Workforce problems at rural public health-centres in India: a WISN retrospective analysis and national-level modelling study.

BACKGROUND: Rural India has a severe shortage of human resources for health (HRH). The National Rural Health Mission (NRHM) deploys HRH in the rural public health system to tackle shortages. Sanctioning under NRHM does not account for workload resulting in inadequate and inequitable HRH allocation. The Workforce Indicators of Staffing Needs (WISN) approach can identify shortages and inform appropriate sanctioning norms. India currently lacks nationally relevant WISN estimates. We used existing data and modelling techniques to synthesize such estimates. METHODS: We conducted a retrospective analysis of existing survey data for 93 facilities from 5 states over 8 years to create WISN calculations for HRH cadres at primary and community health centres (PHCs and CHCs) in rural areas. We modelled nationally representative average WISN-based requirements for specialist doctors at CHCs, general doctors and nurses at PHCs and CHCs. For 2019, we calculated national and state-level overall and per-centre WISN differences and ratios to depict shortage and workload pressure. We checked correlations between WISN ratios for cadres at a given centre-type to assess joint workload pressure. We evaluated the gaps between WISN-based requirements and sanctioned posts to investigate suboptimal sanctioning through concordance analysis and difference comparisons. RESULTS: In 2019, at the national-level, WISN differences depicted workforce shortages for all considered HRH cadres. WISN ratios showed that nurses at PHCs and CHCs, and all specialist doctors at CHCs had very high workload pressure. States with more workload on PHC-doctors also had more workload on PHC-nurses depicting an augmenting or compounding effect on workload pressure across cadres. A similar result was seen for CHC-specialist pairs-physicians and surgeons, physicians and paediatricians, and paediatricians and obstetricians-gynaecologists. We found poor concordance between current sanctioning norms and WISN-based requirements with all cadres facing under-sanctioning. We also present across-state variations in workforce problems, workload pressure and sanctioning problems. CONCLUSION: We demonstrate the use of WISN calculations based on available data and modelling techniques for national-level estimation. Our findings suggest prioritising nurses and specialists in the rural public health system and updating the existing sanctioning norms based on workload assessments. Workload-based rural HRH deployment can ensure adequate availability and optimal distribution.

Quantifying Intracellular Particle Flows by DIC Object Tracking.

Label-free imaging techniques such as differential interference contrast (DIC) allow the observation of cells and large subcellular structures in their native, unperturbed states with minimal exposure to light. The development of robust computational image-analysis routines is vital to quantitative label-free imaging. The reliability of quantitative analysis of time-series microscopy data based on single-particle tracking relies on accurately detecting objects as distinct from the background, i.e., segmentation. Typical approaches to segmenting DIC images either involve converting images to those resembling phase contrast, mimicking the optics of DIC object formation, or using the morphological properties of objects. Here, we describe MATLAB based, single-particle tracking tool with a GUI for mobility analysis of objects from in vitro and in vivo DIC time-series microscopy. The tool integrates contrast enhancement with multiple modified Gaussian filters, automated threshold detection for segmentation and minimal distance-based two-dimensional single-particle tracking. We compare the relative performance of multiple filters and demonstrate the utility of the tool for DIC object tracking (DICOT). We quantify subcellular dynamics of a time series of Caenorhabditis elegans embryos in the one-celled stage by detecting birefringent yolk granules in the cytoplasm with high precision. The resulting two-dimensional map of oscillatory dynamics of granules quantifies the cytoplasmic flows driven by anaphasic spindle oscillations. The frequency of oscillations across the anterior-posterior (A-P) and transverse axes of the embryo correspond well with the reported frequency of spindle oscillations. We validate the quantitative accuracy of our method by tracking the in vitro diffusive mobility of micron-sized beads in glycerol solutions. Estimates of the diffusion coefficients of the granules are used to measure the viscosity of a dilution series of glycerol. Thus, our computational method is likely to be useful for both intracellular mobility and in vitro microrheology.

Modeling the tunability of the dual-feedback genetic oscillator.

Oscillatory gene circuits are ubiquitous to biology and are involved in fundamental processes of cell cycle, circadian rhythms, and developmental systems. The synthesis of small, non-natural oscillatory genetic circuits has been increasingly used to test the fundamental principles of genetic network dynamics. While the "repressilator" was used to first demonstrate the proof of principle, a more recently developed dual-feedback, fast, tunable genetic oscillator has demonstrated a greater degree of robustness and control over oscillatory behavior by combining positive- and negative-feedback loops. This oscillator, combining lacI (negative-) and araC (positive-) feedback loops, was, however, modeled using multiple layers of differential equations to capture the molecular complexity of regulation, in order to explain the experimentally measured oscillations. In the search for design principles of such minimal oscillatory circuits, we have developed a reduced model of this dual-feedback loop oscillator consisting of just six differential equations, two of which are delay differential equations. The delay term is optimized, as the only free parameter, to fit the experimental dynamics of the oscillator period and amplitude tunability by the two inducers isopropyl ß-D-1-thiogalactopyranoside (IPTG) and arabinose. We proceed to use our reduced and experimentally validated model to redesign the network by comparing the effect of asymmetry in gene expression at the level of (a) DNA copy numbers and the rates of (b) mRNA translation and (c) degradation, since experimental and theoretical work had predicted a need for an asymmetry in the copy numbers of activator (araC) and repressor (lacI) genes encoded on plasmids. We confirm that the minimal period of the oscillator is sensitive to DNA copy number asymmetry, and can demonstrate that while the asymmetry in the translation rate has an identical effect as the plasmid copy numbers, modulating the asymmetry in mRNA degradation can improve the tunability of the period and amplitude of the oscillator. Thus, our model predicts control at the level of translation can be used to redesign such networks, for improved tunability, while at the same time making the network robust to replication "noise" and the effects of the host cell cycle. Thus, our model predicts experimentally testable principles to redesign a potentially more robust oscillatory genetic network.