cognitive or behavioral disturbances. HLDs have been classified according to gene defects or associated signs. The X-linked HDL1 (PMD and SPG2) is related to the gene that controls the production of the major CNS myelin proteins, the proteolipid proteins (PLP). The gap junction protein, gamma 2 gene (GJC2) encoding oligodendrocyte-specific connexin, has been shown to be involved in the autosomal recessive HLD2 (PMLD1 and SPG44).


[Skin aging-cellular senescence : What is the future?]
Cellular senescence is the main cause of skin and organ aging and is associated with a wide range of aging-related diseases. To understand which senolytics, senomorphics, and cell-based therapies have been developed to alleviate and even rejuvenate skin aging and reduce cellular senescence. Basic literature for the mode of action of senolytics and senomorphics and their clinical perspectives in daily routine are discussed. Various causes lead to mitochondrial dysfunction and the activation of pro-aging signaling pathways, which eventually lead to cellular senescence with degradation of structural proteins of the dermal connective tissue and severe suppression of regenerative stem cell niches of the skin. Depletion of senescent cells suppress skin aging and enforce rejuvenation of skin and other organs and their function. The removal of senescent cells by cells of the native immune system is severely disturbed during aging. Selected senolytics and senomorphics are approved and are already on the market.


Hexokinases link DJ-1 to the PINK1/parkin pathway.
Early onset Parkinson's disease is caused by variants in PINK1, parkin, and DJ-1. PINK1 and parkin operate in pathways that preserve mitochondrial integrity, but the function of DJ-1 and how it relates to PINK1 and parkin is poorly understood. A series of unbiased high-content screens were used to analyze changes at the protein, RNA, and metabolite level in rodent brains lacking DJ-1. Results were validated using targeted approaches, and cellular assays were performed to probe the mechanisms involved. We find that in both rat and mouse brains, DJ-1 knockout results in an age-dependent accumulation of hexokinase 1 in the cytosol, away from its usual location at the mitochondria, with subsequent activation of the polyol pathway of glucose metabolism in vivo. Both in the brain and in cultured cells, DJ-1 deficiency is associated with accumulation of the phosphatase PTEN that antagonizes the kinase AKT. In cells, addition of an inhibitor of AKT (MK2206) or addition of a peptide to dissociate association of hexokinases from mitochondria both inhibit the PINK1/parkin pathway, which works to maintain mitochondrial integrity. Hexokinases are an important link between three major genetic causes of early onset Parkinson's disease. Because aging is associated with deregulated nutrient sensing, these results help explain why DJ-1 is associated with age-dependent disease.


[Genetic control of chromosome synapsis in mice heterozygous for a paracentric inversion].
Frequencies of formation of inversion loops and their relative sizes were studied in laboratory mice heterozygous at paracentric inversion In1(1)Rk in chromosome 1, depending on the genetic background. Homozygotes In1/In1 were crossed with mice from five inbred strains (A/HeJ, BALB/cJ, C3H/HeJ, C57BL/6J, DBA2/J). The frequency of formation of inversion loops, their relative sizes, and the dependence of these parameters on the stage of pachitene were analyzed on electron-microscopic slides of spread spermatocytes in first-generation hybrids. It was shown that the genetic background and cross direction statistically significantly influenced the duration of individual pachitene stages and the frequency of inversion loops, but not relative loop size. Using a database on SNP distribution in the inbred strains examined, we carried out in silico mapping of genes affecting the genotype-dependent characters. We have found that the efficiency of synapsis in the inversion does not depend on interstrain differences in homology of the chromosome 1 region involved in the inversion. Genes controlling the inversion loop frequency in the inversion heterozygotes were mapped to chromosome 7, and genes controlling the duration of individual pachitene stages, to chromosomes 2 and 5.


Preliminary investigation of polycyclic aromatic hydrocarbons (PAHs) concentration, compositional pattern, and ecological risk in crude oil-impacted soil from Niger delta, Nigeria.
Crude oil contamination could serve as an important source of polycyclic aromatic hydrocarbons (PAHs) in the environment. Determining the concentration and distribution of PAHs and their ecological risk could provide clues for appropriate remediation. The present study investigated the concentrations, composition pattern, and ecological risk of PAHs in crude oil-contaminated soil collected from the Niger Delta, Nigeria. The concentrations of Σ29PAHs and 16 priority US-EPA PAHs (Σ16PAHs) in the soil ranged from 24230.68 to 40845.32 ng/g (average: 29953.47 ng/g) and 7361.66-14141.49 ng/g (average: 9819.96 ng/g), respectively. The concentrations of Σ16PAHs US-EPA in all the studied locations far exceeded the safety value of 1000 ng/g set by the soil quality guidelines of Switzerland and above which is regarded as being highly contaminated for Agricultural soils of Poland. The percentage composition of 2-, 3-,4-, 5-, and 6-ring in the soils were 33.69%, 56.31%, 9.47%, 0.52%, and 0.02% of the total PAHs, respectively, indicating the predominance of low molecular weight PAHs typical of petrogenic origin. Ecological risk assessment indicated a high risk to the soil biota and ecosystem in the studied soil samples as presented by individual PAH and total PAHs. 2-, 3- and 4-ring PAHs in the soil contributed significantly to the ecological risk burden in the soils. This research work provides useful information on the ecological risk associated with crude oil contamination and consequently would assist the government in formulating precise, targeted, and effective remediation measures for soil contaminated with crude oil.


Palliative Care Needs Assessment for Pediatric Complex Care Providers.
Children with medical complexity (CMC) are often cared for by both complex care and palliative care pediatric teams. No prior research has investigated the relationship between these two disciplines. The purpose of this article is to investigate challenges that complex care programs face in caring for children with medical complexity (CMC), as well as to explore whether identified challenges could be met through collaboration with pediatric palliative care or additional training for complex care teams. Medical providers who self-identified as providing clinical care to children with medical complexity were asked to complete an online anonymous survey. Subjects were recruited through a Complex Care listerv. Data were analyzed using descriptive statistics. 85 subjects completed the survey, of whom 87.1% (n=74) were physicians, and 12.0% (n=11) were nurse practitioners. Subjects reported several challenges in caring for CMC, including symptom management, establishing goals of care, advance care planning, and coordination of care. A majority of subjects reported benefitting from palliative care consultative assistance in each subject area. Most subjects described their relationship with palliative care as a close partnership with frequent overlap. The evolving field of pediatric complex care is associated with an array of challenges in caring for CMC. Many of these challenges include competency areas where palliative care providers receive concerted training. Our research suggests greater palliative care involvement in the CMC population can benefit complex care teams and patients, given the expertise palliative providers can bring to the population and the discipline of complex care.