Circadian Behavioral Responses to Light and Optic Chiasm-Evoked Glutamatergic EPSCs in the Suprachiasmatic Nucleus of ipRGC Conditional vGlut2 Knock-Out Mice

Intrinsically photosensitive retinal ganglion cells (ipRGCs) innervate the hypothalamic suprachiasmatic nucleus (SCN), a circadian oscillator that functions as a biological clock. ipRGCs use vesicular glutamate transporter 2 (vGlut2) to package glutamate into synaptic vesicles and light-evoked resetting of the SCN circadian clock is widely attributed to ipRGC glutamatergic neurotransmission. Pituitary adenylate cyclase-activating polypeptide (PACAP) is also packaged into vesicles in ipRGCs and PACAP may be coreleased with glutamate in the SCN. vGlut2 has been conditionally deleted in ipRGCs in mice [conditional knock-outs (cKOs)] and their aberrant photoentrainment and residual attenuated light responses have been ascribed to ipRGC PACAP release. However, there is no direct evidence that all ipRGC glutamatergic neurotransmission is eliminated in vGlut2 cKOs. Here, we examined two lines of ipRGC vGlut2 cKO mice for SCN-mediated behavioral responses under several lighting conditions and for ipRGC glutamatergic neurotransmission in the SCN. Circadian behavioral responses varied from a very limited response to light to near normal photoentrainment. After collecting behavioral data, hypothalamic slices were prepared and evoked EPSCs (eEPSCs) were recorded from SCN neurons by stimulating the optic chiasm. In cKOs, glutamatergic eEPSCs were recorded and all eEPSC parameters examined (stimulus threshold, amplitude, rise time or time-to-peak and stimulus strength to evoke a maximal response) were similar to controls. We conclude that a variable number but functionally significant percentage of ipRGCs in two vGlut2 cKO mouse lines continue to release glutamate. Thus, the residual SCN-mediated light responses in these cKO mouse lines cannot be attributed solely to ipRGC PACAP release

Variation in Organophosphate Pesticide Metabolites in Urine of Children Living in Agricultural Communities

Children of migrant farmworkers are at increased risk of exposure to organophosphate pesticides because of “carry-home” transport processes and residential location. Although this at-risk status is generally recognized, few available reports describe the extent of this exposure among agricultural communities. We quantified dialkyl phosphate (DAP) levels in serial samples of urine from 176 children, 2–6 years of age, in three Oregon communities hosting differing agricultural industries: pears, cherries, and fruit berries. Up to three spot samples of urine were collected from children at the beginning, mid-point, and end of their parents’ work seasons. The median levels of dimethylthiophosphate (DMTP), the most commonly detected metabolite, was significantly higher in urine samples from children in each of the three agricultural communities (17.5, 19.0, and 41.0 ng/mL) relative to a reference group of children who lived in an urban community and whose parents did not work in agriculture (6.5 ng/mL; Kruskal-Wallis, p < 0.001). After controlling for age, sex, and weight, the median level of DMTP in children in the pear community was 1.92 times higher than the level in children of the berry community [95% confidence interval (CI), 1.14–3.23] and 1.75 times higher than the level in children of the cherry community (95% CI, 0.95–3.23). We observed increasing levels of DMTP across the work season only within the berry community. Levels decreased in the cherry community and remained constant in the pear community. Substantial temporal variation within the children followed demonstrates the need for multiple urine samples to most accurately characterize longer term and/or cumulative exposure. The observed variability in urinary DAP levels, between communities and over time, could be attributed to the types and amounts of organophosphate pesticides used, the timing of applications and degradation of residues in the environment, work operations and hygiene practices, the proximity of housing to orchards and fields, or the movement of these working families. Additional studies of variation in pesticide exposure across agricultural regions are needed

Near Earth space sporadic radio emission busts occurring during sunrise

During the period of low solar activity at sunrise the effect of sporadic high frequency near Earth space radio emission was experimentally discovered at middle latitudes. The possible mechanism of its origin is discussed

In vivo fluorescence lifetime imaging of macrophage intracellular metabolism during wound responses in zebrafish

The function of macrophages in vitro is linked to their metabolic rewiring. However, macrophage metabolism remains poorly characterized in situ. Here, we used two-photon intensity and lifetime imaging of autofluorescent metabolic coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD), to assess the metabolism of macrophages in the wound microenvironment. Inhibiting glycolysis reduced NAD(P)H mean lifetime and made the intracellular redox state of macrophages more oxidized, as indicated by reduced optical redox ratio. We found that TNFα+ macrophages had lower NAD(P)H mean lifetime and were more oxidized compared to TNFα− macrophages. Both infection and thermal injury induced a macrophage population with a more oxidized redox state in wounded tissues. Kinetic analysis detected temporal changes in the optical redox ratio during tissue repair, revealing a shift toward a more reduced redox state over time. Metformin reduced TNFα+ wound macrophages, made intracellular redox state more reduced and improved tissue repair. By contrast, depletion of STAT6 increased TNFα+ wound macrophages, made redox state more oxidized and impaired regeneration. Our findings suggest that autofluorescence of NAD(P)H and FAD is sensitive to dynamic changes in intracellular metabolism in tissues and can be used to probe the temporal and spatial regulation of macrophage metabolism during tissue damage and repair

АУТОТРАНСПЛАНТАЦИЯ АНТЕРОЛАТЕРАЛЬНОГО БЕДРЕННОГО ЛОСКУТА — МЕТОД ВЫБОРА В НЕОТЛОЖНОЙ РЕКОНСТРУКТИВНОЙ ХИРУРГИИ НИЖНЕЙ КОНЕЧНОСТИ (АНАЛИЗ КЛИНИЧЕСКИХ НАБЛЮДЕНИЙ)

BACKGROUND. Requirements for the graft used in microsurgery are simple retrieval, minimal anatomic variability, the possibility to operate on one surgical area, great length and diameter of flap vessels.PURPOSE OF STUDY. Evaluation of the results and advantages of revascularized free anterolateral muscle flap usage in emergency surgery.MATERIALS AND METHODS. Free muscle flap of the lateral vastus muscle on a vascular pedicle of the descending branch of the lateral femoral circumflex artery (anterolateral flap) was used to replace the defect in 2 patients. In one case, a patient had open fractures of the lower leg, complicated with primary defects of soft tissue, and in the other case a patient had incomplete traumatic amputation of the left foot. All the victims underwent soft tissue defects restoration within the first hours after the injury, next to fixation of the fracture.RESULTS. All grafts have completely healed, total necrosis of muscle flaps hasn’t been observed. All patients had primary wound healing after the transfer. Cases of deep purulent infection after the surgery haven’t been noted.CONCLUSION. The transfer of a free anterolateral muscle flap is the best method for emergency plastic and reconstructive surgery of the lower limbs. The advantages are simple and prompt retreival, no need to turn the patient to the lateral position, large amount of the flap, great length and caliber of vessels. АКТУАЛЬНОСТЬ. Требования, предъявляемые к трансплантату, используемому в экстренной микрохирургии — это простота забора, минимальная вариабельность анатомии, возможность работы на одном операционном поле, большие длина и диаметр сосудов.ЦЕЛЬ ИССЛЕДОВАНИЯ. Оценить результаты использования и преимущества свободного реваскуляризированного мышечного антеролатерального бедренного трансплантата в экстренной микрохирургии.МАТЕРИАЛ И МЕТОДЫ. Свободный мышечный лоскут из латеральной широкой мышцы бедра на сосудистой ножке из нисходящей ветви латеральной артерии, огибающей бедренную кость (антеролатеральный лоскут), был применен для замещения дефекта 2 пострадавшим. В одном случае это был пациент с открытым переломом костей голени, осложненным первичными дефектами мягких тканей, в другом — неполная травматическая ампутация левой стопы. Всем пострадавшим замещение дефекта мягких тканей выполнено в первые часы после травмы, после фиксации перелома.РЕЗУЛЬТАТЫ. Все трансплантаты прижились полностью, тотальных некрозов мышечных лоскутов не установлено. У всех пациентов наблюдалось первичное заживление ран после пересадки лоскута. Случаев глубокой гнойной инфекции после закрытия дефекта лоскутом не отмечено.ВЫВОДЫ. Пересадка свободного антеролатерального бедренного мышечного лоскута — это оптимальный метод для экстренной пластической и реконструктивной хирургии нижних конечностей. Его преимуществами являются простота и быстрота забора, отсутствие необходимости поворота пострадавшего на бок, большой объем лоскута, большие длина и калибр сосудов.

ВОССТАНОВЛЕНИЕ ПОКРОВНЫХ ТКАНЕЙ У ПОСТРАДАВШИХ С ТЯЖЕЛЫМИ ОТКРЫТЫМИ ПЕРЕЛОМАМИ КОСТЕЙ ГОЛЕНИ

RESUME. The aim of our study is to develop and introduce the algorithm of the early soft tissue reconstruction for the patients with severe open tibia fractures. We analyzed the treatment results retrospectively and prospectively 84 patients with severe open tibia fractures, complicated by soft tissue defects. In the comparison group (56 patients) we apply late soft tissue reconstruction. In the study group (28 patients) we applied the algorithm of early soft tissue reconstruction by plastic surgery methods. Surgery tactic depends on the patient’s condition, trauma mechanism, soft tissue defects size and localization. The algorithm of early soft tissue reconstruction was used for patients with severe open tibia fractures decreases duration of hospital treatment, rate of deep wound infection and partial necrosis of tibia bone. РЕЗЮМЕ. Целью исследования являлась разработка и применение алгоритма ранней реконструкции мягких тканей у пациентов с тяжелыми открытыми переломами костей голени. Мы ретроспективно и проспективно изучили результаты лечения 84 пострадавших с открытыми переломами костей голени, осложненными дефектами мягких тканей. В группе сравнения (56 пострадавших) применяли тактику отсроченного восстановления мягких тканей. В исследуемой группе (28 пострадавших) применили алгоритм раннего восстановления покровных тканей методами пластической хирургии. Выбор хирургической тактики проводили в зависимости от тяжести состояния пострадавшего, механизма образования, величины и локализации дефекта мягких тканей. При применении алгоритма раннего восстановления покровных тканей у пострадавших с открытыми переломами костей голени отмечено значительное сокращение сроков стационарного лечения, частоты глубокой раневой инфекции и частоты некроза участков большеберцовой кости.

The experience of levetiracetam (keppra) application for the treatment of pain syndromes in oncologic patients on the background of chemotherapy and patients with facial neuralgia

Antiepileptic preparation Levetiracetam (Keppra) possesses minimal medicinal interaction with other drugs which are used for epileptic patients with severe concomitant diseases. We have studied Levetiracepam efficacy in patients with tumours of various localization, with marked pain syndrome generalized after a complex treatment (operative, radiation) on the background of chemotherapy. The second group included the patients with facial neuralgia. Analyzing the results of the preparation application we have determined that Levetiracetam is mostly effective with the dose of 2500-3000mg for 60-65old patients having metastases, with the dose reaching 2000mg per day for 70-80old patients having facial neuralgia.Противоэпилептический препарат леветирацетам (кеппра) обладает минимальным лекарственным взаимодействием с другими препаратами, которые применяются у больных эпилепсией с тяжелыми сопутствующими заболеваниями. Изучена эффективность леветирацетама у пациентов с различной локализацией опухолевого процесса, с выраженным болевым синдромом, генерализовавшимся после проведенного ранее комплексного лечения (оперативное, лучевое) на фоне химиотерапии. Вторую группу составили пациенты с невралгией тройничного нерва. Анализируя результаты применения препарата, установлено, что леветирацетам наиболее эффективен у больных с метастазами в 60-65 лет в дозе - 2500-3000 мг, с НТН в 70-80 лет при достижении дозы 2000 мг в сутки

The Cycad Genotoxin MAM Modulates Brain Cellular Pathways Involved in Neurodegenerative Disease and Cancer in a DNA Damage-Linked Manner

Methylazoxymethanol (MAM), the genotoxic metabolite of the cycad azoxyglucoside cycasin, induces genetic alterations in bacteria, yeast, plants, insects and mammalian cells, but adult nerve cells are thought to be unaffected. We show that the brains of adult C57BL6 wild-type mice treated with a single systemic dose of MAM acetate display DNA damage (O6-methyldeoxyguanosine lesions, O6-mG) that remains constant up to 7 days post-treatment. By contrast, MAM-treated mice lacking a functional gene encoding the DNA repair enzyme O6-mG DNA methyltransferase (MGMT) showed elevated O6-mG DNA damage starting at 48 hours post-treatment. The DNA damage was linked to changes in the expression of genes in cell-signaling pathways associated with cancer, human neurodegenerative disease, and neurodevelopmental disorders. These data are consistent with the established developmental neurotoxic and carcinogenic properties of MAM in rodents. They also support the hypothesis that early-life exposure to MAM-glucoside (cycasin) has an etiological association with a declining, prototypical neurodegenerative disease seen in Guam, Japan, and New Guinea populations that formerly used the neurotoxic cycad plant for food or medicine, or both. These findings suggest environmental genotoxins, specifically MAM, target common pathways involved in neurodegeneration and cancer, the outcome depending on whether the cell can divide (cancer) or not (neurodegeneration). Exposure to MAM-related environmental genotoxins may have relevance to the etiology of related tauopathies, notably, Alzheimer's disease

Circadian Behavioral Responses to Light and Optic Chiasm-Evoked Glutamatergic EPSCs in the Suprachiasmatic Nucleus of ipRGC Conditional vGlut2 Knock-Out Mice

Intrinsically photosensitive retinal ganglion cells (ipRGCs) innervate the hypothalamic suprachiasmatic nucleus (SCN), a circadian oscillator that functions as a biological clock. ipRGCs use vesicular glutamate transporter 2 (vGlut2) to package glutamate into synaptic vesicles and light-evoked resetting of the SCN circadian clock is widely attributed to ipRGC glutamatergic neurotransmission. Pituitary adenylate cyclase-activating polypeptide (PACAP) is also packaged into vesicles in ipRGCs and PACAP may be coreleased with glutamate in the SCN. vGlut2 has been conditionally deleted in ipRGCs in mice [conditional knock-outs (cKOs)] and their aberrant photoentrainment and residual attenuated light responses have been ascribed to ipRGC PACAP release. However, there is no direct evidence that all ipRGC glutamatergic neurotransmission is eliminated in vGlut2 cKOs. Here, we examined two lines of ipRGC vGlut2 cKO mice for SCN-mediated behavioral responses under several lighting conditions and for ipRGC glutamatergic neurotransmission in the SCN. Circadian behavioral responses varied from a very limited response to light to near normal photoentrainment. After collecting behavioral data, hypothalamic slices were prepared and evoked EPSCs (eEPSCs) were recorded from SCN neurons by stimulating the optic chiasm. In cKOs, glutamatergic eEPSCs were recorded and all eEPSC parameters examined (stimulus threshold, amplitude, rise time or time-to-peak and stimulus strength to evoke a maximal response) were similar to controls. We conclude that a variable number but functionally significant percentage of ipRGCs in two vGlut2 cKO mouse lines continue to release glutamate. Thus, the residual SCN-mediated light responses in these cKO mouse lines cannot be attributed solely to ipRGC PACAP release

Analytical Validation of Familial Hypercholesterolemia Biomarkers in Dried Blood Spots

Heterozygous familial hypercholesterolemia (HeFH) is a common, treatable genetic disorder characterized by premature atherosclerosis and cardiovascular disease, yet the majority of affected individuals remain undiagnosed. Newborn screening could play a role in identification of at-risk individuals and provide an opportunity for early intervention, prior to the onset of symptoms. The objective of this study was to develop and validate assays for quantification of candidate HeFH biomarkers in dried blood spots (DBS). Commercially available enzyme assay kits for quantification of serum total cholesterol (TC) and low-density lipoprotein-cholesterol (LDL-C) were modified for high-throughput analysis of DBS. Apolipoprotein B (ApoB) concentrations in DBS were measured using an immunoassay with modifications from published studies. All three assays were validated according to the College of American Pathologists guidelines for clinical laboratories. The performance of TC, LDL-C, and ApoB assays was assessed by precision, recovery, limit of quantification (LOQ) and linearity. Precision studies yielded coefficients of variation (CV) of less than 15%, with recovery greater than 75% for all three assays. The determined LOQ and linearity were comparable to serum-based assays. In a direct comparison between serum and DBS concentrations, positive correlations were demonstrated for TC, LDL-C, and ApoB. Additionally, the initial evaluation of the three biomarker concentrations within the unaffected population was similar to values obtained in previous published studies. This study reports on methods for quantification of TC, LDL-C, and ApoB in DBS. Assay validation results were within acceptable limits for newborn screening. This is an important first step toward the identification of newborns with HeFH