Standardization of rate of sugar addition for the manufacture of Thabdi.

Traditional Indian Dairy Products are manufactured in India using an age old practice which varies from place to place. For manufacture of these products industrially, a standard formulation is required. Thabdi, a region specific, very popular heat desiccated milk product is one of such products which has not been studied scientifically. Sugar plays an important role in physico-chemical, sensory, textural characteristics and also the shelf life of any milk sweet. Hence for process standardization of Thabdi manufacture, different levels of sugar i.e. 4, 6, 8 and 10 (percent of milk) were studied so that an optimum level yielding best organoleptic characteristics in final product can be selected. The product was made from milk standardized to 0.66 Fat:SNF level and added with ghee @ 1.2 % of milk at the time of colour and texture development stage as selected from the earlier phase of study. Based on the results obtained, a level of 8 % sugar addition on the milk basis at the time of manufacture has been selected to have full taste and sensory attributes.

Assessing fall risk in osteoporosis patients: a comparative study of age-matched fallers and nonfallers.

This study aimed to investigate sway parameters and physical activity level of the age/gender-matched older adults with osteoporosis faller and nonfaller patients. By examining these factors, our objective was to understand how these faller and nonfaller groups with osteoporosis differed particularly in terms of balance capabilities and their impact on physical activity levels. We recruited 24 patients with osteoporosis: 12 who reported a fall within a year before recruitment (fallers) and 12 without falls (nonfallers). Given the close association between biochemical markers of musculoskeletal health such as serum calcium, parathyroid hormone (PTH), Vitamin D, and renal function, we compared these markers in both groups. As a result, elderly individuals with osteoporosis and with a history of falls within the preceding year indicated significantly higher sway velocity (P = 0.012*), sway area (P < 0.001*), and sway path length (P = 0.012*). Furthermore, fallers had significantly lower calcium (P = 0.02*) and Parathyroid hormone (PTH) (P = 0.02*), as well as higher Alkaline Phosphatase (ALP) (P = 0.02*) as compared to nonfallers despite similar vitamin D and creatinine levels. In conclusion, diminished biochemical factors in the osteoporosis faller group could possibly cause postural instability resulting in lower physical activity levels in the osteoporosis fall group and increasing the risk of falls.

Electrochemical genosensor based on RNA-responsive human telomeric G-quadruplex DNA: A proof-of-concept with SARS-CoV-2 RNA.

In this report, a facile and label-free electrochemical RNA biosensor is developed by exploiting methylene blue (MB) as an electroactive positive ligand of G-quadruplex. The electrochemical response mechanism of the nucleic acid assay was based on the change in differential pulse voltammetry (DPV) signal of adsorbed MB on the immobilized human telomeric G-quadruplex DNA with a loop that is complementary to the target RNA. Hybridization between synthetic positive control RNA and G-quadruplex DNA probe on the transducer platform rendered a conformational change of G-quadruplex to double-stranded DNA (dsDNA), and increased the redox current of cationic MB π planar ligand at the sensing interface, thereby the electrochemical signal of the MB-adsorbed duplex is proportional to the concentration of target RNA, with SARS-CoV-2 (COVID-19) RNA as the model. Under optimal conditions, the target RNA can be detected in a linear range from 1 zM to 1 µM with a limit of detection (LOD) obtained at 0.59 zM for synthetic target RNA and as low as 1.4 copy number for positive control plasmid. This genosensor exhibited high selectivity towards SARS-CoV-2 RNA over other RNA nucleotides, such as SARS-CoV and MERS-CoV. The electrochemical RNA biosensor showed DPV signal, which was proportional to the 2019-nCoV_N_positive control plasmid from 2 to 200000 copies (R2 = 0.978). A good correlation between the genosensor and qRT-PCR gold standard was attained for the detection of SARS-CoV-2 RNA in terms of viral copy number in clinical samples from upper respiratory specimens.

A histological method for quantifying Plasmodium falciparum in the brain in fatal paediatric cerebral malaria.

BACKGROUND: The sequestration of Plasmodium falciparum-infected erythrocytes in brain microvasculature through cytoadherence to endothelium, is the hallmark of the definitive diagnosis of cerebral malaria and plays a critical role in malaria pathogenesis. The complex pathophysiology, which leads each patient to the final outcome of cerebral malaria, is multifaceted and thus, metrics to delineate specific patterns within cerebral malaria are needed to further parse patients. METHODS: A method was developed for quantification utilizing counts of capillary contents (early-stage parasites, late-stage parasites and fibrin) from histological preparations of brain tissue after death, and compared it to the standard approach, in which the percentage of parasitized vessels in cross-section is determined. RESULTS: Within the initial cohort of 50 patients, two different observers agreed closely on the percentage of vessels parasitized, pigmented parasites and pigment globules (ICC = 0.795-0.970). Correlations between observers for correct diagnostic classification were high (Kendall's tau-b = 0.8779, Kappa = 0.8413). When these methods were applied prospectively to a second set of 50 autopsy samples, they revealed a heterogeneous distribution of sequestered parasites in the brain with pigmented parasites and pigment globules present in the cerebellum > cortex > brainstem. There was no difference in the distribution of early stages of parasites or in the percentage of vessels parasitized across the same sites. The second cohort of cases was also used to test a previously published classification and regression tree (CART) analysis; the quantitative data alone were able to accurately classify and distinguish cerebral malaria from non-cerebral malaria. Classification errors occurred within a subclassification of cerebral malaria (CM1 vs CM2). A repeat CART analysis for the second cohort generated slightly different classification rules with more accurate subclassification, although misclassification still occurred. CONCLUSIONS: The traditional measure of parasite sequestration in falciparum malaria, the percentage of vessels parasitized, is the most reliable and consistent for the general diagnosis of cerebral malaria. Methods that involve quantitative measures of different life cycle stages are useful for distinguishing patterns within the cerebral malaria population; these subclassifications may be important for studies of disease pathogenesis and ancillary treatment.

Supraorbital postmortem brain sampling for definitive quantitative confirmation of cerebral sequestration of Plasmodium falciparum parasites.

BACKGROUND: The conventional clinical case definition of cerebral malaria (CM) is imprecise but specificity is improved by a definitive clinical feature such as retinopathy or confirming sequestration of parasites in a post-mortem examination of the brain. A full autopsy is often not possible, since it is costly and may encounter resistance of the deceased's family. METHODS: We have assessed the use of a cytological smear of brain tissue, obtained post-mortem by supraorbital sampling, for the purpose of quantifying cerebral sequestration in children with fatal malaria in Blantyre, Malawi. We have compared this method to histological quantification of parasites at autopsy. RESULTS: The number of parasites present on cytological smears correlated with the proportion of vessels parasitized as assessed by histology of fixed and stained brain tissue. Use of cytological results in addition to the standard clinical case definition increases the specificity of the clinical case definition alone from 48.3% to 100% with a minimal change in sensitivity. CONCLUSIONS: Post-mortem supraorbital sampling of brain tissue improves the specificity of the diagnosis of fatal cerebral malaria and provides accurate quantitative estimates of cerebral sequestration. This tool can be of great value in clinical, pathogenetic, and epidemiological research studies on cerebral malaria.

Bone turnover markers to monitor oral bisphosphonate therapy.

Bisphosphonates are widely used as first-line therapy to slow bone loss and decrease fracture risk in postmenopausal women with osteoporosis. Nonadherence to oral bisphosphonates diminishes the benefit of reduced bone loss and fracture risk of these medications. Strategies to enhance osteoporosis monitoring and adherence to therapy are crucial to improve outcomes. Dual-energy x-ray absorptiometry (DXA) is the gold standard for monitoring bone mineral density but is slow to detect change after initiation of oral bisphosphonate therapy. Bone turnover markers (BTMs) are by-products released during bone remodeling and are measurable in blood and urine. We review how the rapid change in BTMs can be a useful short-term tool to monitor the effectiveness of oral bisphosphonate therapy, which may ultimately improve adherence to therapy and outcomes.

Assessment of gait and posture characteristics using a smartphone wearable system for persons with osteoporosis with and without falls.

We used smartphone technology to differentiate the gait characteristics of older adults with osteoporosis with falls from those without falls. We assessed gait mannerism and obtained activities of daily living (ADLs) with wearable sensor systems (smartphones and inertial measurement units [IMUs]) to identify fall-risk characteristics. We recruited 49 persons with osteoporosis: 14 who had a fall within a year before recruitment and 35 without falls. IMU sensor signals were sampled at 50 Hz using a customized smartphone app (Lockhart Monitor) attached at the pelvic region. Longitudinal data was collected using MoveMonitor+ (DynaPort) IMU over three consecutive days. Given the close association between serum calcium, albumin, PTH, Vitamin D, and musculoskeletal health, we compared these markers in individuals with history of falls as compared to nonfallers. For the biochemical parameters fall group had significantly lower calcium (P = 0.01*) and albumin (P = 0.05*) and higher parathyroid hormone levels (P = 0.002**) than nonfall group. In addition, persons with falls had higher sway area (P = 0.031*), lower dynamic stability (P < 0.001***), gait velocity (P = 0.012*), and were less able to perform ADLs (P = 0.002**). Thus, persons with osteoporosis with a history of falls can be differentiated by using dynamic real-time measurements that can be easily captured by a smartphone app, thus avoiding traditional postural sway and gait measures that require individuals to be tested in a laboratory setting.

Cigarette smoke extract suppresses the RIG-I-initiated innate immune response to influenza virus in the human lung.

Cigarette smoking is the major cause of chronic obstructive pulmonary disease (COPD) and predisposes subjects to severe respiratory tract infections. Epidemiological studies have shown that cigarette smokers are seven times more likely to contract influenza infection than nonsmokers. The mechanisms underlying this increased susceptibility are poorly characterized. Retinoic acid-inducible gene (RIG)-I is believed to play an important role in the recognition of, and response to, influenza virus and other RNA viruses. Our study focused on how cigarette smoke extract (CSE) alters the influenza-induced proinflammatory response and suppresses host antiviral activity in the human lung using a unique lung organ culture model. We first determined that treatment with 2-20% CSE did not induce cytotoxicity as assessed by LDH release. However, CSE treatment inhibited influenza-induced IFN-inducible protein 10 protein and mRNA expression. Induction of the major antiviral cytokine IFN-ß mRNA was also decreased by CSE. CSE also blunted viral-mediated RIG-I mRNA and protein expression. Inhibition of viral-mediated RIG-I induction by CSE was prevented by the antioxidants N-acetyl-cysteine and glutathione. These findings show that CSE suppresses antiviral and innate immune responses in influenza virus-infected human lungs through oxidative inhibition of viral-mediated induction of the pattern recognition receptor RIG-I. This immunosuppressive effect of CSE may play a role in the enhanced susceptibility of smokers to serious influenza infection in the lung.

Resistance of human alveolar macrophages to Bacillus anthracis lethal toxin.

The etiologic agent of inhalational anthrax, Bacillus anthracis, produces virulence toxins that are important in the disease pathogenesis. Current studies suggest that mouse and human macrophages are susceptible to immunosuppressive effects of one of the virulence toxins, lethal toxin (LT). Thus a paradigm has emerged that holds that the alveolar macrophage (AM) does not play a significant role in the innate immune response to B. anthracis or defend against the pathogen as it is disabled by LT. This is inconsistent with animal models and autopsy studies that show minimal disease at the alveolar surface. We examined whether AM are immunosuppressed by LT. We found that human AM were relatively resistant to LT-mediated innate immune cytokine suppression, MEK cleavage, and induction of apoptosis as compared with mouse RAW 264.7 macrophages. Mouse AM and murine bone marrow-derived macrophages were also relatively resistant to LT-mediated apoptosis despite intermediate sensitivity to MEK cleavage. The binding component of LT, protective Ag, does not attach to human AM, although it did bind to mouse AM, murine bone marrow-derived macrophages, and RAW 264.7 macrophages. Human AM do not produce significant amounts of the protective Ag receptor anthrax toxin receptor 1 (TEM8/ANTXR1) and anthrax toxin receptor 2 (CMG2/ANTXR2). Thus, mature and differentiated AM are relatively resistant to the effects of LT as compared with mouse RAW 264.7 macrophages. AM resistance to LT may enhance clearance of the pathogen from the alveolar surface and explain why this surface is relatively free of B. anthracis in animal models and autopsy studies.

Human lung innate immune cytokine response to adenovirus type 7.

Adenovirus (Ad) type 7 can cause severe infection, including pneumonia, in military recruits and children. The initial inflammation is a neutrophilic interstitial infiltration with neutrophilic alveolitis. Subsequently, monocytes become evident and, finally, there is a predominantly lymphocytic infiltrate. We have established that Ad7 infection of epithelial cells stimulates release of the neutrophil chemotaxin interleukin (IL)-8, and have extended these studies to a human lung tissue model. Here, we studied cytokine responses to Ad7 in human alveolar macrophages (HAM) and our human lung tissue model. Both ELISA and RNase-protection assay (RPA) data demonstrated that, upon Ad7 infection, IP-10 and MIP-1alpha/beta are released from HAM. IP-10 and MIP-1alpha/beta protein levels were induced 2- and 3-fold, respectively, in HAM 24 h after Ad7 infection. We then investigated induction of specific cytokines in human lung tissue by RPA and ELISA. The results showed that IL-8 and IL-6 were induced 8 h after infection and, by 24 h, levels of IL-8, IL-6, MIP-1alpha/beta and MCP-1 were all increased. IP-10, a monocyte and lymphocyte chemokine, was also induced 30-fold, but only 24 h after infection. Immunohistochemistry staining confirmed that IL-8 was only released from the epithelial cells of lung slices and not from macrophages. IP-10 was secreted from both macrophages and epithelial cells. Moreover, full induction of IP-10 is likely to require participation and cooperation of both epithelial cells and macrophages in intact lung. Understanding the cytokine and chemokine induction during Ad7 infection may lead to novel ways to modulate the response to this pathogen.

Protective essential oil attenuates influenza virus infection: an in vitro study in MDCK cells.

BACKGROUND: Influenza is a significant cause of morbidity and mortality. The recent pandemic of a novel H1N1 influenza virus has stressed the importance of the search for effective treatments for this disease. Essential oils from aromatic plants have been used for a wide variety of applications, such as personal hygiene, therapeutic massage and even medical practice. In this paper, we investigate the potential role of an essential oil in antiviral activity. METHODS: We studied a commercial essential oil blend, On Guard™, and evaluated its ability in modulating influenza virus, A/PR8/34 (PR8), infection in Madin-Darby canine kidney (MDCK) cells. Influenza virus was first incubated with the essential oil and infectivity in MDCK cells was quantified by fluorescent focus assay (FFA). In order to determine the mechanism of effects of essential oil in viral infection inhibition, we measured hemagglutination (HA) activity, binding and internalization of untreated and oil-treated virus in MDCK cells by flow cytometry and immunofluorescence microscopy. In addition, the effect of oil treatment on viral transcription and translation were assayed by relative end-point RT-PCR and western blot analysis. RESULTS: Influenza virus infectivity was suppressed by essential oil treatment in a dose-dependent manner; the number of nascent viral particles released from MDCK cells was reduced by 90% and by 40% when virus was treated with 1:4,000 and 1:6,000 dilutions of the oil, respectively. Oil treatment of the virus also decreased direct infection of the cells as the number of infected MDCK cells decreased by 90% and 45% when virus was treated with 1:2,000 and 1:3,000 dilutions of the oil, respectively. This was not due to a decrease in HA activity, as HA was preserved despite oil treatment. In addition, oil treatment did not affect virus binding or internalization in MDCK cells. These effects did not appear to be due to cytotoxicity of the oil as MDCK cell viability was only seen with concentrations of oil that were 2 to 6 times greater than the doses that inhibited viral infectivity. RT-PCR and western blotting demonstrated that oil treatment of the virus inhibited viral NP and NS1 protein, but not mRNA expression. CONCLUSIONS: An essential oil blend significantly attenuates influenza virus PR8 infectivity in vitro without affecting viral binding or cellular internalization in MDCK cells. Oil treated virus continued to express viral mRNAs but had minimal expression of viral proteins, suggesting that the antiviral effect may be due to inhibition of viral protein translation.

The application of empirical mode decomposition for the enhancement of cardiotocograph signals.

Cardiotocograph (CTG) is widely used in everyday clinical practice for fetal surveillance, where it is used to record fetal heart rate (FHR) and uterine activity (UA). These two biosignals can be used for antepartum and intrapartum fetal monitoring and are, in fact, nonlinear and non-stationary. CTG recordings are often corrupted by artifacts such as missing beats in FHR, high-frequency noise in FHR and UA signals. In this paper, an empirical mode decomposition (EMD) method is applied on CTG signals. A recursive algorithm is first utilized to eliminate missing beats. High-frequency noise is reduced using EMD followed by the partial reconstruction (PAR) method, where the noise order is identified by a statistical method. The obtained signal enhancement from the proposed method is validated by comparing the resulting traces with the output obtained by applying classical signal processing methods such as Butterworth low-pass filtering, linear interpolation and a moving average filter on 12 CTG signals. Three obstetricians evaluated all 12 sets of traces and rated the proposed method, on average, 3.8 out of 5 on a scale of 1(lowest) to 5 (highest).

Teriparatide treatment in adult hypophosphatasia in a patient exposed to bisphosphonate: a case report.

We describe the case of a woman with hypophosphatasia previously exposed to bisphosphonate and subsequently treated with teriparatide (recombinant human PTH 1-34).A Caucasian woman sustained bilateral femur stress fractures when she was fifty years old, which widened despite use of calcium, vitamin D and risedronate for 2.5 years and required intramedullary rods for stabilization. Hypophosphatasia was diagnosed in the interim due to low serum alkaline phosphatase (ALP) (ALP 20 IU/L; normal (N), 40-150 IU/L) and high pyridoxal 5' phosphate (3400 nmol/L; N 18-175 nmol/L). She was referred for further management. On presentation, she had significant fracture site pain and generalized bone pain (weight bearing and non-weight bearing) - making her walker dependent at home and wheel chair dependent outside home.She could not sleep at night due to discomfort when she moved. Daily teriparatide injections, 20 mcg subcutaneously were prescribed.At 8-weeks follow-up, fracture site pain, weight-bearing and non weight-bearing pain improved significantly allowing ambulation for prolonged periods without assistance. She slept at night without discomfort. Improvement persisted during her entire treatment period. Radiographs taken at 4 and 16 months of treatment demonstrated healing of femur fractures.Biochemically, mean urine cross-link-N-telopeptide increased 11% as compared to her base-line, while bone specific alkaline phosphatase did not increase as expected.In conclusion, we observed an uncoupling of bone formation and resorption markers during her treatment period in the face of notable clinical and radiological improvement. Off-label use of teriparatide may help patients with hypophosphatasia.

Molecular Imaging in Diagnosis of Tumor-induced Osteomalacia.

Tumor induced osteomalacia (TIO) is a rare paraneoplastic syndrome caused by overproduction of fibroblast growth factor 23 (FGF23) secreted by benign mesenchymal neoplasm. Due to its nonspecific clinical presentation or lack of awareness, the diagnosis of TIO is often significantly delayed resulting in patients' prolonged physical suffering or psychological distress. Successful detection or complete surgical resection of the causative tumor typically leads to rapid resolution of symptoms or reversal of biochemical imbalance. Nuclear medicine and molecular imaging have been playing a promising role as the first-line imaging modalities in the diagnosis and localization of occult FGF23 secreting mesenchymal tumor, especially with the emerging whole-body, head-to-toe Ga68-DOTATATE PET/CT technique. Combined focused diagnostic CT and/or MRI are imperative for accurate delineation of tumor and surgical guidance.

Positive HLA-B27 and sacroiliitis is not always spondyloarthritis.

A 36-year-old man was treated for several years with multiple agents for ankylosing spondylitis based on positive human leukocyte antigen-B27 and sacroiliitis. He was also diagnosed with osteoporosis and hypophosphatemia. Over these years, from being an avid runner, he became dependent on a walker for ambulation. The lack of treatment response and the low phosphorus were clues that eventually led to a diagnosis of tumor-induced osteomalacia. This case discusses the importance of not solely relying on genetic markers and sacroiliitis for diagnosing ankylosing spondylitis as other conditions can cause similar presentations.

Dynamics of perceptual oscillations in form vision.

Certain periodic dot patterns (Marroquin patterns) generate a percept of dynamically oscillating circles, and analogous effects were explored by op artists in the 1960s. Here we show psychophysically that circles are perceived in these patterns only around specific points that are quantitatively predicted by a neural model of configural units hypothesized to reside in cortical area V4. Circles superimposed on the pattern mask perception of illusory circles. A neural model of lateral inhibitory interactions among V4 configural units showing spike-frequency adaptation quantitatively accounts for the human data. The model is consistent with ideas on the neural basis of attention in V4, and it suggests that attention may be biased via neuromodulation of slow hyperpolarizing potentials in cortical neurons.

Pramlintide, the synthetic analogue of amylin: physiology, pathophysiology, and effects on glycemic control, body weight, and selected biomarkers of vascular risk.

Pramlintide is a synthetic version of the naturally occurring pancreatic peptide called amylin. Amylin and pramlintide have similar effects on lowering postprandial glucose, lowering postprandial glucagon and delaying gastric emptying. Pramlintide use in type 1 and insulin requiring type 2 diabetes mellitus (DM) is associated with modest reductions in HbAlc often accompanied by weight loss. Limited data show a neutral effect on blood pressure. Small studies suggest small reductions in LDL-cholesterol in type 2 DM and modest reductions in triglycerides in type 1 DM. Markers of oxidation are also reduced in conjunction with reductions in postprandial glucose. Nausea is the most common side effect. These data indicate that pramlintide has a role in glycemic control of both type 1 and type 2 DM. Pramlintide use is associated with favorable effects on weight, lipids and other biomarkers for atherosclerotic disease.

Distributed Learning: Revitalizing Anesthesiology Training in Resource-Limited Ethiopia.

BACKGROUND: Ethiopia has a significant paucity of available health-care workers. Despite the increasing number of medical schools, there are not enough physician instructors. Furthermore, availability and standardization of postgraduate training are lacking. Modalities of e-learning have been shown to be successful when used to impart medical education in other resource-limited countries. The Emory University and Addis Ababa University (AAU) Departments of Anesthesiology have formed a collaboration with the intent of improving the AAU Anesthesiology residency program, one of two postgraduate training programs for anesthesiology in Ethiopia. METHODS: An initial educational needs assessment identified areas in the existing training program that required improvement. In this pilot study, we describe how the current classroom-based curriculum is augmented by the introduction of interactive educational sessions and distributed learning in the form of video lectures. Video lectures covered topics based on areas identified by Ethiopian residents and faculty. Interactive sessions included hands-on ultrasound workshops and epidural placement practicums, a journal club, problem-based learning sessions, and a mock code simulation. Assessment of the additions of the newly introduced blended learning technique was conducted via pre- and posttests on the topics presented. RESULTS: Pre- to posttest score averages increased from 54.5% to 83.6%. CONCLUSION: An expansion of educational resources and modes of didactics are needed to fill the gaps that exist in Ethiopian anesthesiology training. Incorporating distributed learning into the existing didactic structure may lead to more efficacious instruction resulting in a higher retention rate of information.