In Situ Synthesis of Horseradish Peroxidase Nanoflower@Carbon Nanotube Hybrid Nanobiocatalysts with Greatly Enhanced Catalytic Activity.

Organic-inorganic hybrid nanoflowers (NFs) consisting of horseradish peroxidase (HRP) and copper II (Cu2+) are successfully synthesized with the involvement of carbon nanotubes (CNTs) by in situ and post-modification methods. Catalytic activities of in situ synthesized HRP-NF@CNT (HRP-NF@CNT-Is) and post-modification-synthesized HRP-NF@CNTs (HRP-NF@CNT-Pm) are systematically examined. The 30 mg CNTs incorporated HRP-NF@CNT-Is (HRP-NF@CNT-30Is) exhibits greatly increased catalytic activity and stability toward 3,3',5,5'-tetramethylbenzidine (TMB), thanks to the synergistic effect between HRP-NF and CNTs and the peroxidase-like activity of CNTs in the presence of hydrogen peroxide (H2O2). While HRP-NF@CNT-30Is retains almost 85% of its initial activity even after 10 cycles, HRP-NF (without CNTs) loses half of its initial activity at the same experimental conditions. We study how two experimental parameters, the pH values and temperatures, influence the catalytic activity of HRP-NF@CNT-30Is, in addition to the fact that HRP-NF@CNT-30Is is employed to detect the presence of H2O2 and glutathione (GSH) with colorimetric and spectrophotometric readouts. For instance, HRP-NF@CNT-30Is is used to sensitively detect H2O2 in the range of 20 to 300 µM with an LOD of 2.26 µM. The catalytic activity of HRP-NF@CNT-30Is is suppressed in the presence of GSH, and then an obvious color change from blue to nearly colorless is observed. Using this strategy, GSH is also sensitively determined in the range of 20-200 µM with an LOD of 11.2 µM. We expect that HRP-NF@CNTs can be used as a promising and novel nanobiocatalyst for various biomedical and industrial applications in the near future.

Hollow microneedle array fabrication using a rational design to prevent skin clogging in transdermal drug delivery.

Microneedle (MN) technology is promising to replace hypodermic needles for practical use and painless drug delivery. However, the complex top-down fabrication process of functional MN arrays is a bottleneck that hinders their widespread use. Here, we fabricate the tapered hollow MN array using a unique bi-level-tip by combining strain-engineering and capillary self-assembly of carbon nanotube (CNT) microstructures. Strain-engineering facilitated by the offset pattern of the catalyst enables the growth of bent, bi-level CNT microstructures while capillary self-assembly helps in constituting the tapered geometry of MNs. The bottom-up fabrication that consists of only two standard photolithography steps and CNT growth to form the scaffold of MNs followed by a polymer (polyimide) reinforcement step to impart mechanical stiffness to MNs provides scalable and fewer processing steps. The tapered shape of the MN allows an 8 times smaller force to pierce and penetrate the skin compared to the straight MN. The liquid delivery rate of the bi-level-tip MN is measured to be 26% better than the flat tip MN of the same lumen size as its geometry reduces skin clogging effect at the needle tip. In addition, cytotoxicity tests verify that the polyimide reinforced CNT-MNs are biocompatible for future in vivo applications.

PLASMA NEPRILYSIN LEVELS IN PATIENTS WITH POLYCYSTIC OVARY SYNDROME.

Context: Insulin resistance has been detected in a majority of patients with polycystic ovary syndrome (PCOS). Elevated neprilysin levels are associated with insulin resistance. Objective: The present study aims to investigate plasma neprilysin and its relationship with endocrine and metabolic characteristics in patients with PCOS. Subjects and Methods: Thirty-five premenopausal PCOS patients and 35 healthy volunteers of similar age were included in the study. Demographic characteristics, biochemical and hormonal findings and also plasma neprilysin levels were determined in these patients and healthy controls. Results: In our study, HOMA-IR values were significantly higher in PCOS patients (3.3 ± 1.8) compared with the controls [(1.6 ± 1), p<0.01]. Plasma neprilysin levels were significantly higher in the PCOS group compared to the control group (1502.1 ± 1641.2 vs. 764.6 ± 562.6 pg/mL). There was no difference in plasma neprilysin levels when PCOS patients were classified as overweight-obesity (BMI≥25kg/m2) or non-obesity (BMI<25kg/m2). Conclusion: Our findings revealed significantly higher levels for plasma neprilysin and HOMA-IR values in PCOS patients when compared to controls. No significant differences were noted between obese PCOS patients and non-obese PCOS patients in terms of plasma neprilysin levels.

Assessment of mental health issues in pregnant women with fetal complications: Relation to attachment and anxiety.

PURPOSE: Pregnancy period is important for all women. The aim of this study was to evaluate mental status, antenatal depression, attachment, and anxiety in pregnant women with fetal complications. DESIGN AND METHODS: Mothers who had been diagnosed with fetal complications during pregnancy were interviewed after the delivery of their babies. FINDINGS: The results showed that anxiety and depression scores were significantly higher in the mothers who had received psychiatric support treatment. Maternal attachment scores were found low, depression scores were found high in mothers of children with congenital anomalies. CONCLUSION: According to the results of the study, a maternal attachment was low and depression was high in mothers with congenital anomalies. PRACTICE IMPLICATIONS: Possible psychological problems after birth can be prevented by psychotherapeutic nursing interventions during pregnancy to pregnant women who have babies with fetal anomalies.

Unconventional supercurrent phase in Ising superconductor Josephson junction with atomically thin magnetic insulator.

In two-dimensional (2D) NbSe2 crystal, which lacks inversion symmetry, strong spin-orbit coupling aligns the spins of Cooper pairs to the orbital valleys, forming Ising Cooper pairs (ICPs). The unusual spin texture of ICPs can be further modulated by introducing magnetic exchange. Here, we report unconventional supercurrent phase in van der Waals heterostructure Josephson junctions (JJs) that couples NbSe2 ICPs across an atomically thin magnetic insulator (MI) Cr2Ge2Te6. By constructing a superconducting quantum interference device (SQUID), we measure the phase of the transferred Cooper pairs in the MI JJ. We demonstrate a doubly degenerate nontrivial JJ phase (ϕ), formed by momentum-conserving tunneling of ICPs across magnetic domains in the barrier. The doubly degenerate ground states in MI JJs provide a two-level quantum system that can be utilized as a new dissipationless component for superconducting quantum devices. Our work boosts the study of various superconducting states with spin-orbit coupling, opening up an avenue to designing new superconducting phase-controlled quantum electronic devices.

Retrospective analysis of vitamin D status on inflammatory markers and course of the disease in patients with COVID-19 infection.

PURPOSE: The aim of the study was to investigate the association between serum 25-hydroxyvitamin D status within the last 6 months prior to COVID-19 infection and parameters of immune function and clinical outcomes. METHODS: Fifty-six patients, who were admitted to the emergency clinic and diagnosed with COVID-19 infection, were included in the study. Data on clinical characteristics, inflammatory parameters and vitamin D status were recorded for each patient. All the participants had data on 25-hydroxyvitamin D status within the last 6 months prior to COVID-19 infection. RESULTS: The patients were stratified as those with vitamin D status less than 20 ng/mL and higher than 20 ng/mL. A group with vitamin D status less than 20 ng/mL had lower lymphocyte counts and lower haemoglobin levels that was statistically significant (respectively; p = 0.021, p = 0.035). Higher C-reactive protein (CRP) levels were seen in the vitamin D-deficient group (p = 0.013). It was observed that vitamin D status of the patients who required oxygen therapy were lower than those who did not require oxygen therapy, not statistically significant (p = 0.05). Patients who did not use vitamin D supplementation within 6 months prior to COVID-19 infection had more likely to be diagnosed with pneumonia (p = 0.004). CONCLUSION: Cases with lower vitamin D status had increased inflammatory markers and worse clinical outcomes than patients with higher vitamin D status. This study suggests that vitamin D status can be used as a prognostic factor in COVID-19 patients, and vitamin D supplementation can be recommended to improve the clinical outcomes in COVID-19 infection.

Association Between Resistance to Cinacalcet and Parathyroid Gland Hyperplasia in Kidney Transplant Recipients with Persistent Hypercalcemia.

BACKGROUND: Persistent hypercalcemia and hyperparathyroidism after successful kidney transplantation can be detrimental in some recipients and should be ameliorated. OBJECTIVE: To point out the concerns regarding resistance to cinacalcet in kidney transplant recipients with persistent hypercalcemia. METHODS: 14 renal transplant recipients who received cinacalcet treatment because of persistent hypercalcemia were included in the study. Serum creatinine, estimated glomerular filtration rate (eGFR), calcium, phosphorus, and intact parathyroid hormone (PTH) levels at the baseline and throughout the treatment, and ultrasonography and parathyroid scintigraphy findings were recorded. RESULTS: Cinacalcet treatment was initiated after a mean±SD of 20.7±19.7 months of transplantation and maintained for 16.9±7.9 months. Serum calcium levels were significantly decreased with the cinacalcet treatment. There were no significant changes in serum creatinine, eGFR, phosphorus, and PTH levels. In all participants, serum calcium levels were increased from 9.8±0.6 to 11.1±0.6 mg/dL (p<0.001) within 1 month of cessation of cinacalcet. 7 recipients with adenoma-like hyperplastic glands underwent parathyroidectomy (PTx) due to failure with cinacalcet. CONCLUSION: Cinacalcet may be an appropriate treatment for a group of recipients with hypercalcemia without adenoma-like hyperplastic glands or who had a contraindication for surgery. Recipients with enlarged parathyroid gland may resist to cinacalcet-induced decrease in serum PTH, although the concomitant hypercalcemia may be corrected.

Evaluation of upper oesophageal sphincter in unilateral vocal fold paralysis.

OBJECTIVE: To evaluate dysphagia and manometric changes in the upper oesophageal sphincter in patients with unilateral vocal fold paralysis. METHODS: Thirty patients with unilateral vocal fold paralysis due to vagal nerve paralysis scheduled for evaluation were enrolled in the study group; 24 healthy subjects were included in the control group. Upper oesophageal sphincter basal and residual pressure, relaxation time, and pharyngeal pressure values were evaluated by manometry. All patients completed the Turkish Eating Assessment Tool 10 questionnaire, the MD Anderson dysphagia questionnaire and the reflux symptom index form. RESULTS: Swallowing assessment questionnaires and reflux symptom index results were significantly higher in the study group. Upper oesophageal sphincter basal and relaxation pressures were lower in the study group. Upper oesophageal sphincter relaxation time was shorter in the study group, but pressure values recorded from the pharynx were higher. CONCLUSION: Upper oesophageal sphincter manometric pressure was lower in patients with unilateral vocal fold paralysis. A hypotonic sphincter likely contributes to dysphagia and aspiration.

SHORT TERM EFFECT OF LAPAROSCOPIC SLEEVE GASTRECTOMY ON CLINICAL, RENAL PARAMETERS AND URINARY NGAL LEVELS IN DIABETIC AND NON DIABETIC OBESITY.

BACKGROUND: Although diseases such as diabetes, hypertension, obstructive sleep apnea and hyperlipidemia are clearly documented as obesity associated diseases, it is not well-known whether obesity causes renal pathologies. The aim of the present study was to evaluate the effect of weight loss following laparoscopic sleeve gastrectomy on clinical, renal parameters and urinary Neutrophil gelatinase-associated lipocalin (NGAL) levels in diabetic and non-diabetic obese patients. METHODS: Nineteen morbidly obese patients (10 diabetic and 9 non diabetic) who underwent laparoscopic sleeve gastrectomy were evaluated clinically (anthropometric measurements) and biochemically before surgery and at 6 months from surgery. RESULTS: Significant decreases in weight, BMI, FPG, PPG and HbA1c levels were observed in the diabetic group when the baseline and 6th month parameters of the patients were compared. There was also a significant decrease in SBP and DBP. At 6th month following laparoscopic sleeve gastrectomy, renal parameters such as creatinine, mAlb/creatinine, NGAL/creatinine did not differ in the diabetic group. In the nondiabetic group, serum creatinine levels were significantly decreased, but other renal parameters such as mAlb/creatinine and NGAL/creatinine were not significantly different. CONCLUSIONS: Our findings revealed significant decreases in weight, body mass index and glycemic parameters after sleeve gastrectomy in diabetic and non-diabetic patients, while no significant alteration was noted in renal functions, urinary NGAL and microalbumin levels.

The effect of primary hyperparathyroidism on pancreatic exocrine function.

BACKGROUND: Elastase-1 is a proteolytic enzyme secreted by pancreatic acinar cells, and measurements of the concentration this enzyme are used to evaluate pancreatic exocrine function. We aimed to determine whether pancreatic exocrine function declines due to chronic hypercalcemia by measuring fecal elastase levels. METHODS: 75 patients with primary hyperparathyroidism (18 men and 47 women) and 30 healthy subjects (11 men and 19 women) participated in this study. Renal function tests, lipid parameters, bone mineral density, and serum calcium, phosphorus, vitamin D, parathormone, glucose, and thyroid stimulating hormone levels as well as fecal elastase concentrations, were determined in these patients and controls. RESULTS: The mean fecal elastase level was 335.3 ± 181.4 µg/g in the PHPT group and 317.4 ± 157.3 µg/g in the control group. There was no significant difference in fecal elastase levels between the two groups (p = 0.5). CONCLUSIONS: Chronic hypercalcemia in primary hyperparathyroidism did not decrease the fecal elastase level, which is an indirect indicator of chronic pancreatitis; therefore, chronic hypercalcemia in PHPT may not cause chronic pancreatitis.

Formation of functional nanobiocatalysts with a novel and encouraging immobilization approach and their versatile bioanalytical applications.

The discovery of functional organic-inorganic hybrid nanoflowers (FNFs) consisting of proteins/enzymes as the organic components and Cu(ii) ion as the inorganic component has made an enormous impact on enzyme immobilization studies. The FNFs synthesized by an encouraging and novel approach not only showed high stabilities but also much enhanced catalytic activities as compared to free and conventionally immobilized enzymes. A recent development demonstrated that FNF formation has moved beyond the initial discovery in which enzymes and Cu2+ ions used as the organic and inorganic parts, respectively, are replaced with new organic (chitosan, amino acid and plant extracts) and inorganic (Cu2+ and Fe2+) materials. The new organic materials incorporated into FNFs act as Fenton-like agents and then show peroxidase-like activity owing to the metal ions and the porous structure of FNFs in the presence of hydrogen peroxide (H2O2). All FNFs have been widely utilized in many different scientific and industrial fields due to their greatly enhanced activities and stabilities. This review focuses primarily on the preparation, characterization, and bioanalytical applications of FNFs and explains the mechanisms of their formation and enhanced activities and stabilities.

Conductance through a helical state in an Indium antimonide nanowire.

The motion of an electron and its spin are generally not coupled. However in a one-dimensional material with strong spin-orbit interaction (SOI) a helical state may emerge at finite magnetic fields, where electrons of opposite spin will have opposite momentum. The existence of this helical state has applications for spin filtering and cooper pair splitter devices and is an essential ingredient for realizing topologically protected quantum computing using Majorana zero modes. Here, we report measurements of a quantum point contact in an indium antimonide nanowire. At magnetic fields exceeding 3 T, the 2 e 2/h conductance plateau shows a re-entrant feature toward 1 e 2/h which increases linearly in width with magnetic field. Rotating the magnetic field clearly attributes this experimental signature to SOI and by comparing our observations with a numerical model we extract a spin-orbit energy of approximately 6.5 meV, which is stronger than the spin-orbit energy obtained by other methods.Indium antimonide nanowires have large spin-orbit coupling, which can give rise to helical states that are an important part of proposals for topological quantum computing. Here the authors measure conductance through the helical states and extract a larger spin-orbit energy than obtained before.

Single Molecule Bioelectronics and Their Application to Amplification-Free Measurement of DNA Lengths.

As biosensing devices shrink smaller and smaller, they approach a scale in which single molecule electronic sensing becomes possible. Here, we review the operation of single-enzyme transistors made using single-walled carbon nanotubes. These novel hybrid devices transduce the motions and catalytic activity of a single protein into an electronic signal for real-time monitoring of the protein's activity. Analysis of these electronic signals reveals new insights into enzyme function and proves the electronic technique to be complementary to other single-molecule methods based on fluorescence. As one example of the nanocircuit technique, we have studied the Klenow Fragment (KF) of DNA polymerase I as it catalytically processes single-stranded DNA templates. The fidelity of DNA polymerases makes them a key component in many DNA sequencing techniques, and here we demonstrate that KF nanocircuits readily resolve DNA polymerization with single-base sensitivity. Consequently, template lengths can be directly counted from electronic recordings of KF's base-by-base activity. After measuring as few as 20 copies, the template length can be determined with <1 base pair resolution, and different template lengths can be identified and enumerated in solutions containing template mixtures.

Processive Incorporation of Deoxynucleoside Triphosphate Analogs by Single-Molecule DNA Polymerase I (Klenow Fragment) Nanocircuits.

DNA polymerases exhibit a surprising tolerance for analogs of deoxyribonucleoside triphosphates (dNTPs), despite the enzymes' highly evolved mechanisms for the specific recognition and discrimination of native dNTPs. Here, individual DNA polymerase I Klenow fragment (KF) molecules were tethered to a single-walled carbon nanotube field-effect transistor (SWCNT-FET) to investigate accommodation of dNTP analogs with single-molecule resolution. Each base incorporation accompanied a change in current with its duration defined by τclosed. Under Vmax conditions, the average time of τclosed was similar for all analog and native dNTPs (0.2 to 0.4 ms), indicating no kinetic impact on this step due to analog structure. Accordingly, the average rates of dNTP analog incorporation were largely determined by durations with no change in current defined by τopen, which includes molecular recognition of the incoming dNTP. All α-thio-dNTPs were incorporated more slowly, at 40 to 65% of the rate for the corresponding native dNTPs. During polymerization with 6-Cl-2APTP, 2-thio-dTTP, or 2-thio-dCTP, the nanocircuit uncovered an alternative conformation represented by positive current excursions that does not occur with native dNTPs. A model consistent with these results invokes rotations by the enzyme's O-helix; this motion can test the stability of nascent base pairs using nonhydrophilic interactions and is allosterically coupled to charged residues near the site of SWCNT attachment. This model with two opposing O-helix motions differs from the previous report in which all current excursions were solely attributed to global enzyme closure and covalent-bond formation. The results suggest the enzyme applies a dynamic stability-checking mechanism for each nascent base pair.

One-Dimensional Poole-Frenkel Conduction in the Single Defect Limit.

A single point defect surrounded on either side by quasi-ballistic, semimetallic carbon nanotube is a nearly ideal system for investigating disorder in one-dimensional (1D) conductors and comparing experiment to theory. Here, individual single-walled nanotubes (SWNTs) are investigated before and after the incorporation of single point defects. Transport and local Kelvin Probe force microscopy independently demonstrate high-resistance depletion regions over 1.0 µm wide surrounding one point defect in semimetallic SWNTs. Transport measurements show that conductance through such wide depletion regions occurs via a modified, 1D version of Poole-Frenkel field-assisted emission. Given the breadth of theory dedicated to the possible effects of disorder in 1D systems, it is surprising that a Poole-Frenkel mechanism appears to describe defect scattering and resistance in this semimetallic system.

Observing lysozyme's closing and opening motions by high-resolution single-molecule enzymology.

Single-molecule techniques can monitor the kinetics of transitions between enzyme open and closed conformations, but such methods usually lack the resolution to observe the underlying transition pathway or intermediate conformational dynamics. We have used a 1 MHz bandwidth carbon nanotube transistor to electronically monitor single molecules of the enzyme T4 lysozyme as it processes substrate. An experimental resolution of 2 µs allowed the direct recording of lysozyme's opening and closing transitions. Unexpectedly, both motions required 37 µs, on average. The distribution of transition durations was also independent of the enzyme's state: either catalytic or nonproductive. The observation of smooth, continuous transitions suggests a concerted mechanism for glycoside hydrolysis with lysozyme's two domains closing upon the polysaccharide substrate in its active site. We distinguish these smooth motions from a nonconcerted mechanism, observed in approximately 10% of lysozyme openings and closings, in which the enzyme pauses for an additional 40-140 µs in an intermediate, partially closed conformation. During intermediate forming events, the number of rate-limiting steps observed increases to four, consistent with four steps required in the stepwise, arrow-pushing mechanism. The formation of such intermediate conformations was again independent of the enzyme's state. Taken together, the results suggest lysozyme operates as a Brownian motor. In this model, the enzyme traces a single pathway for closing and the reverse pathway for enzyme opening, regardless of its instantaneous catalytic productivity. The observed symmetry in enzyme opening and closing thus suggests that substrate translocation occurs while the enzyme is closed.

Manufacture and characterization of kefir made from cow and buffalo milk, using kefir grain and starter culture.

The microbiological and chemical characteristics as well as organic and amino acid profiles of kefir samples made from cow and buffalo milks fermented by kefir grains and starter culture were investigated during storage for 21 d at 4°C. After incubation, lactic, acetic, and citric acid concentrations showed a difference among the samples due to milk type and production methods. Storage time had little effect on the organic acid values of kefir samples. As compared with cow milk kefir, buffalo milk kefir had higher numbers of microorganisms, except lactobacilli, at the end of storage. Whereas pH and titratable acidity exhibited similar changes during storage in all kefir samples, ethanol levels were significantly increased in buffalo milk kefir samples. Glutamic acid was the major amino acid at all sampling times for all samples. Tyrosine, serine, histidine, alanine, methionine, and lysine concentrations were determined to be different in all samples depending on milk type. In general, due to the higher microbial population (especially yeast), kefir made from buffalo milk may be preferred.

Giant magnetoconductance oscillations in hybrid superconductor-semiconductor core/shell nanowire devices.

The magnetotransport of GaAs/InAs core/shell nanowires contacted by two superconducting Nb electrodes is investigated, where the InAs shell forms a tube-like conductive channel around the highly resistive GaAs core. By applying a magnetic field along the nanowire axis, regular magnetoconductance oscillations with an amplitude in the order of e(2)/h are observed. The oscillation amplitude is found to be larger by 2 orders of magnitude compared to the measurements of a reference sample with normal metal contacts. For the Nb-contacted core/shell nanowire the oscillation period corresponds to half a flux quantum Φ0/2 = h/2e in contrast to the period of Φ0 of the reference sample. The strongly enhanced magnetoconductance oscillations are explained by phase-coherent resonant Andreev reflections at the Nb-core/shell nanowire interface.

Electrochemical charge-transfer resistance in carbon nanotube composites.

Using a model system of single, isolated carbon nanotubes loaded with high-capacitance metal-oxide films, we have quantitatively investigated electrochemical composites on the single-nanotube scale. Electrochemical charging and discharging of a model MnO2 storage material was used to probe interfacial charge transfer and surface impedances at the nanotube interface. We found that one single-walled carbon nanotube has an apparent surface resistivity of 30 mΩ cm(2), approximately 4 times smaller than for a multiwalled carbon nanotube and 50 times smaller than the 1.5 Ω cm(2) resistivity of Pt or graphite films. The improvement originates in the electrochemical-transport properties of microelectrodes shrunk to a nanotube's dimensions rather than any unique nanotube property like curvature, bandstructure, or surface chemistry. In explaining the enhanced performance of certain nanotube-containing composites, the results overturn widely held assumptions about nanotubes' roles while also providing guidelines for optimizing effective composites.

Electronic measurements of single-molecule catalysis by cAMP-dependent protein kinase A.

Single-molecule studies of enzymes open a window into their dynamics and kinetics. A single molecule of the catalytic domain of cAMP-dependent protein kinase A (PKA) was attached to a single-walled carbon nanotube device for long-duration monitoring. The electronic recording clearly resolves substrate binding, ATP binding, and cooperative formation of PKA's catalytically functional, ternary complex. Using recordings of a single PKA molecule extending over 10 min and tens of thousands of binding events, we determine the full transition probability matrix and conversion rates governing formation of the apo, intermediate, and closed enzyme configurations. We also observe kinetic rates varying over 2 orders of magnitude from one second to another. Anti-correlation of the on and off rates for PKA binding to the peptide substrate, but not ATP, demonstrates that regulation of enzyme activity results from altering the stability of the PKA-substrate complex, not its binding to ATP. The results depict a highly dynamic enzyme offering dramatic possibilities for regulated activity, an attribute useful for an enzyme with crucial roles in cell signaling.