Regulatory mode shift of Tbc1d1 is required for acquisition of insulin-responsive GLUT4-trafficking activity.
Tbc1d1 is key to skeletal muscle GLUT4 regulation. By using GLUT4 nanometry combined with a cell-based reconstitution model, we uncover a shift in the regulatory mode of Tbc1d1 by showing that Tbc1d1 temporally acquires insulin responsiveness, which triggers GLUT4 trafficking only after an exercise-mimetic stimulus such as aminoimidazole carboxamide ribonucleotide (AICAR) pretreatment. The functional acquisition of insulin responsiveness requires Ser-237 phosphorylation and an intact phosphotyrosine-binding (PTB) 1 domain. Mutations in PTB1, including R125W (a natural mutant), thus result in complete loss of insulin-responsiveness acquisition, whereas AICAR-responsive GLUT4-liberation activity remains intact. Thus our data provide novel insights into temporal acquisition/memorization of Tbc1d1 insulin responsiveness, relying on the PTB1 domain, possibly a key factor in the beneficial effects of exercise on muscle insulin potency.


Riboflavin deficiency and esophageal cancer: a case control-household study in the Caspian Littoral of Iran.
In a case-household-control-household study in two very high and low esophageal cancer (EC) risk regions of the Caspian Littoral of Iran, a total of 21 cases (12 subjects from the high risk and 9 subjects from the low-risk region) with a total of 91 household members (57 subjects from the high risk and 34 subjects from the low-risk region) were investigated. Cases were matched for sex and age (+/-5 years) with non-blood relative controls. A standard 24-h dietary recall questionnaire was used to estimate riboflavin intake. The erythrocyte glutathione reductase activity coefficient (EGR-AC) was measured to assess riboflavin status. The Student t-test was used to test differences, and chi2 analysis was applied to test associations. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were also calculated. Results indicated that in both regions, the mean daily intake of riboflavin for cases was less than that of the controls (0.66+/-0.43 mg/day versus 0.82+/-0.37 mg/day) whereas for their households, it was virtually the same. Both cases and control households showed riboflavin deficiency in two regions, with higher deficiency in the high risk area. Statistical analysis revealed significant differences between the two regions for EGR-AC (P<0.001). Odd ratios indicated that the risk of developing EC for persons living in riboflavin-deficient households was more than twice of non-deficient households. Therefore, this study suggests that riboflavin deficiency may play an important role in the etiology of esophageal cancer in the Caspian Littoral of Iran.


Long-term patterns in Iberian hare population dynamics in a protected area (Doñana National Park) in the southwestern Iberian Peninsula: Effects of weather conditions and plant cover.
The Iberian hare (Lepus granatensis) is a widely distributed endemic species in the Iberian Peninsula. To improve our knowledge of its population dynamics, the relative abundance and population trends of the Iberian hare were studied in the autumns of 1995-2012 in a protected area (Doñana National Park) by spotlighting in 2 different habitats: marshland and ecotones. The average relative abundance was 0.38 hare/km (SD = 0.63) in the marshland and 3.6 hares/km (SD = 4.09) in ecotones. The Iberian hare population exhibited local interannual fluctuations and a negative population trend during the study period (1995-2012). The results suggest that its populations are in decline. The flooding of parts of the marshland in June, July and October favor hare abundance in the ecotone. Hare abundance in the marshland increases as the flooded surface area increases in October. These effects are more pronounced if the rains are early (October) and partially flood the marsh. By contrast, when marsh grasses and graminoids are very high and thick (as measured using the aerial herbaceous biomass [biomass marshland] as a proxy), the abundance of hares decreases dramatically as does the area of the marsh that is flooded (in November).


Determination of intracellular pH using sensitive, clickable fluorescent probes.
We synthesized and evaluated a series of acidic fluorescent pH probes exhibiting robust pH dependence, high sensitivity and photostability, and excellent cell membrane permeability. Titration analyses indicated that probe 3 could increase its fluorescence intensity 800-fold between pH 8.0 and 4.1. Additionally, its pK(a) value is optimal for intracellular probing of acidic organelles. Fluorescent imaging of HepG2 and Hela cells further revealed that probe 3 demonstrates outstanding capacity for monitoring of intracellular [H(+)] levels. The easily accessible terminal alkyne/azido function groups of these probes offer the possibility of rapidly constructing sensor molecule libraries using 'click' chemistry.


MicroRNA125b-mediated Hedgehog signaling influences liver regeneration by chorionic plate-derived mesenchymal stem cells.
Although chorionic plate-derived mesenchymal stem cells (CP-MSCs) were shown to promote liver regeneration, the mechanisms underlying the effect remain unclear. Hedgehog (Hh) signaling orchestrates tissue reconstruction in damaged liver. MSCs release microRNAs mediating various cellular responses. Hence, we hypothesized that microRNAs from CP-MSCs regulated Hh signaling, which influenced liver regeneration. Livers were obtained from carbon tetrachloride (CCl4)-treated rats transplanted with human CP-MSCs (Tx) or saline (non-Tx). Sonic Hh, one of Hh ligands, increased in CCl4-treated liver, whereas it decreased in CP-MSC-treated liver with CCl4. The expression of Hh-target genes was significantly downregulated in the Tx. Reduced expansion of progenitors and regressed fibrosis were observed in the liver of the Tx rats. CP-MSCs suppressed the expression of Hh and profibrotic genes in co-cultured LX2 (human hepatic stellate cell) with CP-MSCs. MicroRNA-125b targeting smo was retained in exosomes of CP-MSCs. CP-MSCs with microRNA-125b inhibitor failed to attenuate the expression of Hh signaling and profibrotic genes in the activated HSCs. Therefore, these results demonstrated that microRNA-125b from CP-MSCs suppressed the activation of Hh signaling, which promoted the reduced fibrosis, suggesting that microRNA-mediated regulation of Hh signaling contributed to liver regeneration by CP-MSCs.


Prevalence and predictors of lipid abnormalities in patients treated with statins in the UK general practice.
To identify the prevalence and predictors of lipid abnormalities in statin-treated patients in the UK. A retrospective cohort study was performed using the UK General Practice Research Database. Patients >or=35 years of age were included if they received first-ever statin between January 2000 and December 2004, received statins for at least 6 weeks, had >2 years of pre- and 1 year of post-statin initiation database history, received no concomitant lipid lowering drugs and had at least one complete lipid profile conducted within 1 year before and after initiating statins. Predictors of each lipid abnormality were determined using random effects logistic regression. Within 1 year of statin initiation, 34.7%, 27.4%, 68.2% and 57.6% of patients did not reach optimal levels of TC, LDL-C, HDL-C and TG, respectively. Failure to attain TC goal was explained by smoking (odds ratio=1.13, 95% confidence interval [1.02-1.23]) and baseline TC >6.2mmol/L (5.01, [4.58-5.48]). Failure to attain LDL-C goal was associated with smoking (1.28, [1.14-1.43]), LDL-C >or=4.1 to <4.9mmol/L (2.72, [2.45-3.03]) and LDL-C >or=4.9mmol/L (8.54, [7.62-9.54]). High CHD risk was associated with low HDL-C in women at follow-up (1.94, [1.51-2.48]). Elevated TG was associated with baseline TG >or=2.2 to <5.6mmol/L (4.19, [3.81-4.59]), TG >or=5.6mmol/L (16.10, [3.67-70.57]), smoking (1.26, [1.11-1.42]) and hypertension (1.12, [1.01-1.23]). While TC and LDL-C management appear appropriate in the UK, attainment of recommended levels of HDL-C and TG remains poor. The latter remains a concern, as poor attainment of recommended levels of HDL-C and TG is associated with high CHD risk in women, smoking and hypertension.


Synthetic applications with use of a silica-supported alkyne metathesis catalyst.
A highly active and durable fumed silica-supported heterogeneous molybdenum(VI) catalyst was applied to ring closing alkyne metathesis and cyclooligomerization reactions to give high yields of metathesis products near room temperature conditions.


Identification of calcium release-triggering and blocking regions of the II-III loop of the skeletal muscle dihydropyridine receptor.
In an attempt to identify and characterize functional domains of the rabbit skeletal muscle dihydropyridine receptor alpha 1 subunit II-III loop, we synthesized several peptides corresponding to different regions of the loop: peptides A, B, C, C1, C2, D (cf. Fig. 1). Peptide A (Thr671-Leu690) activated [3H]ryanodine binding to, and induced Ca2+ release from, rabbit skeletal muscle triads, but none of the other peptides had such effects. Peptide A-induced Ca2+ release and activation of ryanodine binding were partially suppressed by an equimolar concentration of peptide C (Glu724-Pro760) but were not affected by the other peptides. These results suggest that the short stretch in the II-III loop, Thr671-Leu690, is responsible for triggering SR Ca2+ release, while the other region, Glu724-Pro760, functions as a blocker of the release trigger. A hypothesis is proposed to account for how these subdomains interact with the sarcoplasmic reticulum Ca2+ release channel protein during excitation-contraction coupling.


Effects of Mechanical Complications on Radiation Exposure During Fluoroscopically Guided Gastrojejunostomy Exchange in the Pediatric Population.
The purpose of the article is to evaluate the effects of mechanical complications, such as clogging or coiling, of gastrojejunostomy tubes on radiation exposure during exchange in the pediatric population. In this HIPAA-compliant and IRB-approved study, we retrospectively reviewed procedural records for patients undergoing gastrojejunostomy (GJ) tube exchange during a 4-month period in 2014. Success of the procedure, specifications of the tube, age, and sex of the patient as well as radiation exposure during the procedure were included. Radiation exposure was measured in fluoroscopy time and cumulative air kerma. Complications encountered during exchange were also recorded, if available. Patients presenting for gastrostomy to GJ conversions or combined procedures were excluded from the study. Ordinary and mixed effect linear regression models were used to test associations between GJ tube parameters, presence of mechanical complications, and fluoroscopy time and radiation dose. 146 patients undergoing 285 GJ exchanges met inclusion criteria over the 4-month study period (M:F 82:64). All exchanges were successful with 85 demonstrating a form of mechanical complication (44 coiled, 41 clogged). Of the reported GJ tube specifications, only tube length was significantly associated with mechanical complications (p < 0.001). The presence of mechanical complication was significantly associated with increased radiation exposure and fluoroscopy time (p < 0.0001). Mechanical complications of gastrojejunostomy tubes, such as clogging or coiling, are associated with increased radiation exposure during exchange. Strategies to decrease these complications, including re-siting the gastrostomy tract or placement of a surgical jejunostomy in the event of repeated coiling of a tube should be strongly considered.


Molecular staging of lymph node-negative colon carcinomas by one-step nucleic acid amplification (OSNA) results in upstaging of a quarter of patients in a prospective, European, multicentre study.
Current histopathological staging procedures in colon carcinomas depend on midline division of the lymph nodes with one section of haematoxylin & eosin (H&E) staining only. By this method, tumour deposits outside this transection line may be missed and could lead to understaging of a high-risk group of stage UICC II cases, which recurs in ∼20% of cases. A new diagnostic semiautomated system, one-step nucleic acid amplification (OSNA), detects cytokeratin (CK) 19 mRNA in lymph node metastases and enables the investigation of the whole lymph node. The objective of this study was to assess whether histopathological pN0 patients can be upstaged to stage UICC III by OSNA. Lymph nodes from patients who were classified as lymph node negative after standard histopathology (single (H&E) slice) were subjected to OSNA. A result revealing a CK19 mRNA copy number >250, which makes sure to detect mainly macrometastases and not isolated tumour cells (ITC) or micrometastases only, was regarded as positive for lymph node metastases based on previous threshold investigations. In total, 1594 pN0 lymph nodes from 103 colon carcinomas (median number of lymph nodes per patient: 14, range: 1-46) were analysed with OSNA. Out of 103 pN0 patients, 26 had OSNA-positive lymph nodes, resulting in an upstaging rate of 25.2%. Among these were 6/37 (16.2%) stage UICC I and 20/66 (30.3%) stage UICC II patients. Overall, 38 lymph nodes were OSNA positive: 19 patients had one, 3 had two, 3 had three, and 1 patient had four OSNA-positive lymph nodes. OSNA resulted in an upstaging of over 25% of initially histopathologically lymph node-negative patients. OSNA is a standardised, observer-independent technique, allowing the analysis of the whole lymph node. Therefore, sampling bias due to missing investigation of certain lymph node tissue can be avoided, which may lead to a more accurate staging.


Host susceptibility to tuberculosis: insights from a longitudinal study of gene expression in diabetes.
Tuberculosis (TB) remains a major global disease, and diabetes, which is documented to increase susceptibility to TB threefold, is also becoming pandemic. This susceptibility has been attracting extensive research interest. The increased risk of TB in diabetes may serve as a unique model to understand host susceptibility to specific pathogens in humans. To examine this rationale, we investigated the expression of reported TB candidate genes in a longitudinal diabetes study. Two genes, HK2 and CD28, emerged as potential culprits in diabetes-increased TB susceptibility.


Recent Advances in Lipid-based Nanodrug Delivery Systems in Cancer Therapy.
Cancer is the second leading cause of death globally, with every sixth death being attributable to cancer. Nevertheless, the efficacy of conventional chemotherapeutic drugs is often limited due to their poor solubility, unfavorable pharmacokinetic profile, and lack of tumor selectivity. The use of nanotechnology provides an opportunity to enhance the efficacy of a chemotherapeutic drug by improving its bioavailability and pharmacokinetic profile while facilitating preferential accumulation at the tumor tissue. To date, a variety of platforms have been investigated as nanocarriers in oncology, which include lipid-based, polymer-based, inorganic materials, and even viruses. Among different nanocarriers, lipid-based delivery systems have been extensively used in oncology because of their biocompatibility, biodegradability, ability to encapsulate diverse drug molecules, high temporal and thermal stability, and offer prolonged and controlled drug release. This review discusses the current status of the lipid-based nanocarriers and their applications in cancer treatment as well as an overview of the different liposomal formulations commercially available for cancer therapy.


Ecological and Convergent Validity of Experimentally and Dynamically Assessed Capacity for Social Contingency Detection Using the Perceptual Crossing Experiment in Adolescence.
The Perceptual Crossing Experiment (PCE) captures the capacity for social contingency detection using real-time social interaction dynamics but has not been externally validated. We tested ecological and convergent validity of the PCE in a sample of 208 adolescents from the general population, aged 11 to 19 years. We expected associations between PCE performance and (a) quantity and quality of social interaction in daily life, using Experience Sampling Methodology (ESM; ecological validity) and (b) self-reported social skills using a questionnaire (convergent validity). We also expected PCE performance to better explain variance in ESM social measures than self-reported social skills. Multilevel analyses showed that only self-reported social skills were positively associated with social experience of company in daily life. These initial results do not support ecological and convergent validity of the PCE. However, fueled by novel insights regarding the complexity of capturing social dynamics, we identified promising methodological advances for future validation efforts.


[The functional status of natural killer cells in patients with cancer of the digestive organs].
The author investigated the state of functional activity of the system of natural resistance in patients suffering of cancer of the gastrointestinal tract organs. It was established that in the course of development of the disease natural resistance reduced. The obtained data may be used for immunomonitoring in oncological clinical practice.


Enhancing functional recovery following postpartum femoral neuropathy: early neurorehabilitation and multidisciplinary obstetric care.
Postpartum femoral neuropathy has a reported incidence of less than 1% and its total recovery time extends up to 6 months to a year. A multidisciplinary approach is vital to rule out permanent disability and to assure a correct diagnosis and earlier rehabilitation. We report a case of a 37-year-old puerperal woman with a history of intrapartum epidural analgesia, who presented post-labor unilateral lower-limb motor weakness and sensory loss, with functional compromise on independent gait. A multidisciplinary team consisting of an anesthesiologist, a physiatrist, a neurologist, and an obstetrician was then established. In the initial physiatry and neurology assessment, the patient reported pain (numerical rating scale 7/10) over the inguinal ligament, lower limb hypoesthesia, and muscle weakness. Femoral neuropathy was suspected. Magnetic resonance imaging ruled out potential complications related to the anesthetic procedure. The patient was then enrolled in a supervised rehabilitation program and, 3 weeks later, electrodiagnostic studies confirmed the initial suspicion. Two months later, the patient had regained lower-limb active range of motion and no pain nor paresthesia was reported. Our case report describes how an early multimodal rehabilitation program within a multidisciplinary framework allows for sooner neuromotor function improvement and activities of daily living independence.


Effective communication: Core to promoting respectful maternity care for disabled women.
Previous research highlights that disabled women have less choice, control and respect of their dignity during pregnancy, childbirth and parenting. The experience of dignity and respect during pregnancy and childbirth for those with physical and sensory disability in the UK and Ireland is explored. Narrative, semi-structured telephone and Skype interviews were conducted with ten disabled women in the UK and Ireland. Interviews were audio-recorded and transcribed verbatim. Intra- and inter-thematic analysis was undertaken, beginning with a process of open coding. As themes developed, a process of constant comparison was used. The key finding is that effective communication with women was perceived to best respect the women's dignity in childbirth. This meant enabling women to feel heard, enabling women to make informed decisions about their care and providing individualized care. The single most important factor that enabled this communication was continuity of carer. The factor that most promoted maternity care that was perceived to be respectful was effective communication. This is not so different in other parts of the world, or for non-disabled women, however care providers should be particularly careful to enable good communication with disabled women.


Expression of galactose-1-p-uridyltransferase in Chinese hamster x human galactosemia somatic cell hybrids.
Lymphocytes from a patient with classic galactosemia (GALT deficiency) were hybridized with a Chinese hamster cell line. Electrophoretic valuation of GALT in 31 independently derived interspecific hybrid clones failed to demonstrate expression of the human GALT gene even when human chromosome 9 was present. Possible mechanisms for this lack of expression are presented.


Lack of methane hotspot in the upstream dam: Case study in a tributary of the Three Gorges Reservoir, China.
River damming has seen a growing trend in demand worldwide and the impounded reaches are considered hotspots of greenhouse gas emissions. However, it remains unclear how the spatial distribution of C-gas in sediments and methane (CH4) emissions of dammed tributary changes under different operation periods of the Three Gorges reservoir (TGR). We measured CH4 and carbon dioxide (CO2) concentrations in sediment and CH4 emissions from a dammed river of the TGR, and evaluated the effect of damming on the spatial variability of carbon in the sediment and on CH4 flux. It was found that damming led to a distinct spatial pattern of total organic carbon (TOC) in the sediment, which resulted in higher CH4 and CO2 in upstream sediment compared to the downstream. During the TGR impounding period, the upstream CH4 diffusive flux (0.253-0.427 mg m-2 h-1) across the water-air interface was higher than in the downstream (0.093 mg m-2 h-1), which was consistent with the spatial variation of CH4 in the sediments. However, the CH4 emission was predominantly by ebullition and the flux in the downstream (169.173 mg m-2 h-1) was significantly higher than upstream (12.23-123.05 mg m-2 h-1) in the discharging period. This can be attributed to a sharp increase in TOC in downstream sediment due to riparian zone soil erosion on both banks, which was caused by periodic large fluctuation in the water level, and a shallow water depth in the downstream. This study adds to our understanding of effects of the TGR's operation on CH4 emissions from a dammed tributary and suggests that the water level fluctuation of tributaries which has direct influence on ebullition and methane oxidation caused by manipulation of the TGR should not be overlooked.


Engineering Metallic Heterostructure Based on Ni3 N and 2M-MoS2 for Alkaline Water Electrolysis with Industry-Compatible Current Density and Stability.
Alkaline water electrolysis is commercially desirable to realize large-scale hydrogen production. Although nonprecious catalysts exhibit high electrocatalytic activity at low current density (10-50 mA cm-2 ), it is still challenging to achieve industrially required current density over 500 mA cm-2 due to inefficient electron transport and competitive adsorption between hydroxyl and water. Herein, the authors design a novel metallic heterostructure based on nickel nitride and monoclinic molybdenum disulfide (Ni3 N@2M-MoS2 ) for extraordinary water electrolysis. The Ni3 N@2M-MoS2 composite with heterointerface provides two kinds of separated reaction sites to overcome the steric hindrance of competitive hydroxyl/water adsorption. The kinetically decoupled hydroxyl/water adsorption/dissociation and metallic conductivity of Ni3 N@2M-MoS2 enable hydrogen production from Ni3 N and oxygen evolution from the heterointerface at large current density. The metallic heterostructure is proved to be imperative for the stabilization and activation of Ni3 N@2M-MoS2 , which can efficiently regulate the active electronic states of Ni/N atoms around the Fermi-level through the charge transfer between the active atoms of Ni3 N and MoMo bonds of 2M-MoS2 to boost overall water splitting. The Ni3 N@2M-MoS2 incorporated water electrolyzer requires ultralow cell voltage of 1.644 V@1000 mA cm-2 with ≈100% retention over 300 h, far exceeding the commercial Pt/C║RuO2 (2.41 V@1000 mA cm-2 , 100 h, 58.2%).


Dynorphin acts as a neuromodulator to inhibit itch in the dorsal horn of the spinal cord.
Menthol and other counterstimuli relieve itch, resulting in an antipruritic state that persists for minutes to hours. However, the neural basis for this effect is unclear, and the underlying neuromodulatory mechanisms are unknown. Previous studies revealed that Bhlhb5(-/-) mice, which lack a specific population of spinal inhibitory interneurons (B5-I neurons), develop pathological itch. Here we characterize B5-I neurons and show that they belong to a neurochemically distinct subset. We provide cause-and-effect evidence that B5-I neurons inhibit itch and show that dynorphin, which is released from B5-I neurons, is a key neuromodulator of pruritus. Finally, we show that B5-I neurons are innervated by menthol-, capsaicin-, and mustard oil-responsive sensory neurons and are required for the inhibition of itch by menthol. These findings provide a cellular basis for the inhibition of itch by chemical counterstimuli and suggest that kappa opioids may be a broadly effective therapy for pathological itch.


Addition of indapamide to frusemide increases natriuresis and creatinine clearance, but not diuresis, in fluid overloaded ICU patients.
Fluid and sodium overload are a common problem in critically ill patients. Frusemide may result in diuresis in excess of natriuresis. The addition of indapamide may achieve a greater natriuresis, and also circumvent some of the problems associated with frusemide. The objective of this study was to examine the effect of adding indapamide to frusemide on diuresis, natriuresis, creatinine clearance and serum electrolytes. Fluid overloaded ICU patients were randomised to either intravenous frusemide (Group F) or intravenous frusemide and enteral indapamide (Group F + I). Comprehensive exclusion criteria were applied to address confounders. 24 hour urine was analysed for electrolytes and creatinine. Serum electrolytes were measured before and 24 hours after administration of diuretics. Forty patients (20 in each group) were included in the study. The groups were similar in their baseline characteristics. Over the 24 h study period, patients in Group F + I, had a larger natriuresis (P = 0.01), chloride loss (P = 0.01) and kaliuresis (P = 0.047). Patients in Group F + I also had a greater 24 hour urinary creatinine clearance (P = 0.01). The 24 hour urine volume and fluid balance was similar between the groups. Patients in Group F had an increase in serum sodium (P = 0.04), while patients in Group F + I had a decrease in both serum chloride (P = 0.01) and peripheral oedema (P < 0.001) during the study duration. In fluid overloaded ICU patients, addition of indapamide to frusemide led to a greater natriuresis and creatinine clearance. Such a strategy might be utilised in optimising sodium balance in ICU patients.


A dynamometer for evaluation of dynamic muscle work.
The validation of a new dynamometer for evaluation of dynamic muscle work is presented. The device was based on a precise measurement of load displacements of any machine using gravitational loads as external resistance. It allowed, through a sensor consisting of an infrared photo interrupter, the calculation of velocity, force and power during concentric, eccentric and stretch-shortening cycle activity. To validate the dynamometer 33 male and female track and field athletes (12 throwers and 21 jumpers) participated in the study. The throwers (4 women and 8 men) were asked to perform half-squat exercises on a slide machine with a load of 100% of the subject's body mass. The day-to-day reproducibility of half-squat exercises gave a correlation coefficient of r = 0.88, 0.97 and 0.95 for average push-off force (AF), average push-off velocity (AV), and average push-off power (AP) respectively. Comparison of half-squat measurements was performed against jumping and running test evaluation by the jumpers (7 women and 14 men). The interrelationships among the different variables studied demonstrated a strong correlation between AF, AV and AP and sprinting and jumping parameters (r = 0.53-0.97; P < 0.05-0.001). Using values of AF, AV and AP developed in half-squat exercises executed with different loads, ranging from 35% to 210% of the subject's body mass, it was also possible to establish the force-velocity and power-velocity relationships for both male and female jumpers. In any individual case, the maximal error due to the measurement system was calculated to be less than 0.3%, 0.9% and 1.2% for AF, AV, and AP respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Improving quality of life in patients with metastatic prostate cancer following one cycle of 177Lu-PSMA-617 radioligand therapy: a pilot study.
To evaluate the clinical therapeutic response of PSMA targeted radioligand therapy with 177Lu-PSMA-617 in patients with metastatic castration-resistant prostate cancer. The current study analyzed disease-related quality of life (QoL) in patients undergoing PSMA therapy with a special focus on the association with simultaneous PSA response. Thirty patients (age range 50-87 years, median 73.5 years) undergoing 177Lu-PSMA-617 therapy from 2014 to 2016 at our institution were included in this pilot study. Health-related QoL was assessed by EORTC QLQ-C30 questionnaire filled in at baseline and two months after initializing the PSMA-therapy. The treatment response was evaluated under three categories with regard to changes in (a) global health status and other functional scales, (b) disease-related symptoms, and (c) effects of PSA values. Most patients underwent three treatment cycles (n = 12); at least 2 cycles (n = 6) or at most 8 cycles (n = 1) were performed. Out of 30 cases, PSA response after the first cycle was observed in 73 % (n = 22). Compared to baseline, QoL was significantly improved at 2-month follow-up revealing increase in global health status (p = 0.025), role functioning (p = 0.017) and emotional functioning (0.010), and decrease in pain (p = 0.033). Global health status variation can be explained up to 20.5 % by response in PSA (p = 0.012), this improved with PSA reduction. PSMA radioligand therapy seems to be an effective treatment option of metastatic castration-resistant prostate cancer patients as it improves their QoL in terms of increasing global health and mitigation of disease-related pain.


Updates on the Diagnosis and Management of Glaucoma.
Glaucoma is the leading cause of blindness throughout the world (after cataracts); therefore, general physicians should be familiar with the diagnosis and management of affected patients. Glaucomas are usually categorized by the anatomy of the anterior chamber angle (open vs narrow/closed), rapidity of onset (acute vs chronic), and major etiology (primary vs secondary). Most glaucomas are primary (ie, without a contributing comorbidity); however, several coexisting ophthalmic conditions may serve as the underlying etiologies of secondary glaucomas. Chronic glaucoma occurs most commonly; thus, regular eye examinations should be performed in at-risk patients to prevent the insidious loss of vision that can develop before diagnosis. Glaucoma damages the optic nerve and retinal nerve fiber layer, leading to peripheral and central visual field defects. Elevated intraocular pressure (IOP), a crucial determinant of disease progression, remains the only modifiable risk factor; thus, all current treatments (medications, lasers, and operations) aim to reduce the IOP. Pharmacotherapy is the usual first-line therapy, but noncompliance, undesirable adverse effects, and cost limit effectiveness. Laser and surgical treatments may lower IOP significantly over long periods and may be more cost effective than pharmacotherapy, but they are plagued by greater procedural risks and frequent treatment failures. Traditional incisional procedures have recently been replaced by several novel, minimally invasive glaucoma surgeries with improved safety profiles and only minimal decreases in efficacy. Minimally invasive glaucoma surgeries have dramatically transformed the surgical management of glaucoma; nevertheless, large, randomized trials are required to assess their long-term efficacy.


Pyridine carboxylic acids as inhibitors and substrates of the Escherichia coli gab permease encoded by gabP.
Although considered selective for its natural substrate, 4-aminobutyrate, gab permease was inhibited by 1,2,3,6-tetrahydro-3-pyridinecarboxylate and 1,2,3,6-tetrahydro-4-pyridinecarboxylate. The former is a transported substrate, since its preloading into metabolically poisoned cells stimulated transient accumulation of 4-aminobutyrate via counterflow.


A comparative study of immunomagnetic methods used for separation of human natural killer cells from peripheral blood.
Immunomagnetic sorting of natural killer (NK) cells from the peripheral blood of healthy donors has been evaluated in a comparative study of composition, yield and activation of target cells obtained by positive (Dynabeads, Microbeads) and negative (Microbeads) sorting procedures. Positively sorted target cells were selected by expression of the NK cell marker CD56, whereas NK cells obtained by negative sorting were those remaining after steps to remove all non-NK cell leukocyte populations were accomplished. In positive sorting, both CD56+CD3- NK cells and CD56+CD3+ natural killer T (NKT) cells were included. The NKT cell fraction differed between individuals, but not between the positive sorting methods. Whereas 20-30% of positively sorted target cells were NKT cells, only approximately 3% of negatively sorted cells were CD3+. Contamination with monocytes and B cells was low (1-3%) in all methods studied. Sorting with Microbeads (both positive and negative) gave higher cell yields than those obtained with Dynabeads (14% vs. 5% of total leukocyte numbers). A higher CD56 fluorescence intensity of NK cells and a better discrimination between the CD56bright and CD56dim NK cell subpopulations was obtained after negative sorting. Dynabeads-separated cells had, shortly after separation, a significantly higher expression (approximately 30%) of the early activation marker CD69 than cells either positively or negatively separated by Microbeads (approximately 8%). CD56+ cells positively sorted by Microbeads demonstrated a significantly higher production of TNF-alpha and IFN-gamma after IL-2 stimulation than Dynabeads-sorted cells. However, the cytotoxicity of cells obtained by the two positive sorting procedures did not differ. In conclusion, positive selection of CD56+ cells by Microbeads is better than Dynabeads, as determined from cell yield and procedure-associated cell activation and should be chosen for in vitro studies of NK/NKT cells. However, when pure NK cells and phenotypic subtypes are to be studied, negative sorting seems most appropriate.


Use of Supplemented or Human Material to Simulate PD Behavior of Antibiotics at the Target Site In Vitro.
In antimicrobial drug development, in vitro antibiotic susceptibility testing is conducted in standard growth media, such as Mueller-Hinton broth (MHB). These growth media provide optimal bacterial growth, but do not consider certain host factors that would be necessary to mimic the in vivo bacterial environment in the human body. The present review aimed to include relevant data published between 1986 and 2019. A database search (PubMed) was done with text keywords, such as "MIC" (minimal inhibitory concentration), "TKC" (time kill curve), "blood", "body fluid", "PD" (pharmacodynamic), and "in vitro", and 53 papers were ultimately selected. Additionally, a literature search for physiologic characteristics of body fluids was conducted. This review gives an excerpt of the complexity of human compartments with their physiologic composition. Furthermore, we present an update of currently available in vitro models operated either with adapted growth media or body fluids themselves. Moreover, the feasibility of testing the activity of antimicrobials in such settings is discussed, and pro and cons for standard practice methods are given. The impact on bacterial killing varies between individual adapted microbiological media, as well as direct pharmacodynamic simulations in body fluids, between bacterial strains, antimicrobial agents, and the compositions of the adjuvants or the biological fluid itself.


Expression of Cyr61 (CCN1) in human oral squamous cell carcinoma: An independent marker for poor prognosis.
Cysteine-rich 61 (Cyr61 [CCN1]) has disparate functions in tumorigenesis that are dependent on the cell types. The aim of the study was to investigate its role in the growth of oral squamous cell carcinoma (SCC). The study used immunohistochemistry to examine Cyr61 expression in 93 oral SCC specimens and assessed the effect of Cyr61 overexpression on proliferation and migration of oral SCC cells in vitro and xenograft growth in severe combined immunodeficient (SCID) mice. High expression of Cyr61 significantly correlated with large tumor size (p = .009) and advanced tumor stage (p = .036). Multivariate analysis revealed that high Cyr61 (relative risk [RR] 2.44, 95% confidence interval [CI] 1.209-4.95, p = .010) significantly correlated with mortality. Forced expression of Cyr61 stimulated the motility and growth of Ca9-22 cells in vitro and enhanced xenograft growth in SCID mice. Cyr61 is a positive growth modulator of oral SCC and Cyr61 overexpression is an independent prognostic indicator for patients with oral SCC.


[Ophthalmological aspects of the Gutenberg Health Study (GHS): an interdisciplinary prospective population-based cohort study].
The Gutenberg Health Study (GHS) is a prospective, single center, population-based cohort study. It is an interdisciplinary project for investigation of ocular, cardiovascular, psychosomatic and immune diseases in the population of the City of Mainz and the district of Mainz-Bingen in central Germany. The main goals of the ophthalmological branch of GHS are determination of the prevalence and incidence of major ocular risk factors and diseases, exploring the genetic determinants and assessing complex interdisciplinary associations. The study cohort includes 15,010 participants aged between 35 and 74 years at the time of inclusion. After completing the baseline investigations the 5-year follow-up of the whole study cohort started in April 2012. The GHS is the most extensive data set of major ophthalmological conditions and risk factors in Germany and will help researchers in understanding complex medical associations.


Does geographical variability influence five-year MACCE rates in the multicentre SYNTAX revascularisation trial?
The use of multiple geographical sites for randomised cardiovascular trials may lead to important heterogeneity in treatment effects. This study aimed to determine whether treatment effects from different geographical recruitment regions impacted significantly on five-year MACCE rates in the SYNTAX trial. Five-year SYNTAX results (n=1,800) were analysed for geographical variability by site and country for the effect of treatment (CABG vs. PCI) on MACCE rates. Fixed, random, and linear mixed models were used to test clinical covariate effects, such as diabetes, lesion characteristics, and procedural factors. Comparing five-year MACCE rates, the pooled odds ratio (OR) between study sites was 0.58 (95% CI: 0.47-0.71), and countries 0.59 (95% CI: 0.45-0.73). By homogeneity testing, no individual site (X2=93.8, p=0.051) or country differences (X2=25.7, p=0.080) were observed. For random effects models, the intraclass correlation was minimal (ICC site=5.1%, ICC country=1.5%, p<0.001), indicating minimal geographical heterogeneity, with a hazard ratio of 0.70 (95% CI: 0.59-0.83). Baseline risk (smoking, diabetes, PAD) did not influence regional five-year MACCE outcomes (ICC 1.3%-5.2%), nor did revascularisation of the left main vs. three-vessel disease (p=0.241), across site or country subgroups. For CABG patients, the number of arterial (p=0.49) or venous (p=0.38) conduits used also made no difference. Geographic variability has no significant treatment effect on MACCE rates at five years. These findings highlight the generalisability of the five-year outcomes of the SYNTAX study.


Effects of tyrosine kinase inhibitor STI571 on human mast cells bearing wild-type or mutated c-kit.
STI571 is a tyrosine kinase inhibitor which inhibits the kinase activity of kit, the receptor for stem cell factor (SCF). Because activating mutations of c-kit affecting codon 816 are associated with human mast cell neoplasms, we determined whether STI571 exerted a similar cytotoxic effect on neoplastic and normal human mast cells. We investigated the effect of addition of STI571 in increasing concentrations (0.01 to 10 micromolar) to two HMC-1 human mast cell leukemia cell lines carrying two different activating c-kit mutations in codons 816 or 560, as well as the effect of the drug on short-term bone marrow cultures obtained from patients who carry a mutated codon 816 or wild-type c-kit. STI571 failed to inhibit the growth of HMC-1(560,816) cells bearing a codon 816 mutation but effectively suppressed the proliferation of HMC-1(560) carrying c-kit with the wild-type codon 816. STI571 did not induce preferential killing of neoplastic bone marrow mast cells in short-term cultures from patients bearing a codon 816 c-kit mutation. In contrast, STI571 caused a dramatic reduction in mast cells in patients without codon 816 c-kit mutations. These results suggest that STI571, while effectively killing mast cells with wild-type c-kit, did not show preferential cytotoxicity to neoplastic human mast cells and thus may not be effective in the treatment of human systemic mastocytosis associated with codon 816 c-kit mutations.


Comparison of aconitine-modified human heart (hH1) and rat skeletal (mu1) muscle Na+ channels: an important role for external Na+ ions.
Neurotoxins such as aconitine (AC) bind to receptor site 2 on voltage-gated sodium channels and modify channel kinetics. Although AC modification typically induces hyperpolarizing shifts in sodium channel activation, the effects on channel inactivation seem to vary depending on the tissue origin of the channel. In the present study, the alpha subunits of human heart (hH1) and rat skeletal muscle (mu1) sodium channels were transiently expressed in human embryonic kidney (HEK293t) cells. Whole-cell currents were examined before and after AC modification of the channels to determine whether the toxin had isoform-specific effects on channel kinetics. The magnitudes of the hyperpolarizing shifts in steady-state current activation and inactivation were similar for AC-modified hH1 and mu1 channels, and AC modification did not alter the voltage dependence of macroscopic current decay of either channel subtype. There were two notable differences between hH1 and mu1 channels after AC modification. First, the steady-state availability of AC-modified mu1 channels decreased by 5-10% after very negative conditioning pulses. Second, AC-modified mu1 channels inactivated completely at all voltages, whereas AC-modified hH1 channels exhibited sustained inward currents at voltages near the threshold of current activation. Interestingly, AC-modified hH1 channels inactivated completely if the external solution did not contain sodium ions. The data demonstrate that AC modification affects the activation of hH1 and mu1 channels similarly but affects inactivation of the two channels distinctly. The results also imply that the reduced inactivation of AC-modified hH1 channels at least partially depends on the presence of extracellular sodium.


Perceived barriers to a career in pain medicine in the Republic of Ireland.
As the population ages, musculoskeletal disorders are a prominent contributory factor to loss of mobility and autonomy. Pain is a predictor of disability and worsening frailty and the role of the chronic pain specialist in the management of this cohort cannot be overstated. With growing requirements for pain specialists, we sought to identify barriers to recruitment within this specialty. Establish baseline attitudes and perceived barriers towards a career in pain medicine among Irish anaesthesiology trainees. Suggest a framework to improve recruitment to the specialty. Ethical approval was obtained. A web-based questionnaire was sent to all anaesthesiologists in training in the Republic of Ireland. Data was analyzed using SPSS. Two hundred forty-eight trainees received the questionnaire, 59 responded. 54.2% male, 45.8% female. 79.7% had previous clinical exposure to pain medicine, most of whom had spent over one month with a service. Only 10.2% of respondents were considering a career in pain medicine. Factors that attracted trainees to the subspeciality included: interventional practice (81%), variation in clinical practice (66.7%), autonomy of practice (61.9%) and perceived good work-life balance (42.9%). Deterrents from the subspecialty included a psychologically challenging patient cohort (69.5%), frequency of clinics (50.8%), and additional exams (32.2%). When asked how to improve engagement with the speciality, 62% suggested earlier exposure to the speciality and 32.2% suggested increased frequency of formal teaching and workshops. Increased exposure of trainees to the specialty during early stages of training may improve future recruitment to the subspecialty in Ireland.


Non-invasive assessment of muscle compartment elasticity by pressure-related ultrasound in pediatric trauma: a prospective clinical study in 25 cases of forearm shaft fractures.
Soft-tissue swelling after limb fractures in pediatric patients is well known to be a risk factor for developing acute compartment syndrome (ACS). Clinical assessment alone is uncertain in specific cases. Recently, we proposed a non-invasive ultrasound-based method to objectify muscle compartment elasticity for monitoring. We hypothesize a strong correlation between the soft-tissue swelling after stabilization of upper limb fractures and the compartment elasticity objectified with a novel ultrasound-based approach in pediatric trauma. In a prospective clinical study, children suffering forearm fractures but not developing an ACS were included. The muscle compartment elasticity of the m. flexor carpi ulnaris was assessed after surgical intervention by a non-invasive, ultrasound-based method resulting in a relative elasticity (RE in %) in both the control (healthy limb) and study group (fractured limb). Soft-tissue swelling was categorized in four different levels (0-3) and correlated with the resulting RE (%). The RE in the study group (15.67%, SD ± 3.06) showed a significantly decreased level (p < 0.001) compared with the control (22.77%, SD ± 5.4). The categorized grade of soft-tissue swelling resulted in a moderate correlation with the RE (rs = 0.474). The presented study appears to represent a novel approach to assess the posttraumatic pressure changes in a muscle compartment after fracture stabilization non-invasively. In this first clinical study in pediatric cases, our measurement method represents a low-cost, easy, and secure approach that has the potential to substitute invasive measurement of suspected ACS in muscle compartment conditions. Further investigations in lager cohorts are required to prove its daily clinical practicability and to confirm the expected reliability.


Protocol to establish a genetically engineered mouse model of IDH1-mutant astrocytoma.
Lower-grade gliomas exhibit a high prevalence of isocitrate dehydrogenase 1 (IDH1) mutations, but faithful models for studying these tumors are lacking. Here, we present a protocol to establish a genetically engineered mouse (GEM) model of grade 3 astrocytoma driven by the Idh1R132H oncogene. We describe steps for breeding compound transgenic mice and intracranially delivering adeno-associated virus particles, followed by post-surgical surveillance via magnetic resonance imaging. This protocol enables the generation and use of a GEM to study lower-grade IDH-mutant gliomas. For complete details on the use and execution of this protocol, please refer to Shi et al. (2022).1.


The effect of hydrocortisone and adrenocorticotrophic hormone on monoamine oxidase and tyrosine hydroxylase in explant cultures of embryonic chick sympathetic ganglia.
Sympathetic ganglia from 13- to 15-day-old embryonic chicks were cultured for up to 2 days in Leighton tubes. The influence of hydrocortisone and ACTH added to the culture medium on the enzymes monoamine oxidase (MAO) and tyrosine hydroxylase was studied. Hydrocortisone (5 times 10(-5)M) had no effect on tyrosine hydroxylase but increased MAO activity by up to 46 percent over control values under conditions of low or zero nerve growth factor (NGF) concentration. ACTH also increased ganglionic MAO activity, the effect again depending on NGF concentration. This time the maximal response (an increase of 50 percent over controls) was seen at high NGF concentrations. This response was similar to the effect of 1 mM dibutyryl cyclic AMP, and was blocked by 1 times 10-5 M propranolol and 10 muM prostaglandin E(1). ACTH only slightly increased tyrosine hydroxylase activity and this effect was due to a small (18 percent) increase in sympathetic neurone number. Guanosine 5-diphosphate (0.5 mM) was found to increase tyrosine hydroxylase activity by 57 percent and this effect was blocked by the presence of ACTH.


The effect of a specialist seating assessment clinic on the skin management of individuals with spinal cord injury.
To evaluate the specialist seating clinic's effectiveness in improving skin management knowledge and independence, represented by the Needs Assessment Checklist (NAC). Longitudinal, between subjects design, with two intervention groups and one control. Tertiary care, spinal cord injury centre (National Spinal Injuries Centre), Stoke Mandeville Hospital, United Kingdom. This study assessed the skin management ability of three groups. Group 1 consisted of individuals who had attended a specialist seating assessment (SSA) clinic before their first needs assessment, group 2 had attended SSA between their first and second needs assessment, and group 3 (control) had not attended at all. Patient skin management ability was assessed using the skin management subscale of the NAC, a measure of rehabilitation outcome, at two time points. Significant differences were identified between group 3 and group 1 at both the first (t = 2.36, degrees of freedom (df) = 37, p < 0.05) and second (t = 2.84, df = 37, p < 0.01) needs assessment. Significant improvements were also observed within each group between the first and second needs assessment time points in all seating assessment categories. Skin management achievement scores were significantly higher for patients who had attended a SSA clinic before their first NAC (group 1) at both time points, supporting the use of SSA as a proactive intervention to improve patient independence, knowledge and awareness, and potentially reduce pressure ulcer incidence.


Cadmium induces the activation of cell wall integrity pathway in budding yeast.
MAP kinases are important signaling molecules regulating cell survival, proliferation and differentiation, and can be activated by cadmium stress. In this study, we demonstrate that cadmium induces phosphorylation of the yeast cell wall integrity (CWI) pathway_MAP kinase Slt2, and this cadmium-induced CWI activation is mediated by the cell surface sensor Mid2 through the GEF Rom1, the central regulator Rho1 and Bck1. Nevertheless, cadmium stress does not affect the subcellular localization of Slt2 proteins. In addition, this cadmium-induced CWI activation is independent on the calcium/calcineurin signaling and the high osmolarity glycerol (HOG) signaling pathways in yeast cells. Furthermore, we tested the cadmium sensitivity of 42 paired double-gene deletion mutants between six CWI components and seven components of the HOG pathway. Our results indicate that the CWI pathway is epistatic to the HOG pathway in cadmium sensitivity. However, gene deletion mutations for the Swi4/Swi6 transcription factor complex show synergistic effects with mutations of HOG components in cadmium sensitivity.


Comprehensive modified approaches to reducing the interference of moisture from an FTIR spectrum and the corresponding second derivative spectrum.
We proposed a modified and improved approach to removing the interference of moisture from an IR spectrum and the corresponding second derivative spectrum. The temperature fluctuation in the air of the optical path and baseline-drift lead to the small but persistent presence of the interference of moisture. The problem has been successfully addressed by adopting a double-matching strategy. Additionally, two-dimensional correlationspectra (2D-COS) are generated using the second derivative or third derivative spectrum of the negative base 10 logarithms of the single-beam spectra, thereby removing the linear slope or quadratic portion of baseline-drift. Using the newly adopted approach, the residual interferences of moisture are attenuated. We applied the new approach to the IR spectra and the second derivative spectra of neat hexadecanol and biaxially oriented polypropylene (BOPP) film, and some promising preliminary results are obtained. In hexadecanol, two highly overlapping peaks at 1464 and 1463 cm-1 are revealed. In BOPP, the envelope at 1458 cm-1 is found to be composed of a number of sub-peaks.


How addictive are gabapentin and pregabalin? A systematic review.
In the last ten years, gabapentin and pregabalin have been becoming dispensed broadly and sold on black markets, thereby, exposing millions to potential side-effects. Meanwhile, several pharmacovigilance-databases have warned for potential abuse liabilities and overdose fatalities in association with both gabapentinoids. To evaluate their addiction risk in more detail, we conducted a systematic review on PubMed/Scopus and included 106 studies. We did not find convincing evidence of a vigorous addictive power of gabapentinoids which is primarily suggested from their limited rewarding properties, marginal notes on relapses, and the very few cases with gabapentinoid-related behavioral dependence symptoms (ICD-10) in patients without a prior abuse history (N=4). In support, there was no publication about people who sought treatment for the use of gabapentinoids. Pregabalin appeared to be somewhat more addictive than gabapentin regarding the magnitude of behavioral dependence symptoms, transitions from prescription to self-administration, and the durability of the self-administrations. The principal population at risk for addiction of gabapentinoids consists of patients with other current or past substance use disorders (SUD), mostly opioid and multi-drug users, who preferred pregabalin. Pure overdoses of gabapentinoids appeared to be relative safe but can become lethal (pregabalin > gabapentin) in mixture with other psychoactive drugs, especially opioids again and sedatives. Based upon these results, we compared the addiction risks of gabapentin and pregabalin with those of traditional psychoactive substances and recommend that in patients with a history of SUD, gabapentinoids should be avoided or if indispensable, administered with caution by using a strict therapeutic and prescription monitoring.


Enterococcus faecalis pheromone-responding plasmid pAD1 gives rise to an aggregation (clumping) response when cells are exposed to subinhibitory concentrations of chloramphenicol, erythromycin, or tetracycline.
The Enterococcus faecalis conjugative cytolysin plasmid pAD1 encodes a specific aggregation (clumping) response to the peptide sex pheromone cAD1 secreted by plasmid-free strains. Here it is shown that, in the absence of cAD1, exposure of E. faecalis cells harboring pAD1 to subinhibitory concentrations of chloramphenicol, erythromycin, or tetracycline also results in an aggregation response that appears related to a stress-sensitive mechanism associated with a component of the pheromone response.


Percutaneous cutting balloon angioplasty for the treatment of renovascular hypertension in children and adolescents.
Percutaneous transluminal renal angioplasty (PTRA), the recommended treatment in children with renovascular hypertension (RVH), often has unsatisfactory outcomes. Cutting balloons may improve the results of angioplasty in different vascular beds with complex and resistant lesions. We retrospectively analysed the effects of percutaneous cutting balloon angioplasty (PCBA) on blood pressure, cardiac mass and renal artery acceleration time in children/adolescents referred to our centre for RVH. Thirteen patients (aged 9-19 years) with renal artery stenosis (RAS) and severe hypertension were identified. RASs were focal fibromuscular (FMD) or FMD-like dysplasia (in six cases bilateral, in five associated with mid aortic syndrome). Ten patients had uncontrolled hypertension, in nine cases associated with left ventricular hypertrophy (LVH). Acceleration time was abnormal in all stenotic arteries. Eighteen PCBA were performed, in three arteries associated with stent implantation. PCBA was technically successful in all individuals without major complications. In one patient, an intra-stent restenosis occurred, successfully redilated with conventional angioplasty without recurrence at 4 years distance. One year after PCBA, mean SBP and DBPs were markedly reduced from 146 ± 25 to 121 ± 10 mmHg and from 87 ± 11 to 65 ± 12 mmHg, respectively ( P < 0.001 for both). At that time, hypertension was cured in seven children and controlled in five individuals. This favourable outcome was confirmed with ambulatory blood pressure measurement in four patients. At the latest follow-up, left ventricular mass and acceleration time were normal in all patients. PCBA proved to be a well tolerated and effective procedure that can be considered as an alternative to PTRA to treat hypertensive children/adolescents with recurrent or resistant RAS.


Epidemiology of Oral and Maxillofacial Infections.
Dental caries and periodontal disease are the most common dental infections and are constantly increasing worldwide. Distribution, occurrence of dental caries, gingivitis, periodontitis, odontogenic infections, antibiotic resistance, oral mucosal infections, and microbe-related oral cancer are important to understand the public impact and methods of controlling such disease. Distribution of human papilloma virus and human immunodeficiency virus -related oral cancers in the US population is presented.


[Complex evaluation of the long-term results of resection of postinfarct aneurysms of the left ventricle].
Late results of left ventricular postinfarction aneurysm resection were studied in 112 patients. The results were good or satisfactory in 84.7% of patients, who mostly showed no hemodynamically significant coronary arterial stenoses or with one major branch affected, and a dyskinetic aneurysm. Clinical improvement was accompanied by a significant increase in stress tolerance, an increase in maximum oxygen utilization per 1 kg body weight, a decrease in the proportion of oxygen debt to oxygen want and better myocardial contractility, evidence of increased physical working capacity of the postoperative patients. The results were poor in patients with multiple coronary arterial stenoses and heart failure uncontrollable by medication.


Diagnostic laparoscopy for the evaluation of occult diaphragmatic injury following penetrating thoracoabdominal trauma.
Occult diaphragmatic injury following penetrating thoracoabdominal trauma can be difficult to diagnose. Radiographic findings are often non-specific or absent. Undetected injuries may remain clinically silent, only to present later with life-threatening complications associated with diaphragmatic herniation. Diagnostic laparoscopy allows for the evaluation of trauma patients lacking clinical indications for a formal laparotomy. The purpose of this study was to evaluate the incidence of occult diaphragmatic injury and investigate the role of laparoscopy in patients with penetrating thoracoabdominal trauma who lack indications for exploratory laparotomy except the potential for a diaphragmatic injury. Haemodynamically stable patients with penetrating thoracoabdominal trauma without indications for laparotomy (haemodynamic instability, evisceration, or peritonitis on exam) and evaluated with diagnostic laparoscopy to determine the presence of a diaphragmatic injury were retrospectively reviewed. Thoracoabdominal wounds were defined as wounds bounded by the nipple line over the anterior and posterior chest superiorly and the costal margin inferiorly. One hundred and eight patients were evaluated for penetrating thoracoabdominal injuries (80 stabs and 28 gunshots) over the study period. 22 (20%) diaphragmatic injuries were identified. These were associated with injuries to the spleen (5), stomach (3) and liver (2). There was a greater incidence of haemopneumothorax (HPTX) in patients with diaphragmatic injury (32%) compared to those without injury (20%). 29% of patients with a HPTX had a diaphragmatic injury. However, 18% of patients with a normal chest radiograph were also found to have a diaphragmatic injury. The incidence of diaphragmatic injury associated with penetrating thoracoabdominal trauma is high. Clinical and radiographic findings can be unreliable for detecting occult diaphragmatic injury. Diagnostic laparoscopy provides a vital tool for detecting occult diaphragmatic injury among patients who have no other indications for formal laparotomy.


[Oxygen balance in the normal dog myocardium with increased concentrations of inhaled anesthetics].
Myocardial oxygen supply and demand in the normal dog were evaluated as the concentration of halothane, enflurane or isoflurane was increased. Although coronary blood flow decreased as the anaesthetic gas concentration increased, the O2 supply-to-demand ratio remained stable owing to a decrease in the myocardial O2 consumption.


The biochemistry of aging muscle.
Between the ages of 20 and 80, humans lose approximately 20-30% of their skeletal muscle mass. This age-related loss of muscle mass, sometimes described as 'sarcopenia of old age', is the consequence of complicated multifactorial processes and is commonly associated with osteopenia or osteoporosis. Consequences of the aging changes in muscle are declining physiological function and loss of muscle strength, typically associated with reduced physical activities. Consequently, falls and subsequent serious injuries are prevalent in the elderly. Thus, it is imperative to try and understand the processes, leading to age-related muscle loss, in order to develop means to retard this phenomenon leading to improved quality of life in the elderly. It is possible to divide the causes of muscle aging to intrinsic factors, involving changes at the molecular and cellular levels, and to extrinsic or environmental factors. The purpose of this review is to describe some of the biochemical processes and the possible mechanisms of muscle aging and to evaluate the importance of various extrinsic factors such as nutrition, exercise and limb immobilization. Changes in the aging skeletal muscle are reviewed with regard to: (a) enzyme activities, protein turnover and repair capacities (b) mitochondrial functioning and energy reserve systems (c) ion content and regulation (d) oxidative stress and free radicals (e) nutrition and caloric restriction (f) exercise and limb immobilization.


The pathobiology of Paracoccidioides brasiliensis.
Paracoccidioides brasiliensis causes one of the most prevalent systemic mycoses in Latin America--paracoccidioidomycosis. It is a dimorphic fungus that undergoes a complex transformation in vivo, with mycelia in the environment producing conidia, which probably act as infectious propagules upon inhalation into the lungs, where they transform to the pathogenic yeast form. This transition is readily induced in vitro by temperature changes, resulting in modulation of the composition of the cell wall. Notably, the polymer linkages change from beta-glucan to alpha-glucan, possibly to avoid beta-glucan triggering the inflammatory response. Mammalian oestrogens inhibit this transition, giving rise to a higher incidence of disease in males. Furthermore, the susceptibility of individuals to paracoccidioidomycosis has a genetic basis, which results in a depressed cellular immune response in susceptible patients; resistance is conferred by cytokine-stimulated granuloma formation and nitric oxide production. The latency period and persistence of the disease and the apparent lack of efficacy of humoral immunity are consistent with P. brasiliensis existing as a facultative intracellular pathogen.


Sleep and breathing disturbances in infancy and early childhood.
The nature of sleep-related breathing problems varies with age. Ventilatory function may be compromised at the level of the brainstem, the autonomic nerves, or the upper airway musculature. These disorders are state-dependent, being impacted by both rapid eye movement and non-rapid eye movement sleep, and also by the degree of maturation of the nervous system. Many are treatable. An understanding of the molecular basis of these disorders is just now becoming apparent.


Organochlorines (PCBs and pesticides) in the bivalves Anadara (Senilis) senilis, Crassostrea tulipa and Perna perna from the lagoons of Ghana.
The bivalves, Anadara (Senilia) senilis (n=95), Crassostrea tulipa (n=75) and Perna perna (n=30) from Ghana, were analyzed for their organochlorines (pesticides and PCBs) concentrations and the body burden. A comparison was done based on two different standard PCB mixtures (Aroclor 1254 and 1260) with the percentage individual congener contribution and that of the sample. From these comparisons, it appeared that the pattern of PCB residues in the samples was neither Aroclor 1254 nor 1260. On the other hand, there was seasonal qualitative difference in the pattern that is purely marine (in the dry season) versus terrestrial input (in the wet season). SigmaPCB was 0.10 microg/g dw or 2.2 microg/g lw (median values). There was no correlation between PCB concentration and lipid content reflecting the importance of indirect contamination of the bivalves. The pesticides, aldrin, dieldrin, endrin, lindane, DDTs, HCH, HCB, endosulfan, heptachlor epoxide and heptachlor, were detected. The most abundant pesticides were the SigmaDDT: 73 and SigmaHCH: 29 ng/g dw. DDT/DDE and hept./heptachlor epoxide ratios reflect their recent applications, while the ratio for aldrin/dieldrin suggests that the use of aldrin has been discontinued, at least on the coast of Ghana.


Enhanced reduction of phenol content and toxicity in olive mill wastewaters by a newly isolated strain of Coriolopsis gallica.
The search for novel microorganisms able to degrade olive mill wastewaters (OMW) and withstand the toxic effects of the initially high phenolic concentrations is of great scientific and industrial interest. In this work, the possibility of reducing the phenolic content of OMW using new isolates of fungal strains (Coriolopsis gallica, Bjerkandera adusta, Trametes versicolor, Trichoderma citrinoviride, Phanerochaete chrysosporium, Gloeophyllum trabeum, Trametes trogii, and Fusarium solani) was investigated. In vitro, all fungal isolates tested caused an outstanding decolorization of OMW. However, C. gallica gave the highest decolorization and dephenolization rates at 30 % v/v OMW dilution in water. Fungal growth in OMW medium was affected by several parameters including phenolic compound concentration, nitrogen source, and inoculum size. The optimal OMW medium for the removal of phenolics and color was with the OMW concentration (in percent)/[(NH4)2SO4]/inoculum ratio of 30:6:3. Under these conditions, 90 and 85 % of the initial phenolic compounds and color were removed, respectively. High-pressure liquid chromatography analysis of extracts from treated and untreated OMW showed a clear and substantial reduction in phenolic compound concentrations. Phytotoxicity, assessed using radish (Raphanus sativus) seeds, indicated an increase in germination index of 23-92 % when a 30 % OMW concentration was treated with C. gallica in different dilutions (1/2, 1/4, and 1/8).


Distinguishing clinical characteristics of central nervous system tuberculosis in immunodeficient and non-immunodeficient individuals: a 12-year retrospective study.
Central nervous system tuberculosis (CNS TB) is a severe Mycobacterium tuberculosis (MTB) infection. It is unclear whether a patient's immune status alters the clinical manifestations and treatment outcomes of CNS TB. Between January 2007-December 2018, chart reviews of CNS TB, including tuberculous meningitis (TBM), tuberculoma/abscess, and TB myelitis, were made. Subjects were categorized as immunodeficient (ID) and non-immunodeficient (NID). Of 310 subjects, 160 (51.6%) were in the ID group-132 (42.6%) had HIV and 28 (9.0%) had another ID, and 150 (48.4%) were in the NID group. The mean age was 43.64 ± 16.76 years, and 188 (60.6%) were male. There were 285 (91.9%) TBM, 16 (5.2%) tuberculoma/abscess, and 9 (2.9%) myelitis cases. The TBM characteristics in the ID group were younger age (p = 0.003), deep subcortical location of tuberculoma (p = 0.030), lower hemoglobin level (p < 0.001), and lower peripheral white blood cell count (p < 0.001). Only HIV individuals with TBM had an infection by multidrug-resistant MTB (p = 0.013). TBM mortality was varied by immune status -HIV 22.8%, other ID 29.6%, and NID 14.8% (p < 0.001). Factors significantly associated with unfavorable outcomes in TBM also differed between the HIV and NID groups. TBM is the most significant proportion of CNS TB. Some of the clinical characteristics of TBM, such as age, radiographic findings, hematological derangement, and mortality, including factors associated with unfavorable outcomes, differed between ID and non-ID patients.


Evaluation of Genomic Selection for Seven Economic Traits in Yellow Drum (Nibea albiflora).
Yellow drum (Nibea albiflora) is an important maricultural fish in China, and genetic improvement is necessary for this species. This research evaluated the application of genomic selection methods to predict the genetic values of seven economic traits for yellow drum. Using genome-wide single-nucleotide polymorphisms (SNPs), we estimated the genetic parameters for seven traits, including body length (BL), swimming bladder index (SBI), swimming bladder weight (SBW), body thickness (BT), body height (BH), body length/body height ratio (LHR), and gonad weight index (GWI). The heritability estimates ranged from 0.309 to 0.843. We evaluated the prediction performance of various statistical methods, and no one method provided the highest predictive ability for all traits. We then evaluated and compared the use of genome-wide association study (GWAS)-informative SNPs and random SNPs for prediction and found that GWAS-informative SNPs obviously increased. It only needed 5 and 100 informative SNPs for LHR and BT to achieve almost the same predictive abilities as using genome-wide SNPs, and for BL, SBI, SBW, BH, and GWI, about 1000 to 3000 informative SNPs were needed to achieve whole-genome level predictive abilities. It can be concluded from the test results that breeders can use fewer SNPs to save the breeding costs of genomic selection for some traits.


Acetylcholinesterase activity in neural crest cells of the early chick embryo.
The appearance and distribution of AChE activity in the neural crest cells of the chick embryo were histochemically investigated. Prior to closure of the neural tube, neural crests were not demonstrated and most of the cells constituting the neural plate and the more lateral ectoderm were AChE-negative. With the closure of the neural tube, the neural crests assumed the form of a cell mass in its mid-dorsal portion and AChE activity was demonstrated in some elements of both tube and crests. The neural crest cells beginning to migrate ventrally or laterally were AChE-positive, and some showed intense enzymatic activity. Electron microscopically, the neural crest cells and the cells migrating from the neural crest displayed AChE activity in the cisternae of the nuclear envelope and in a few r-ER profiles, but were morphologically undifferentiated. As assessed by 3H-thymidine autoradiography, these cells possessed the potential to proliferate. These findings indicate that with the formation of the neural tube and neural crest, cells constituting these structures begin to differentiate with respect to AChE activity and that the enzyme appears in the neural crest cells before the onset of neuronal differentiation.


Natural killer-92 cells maintain cytotoxic activity after long-term cryopreservation in novel DMSO-free media.
Natural killer (NK) cells are a critical part of the innate immune system, and have emerged as attractive targets for immunotherapies for various malignancies. Alongside the need for expansion of NK cells to reach clinically useful numbers, a critical component in the availability of NK cells for allogeneic therapy is cryopreservation. While a continuously-growing cell line such as NK-92 can avoid issues associated with isolating, activating, expanding, and manufacturing large numbers of peripheral blood-derived NKs, cryopreservation of these cells has not made much progress. NK cells are highly sensitive to freezing and thawing, while the use of DMSO during cryopreservation raises serious safety concerns. In this work, we evaluated a number of cryoprotectants that do not contain DMSO for their capacity to cryopreserve NK-92 cells over long-term while retaining their cytotoxic activity and viability, with the aim of identifying potential replacements to DMSO for safe clinical use of these cells.


RNA-Seq transcriptomic analysis with Bag2D software identifies key pathways enhancing lipid yield in a high lipid-producing mutant of the non-model green alga Dunaliella tertiolecta.
For many years, increasing demands for fossil fuels have met with limited supply. As a potential substitute and renewable source of biofuel feedstock, microalgae have received significant attention. However, few of the current algal species produce high lipid yields to be commercially viable. To discover more high yielding strains, next-generation sequencing technology is used to elucidate lipid synthetic pathways and energy metabolism involved in lipid yield. When subjected to manipulation by genetic and metabolic engineering, enhancement of such pathways may further enhance lipid yield. In this study, transcriptome profiling of a random insertional mutant with enhanced lipid production generated from a non-model marine microalga Dunaliella tertiolecta is presented. D9 mutant has a lipid yield that is 2- to 4-fold higher than that of wild type. Using novel Bag2D-workflow scripts developed and reported here, the non-redundant transcripts from de novo assembly were annotated based on the best hits in five model microalgae, namely Chlamydomonas reinhardtii, Coccomyxa subellipsoidea C-169, Ostreococcus lucimarinus, Volvox carteri, Chlorella variabilis NC64A and a high plant species Arabidopsis thaliana. The assembled contigs (~181 Mb) includes 481,381 contigs, covering 10,185 genes. Pathway analysis showed that a pathway from inositol phosphate metabolism to fatty acid biosynthesis is the most significantly correlated with higher lipid yield in this mutant. Herein, we described a pipeline to analyze RNA-Seq data without pre-existing transcriptomic information. The draft transcriptome of D. tertiolecta was constructed and annotated, which offered useful information for characterizing high lipid-producing mutants. D. tertiolecta mutant was generated with an enhanced photosynthetic efficiency and lipid production. RNA-Seq data of the mutant and wild type were compared, providing biological insights into the expression patterns of contigs associated with energy metabolism and carbon flow pathways. Comparison of D. tertiolecta genes with homologs of five other green algae and a model high plant species can facilitate the annotation of D. tertiolecta and lead to a more complete annotation of its sequence database, thus laying the groundwork for optimization of lipid production pathways based on genetic manipulation.


Effect of in vivo growth conditions upon expression of surface protein antigens in Enterococcus faecalis.
Western blotting of whole-cell preparations of Enterococcus faecalis showed the protein-antigen profiles to be markedly influenced by growth conditions. The E. faecalis-specific antigens of 40 and 37 kDa, which are prominent in endocarditis, were strongly expressed following growth in serum or brain heart infusion, but not after growth in a chemically defined medium. The expression of these antigens in vivo was demonstrated in cells grown as a biofilm on silastic discs in the peritoneum of rabbits. These in vivo culture conditions may be useful in studying the pathogenesis of E. faecalis infections and the effectiveness of antibiotic therapy.


Residual fluxes of water and nutrient transport through the main inlet of a tropical estuary, Cochin estuary, West Coast, India.
Determining robust values for estuarine material fluxes has been a complex task and an interdisciplinary research challenge. With the advent of acoustic Doppler profilers (ADPs) having bottom-track capability and which provides three-dimensional current velocity profiles, more accurate estimation of cross-sectional fluxes is far accomplished in unsteady and bidirectional flow conditions of estuaries. This paper reports for the first time the discharge measurements conducted across Cochin inlet using ADP to examine the spring-neap variability in residual fluxes of water and nutrients during dry season. Cross-sectional current velocity profiles and salinity profiles were captured using ADP and conductivity temperature depth (CTD) instrumentation. Samples of surface and bottom water were also collected at 3-h intervals. The results indicated that there is a distinct transition from the neap to spring tides related to flow and salinity structure. The neap tide was partially mixed with large diurnal inequalities whereas the spring tide was well-mixed with symmetric tides. During ebb, an increase in the concentrations of nitrate, nitrite, phosphate, and silicate was noticed indicating upstream sources for their inputs. In contrast, elevated levels of ammonia were found in the estuary throughout the period of observation. There was net residual outflow during both tides, and the computed residual water fluxes of neap doubled that of spring. The strong ebb currents and the increased nutrient concentrations during ebb resulted in the export of all nutrients (except ammonia during spring) into the sea. The findings of this study highlight the consequences of anthropogenic interventions in the estuary and their effects on the fluxes of ecologically relevant substances.


Barotolerance of Acid-adapted and Cold-adapted Bacterial Isolates of E. coli O157:H7, Salmonella spp., and L. monocytogenes in an Acidic Buffer Model.
The fruit and vegetable juice industry has shown a growing trend in minimally processed juices. A frequent technology used in the production of functional juices is cold pressure, which refers to the application of high pressure processing (HPP) at low temperatures to inactivate foodborne pathogens. HPP juice manufacturers are required to demonstrate a 5-log reduction of the pertinent microorganism to comply with FDA Juice HACCP. However, there is no consensus on validation study approaches for bacterial strain selection or their preparation. Individual bacterial strains were grown using three different growth conditions: neutral, cold-adapted, and acid-adapted. Approximately 6.0-7.0 log CFU/mL of the matrix-adapted bacterial strains were inoculated individually into buffered peptone water (BPW) at pH 3.50 ± 0.10 (HCl adjusted) and treated at sublethal pressures of 500 MPa for Escherichia coli O157:H7 and 200 MPa for Salmonella spp. and Listeria monocytogenes (180 s, 4°C). Analyses were conducted at 0, 24, and 48 h (4°C storage) post-HPP on nonselective media. E. coli O157:H7 exhibited greater barotolerance than Salmonella spp. and L. monocytogenes. In neutral growth conditions, E. coli O157:H7 strain TW14359 demonstrated the greatest resistance (2.94 ± 0.64 log reduction), and E. coli O157:H7 strain SEA13B88 was significantly more sensitive (P < 0.05). Salmonella isolates, neutral and acid-adapted, expressed similar barotolerance to one another. Cold-adapted S. Cubana and S. Montevideo showed greater resistance compared to other cold-adapted strains. Acid-adapted L. monocytogenes strain MAD328 had <1.00 ± 0.23 log reduction while acid-adapted L. monocytogenes strains CDC and Scott A were significantly more sensitive (P < 0.05) with reductions of 2.13 ± 0.48 and 3.43 ± 0.50 log CFU/mL, respectively. These results suggested, under the conditions tested, bacterial strain and preparation methods influence HPP efficacy and should be considered when conducting validation studies.


Indian Ocean salinity build-up primes deglacial ocean circulation recovery.
The Indian Ocean provides a source of salt for North Atlantic deep-water convection sites, via the Agulhas Leakage, and may thus drive changes in the ocean's overturning circulation1-3. However, little is known about the salt content variability of Indian Ocean and Agulhas Leakage waters during past glacial cycles and how this may influence circulation. Here we show that the glacial Indian Ocean surface salt budget was notably different from the modern, responding dynamically to changes in sea level. Indian Ocean surface salinity increased during glacial intensification, peaking in glacial maxima. We find that this is due to rapid land exposure in the Indonesian archipelago induced by glacial sea-level lowering, and we suggest a mechanistic link via reduced input of relatively fresh Indonesian Throughflow waters into the Indian Ocean. Using climate model results, we show that the release of this glacial Indian Ocean salinity via the Agulhas Leakage during deglaciation can directly impact the Atlantic Meridional Overturning Circulation and global climate.


Segmentation of three-dimensional retinal image data.
We have combined methods from volume visualization and data analysis to support better diagnosis and treatment of human retinal diseases. Many diseases can be identified by abnormalities in the thicknesses of various retinal layers captured using optical coherence tomography (OCT). We used a support vector machine (SVM) to perform semi-automatic segmentation of retinal layers for subsequent analysis including a comparison of layer thicknesses to known healthy parameters. We have extended and generalized an older SVM approach to support better performance in a clinical setting through performance enhancements and graceful handling of inherent noise in OCT data by considering statistical characteristics at multiple levels of resolution. The addition of the multi-resolution hierarchy extends the SVM to have "global awareness." A feature, such as a retinal layer, can therefore be modeled.


[Synthetic evaluation of the clinical effect of the Shengmai capsule for treatment of chronic congestive heart failure using analytic hierarchy process].
To explore the approaches and techniques for synthetic evaluation of the clinical therapeutic effect of new Chinese herbal medicine in clinical trials. In a double-blind, randomized, and placebo-controlled clinical trail, analytic hierarchy process (AHP) was applied to evaluate the clinical therapeutic effect of Shengmai capsule in the treatment of chronic congestive heart failure. Shengmai capsule produced positive therapeutic effect on chronic congestive heart failure. A feasible method is established for evaluating and grading the clinical therapeutic effect of Chinese herbal medicine.


Comparative Proteomic Insights into the Lactate Responses of Halophilic Salinicoccus roseus W12.
Extremophiles use adaptive mechanisms to survive in extreme environments, which is of great importance for several biotechnological applications. A halophilic strain, Salinicoccus roseus W12, was isolated from salt lake in Inner Mongolia, China in this study. The ability of the strain to survive under high sodium conditions (including 20% sodium lactate or 25% sodium chloride, [w/v]) made it an ideal host to screen for key factors related to sodium lactate resistance. The proteomic responses to lactate were studied using W12 cells cultivated with or without lactate stress. A total of 1,656 protein spots in sodium lactate-treated culture and 1,843 spots in NaCl-treated culture were detected by 2-dimensional gel electrophoresis, and 32 of 120 significantly altered protein spots (fold change > 2, p < 0.05) were identified by matrix-assisted laser-desorption ionization time-of-flight mass spectrometry. Among 21 successfully identified spots, 19 proteins were upregulated and 2 were downregulated. The identified proteins are mainly involved in metabolism, cellular processes and signaling, and information storage and processing. Transcription studies confirmed that most of the encoding genes were upregulated after the cells were exposed to lactate in 10 min. Cross-protecting and energy metabolism-related proteins played an important role in lactate tolerance for S. roseus W12.


Electron microscopic study of the rubrocerebellar projection in the cat.
Rubral neurons sending axons to the cerebellar anterior interpositus nucleus (AIN) in the cat were identified light microscopically by labeling them with horseradish peroxidase (HRP). The synaptic organization of these rubral neurons and of their afferents from the cerebral motor cortex and the AIN was also analyzed electron microscopically by combined anterograde degeneration and retrograde HRP-labeling techniques. In the light microscopic study, either HRP or a mixture of HRP and kainic acid was injected into the AIN. Both of the injections resulted in retrograde labeling of rubrocerebellar projection neurons in the red nucleus on the contralateral side. The labeled neurons were distributed throughout the rostrocaudal extent of the red nucleus: some lay in clusters. Most labeled neurons were small to medium-sized, although some were large. The injection of HRP into the AIN also resulted in anterograde labeling of cerebellorubral projection fibers terminating in a wider area of the red nucleus on the contralateral side of the injection, whereas the injection of a mixture of HRP and kainic acid showed no anterograde labeling of fibers or terminals. In one set of electron microscopic observations, HRP injections into the AIN were combined with ablation of the motor cortex. Degenerating axon terminals were occasionally found to synapse with both dendrites and neuronal somata labeled with HRP retrogradely. In another set of electron microscopic observations, a mixture of HRP and kainic acid was injected into the AIN in order to label rubrocerebellar projection neurons retrogradely and to bring about degeneration in the cerebellorubral projection fibers anterogradely. Abundant degenerating axon terminals were observed to make axosomatic synaptic contacts with rubral neurons labeled with HRP retrogradely and also with unlabeled rubral neurons. These results indicate that cerebrorubrocerebellar and rubrocerebellorubral monosynaptic circuitries exist which constitute one of the cerebrocerebellar linkages, as well as those linkages via the inferior olivary complex and the pontine nuclei.


High expression of shMDG1 gene is associated with low metastatic potential of tumor cells.
Metastasis is the primary cause of mortality associated with cancer. Molecular mechanisms leading to metastatic spread are poorly studied. To get a better understanding of this process, we compared the gene expression pattern of two isogenic cell lines, HET-SR and HET-SR1 (Rous Sarcoma Virus-transformed embryo hamster fibroblasts) with different metastatic activity using the differential display technique. A novel cDNA of hamster gene shMDG1 (Syrian hamster homologue of microvascular differentiation gene 1), which had 94% homology with rat MDG1 gene, was identified. Expression of shMDG1 was increased in low metastatic HET-SR cell line in comparison to high metastatic HET-SR1. Sequence analysis of the ORF of shMDG1 gene showed that it belongs to the DnaJ/heat-shock proteins of 40 kDa (HSP40) chaperones family, considered to function as a cochaperone of HSP70 family. In order to confirm involvement of shMDG1 in metastasis, we injected parental and shMDG1 overexpressed cells into animals. We showed that overexpression of the shMDG1 gene significantly diminished the metastatic activity of both HET-SR and HET-SR1 cells. The shMDG1-induced repression of metastasis was not connected with alterations in cell proliferation and motility in vitro, but correlated well with a decrease in content of the Asn-linked beta1-6 branched oligosaccharides on cell surface.


Serum neutrophil gelatinase-associated lipocalin concentration reflects severity of coronary artery disease in patients without heart failure and chronic kidney disease.
Serum neutrophil gelatinase-associated lipocalin (NGAL) is recognized as a useful biomarker for acute kidney injury. Recently, elevated NGAL levels were reported in patients with heart failure and cardiac events, but the association between serum NGAL and severity of coronary artery disease (CAD) has not been investigated adequately. This study aimed to evaluate the association between serum NGAL concentration and CAD severity in patients without heart failure and chronic kidney disease. Two-hundred thirteen patients [mean age: 66.2 ± 9.2 (SD)] without heart failure and chronic kidney disease (estimated glomerular filtration rate >60 mL/min/1.73 m(2)) who underwent coronary angiography were retrospectively analyzed using the SYNTAX score. The mean concentration of serum NGAL was 134.3 ± 111.3 ng/mL. A statistically significant correlation was observed between serum NGAL levels and the SYNTAX score (R = 0.18, P = 0.0091). Multivariable analysis also showed elevated serum NGAL as an independent risk factor for a high SYNTAX score (P < 0.01). Moreover, we evaluated the association of serum NGAL and brain natriuretic peptide (BNP) with the SYNTAX score. Patients with high levels of serum NGAL (>100 ng/mL) and high levels of BNP (>25 pg/mL) had a higher SYNTAX score (low-low vs. high-high: 13.8 ± 13.4 vs. 20.8 ± 18.9, P < 0.05). Serum NGAL levels were positively and significantly associated with CAD severity, and the evaluation of both serum NGAL and BNP was useful for predicting CAD in patients without renal dysfunction and heart failure. Serum NGAL might be a biomarker for CAD severity.


Antioxidant, antimicrobial, and theoretical studies of the thiosemicarbazone derivative Schiff base 2-(2-imino-1-methylimidazolidin-4-ylidene)hydrazinecarbothioamide (IMHC).
Adverse antimicrobial activities of thiosemicarbazone (TSC) and Schiff base derivatives have widely been studied by using different kinds of microbes, in addition different methods were used to assay the antioxidant activities using DPPH, peroxids, or ntrosyl methods. However, there are no studies describing the synthesis of TSC derived from creatinine. In this study, 2-(2-imino-1-methylimidazolidin-4-ylidene)hydrazinecarbothioamide (IMHC) was synthesized by the reaction of creatinine with thiosemicarbazide. The novel molecule was characterized by FT-IR, UV-VIS, and NMR spectra in addition of the elemental analysis. The free radical scavenging ability of the IMHC was determined by it interaction with the stable-free radical 2,2"-diphenyl-1-picrylhydrazyl (or nitric oxide or hydrogen peroxide) and showed encouraging antioxidant activities. Density functional theory calculations of the IMHC performed using molecular structures with optimized geometries. Molecular orbital calculations provide a detailed description of the orbitals, including spatial characteristics, nodal patterns, and the contributions of individual atoms. Highest occupied molecular orbital-lowest unoccupied molecular orbital energies and structures are shown. IMHC shows considerable antibacterial and antifungal activities. The free radical scavenging activity of synthesized compound was screened for in vitro antioxidant activity.


EMBED: Essential MicroBiomE Dynamics, a dimensionality reduction approach for longitudinal microbiome studies.
Dimensionality reduction offers unique insights into high-dimensional microbiome dynamics by leveraging collective abundance fluctuations of multiple bacteria driven by similar ecological perturbations. However, methods providing lower-dimensional representations of microbiome dynamics both at the community and individual taxa levels are not currently available. To that end, we present EMBED: Essential MicroBiomE Dynamics, a probabilistic nonlinear tensor factorization approach. Like normal mode analysis in structural biophysics, EMBED infers ecological normal modes (ECNs), which represent the unique orthogonal modes capturing the collective behavior of microbial communities. Using multiple real and synthetic datasets, we show that a very small number of ECNs can accurately approximate microbiome dynamics. Inferred ECNs reflect specific ecological behaviors, providing natural templates along which the dynamics of individual bacteria may be partitioned. Moreover, the multi-subject treatment in EMBED systematically identifies subject-specific and universal abundance dynamics that are not detected by traditional approaches. Collectively, these results highlight the utility of EMBED as a versatile dimensionality reduction tool for studies of microbiome dynamics.


Characterization of a human bladder cancer cell line selected for resistance to BMY 25067, a novel analogue of mitomycin C.
This study describes characteristics of a human bladder cancer cell line, SCaBER/R, selected for resistance to a mitomycin C (MMC) analogue BMY 25067. The SCaBER/R cell line was isolated by repeated 24 h exposures of the parental cells to 0.09 microM BMY 25067 (IC90, 24 h drug exposure) over a period of about 180 days. Approximately 2.2-fold higher concentration of BMY 25067 was required to kill 50% of the SCaBER/R cell line compared with parental cells (p < 0.001). The IC20 and IC90 values for BMY 25067 were also significantly higher in the SCaBER/R cell line than in SCaBER. Unlike most MMC resistant cell lines, the SCaBER/R cell line displayed a marked cross-resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and lacked cross-resistance to cisplatin, doxorubicin or VP-16. The SCaBER/R cell line also displayed a marked cross-resistance to the parent drug (MMC) and BMY 25282, another analogue of MMC. NADPH cytochrome P450 reductase activity, an enzyme implicated in bio-reductive activation of MMC, did not differ significantly in these cells. DT-diaphorase activity, another MMC activation enzyme, was significantly lower in the SCaBER/R cell line when compared to the SCaBER cells. These results suggest that relatively lower sensitivity of SCaBER/R cell line to MMC and BMY 25067 may result from impaired drug activation. Cellular levels of glutathione (GSH) and GSH-transferase (GST), which have been suggested to affect the cytotoxicity of MMC, were comparable in SCaBER and SCaBER/R cell lines. BMY 25067 induced DNA interstrand cross-links (DNA-ISC) could not be detected in either of the cell lines even at drug concentrations which produced a significant cell kill. These findings suggest that (a) cellular resistance to BMY 25067 in the SCaBER/R cell line may be due to impaired drug activation, and (b) the nature of the cytotoxic produced by BMY 25067 may be different from that of MMC.


Enzymatic fucosylations of non-natural sialylated type-I trisaccharides with recombinant fucosyl-transferase-III.
Recombinant fucosyl-transferase-III (Lewis type enzyme) is used to prepare a series of non-natural sialyl-Lewis derivatives on a preparative scale. The enzyme tolerates a wide range of acceptors which have the natural N-acetyl group of the glucosamine moiety replaced by substituted aromatic and heteroaromatic amides.


Biomimetic surgical mesh to replace fascia with tunable force-displacement.
Here we mimic the mechanical properties of native fascia to design surgical mesh for fascia replacement. Despite the widespread acceptance of synthetic materials as tissue scaffolds for pelvic floor disorders, mechanical property mismatch between mesh and adjacent native tissue drives fibrosis and erosion, leading the FDA to remove several surgical meshes from the market. However, autologous tissue does not induce either fibrosis or adjacent tissue erosion, suggesting the potential for biomimetic surgical mesh. In this study, we determined the design rules for mesh that mimics native fascia by mathematically modeling multi-component polymer networks, composed of elastin-like and collagen-like fibers, using a spring-network model. To validate the model, we measured the stress-strain curves of native bovine and nonhuman primate (Macaca mulatta) abdominal fascia in both toe and linear regions. We find that the stiffer collagen-like fibers must remain limp until the elastin-like fibers extend to the initial length of spanning collagen-like fibers under uniaxial tension. Comparing model results to experiment determines the product of fiber volume fraction and elastic modulus, a critical design parameter. Dual fiber mesh with mechanical properties that mimic fascia are feasible. These results have broad application to a wide range of soft tissue replacements including ~200,000 surgeries/year for pelvic floor disorders, because standard-of-care mesh contain only stiffer polymers that behave more like collagen than native tissue.