Various Forms of Programmed Cell Death Are Concurrently Activated in the Population of Retinal Ganglion Cells after Ischemia and Reperfusion.

Retinal ischemia-reperfusion (IR)-which ultimately results in retinal ganglion cell (RGC) death-is a common cause of visual impairment and blindness worldwide. IR results in various types of programmed cell death (PCD), which are of particular importance since they can be prevented by inhibiting the activity of their corresponding signaling cascades. To study the PCD pathways in ischemic RGCs, we used a mouse model of retinal IR and a variety of approaches including RNA-seq analysis, knockout animals, and animals treated with an iron chelator. In our RNA-seq analysis, we utilized RGCs isolated from retinas 24 h after IR. In ischemic RGCs, we found increased expression of many genes that regulate apoptosis, necroptosis, pyroptosis, oxytosis/ferroptosis, and parthanatos. Our data indicate that genetic ablation of death receptors protects RGCs from IR. We showed that the signaling cascades regulating ferrous iron (Fe2+) metabolism undergo significant changes in ischemic RGCs, leading to retinal damage after IR. This data suggests that the activation of death receptors and increased Fe2+ production in ischemic RGCs promote the simultaneous activation of apoptosis, necroptosis, pyroptosis, oxytosis/ferroptosis, and parthanatos pathways. Thus, a therapy is needed that concurrently regulates the activity of the multiple PCD pathways to reduce RGC death after IR.

Molecular Dynamics Modeling of Pulsed Laser Fragmentation of Solid and Porous Si Nanoparticles in Liquid Media.

The production of non-toxic and homogeneous colloidal solutions of nanoparticles (NPs) for biomedical applications is of extreme importance nowadays. Among the various methods for generation of NPs, pulsed laser ablation in liquids (PLAL) has proven itself as a powerful and efficient tool in biomedical fields, allowing chemically pure silicon nanoparticles to be obtained. For example, laser-synthesized silicon nanoparticles (Si NPs) are widely used as contrast agents for bio visualization, as effective sensitizers of radiofrequency hyperthermia for cancer theranostics, in photodynamic therapy, as carriers of therapeutic radionuclides in nuclear nanomedicine, etc. Due to a number of complex and interrelated processes involved in the laser ablation phenomenon, however, the final characteristics of the resulting particles are difficult to control, and the obtained colloidal solutions frequently have broad and multimodal size distribution. Therefore, the subsequent fragmentation of the obtained NPs in the colloidal solutions due to pulsed laser irradiation can be utilized. The resulting NPs' characteristics, however, depend on the parameters of laser irradiation as well as on the irradiated material and surrounding media properties. Thus, reliable knowledge of the mechanism of NP fragmentation is necessary for generation of a colloidal solution with NPs of predesigned properties. To investigate the mechanism of a laser-assisted NP fragmentation process, in this work, we perform a large-scale molecular dynamics (MD) modeling of FS laser interaction with colloidal solution of Si NPs. The obtained NPs are then characterized by their shape and morphological properties. The corresponding conclusion about the relative input of the properties of different laser-induced processes and materials to the mechanism of NP generation is drawn.

COVID-19 in Children: Molecular Profile and Pathological Features.

Although the World Health Organization has declared the end of the COVID-19 pandemic, doctors continue to register new cases of the disease among both adults and children. Unfortunately, the course of COVID-19 in children can have a severe form, with death being a potential outcome. The absence of published works discussing the pathological morphology of COVID-19 in children prevents the objective analysis of the disease's pathogenesis, including among the adult population. In this vein, the objective of our study is to identify the morphological features of the lungs' involvement and evaluate virus-host interactions in the case of COVID-19 in patients at a pediatric medical practice. We present the results of the study of the lungs of three children who died due to COVID-19, highlighting the predominant involvement of their respiratory organs at different stages of the disease (5, 21, and 50 days). This article presents data obtained from histopathological and immunohistochemical investigations, taking into account the results of clinical and laboratory indicators and intravital and postmortem SARS-CoV-2 PCR investigations. The common finding of all of the examined COVID-19 cases is the involvement of the endothelium in microcirculation vessels, which are considered to be a primary target of various pathogenic influencing factors. We also discuss both the significance of apoptosis as a result of virus-host interactions and the most likely cause of endothelium cell destruction. The results of this study could be useful for the development of endothelium-protective therapy to prevent the progression of disseminated intravascular coagulation syndrome.

A dynamic approach to supply chain reconfiguration and ripple effect analysis in an epidemic.

The COVID-19 pandemic has illustrated the unprecedented challenges of ensuring the continuity of operations in a supply chain as suppliers' and their suppliers stop producing due the spread of infection, leading to a degradation of downstream customer service levels in a ripple effect. In this paper, we contextualize a dynamic approach and propose an optimal control model for supply chain reconfiguration and ripple effect analysis integrated with an epidemic dynamics model. We provide supply chain managers with the optimal choice over a planning horizon among subsets of interchangeable suppliers and corresponding orders; this will maximize demand satisfaction given their prices, lead times, exposure to infection, and upstream suppliers' risk exposure. Numerical illustrations show that our prescriptive forward-looking model can help reconfigure a supply chain and mitigate the ripple effect due to reduced production because of suppliers' infected workers. A risk aversion factor incorporates a measure of supplier risk exposure at the upstream echelons. We examine three scenarios: (a) infection limits the capacity of suppliers, (b) the pandemic recedes but not at the same pace for all suppliers, and (c) infection waves affect the capacity of some suppliers, while others are in a recovery phase. We illustrate through a case study how our model can be immediately deployed in manufacturing or retail supply chains since the data are readily accessible from suppliers and health authorities. This work opens new avenues for prescriptive models in operations management and the study of viable supply chains by combining optimal control and epidemiological models.

Russian Physicians Burnout during the COVID-19 Pandemic: A Cross-Sectional Survey Study.

Objective: To study burnout of Russian physicians in the conditions of COVID-19 pandemic and how their work with coronavirus-infected patients influenced it. According to a three-factor model of burnout developed by Maslach and Jackson, this syndrome includes emotional exhaustion, depersonalization, and reduction of personal accomplishment.Design: A cross-sectional survey study.Setting: Large medical practice.Participants: Physicians of different specialties.Methods: Data collection was conducted from June 23 to July 12, 2020. We developed a Google form including a questionnaire and psychological inventories and placed it in a medical portal. Maslach Burnout Inventory - Human Services Survey for Medical Personnel was used to study burnout; the Hospital Anxiety and Depression Scale was used to determine anxiety and depression.Results: Of all the physicians who took part in the study (N = 599), 31.2 % worked with coronavirus-infected patients. Of the medical personnel who treated COVID-19 patients, 63.6% noted increased workload during the pandemic. Compared to other physicians, they more often had a high degree of emotional exhaustion (43.3 % vs 33.0 %, φ* = 2.404, P ≤ 0.01) and depersonalization (41.7 % vs 34, 0%, φ* = 1.803, P ≤ 0.05). An overwhelming majority of physicians, without any dependence on work with infected patients, had an absence of anxiety and depression. The identified interrelations between the symptoms of burnout, anxiety, depression; age and career stage in medical personnel were identical, except for weak correlations between age and emotional exhaustion (rs = -0.097, P ≤ 0.05), as well as career stage and personal accomplishment (rs = 0.102, P ≤ 0.05) in those physicians who worked with COVID-19 patients. The structure of burnout was identical in all physicians and did not depend on interaction with the infected patients.Conclusion: Public health authorities should reduce the workload on physicians involved in treating infected patients against the backdrop of the pandemic. Psychotherapeutic measures focused on preventing burnout should reduce its number among physicians interacting with patients infected with the coronavirus.

Melatonin as the Cornerstone of Neuroimmunoendocrinology.

Much attention has been recently drawn to studying melatonin - a hormone whose synthesis was first found in the epiphysis (pineal gland). This interest can be due to discovering the role of melatonin in numerous physiological processes. It was the discovery of melatonin synthesis in endocrine organs (pineal gland), neural structures (Purkinje cells in the cerebellum, retinal photoreceptors), and immunocompetent cells (T lymphocytes, NK cells, mast cells) that triggered the evolution of new approaches to the unifield signal regulation of homeostasis, which, at the turn of the 21st century, lead to the creation of a new integral biomedical discipline - neuroimmunoendocrinology. While numerous hormones have been verified over the last decade outside the "classical" locations of their formation, melatonin occupies an exclusive position with regard to the diversity of locations where it is synthesized and secreted. This review provides an overview and discussion of the major data regarding the role of melatonin in various physiological and pathological processes, which affords grounds for considering melatonin as the "cornerstone" on which neuroimmunoendocrinology has been built as an integral concept of homeostasis regulation.

Adapting supply chain operations in anticipation of and during the COVID-19 pandemic.

This article investigates the impacts of the COVID-19 pandemic and their proactive mediation by adaptive operational decisions in different network design structures in anticipation of and during the pandemic. In generalized terms, we contribute to the understanding of the effect of preparedness and recovery decisions in a pandemic setting on supply chain operations and performance. In particular, we examine the impact of inventory pre-positioning in anticipation of a pandemic and the adaptation of production-ordering policy during the pandemic. Our model combines three levels, which is not often seen jointly in operations management literature, i.e., pandemic dynamics, supply chain design, and operational production-inventory control policies. The analysis is performed for both two- and three-stage supply chains and different scenarios for pandemic dynamics (i.e., uncontrolled propagation or controlled dispersal with lockdowns). Our findings suggest that two-stage supply chains exhibit a higher vulnerability in disruption cases. However, they are exposed to a lower system inertia and show positive effects at the recovery stage. Supply chain adaptation ahead of a pandemic is more advantageous than during the pandemic when specific operational recovery policies are deployed. We show that it is instructive to avoid simultaneous changes in structural network design and operational policies since that can destabilize the production-inventory system and result in higher product shortages.

Nondestructive Femtosecond Laser Lithography of Ni Nanocavities by Controlled Thermo-Mechanical Spallation at the Nanoscale.

We present a new approach to femtosecond direct laser writing lithography to pattern nanocavities in ferromagnetic thin films. To demonstrate the concept, we irradiated 300 nm thin nickel films by single intense femtosecond laser pulses through glass substrate. Using a fluence above the ablation threshold, the process is destructive, leading to the formation of an ablation crater. By progressively lowering the laser fluence, the formation of closed spallation cavities below the ablation threshold is achieved. Systematic studies by the electron and optical interferometric microscopies, supported by molecular dynamics simulations, enabled us to gain an understanding of the thermo-mechanical spallation mechanism at the solid-molten interface. We achieved the fabrication of periodic arrangements of closed spallation nanocavities. Due to their topology, closed magnetic nanocavities can support unique couplings of multiple excitations (magnetic, optical, acoustic, spintronic). Thereby, they offer a unique physics playground for emerging fields in magnetism, magneto-photonic, and magneto-acoustic applications.

Melatonin, Its Beneficial Effects on Embryogenesis from Mitigating Oxidative Stress to Regulating Gene Expression.

Embryogenesis is a complex multi-stage process regulated by various signaling molecules including pineal and extrapineal melatonin (MT). Extrapineal MT is found in the placenta and ovaries, where it carries out local hormonal regulation. MT is necessary for normal development of oocytes, fertilization and subsequent development of human, animal and avian embryos. This review discusses the role of MT as a regulator of preimplantation development of the embryo and its implantation into endometrial tissue, followed by histo-, morpho- and organogenesis. MT possesses pronounced antioxidant properties and helps to protect the embryo from oxidative stress by regulating the expression of the NFE2L2, SOD1, and GPX1 genes. MT activates the expression of the ErbB1, ErbB4, GJA1, POU5F1, and Nanog genes which are necessary for embryo implantation and blastocyst growth. MT induces the expression of vascular endothelial growth factor (VEGF) and its type 1 receptor (VEGF-R1) in the ovaries, activating angiogenesis. Given the increased difficulties in successful fertilization and embryogenesis with age, it is of note that MT slows down ovarian aging by increasing the transcription of sirtuins. MT administration to patients suffering from infertility demonstrates an increase in the effectiveness of in vitro fertilization. Thus, MT may be viewed as a key factor in embryogenesis regulation, including having utility in the management of infertility.

Maternal Melatonin Deficiency Leads to Endocrine Pathologies in Children in Early Ontogenesis.

The review summarizes the results of experimental and clinical studies aimed at elucidating the causes and pathophysiological mechanisms of the development of endocrine pathology in children. The modern data on the role of epigenetic influences in the early ontogenesis of unfavorable factors that violate the patterns of the formation of regulatory mechanisms during periods of critical development of fetal organs and systems and contribute to the delayed development of pathological conditions are considered. The mechanisms of the participation of melatonin in the regulation of metabolic processes and the key role of maternal melatonin in the formation of the circadian system of regulation in the fetus and in the protection of the genetic program of its morphofunctional development during pregnancy complications are presented. Melatonin, by controlling DNA methylation and histone modification, prevents changes in gene expression that are directly related to the programming of endocrine pathology in offspring. Deficiency and absence of the circadian rhythm of maternal melatonin underlies violations of the genetic program for the development of hormonal and metabolic regulatory mechanisms of the functional systems of the child, which determines the programming and implementation of endocrine pathology in early ontogenesis, contributing to its development in later life. The significance of this factor in the pathophysiological mechanisms of endocrine disorders determines a new approach to risk assessment and timely prevention of offspring diseases even at the stage of family planning.

OR-methods for coping with the ripple effect in supply chains during COVID-19 pandemic: Managerial insights and research implications.

The COVID-19 pandemic unveils unforeseen and unprecedented fragilities in supply chains (SC). A primary stressor of SCs and their subsequent shocks derives from disruption propagation (i.e., the ripple effect) through related networks. In this paper, we conceptualize current state and future research directions on the ripple effect for pandemic context. We scrutinize the existing OR (Operational Research) studies published in international journals dealing with disruption propagation and structural dynamics in SCs. Our study pursues two major contributions in relation to two research questions. First, we collate state-of-the-art research on disruption propagation in SCs and identify a methodical taxonomy along with theories displaying their value and applications for coping with the impacts of pandemics on SCs. Second, we reveal and systemize managerial insights from theory used for operating (adapting) amid a pandemic and during times of recovery, along with becoming more resistant to future pandemics. Streamlining the literature allowed us to reveal several new research tensions and novel categorizations and classifications. The outcomes of our study show that methodical contributions and the resulting managerial insights can be categorized into three levels, i.e., network, process, and control. Our analysis reveals that adaptation capabilities play the most crucial role in managing the SCs under pandemic disruptions. Our findings depict how the existing OR methods can help coping with the ripple effect at five pandemic stages (i.e., Anticipation; Early Detection; Containment; Control and Mitigation; and Elimination) following the WHO classification. The outcomes and findings of our study can be used by industry and researchers alike to progress the decision-support systems guiding SCs amid the COVID-19 pandemic and toward recovery. Suggestions for future research directions are offered and discussed.

Ripple effect in the supply chain network: Forward and backward disruption propagation, network health and firm vulnerability.

A local disruption can propagate to forward and downward through the material flow and eventually influence the entire supply chain network (SCN). This phenomenon of ripple effect, immensely existing in practice, has received great interest in recent years. Moreover, forward and backward disruption propagations became major stressors for SCNs during the COVID-19 pandemic triggered by simultaneous and sequential supply and demand disruptions. However, current literature has paid less attention to the different impacts of the directions of disruption propagation. This study examines the disruption propagation through simulating simple interaction rules of firms inside the SCN. Specifically, an agent-based computational model is developed to delineate the supply chain disruption propagation behavior. Then, we conduct multi-level quantitative analysis to explore the effects of forward and backward disruption propagation, moderated by network structure, network-level health and node-level vulnerability. Our results demonstrate that it is practically important to differentiate between forward and backward disruption propagation, as they are distinctive in the associated mitigation strategies and in the effects on network and individual firm performance. Forward disruption propagation generally can be mitigated by substitute and backup supply and has greater impact on firms serving the assembly role and on the supply/assembly networks, whereas backward disruption propagation is normally mitigated by flexible operation and distribution and has bigger impact on firms serving the distribution role and on distribution networks. We further analyze the investment strategies in a dual-focal supply network under disruption propagation. We provide propositions to facilitate decision-making and summarize important managerial implications.

Competitive pricing of substitute products under supply disruption.

There has been an increased interest in optimizing pricing and sourcing decisions under supplier competition with supply disruptions. In this paper, we conduct an analytical game-theoretical study to examine the effects of supply capacity disruption timing on pricing decisions for substitute products in a two-supplier one-retailer supply chain setting. We investigate whether the timing of a disruption may significantly impact the optimal pricing strategy of the retailer. We derive the optimal pricing strategy and ordering levels with both disruption timing and product substitution. By exploring both the Nash and Stackelberg games, we find that the order quantity with the disrupted supplier depends on price leadership and it tends to increase when the non-disrupted supplier is the leader. Moreover, the equilibrium market retail prices are higher under higher levels of disruption for the Nash game, compared to the Stackelberg game. We also uncover that the non-disrupted supplier can always charge the highest wholesale price if a disruption occurs before orders are received. This highlights the critical role of order timing. The insights can help operations managers to proper design risk mitigation ordering strategies and re-design the supply contracts in the presence of product substitution under supply disruptions.

Mitochondrial targeted therapy with elamipretide (MTP-131) as an adjunct to tumor necrosis factor inhibition for traumatic optic neuropathy in the acute setting.

Traumatic optic neuropathy (TON) can occur following blunt trauma to the orbit and can lead to permanent vision loss. In this study, we investigated the effectiveness of elamipretide (MTP-131), a small mitochondrially-targeted tetrapeptide, in conjunction with etanercept, a tumor necrosis factor (TNF) inhibitor, as neuroprotective agents of retinal ganglion cells (RGCs) after optic nerve trauma with sonication-induced TON (SI-TON) in mice. Treatment with intravitreal MTP-131 and subcutaneous etanercept and MTP-131 showed a 21% increase (p < 0.01) in RGC survival rate compared to PBS-treated control eyes. Subcutaneous etanercept and MTP-131 had an 11% increase (p < 0.05) in RGC survival compared to controls. Subcutaneous etanercept only group showed 20% increase (p < 0.01) in RGC survival compared to controls, while subcutaneous MTP-131 alone showed a 17% increase (p < 0.01). Surprisingly, we did not observe a synergistic effect between the two drugs in the group receiving both etanercept and MTP-131. One possible explanation for the absence of a synergistic effect is that MTP-131 and etanercept may be acting on different portions of the same pathway.

Exploring supply chain structural dynamics: New disruptive technologies and disruption risks.

We discuss recent developments in exploring supply chain structural dynamics. We focus on both positive (i.e., new disruptive technologies) and negative (i.e., disruption risks) triggers of the structural dynamics in complex supply chain networks. We discuss papers in the special issue which focus on supply chain structural dynamics using different methods, collating and presenting recent research in the field. In particular, the ripple effect, blockchain, network resilience, data analytics, and service platforms have been identified as the leading research directions.

Bayesian networks for supply chain risk, resilience and ripple effect analysis: A literature review.

In the broad sense, the Bayesian networks (BN) are probabilistic graphical models that possess unique methodical features to model dependencies in complex networks, such as forward and backward propagation (inference) of disruptions. BNs have transitioned from an emerging topic to a growing research area in supply chain (SC) resilience and risk analysis. As a result, there is an acute need to review existing literature to ascertain recent developments and uncover future areas of research. Despite the increasing number of publications on BNs in the domain of SC uncertainty, an extensive review on their application to SC risk and resilience is lacking. To address this gap, we analyzed research articles published in peer-reviewed academic journals from 2007 to 2019 using network analysis, visualization-based scientometric analysis, and clustering analysis. Through this study, we contribute to literature by discussing the challenges of current research, and, more importantly, identifying and proposing future research directions. The results of our survey show that further debate on the theory and application of BNs to SC resilience and risk management is a significant area of interest for both academics and practitioners. The applications of BNs, and their conjunction with machine learning algorithms to solve big data SC problems relating to uncertainty and risk, are also discussed.

Wnt signaling induces neurite outgrowth in mouse retinal ganglion cells.

Wingless-type (Wnt) signaling pathways mediate axonal growth and remodeling in the embryonic optic nerve, brain and spinal cord. Recent studies demonstrated that the canonical Wnt/ß-catenin signaling pathway also induces axonal regeneration after injury in the optic nerve of adult animals. However, the molecular mechanisms of Wnt-mediated axonal growth are not well understood. Additionally, because Wnt signaling is stimulated in neurons as well as neighboring non-neuronal cells, the cell type(s) responsible for Wnt-induced axonal regeneration are not known. The objectives of this study were to investigate potential mechanisms and target cells of Wnt3a stimulated neurite growth using primary retinal ganglion cell (RGC) cultures. We demonstrated that Wnt3a ligand induced dose-dependent increases in average neurite length and number of neurites in RGCs. QPCR analysis of candidate mediators showed that Wnt3a-dependent neurite growth was associated with lower expression of Ripk1 and Ripk3 genes. Additionally, inhibiting Ripk1 signaling with Necrostatin-1s led to increased neurite number per cell but not increased neurite length. Therefore, Ripk signaling may be involved in mediating the effects of Wnt3a on neurite number but Ripk activity does not seem to be required for Wnt3a-dependent regulation of neurite length. This study shows that RGCs are direct cellular targets of Wnt3a-induced axonal growth, and we identified a novel association between Wnt signaling and Rip kinases in neurite formation.

Numerical Investigation of Ultrashort Laser-Ablative Synthesis of Metal Nanoparticles in Liquids Using the Atomistic-Continuum Model.

We present a framework based on the atomistic continuum model, combining the Molecular Dynamics (MD) and Two Temperature Model (TTM) approaches, to characterize the growth of metal nanoparticles (NPs) under ultrashort laser ablation from a solid target in water ambient. The model is capable of addressing the kinetics of fast non-equilibrium laser-induced phase transition processes at atomic resolution, while in continuum it accounts for the effect of free carriers, playing a determinant role during short laser pulse interaction processes with metals. The results of our simulations clarify possible mechanisms, which can be responsible for the observed experimental data, including the presence of two populations of NPs, having a small (5-15 nm) and larger (tens of nm) mean size. The formed NPs are of importance for a variety of applications in energy, catalysis and healthcare.

Mitochondrial DNA Double-Strand Breaks in Oligodendrocytes Cause Demyelination, Axonal Injury, and CNS Inflammation.

Mitochondrial dysfunction has been implicated in the pathophysiology of neurodegenerative disorders, including multiple sclerosis (MS). To date, the investigation of mitochondrial dysfunction in MS has focused exclusively on neurons, with no studies exploring whether dysregulation of mitochondrial bioenergetics and/or genetics in oligodendrocytes might be associated with the etiopathogenesis of MS and other demyelinating syndromes. To address this question, we established a mouse model where mitochondrial DNA (mtDNA) double-strand breaks (DSBs) were specifically induced in myelinating oligodendrocytes (PLP:mtPstI mice) by expressing a mitochondrial-targeted endonuclease, mtPstI, starting at 3 weeks of age. In both female and male mice, DSBs of oligodendroglial mtDNA caused impairment of locomotor function, chronic demyelination, glial activation, and axonal degeneration, which became more severe with time of induction. In addition, after short transient induction of mtDNA DSBs, PLP:mtPstI mice showed an exacerbated response to experimental autoimmune encephalomyelitis. Together, our data demonstrate that mtDNA damage can cause primary oligodendropathy, which in turn triggers demyelination, proving PLP:mtPstI mice to be a useful tool to study the pathological consequences of mitochondrial dysfunction in oligodendrocytes. In addition, the demyelination and axonal loss displayed by PLP:mtPstI mice recapitulate some of the key features of chronic demyelinating syndromes, including progressive MS forms, which are not accurately reproduced in the models currently available. For this reason, the PLP:mtPstI mouse represents a unique and much needed platform for testing remyelinating therapies.SIGNIFICANCE STATEMENT In this study, we show that oligodendrocyte-specific mitochondrial DNA double-strand breaks in PLP:mtPstI mice cause oligodendrocyte death and demyelination associated with axonal damage and glial activation. Hence, PLP:mtPstI mice represent a unique tool to study the pathological consequences of mitochondrial dysfunction in oligodendrocytes, as well as an ideal platform to test remyelinating and neuroprotective agents.

Retrospective surveillance of antibiotic use in maternity wards and neonatal intensive care units in Saint Petersburg, Russia.

Antibiotic overuse in infants is associated with an increased risk of serious adverse events. Development of antibiotic stewardship programs aimed at reducing overall antibiotic consumption requires epidemiological surveillance. Retrospective surveillance and evaluation of all antibiotics provided to every infant admitted to maternal wards or neonatal intensive care units (NICUs) from 01 January 2014 to 31 December 2014 were performed in five medical centers of Saint Petersburg, Russia. Types of antibiotics and dates of administration were recorded. Antibiotic use was quantified by length of therapy (length of therapy, LOT, per 1000 patient-days, PD) and days of therapy (DOT/1000 PD). An additional parameter named "instant DOT/1000 PD" was introduced by authors for assessment of longitudinal patterns of administrations. Antibiotic load was 825.6 DOT/1000 PD in maternity wards and 1425.8 DOT/1000 PD in the NICUs. These levels are two to four times higher than DOTs reported in the USA for a level III NICU (348 DOT/1000PD). Antibiotic load was associated with the length of hospital stay (LOS) and birth weight. These associations were distorted when assessed using the conventional parameters, LOT and DOT, because they do not reflect the longitudinal component of treatment and underestimate antibiotic load when a patient stays in hospital without treatment. The proposed additional parameter successfully overcame these flaws and uncovered hidden associations. Severe overuse of antibiotics may be taking place in Russia and antibiotic stewardship development should be urged. Instant DOT/1000 PD is a more powerful tool in assessing treatment patterns than DOT/1000 PD.