In situ brain tumor detection using a Raman spectroscopy system-results of a multicenter study.

Safe and effective brain tumor surgery aims to remove tumor tissue, not non-tumoral brain. This is a challenge since tumor cells are often not visually distinguishable from peritumoral brain during surgery. To address this, we conducted a multicenter study testing whether the Sentry System could distinguish the three most common types of brain tumors from brain tissue in a label-free manner. The Sentry System is a new real time, in situ brain tumor detection device that merges Raman spectroscopy with machine learning tissue classifiers. Nine hundred and seventy-six in situ spectroscopy measurements and colocalized tissue specimens were acquired from 67 patients undergoing surgery for glioblastoma, brain metastases, or meningioma to assess tumor classification. The device achieved diagnostic accuracies of 91% for glioblastoma, 97% for brain metastases, and 96% for meningiomas. These data show that the Sentry System discriminated tumor containing tissue from non-tumoral brain in real time and prior to resection.

High-resolution fleezers reveal duplex opening and stepwise assembly by an oligomer of the DEAD-box helicase Ded1p.

DEAD-box RNA helicases are ubiquitous in all domains of life where they bind and remodel RNA and RNA-protein complexes. DEAD-box helicases unwind RNA duplexes by local opening of helical regions without directional movement through the duplexes and some of these enzymes, including Ded1p from Saccharomyces cerevisiae, oligomerize to effectively unwind RNA duplexes. Whether and how DEAD-box helicases coordinate oligomerization and unwinding is not known and it is unclear how many base pairs are actively opened. Using high-resolution optical tweezers and fluorescence, we reveal a highly dynamic and stochastic process of multiple Ded1p protomers assembling on and unwinding an RNA duplex. One Ded1p protomer binds to a duplex-adjacent ssRNA tail and promotes binding and subsequent unwinding of the duplex by additional Ded1p protomers in 4-6 bp steps. The data also reveal rapid duplex unwinding and rezipping linked with binding and dissociation of individual protomers and coordinated with the ATP hydrolysis cycle.

Impaired gastric accommodation in patients with postprandial distress syndrome type of functional dyspepsia assessed by 2D ultrasonography.

BACKGROUND AND PURPOSE: The pathophysiology of postprandial distress syndrome includes impaired gastric accommodation, hypersensitivity to gastric distension and delayed gastric emptying. 2D-ultrasonography is one of the methods to assess gastric accommodation by measuring proximal gastric area and we evaluated its role in calculating proximal gastric area and thus assessing gastric accommodation in Indian patients with postprandial distress syndrome. METHODS: In a hospital-based comparative analysis, proximal gastric area was measured with 2D-ultrasonography of postprandial distress syndrome (PDS) patients and compared with healthy controls. Five readings were measured every five minutes till 25 minutes after 400 mL of vegetable soup. The Patient Assessment of Upper Gastrointestinal Disorders-Quality of Life (PAGI-QOL score) and diets aggravating PDS symptoms were studied through detailed questionnaires. Sample size was calculated at 80% study power and alpha error of 0.05 to be 30 subjects in each group. RESULTS: The mean age of patients (18 males) vs. 30 healthy controls (25 males) was 40.8 ± 11.50 years vs. 36.37 ± 7.58, respectively, (p = 0.084). Proximal gastric area was significantly lower in patients versus healthy controls at five minutes (22.54 ± 2.77 vs. 30.66 ± 2.55 cm2), 10 minutes (23.03 ± 2.45 vs. 31.10 ± 2.06 cm2), 15 minutes (23.06 ± 2.27 vs. 30.31 ± 2.11 cm2), 20 minutes (22.21 ± 2.31 vs. 29.73 ± 1.71 cm2) and 25 minutes (22.02 ± 2.33 vs. 28.39 ± 1.55 cm2); p < 0.001 at all intervals of time, indicating impaired gastric accommodation. The QOL was poor in all patients with PDS with mean PAGI-QOL score of 31.30 ± 15.05, median of 30, minimum score of 12 and maximum score of 66. CONCLUSIONS: Measurement of proximal gastric area with 2D-ultrasonography is simple and non-invasive. Proximal gastric area in patients was lower than controls, indicating impaired gastric accommodation. Poor quality of life was universal in patients with postprandial distress syndrome.

Subsecond lung cancer detection within a heterogeneous background of normal and benign tissue using single-point Raman spectroscopy.

Significance: Lung cancer is the most frequently diagnosed cancer overall and the deadliest cancer in North America. Early diagnosis through current bronchoscopy techniques is limited by poor diagnostic yield and low specificity, especially for lesions located in peripheral pulmonary locations. Even with the emergence of robotic-assisted platforms, bronchoscopy diagnostic yields remain below 80%. Aim: The aim of this study was to determine whether in situ single-point fingerprint (800 to 1700 cm-1) Raman spectroscopy coupled with machine learning could detect lung cancer within an otherwise heterogenous background composed of normal tissue and tissue associated with benign conditions, including emphysema and bronchiolitis. Approach: A Raman spectroscopy probe was used to measure the spectral fingerprint of normal, benign, and cancer lung tissue in 10 patients. Each interrogated specimen was characterized by histology to determine cancer type, i.e., small cell carcinoma or non-small cell carcinoma (adenocarcinoma and squamous cell carcinoma). Biomolecular information was extracted from the fingerprint spectra to identify biomolecular features that can be used for cancer detection. Results: Supervised machine learning models were trained using leave-one-patient-out cross-validation, showing lung cancer could be detected with a sensitivity of 94% and a specificity of 80%. Conclusions: This proof of concept demonstrates fingerprint Raman spectroscopy is a promising tool for the detection of lung cancer during diagnostic procedures and can capture biomolecular changes associated with the presence of cancer among a complex heterogeneous background within less than 1 s.

Cost Analysis of Intranatal Care Services at a Tertiary Care Public Sector Hospital in Rajasthan, India.

Introduction India is responsible for the second-highest maternal deaths and the greatest burden of stillbirths worldwide. The cost of intranatal services is an important determining factor, especially in developing countries like India. Most studies report the cost of delivery from the patient's perspective, but there is a lack of studies from the health system's perspective. This present study aimed to bridge this gap by estimating the overall and unit costs of various types of deliveries at a tertiary-level hospital in Rajasthan, India. Methods The cost estimation of intranatal services was conducted in a tertiary-level teaching hospital in Jaipur, Rajasthan. This cost analysis undertook the health system's perspective, using bottom-up costing methodology. Data on all the resources (capital/recurrent) used for the delivery of intranatal care from April 2020 to March 2021 were collected. Sensitivity analysis was done to account for any variability in cost components on overall intranatal service cost. Results The annual cost of intranatal care services at the tertiary care hospital was INR 149,011,957 (USD 1,988,152). The unit cost per vaginal delivery was INR 8,244.4 (USD 109.9) and the unit cost per cesarean section was INR 10,696.2 (USD 142.7). Among various heads of expenditure, 'human resource' costs were predominant, accounting for 47.7% of the total costs, followed by 'building/space' and 'overhead' costs, accounting for 30.59% and 11.1%, respectively. Conclusion The results may help plan and manage intra-natal care services in Rajasthan. Apart from the judicious utilization of resources, the findings of the study may also serve as a basis for future health economic studies.

High-Resolution Optical Tweezers Combined with Multicolor Single-Molecule Microscopy.

We present an instrument that combines high-resolution optical tweezers and multicolor confocal fluorescence spectroscopy. Biological macromolecules exhibit complex conformation and stoichiometry changes in coordination with their motion and activity. To further our understanding of the complex machinery of life, we need methods that can simultaneously probe more than one degree of freedom of single molecules and complexes. Fluorescence optical tweezers, or "fleezers," combine the capabilities of optical tweezers and single-molecule fluorescence microscopy into a single instrument. Here we present the latest generation of a high-resolution fleezers instrument integrated with multicolor fluorescence spectroscopy. The tweezers portion of the instrument can manipulate biological macromolecules with pN scale forces while measuring subnanometer distances. Simultaneous with tweezers measurements, the multicolor fluorescence capability allows the direct observation of multiple molecules or multiple degrees of freedom which allows, for example, the observation of multiple proteins simultaneously within a complex. The instrument incorporates three fluorescence excitation lasers, all sourced from a single-mode optical fiber allowing a reliable alignment scheme, that allows, for example, three independent fluorescent probes or fluorescence resonance energy transfer (FRET) measurements and also increases flexibility in the choice of fluorescent probes. To avoid photobleaching and improve tweezers stability, the instrument implements a timesharing (using a single trap laser to produce a pair of traps via rapid switching between two locations) and interlacing (turning the trapping beam off when the fluorescence excitation beams are on and vice versa) scheme using acousto-optic modulators (AOM) to rapidly and precisely modulate lasers. Our latest "random phase" trap AOM control method obliterates previous residual trap positioning and bead position measurement errors. Here we present the general design principles and detailed construction and testing protocols for the instrument.

Translation and reliability of the caregiver quality of life index-cancer (CQOLC) Scale in Hindi and it's association with the Kessler psychological distress scale (K10) for cancer caregivers at a tertiary radiotherapy centre in India.

BACKGROUND: With increasing incidence of cancer worldwide and increasing burden of treatment over the patients with cancer, the caregivers of those patients with cancer also suffer from psychological distress which ultimately affects their Quality of Life (QOL). This aspect is often overlooked which may ultimately lead to compromised patient care. PURPOSE: To translate the Caregiver Quality of Life Index-Cancer (CQOLC) Scale in Hindi and assess its association with Kesslers's psychological distress scale (K-10) for determining the quality of life (QoL) of caregivers of patients with cancer in India. METHODS: This is a single institute, hospital based cross sectional study performed from July 2020 to March 2021. Caregiver Quality of Life Index-Cancer (CQOLC) Scale, employing standard 'forward-backforward' translation procedure, the English-language version of the questionnaire was translated into Hindi-language by experts and administered to each caregiver. Cronbach's alpha was used for internal consistency. Kesslers's psychological distress scale (K-10) was analysed on interview basis. RESULTS: A total evaluated responders were 264. The hindi versions of the CQOLC was validated by cronbach's alpha method with internal consistency between 0.8 and 0.91. The majority of the caregivers were of the age group 31-40 yrs (42.05%). The male to female ratio was 6.54:1. CQOLC good score(0-45) was obtained in 27.27%, fair(46-90) in 55.30% and poor(91-140) in 17.42%. For K-10 distress scale very mild(10-19), mild(20-24), moderate(25-29) and severe(30-50) was observed in 47.73%, 11.74%, 11.74% and 28.79% respectively. A strong relation was found between the caregivers with poor CQOLC score and severe K-10 score(p<0.001). CONCLUSION: The Hindi version of CQOLC was accepted and its correlation with the point psychological distress predictor (K-10) scale was a valuable method to identify caregivers with poor QOL and severe distress levels.

An anionic ligand snap-locks a long-range interaction in a magnesium-folded riboswitch.

The archetypical transcriptional crcB fluoride riboswitch from Bacillus cereus is an intricately structured non-coding RNA element enhancing gene expression in response to toxic levels of fluoride. Here, we used single molecule FRET to uncover three dynamically interconverting conformations appearing along the transcription process: two distinct undocked states and one pseudoknotted docked state. We find that the fluoride anion specifically snap-locks the magnesium-induced, dynamically docked state. The long-range, nesting, single base pair A40-U48 acts as the main linchpin, rather than the multiple base pairs comprising the pseudoknot. We observe that the proximally paused RNA polymerase further fine-tunes the free energy to promote riboswitch docking. Finally, we show that fluoride binding at short transcript lengths is an early step toward partitioning folding into the docked conformation. These results reveal how the anionic fluoride ion cooperates with the magnesium-associated RNA to govern regulation of downstream genes needed for fluoride detoxification of the cell.

Salter-Harris I Injury of the Proximal Tibial Physis with Vascular Compromise: A Case Report and Review of Literature.

INTRODUCTION: Physeal injuries of proximal tibia are uncommon and constitute only 1% of all physeal injuries. Major vessel injuries are also rare in children and occur in 0.6% trauma cases, out of which, injuries of popliteal artery comprise only 5%. The combination of these is an extremely rare injury pattern but is potentially limb threatening. Most of the cases of physeal injuries of proximal tibia along with popliteal artery compromise, reported in the literature include Salter Harris (SH) type II, III, and IV patterns (type II being most common) with pure type I being extremely rare. CASE REPORT: We describe a rare posteriorly displaced pure SH I injury of proximal tibial physis with compromised vascularity of the limb. Immediate closed reduction and stabilization with Kirschner wires (K-wires) restored the vascularity of the limb and the patient went on to have a good functional recovery at the last follow-up of 1 year. CONCLUSION: Vascular injuries occurring in SH 1 proximal tibial physeal fractures are often overlooked or missed due to the relatively benign appearance of slip with no sharp fragments to injure major vessels. Close monitoring of neurovascular status and a low threshold for fixation, for example, with K-wires, are important to prevent limb threatening ischemia in these relatively simple appearing injuries.

Dynamic competition between a ligand and transcription factor NusA governs riboswitch-mediated transcription regulation.

Cotranscriptional RNA folding is widely assumed to influence the timely control of gene expression, but our understanding remains limited. In bacteria, the fluoride (F-)-sensing riboswitch is a transcriptional control element essential to defend against toxic F- levels. Using this model riboswitch, we find that its ligand F- and essential bacterial transcription factor NusA compete to bind the cotranscriptionally folding RNA, opposing each other's modulation of downstream pausing and termination by RNA polymerase. Single-molecule fluorescence assays probing active transcription elongation complexes discover that NusA unexpectedly binds highly reversibly, frequently interrogating the complex for emerging, cotranscriptionally folding RNA duplexes. NusA thus fine-tunes the transcription rate in dependence of the ligand-responsive higher-order structure of the riboswitch. At the high NusA concentrations found intracellularly, this dynamic modulation is expected to lead to adaptive bacterial transcription regulation with fast response times.

Vertical Strut Ossiculoplasty: A Versatile Alternate to Conventional Techniques-A Randomized study.

INTRODUCTION: Various ossicular reconstruction materials and techniques have been described in literature using autologous ossicle, cortical bone, autologous cartilage, synthetic materials and implants like total/partial ossicular replacement prosthesis (TORP/PORP) etc., but it has always been a topic of controversy in terms of the efficacy, longevity and complications of the material or method used. MATERIAL AND METHODS: This is a prospective, interventional, comparative, double-blind randomized control study which was done at a tertiary care center to compare outcomes of conventional and carved conchal cartilage (vertical strut) type III Tympanoplasty in terms of graft uptake and hearing gain. A total number of 52 cases were enrolled, randomized and allocated to 2 groups (26 each) i.e. group A (conventional type III) and group B (vertical strut technique). RESULTS: Graft uptake was seen in 25 (96.16%) patients in group B while it was observed in 23 (88.5%) cases in group A. Hearing gains were also better in group B. CONCLUSION: This study suggests that Vertical Strut technique can be studied further as it gives better gains in Air Conduction threshold and A-B Gap along with graft uptake as it provides better middle ear space and ossicular / tympanic membrane interface resulting in better hearing.

Is It Wise to Wait for Ear to Become Dry in Indian Scenario?

Tympanoplasty is the standard and well-established procedure for closure of tympanic membrane perforations. Tympanoplasty in wet ear is still a topic of debate among ENT surgeons. This study discusses the balance between wait and watch policy and early intervention in wet ear. It also compares the rate of graft uptake and hearing improvement in Type I tympanoplasty in dry and wet ears. This is a hospital based, observational, descriptive and comparative study. Total 246 patients enrolled in the study. Two groups were created with 123 patients in each group. One included dry ears and another included wet ears. All patients had mucosal type of chronic otitis media. They all underwent Type I tympanoplasty. Graft uptake rate and hearing was compared between both groups. The overall success rate (graft uptake) was 91.06% (224). The success rate in dry ear group was 93.50% (115) and in wet ear group it was 88.62% (109). This study concluded that there is no added advantage of drying the ear rather the delay in treatment increases morbidity and drop outs in Indian scenario.

Is thyroid excision mandatory with laryngectomy in carcinoma larynx?

BACKGROUND: Advanced stage operable cancers of larynx are treated with total laryngectomy including thyroid resection in most of the cases, which may expose patient to hypothyroidism and hypoparathyroidism. The requirement of thyroidectomy during Total Laryngectomy is controversial. METHODS: A cross sectional observational study was set out to review preoperative clinical and radiological assessment; intraoperative and histopathological findings; and follow-up data to predict thyroid gland invasion in the setting of squamous cell carcinoma of the Larynx. RESULTS: 11 (16%) out of 69 patients had thyroid gland involvement on histopathological examination with mean age 63 years. Out of these 11 cases, 8 (72%) underwent primary total laryngectomy. 90% patients with thyroid gland involvement were male. 9 cases with thyroid gland involvement were staged as T4a preoperatively. CONCLUSION: Invasion of thyroid gland by laryngeal cancer is uncommon. Unnecessary hemithyroidectomies lead to hypothyroidism and hypoparathyroidism. The study points out the clear indications of thyroid excision in patients undergoing total laryngectomy. We can suggest that total thyroidectomy should be done with total laryngectomy in cases which have gross clinical, radiological or intraoperative thyroid gland involvement, subglottic extension and thyroid cartilage invasion. This can save the patients from the brunt of unnecessary morbid hypothyroidism and hypoparathyroidism.

Feature engineering applied to intraoperative in vivo Raman spectroscopy sheds light on molecular processes in brain cancer: a retrospective study of 65 patients.

Raman spectroscopy is a promising tool for neurosurgical guidance and cancer research. Quantitative analysis of the Raman signal from living tissues is, however, limited. Their molecular composition is convoluted and influenced by clinical factors, and access to data is limited. To ensure acceptance of this technology by clinicians and cancer scientists, we need to adapt the analytical methods to more closely model the Raman-generating process. Our objective is to use feature engineering to develop a new representation for spectral data specifically tailored for brain diagnosis that improves interpretability of the Raman signal while retaining enough information to accurately predict tissue content. The method consists of band fitting of Raman bands which consistently appear in the brain Raman literature, and the generation of new features representing the pairwise interaction between bands and the interaction between bands and patient age. Our technique was applied to a dataset of 547 in situ Raman spectra from 65 patients undergoing glioma resection. It showed superior predictive capacities to a principal component analysis dimensionality reduction. After analysis through a Bayesian framework, we were able to identify the oncogenic processes that characterize glioma: increased nucleic acid content, overexpression of type IV collagen and shift in the primary metabolic engine. Our results demonstrate how this mathematical transformation of the Raman signal allows the first biological, statistically robust analysis of in vivo Raman spectra from brain tissue.

Seasonal modulation of deep slow-slip and earthquakes on the Main Himalayan Thrust.

The interaction between seasonally-induced non-tectonic and tectonic deformation along the Himalayan plate boundary remains debated. Here, we propose that tectonic deformation along this plate boundary can be significantly influenced by the deformation induced by the non-tectonic hydrological loading cycles. We explore seasonal mass oscillations by continental water storage in Southeast Asia and Himalayan arc region using continuous Global Positioning System measurements and satellite data from the Gravity Recovery and Climate Experiment. We suggest that the substantially higher transient displacements above the base of the seismogenic zone indicate a role of changes in aseismic slip rate on the deep megathrust that may be controlled by seasonal hydrological loading. We invoke modulation of aseismic slip on the megathrust down-dip of the seismogenic zone due to a fault resonance process induced by the seasonal stress changes. This process modulates mid-crustal ramp associated micro-seismicity and influences the timing of Central Himalayan earthquakes.

Revealing dynamically-organized receptor ion channel clusters in live cells by a correlated electric recording and super-resolution single-molecule imaging approach.

The N-methyl-d-aspartate (NMDA) receptor ion-channel is activated by the binding of ligands, along with the application of action potential, important for synaptic transmission and memory functions. Despite substantial knowledge of the structure and function, the gating mechanism of the NMDA receptor ion channel for electric on-off signals is still a topic of debate. We investigate the NMDA receptor partition distribution and the associated channel's open-close electric signal trajectories using a combined approach of correlating single-molecule fluorescence photo-bleaching, single-molecule super-resolution imaging, and single-channel electric patch-clamp recording. Identifying the compositions of NMDA receptors, their spatial organization and distributions over live cell membranes, we observe that NMDA receptors are organized inhomogeneously: nearly half of the receptor proteins are individually dispersed; whereas others exist in heterogeneous clusters of around 50 nm in size as well as co-localized within the diffraction limited imaging area. We demonstrate that inhomogeneous interactions and partitions of the NMDA receptors can be a cause of the heterogeneous gating mechanism of NMDA receptors in living cells. Furthermore, comparing the imaging results with the ion-channel electric current recording, we propose that the clustered NMDA receptors may be responsible for the variation in the current amplitude observed in the on-off two-state ion-channel electric signal trajectories. Our findings shed new light on the fundamental structure-function mechanism of NMDA receptors and present a conceptual advancement of the ion-channel mechanism in living cells.

Direct Observation of Intermediate State(s) in the Mechanistic Investigation of Domain Specific Protein-Surfactant Interaction.

INTRODUCTION: Interaction of surfactants with proteins can decipher important information regarding the stability and behavior of proteins. For multi-domain proteins, these interactions vary domain wise and these details are crucial in understanding the contribution of different domains of the protein in its overall activity. OBJECTIVE: The objective of the present work is to study the interaction of surfactants with domain III of Human Serum Albumin (HSA) and to compare the same with the global interaction. METHODS: Interaction of the anionic Sodium Dodecyl Sulphate (SDS) and the Cationic Cetyltrimethylammonium Bromide (CTAB) surfactants with domain III of Human Serum Albumin (HSA) has been studied using 8-Anilino-1-Naphthalene-Sulphonate (ANS) as a fluorescent marker. Circular Dichroism (CD) spectroscopy has been used to study the protein-surfactant interaction for the overall protein. RESULTS: SDS is found to interact sequentially with domain III of HSA having two detectable intermediate states in the binding process. In case of CTAB, we have observed only one intermediate state for its interaction with domain III. Although Quantum yield measurement can reflect the presence of such intermediate state, the overall conformational change of the HSA on addition of surfactants, studied by Circular Dichroism (CD) spectroscopy, and the ANS-Trp distance measurement by FRET could not resolve the presence of such intermediate states. The esterase activity of HSA in presence of different amount of surfactants is also in accordance with our above observation. CONCLUSION: The interaction of both the surfactants with HSA is found to be sequential in nature. The most important conclusion revealed from our study is that the nature of protein-surfactant interaction is not same throughout the entire protein. Our study reveals that different parts of the multi-domain HSA have different affinity to the surfactant molecules.

Region-Specific Double Denaturation of Human Serum Albumin: Combined Effects of Temperature and GnHCl on Structural and Dynamical Responses.

In this work, we have investigated the effects of denaturing agents, guanidine hydrochloride (GnHCl) and temperature, on the overall structure, domain-I, and domain-III of human serum albumin (HSA) using circular dichroism (CD) spectroscopy and steady-state, time-resolved fluorescence spectroscopy. We have tagged Cys-34 of HSA, located at domain-I, using N-(7-dimethylamino-4-methylcoumarin-3-yl)iodoacetamide and Tyr-411 of HSA, located at domain-III, using p-nitrophenyl coumarin ester, for this purpose. The CD spectroscopy studies reveal the overall denaturation of the protein. The denaturation follows the expected direction in which the protein is denatured with an increase in the concentration of GnHCl or temperature. The α-helicity of the native state of HSA was found to be 64.2%, and the minimum value of α-helicity was found to be 14.8% in the presence of 6 M GnHCl at room temperature. Steady-state emission studies were carried out on domain-I and domain-III of the protein using site-specific fluorescent tags. The degree of folding of the two domains at different combinations of temperature and GnHCl concentration was calculated and was found to follow a slightly different course of denaturation. Solvation dynamics was found to be quite different for these two domains. The domain-I of HSA has a maximum solvation time of 0.39 ns, and the solvation time tends to decrease with the action of either temperature or GnHCl. On the other hand, the domain-III of HSA showed a much higher solvation time (1.42 ns) and does not show any regular change at higher temperatures or in the presence of GnHCl. This difference could be attributed to the different microenvironment inside the protein cores of the two domains.