IRF8 deficiency induces the transcriptional, functional, and epigenetic reprogramming of cDC1 into the cDC2 lineage.

Conventional dendritic cells (cDCs) consist of two major functionally and phenotypically distinct subsets, cDC1 and cDC2, whose development is dependent on distinct sets of transcription factors. Interferon regulatory factor 8 (IRF8) is required at multiple stages of cDC1 development, but its role in committed cDC1 remains unclear. Here, we used Xcr1-cre to delete Irf8 in committed cDC1 and demonstrate that Irf8 is required for maintaining the identity of cDC1. In the absence of Irf8, committed cDC1 acquired the transcriptional, functional, and chromatin accessibility properties of cDC2. This conversion was independent of Irf4 and was associated with the decreased accessibility of putative IRF8, Batf3, and composite AP-1-IRF (AICE)-binding elements, together with increased accessibility of cDC2-associated transcription-factor-binding elements. Thus, IRF8 expression by committed cDC1 is required for preventing their conversion into cDC2-like cells.

An Automated Single-Cell Droplet-Digital Microfluidic Platform for Monoclonal Antibody Discovery.

Monoclonal antibody (mAb) discovery plays a prominent role in diagnostic and therapeutic applications. Droplet microfluidics has become a standard technology for high-throughput screening of antibody-producing cells due to high droplet single-cell confinement frequency and rapid analysis and sorting of the cells of interest with their secreted mAbs. In this work, a new method is described for on-demand co-encapsulation of cells that eliminates the difficulties associated with washing in between consecutive steps inside the droplets and enables the washing and addition of fresh media. The new platform identifies hybridoma cells that are expressing antibodies of interest using antibody-characterization assays to find the best-performing or rare-cell antibody candidates.

Recent findings in methanotrophs: genetics, molecular ecology, and biopotential.

The potential consequences for mankind could be disastrous due to global warming, which arises from an increase in the average temperature on Earth. The elevation in temperature primarily stems from the escalation in the concentration of greenhouse gases (GHG) such as CO2, CH4, and N2O within the atmosphere. Among these gases, methane (CH4) is particularly significant in driving alterations to the worldwide climate. Methanotrophic bacteria possess the distinctive ability to employ methane as both as source of carbon and energy. These bacteria show great potential as exceptional biocatalysts in advancing C1 bioconversion technology. The present review describes recent findings in methanotrophs including aerobic and anaerobic methanotroph bacteria, phenotypic characteristics, biotechnological potential, their physiology, ecology, and native multi-carbon utilizing pathways, and their molecular biology. The existing understanding of methanogenesis and methanotrophy in soil, as well as anaerobic methane oxidation and methanotrophy in temperate and extreme environments, is also covered in this discussion. New types of methanogens and communities of methanotrophic bacteria have been identified from various ecosystems and thoroughly examined for a range of biotechnological uses. Grasping the processes of methanogenesis and methanotrophy holds significant importance in the development of innovative agricultural techniques and industrial procedures that contribute to a more favorable equilibrium of GHG. This current review centers on the diversity of emerging methanogen and methanotroph species and their effects on the environment. By amalgamating advanced genetic analysis with ecological insights, this study pioneers a holistic approach to unraveling the biopotential of methanotrophs, offering unprecedented avenues for biotechnological applications. KEY POINTS: • The physiology of methanotrophic bacteria is fundamentally determined. • Native multi-carbon utilizing pathways in methanotrophic bacteria are summarized. • The genes responsible for encoding methane monooxygenase are discussed.

Living in urban or rural environments affect the sleep quality of the elderly in Bushehr (Southern Iran): emphasizing the active and inactive of the elderly.

INTRODUCTION: Sleep disorders have a significant negative impact on mental and physical health, especially among the elderly. Various factors can affect the sleep quality of elderly people. The aim of this research to investigate the effect of urban and rural environments on the sleep quality of elderly people with emphasis on physical activity. METHOD: Four hundred and thirty-nine elderly people (226 city residents and 213 village residents) in urban and rural areas of Bushehr (Southern Iran), volunteered to participate in the present study. Information was collected via the General information questionnaire and Petersburg Sleep Questionnaire. RESULT: The results showed that active elderly women (p < 0.001), and total active elderly (male + female) (p < 0.001) living in urban areas compared to inactive elderly and also in rural areas active elderly women (p < 0.001), active elderly men (p < 0.001) and total active elderly (male + female) (p < 0.001) had better overall sleep quality in compared to inactive elderly. Also, elderly men (p < 0.001) and the total elderly (male + female) (p < 0.001) living in urban areas had better sleep quality than the elderly in rural areas. CONCLUSION: Based on the findings, it can be concluded that the way of life (being active) as well as the living environment can affect the sleep quality of elderly people, so that active elderly people and also elderly people living in urban environments had better sleep quality.

Experiences and attitudes of clinical and academic nurses about infection prevention and control nursing curriculum and duties - a qualitative study.

BACKGROUND: Infectious diseases are becoming more widespread and re-emerging, causing psychological, social, economic, and health effects at both national and international levels. Specialist nurses can help prevent and control these infections. However, in Iran, there are currently no specialist infection prevention and control (IPC) nurses to manage and control infections. This study aims to explore clinical and academic nurses' attitudes toward IPC nursing curriculum and duties. METHODS: The study used a qualitative content analysis approach. Thirty-six participants, including clinical and academic nurses, were selected using a purposeful sampling method. Data was collected through seven focused group discussions. The accuracy and validity of the research tools were measured using the Four-Dimension Criteria developed by Lincoln and Guba. Data analysis was conducted using directed content analysis. RESULTS: The data analysis of the discussions held in the seven focus groups extracted 628 codes. Three themes were developed from the qualitative analysis: "Core characteristics of the curriculum", "Expected competencies and skills", and "Evaluation." These themes were derived from nine main categories and 25 subcategories. CONCLUSIONS: Specialist IPC nurses can play important roles in various positions and environments. Therefore, educational policymakers in Iran should consider establishing IPC nursing courses. It is also recommended that policymakers and decision-makers in the nursing field of other less developed countries should prioritize this issue.

Bacterial profiles and their antibiotic resistance background in superinfections caused by multidrug-resistant bacteria among COVID-19 ICU patients from southwest Iran.

This study investigated the bacterial causes of superinfections and their antibiotic resistance pattern in severe coronavirus disease 2019 (COVID-19) patients admitted to the intensive care unit (ICU) of Razi Hospital in Ahvaz, southwest Iran. In this cross-sectional study, endotracheal tube (ETT) secretion samples of 77 intubated COVID-19 patients, confirmed by reverse transcription-quantitative polymerase chain reaction, were investigated by standard microbiology test and analytical profile index kit. Antibiotic susceptibility testing was performed by disc diffusion. The presence of Haemophilus influenzae and Mycoplasma pneumoniae was investigated by the polymerase chain reaction (PCR). Using culture and PCR methods, 56 (72.7%) of the 77 COVID-19 patients (mean age of 55 years, 29 male and 27 female) had superinfections. Using culture, 67 isolates including 29 (43.2%) Gram-positive and 38 (56.7%) Gram-negative bacteria (GNB) were identified from 49 COVID-19 patients. The GNB were more predominant than the Gram-positive pathogens. Klebsiella pneumoniae (28.4%, n = 19/67) was the most common isolate followed by Staphylococcus aureus (22.4%, n = 15/67). Using PCR, 10.4% (8/77) and 11.7% (9/77) of ETT secretion specimens had H. influenzae and M. pneumoniae amplicons, respectively. Gram-positive and Gram-negative isolates showed high resistance rates (>70.0%) to majority of the tested antibiotics including fluoroquinolone, carbapenems, and cephalosporins and 68.7% (46/67) of isolates were multidrug-resistant (MDR). This study showed a high frequency rate of superinfections by MDR bacteria among COVID-19 patients in southwest Iran. The prevention of long-term consequences caused by COVID-19, demands continuous antibiotic surveillance particularly in management of bacterial superinfections.

Recent advances in various bio-applications of bacteria-derived outer membrane vesicles.

Outer membrane vesicles (OMVs) are spherical nanoparticles released from gram-negative bacteria. OMVs were originally classified into native 'nOMVs' (produced naturally from budding of bacteria) and non-native (produced by mechanical means). nOMVs and detergent (dOMVs) are isolated from cell supernatant without any detergent cell disruption techniques and through detergent extraction, respectively. Growth stages and conditions e.g. different stress factors, including temperature, nutrition deficiency, and exposure to hazardous chemical agents can affect the yield of OMVs production and OMVs content. Because of the presence of bacterial antigens, pathogen-associated molecular patterns (PAMPs), various proteins and the vesicle structure, OMVs have been developed in many biomedical applications. OMVs due to their size can be phagocytized by APCs, enter lymph vessels, transport antigens efficiently, and induce both T and B cells immune responses. Non-engineered OMVs have been frequently used as vaccines against different bacterial and viral infections, and various cancers. OMVs can also be used in combination with different antigens as an attractive vaccine adjuvant. Indeed, foreign antigens from target microorganisms can be trapped in the lumen of nonpathogenic vesicles or can be displayed on the surface through bacterial membrane protein to increase the immunogenicity of the antigens. In this review, different factors affecting OMV production including time of cultivation, growth media, stress conditions and genetic manipulations to enhance vesiculation will be described. Furthermore, recent advances in various biological applications of OMVs such as vaccine, drug delivery, cancer therapy, and enzyme carrier are discussed. Generally, the application of OMVs as vaccine carrier in three categories (i.e., non-engineered OMVs, OMVs as an adjuvant, recombinant OMVs (rOMVs)), as delivery system for small interfering RNA and therapeutic agents, and as enzymes carrier will be discussed.

Platelets and platelet-derived vesicles as an innovative cellular and subcellular platform for managing multiple sclerosis.

INTRODUCTION: Multiple sclerosis (MS) is a progressive inflammatory autoimmune disease that involves young individuals. The drug delivery systems now are available for this disease have chronic and non-targeted effects on the patients. Because of the presence of BBB (blood-brain-barrier), their concentration in the CNS (central nervous system) is low. Because of this flaw, it is critical to use innovative active targeted drug delivery methods. RESULT: Platelets are blood cells that circulate freely and play an important role in blood hemostasis. In this review, we emphasize the various roles of activated platelets in the inflammatory condition to recruit other cells to the injured area and limit inflammation. Besides, the activated platelets in the different stages of the MS disease play a significant role in limiting the progression of inflammation in the peripheral area and CNS. DISCUSSION: This evidence indicates that a platelet-based drug delivery system can be an efficient biomimetic candidate for drug targeting to the CNS and limiting the inflammation in the peripheral and central areas for MS therapy.

Carcinogenic and non-carcinogenic risk assessment of exposure to trace elements in groundwater resources of Sari city, Iran.

The daily intake of trace elements through water resources and their adverse health effects is a critical issue. The purpose of this research was to assess the carcinogenic and non-carcinogenic risks of exposure to iron (Fe), copper (Cu), manganese (Mn), zinc (Zn), chromium (Cr), lead (Pb), and arsenic (As) in groundwater resources of Sari city, Iran. The concentrations of the trace elements in a total number of 66 samples from the groundwater sources were measured using inductively coupled plasma mass spectrometry (ICP-MS). The hazard index (HI) levels of exposure to the trace elements from the groundwater sources for adults, teenagers, and children were 0.65, 0.83, and 1.08, respectively. The carcinogenic risk values of Cr and As in the groundwater sources for children, teenagers, and adults were 0.0001, 0.00009, 0.00007, 0.0003, 0.0002, and 0.0001, respectively, causing a total carcinogenic risk value higher than the acceptable range, and removing Cr and As from the groundwater resources is recommended for safe community water supply.

Replacing biochar with mineral premix and its interaction with vitamin C on laying hen production and egg quality factors.

This researchers focused at how adding vitamin C (VC) to biochar and replacing it with a mineral supplement affected egg quality and laying hen performance. 50 experimental units were created from 400 laying hens using a 5 × 2 factorial treatment design (10 treatments, 5 repeats, and 8 laying hens per repetition). Biochar levels (0, 25, 50, 75, and 100% replacement with mineral supplements of diet) and VC levels (0 and 100 mg/kg of diet) were some of the studied variables. The results showed that different experimental diets had no significant effect on performance parameters (feed intake, feed conversion ratio, daily weight gain, egg weight, egg production, and egg mass) of laying hens. In the whole of experiment (50-62 weeks of age), dietary treatments had no influence on egg albumen %, Haugh unit, albumen index, yolk %, yolk index, yolk color, egg shell thickness, or egg shell ash. The results revealed that biochar, due to its availability and easy production, can replace mineral supplements in laying hens' diet, with no adverse effects on productive performance and egg quality traits.

Exploring nursing students' perceptions from nursing role function (SP-NRF) during the COVID-19 pandemic in Ardabil Province: a cross sectional study from Iran.

BACKGROUND: Perception of nursing roles among nursing students significantly influences their active engagement in nursing processes and care delivery. However, there is evidence to suggest that students' interest in and perceptions from the nursing profession at the undergraduate level are often insufficient. OBJECTIVE: This study aimed to assess nursing students' perceptions of nursing role function and identify areas that require improvement. METHODS: A cross-sectional study was conducted in 2021 among nursing students in the third- and fourth-years from three faculties in the Ardabil Province. The participants were selected through census sampling. The data were collected through interviews with the Standardized Professional Nursing Role Function (SP-NRF) questionnaire. Statistical analysis was performed using the SPSS-18 software at a significance level of less than 0.05. RESULTS: A total of 320 nursing students participated in this study. The mean score for nursing role perception was 223.1 ± 20.3 out of 255. The results indicated significant gender differences in the mean scores of perception of the nursing role function, particularly in the supportive, professional-moral care, and professional-educational dimensions. Women scored significantly higher than men did (P < .05). Additionally, students who obtained a mean score of 19 to 20 (A) had significantly higher total scores in perception of the nursing role function than other students. Furthermore, a positive correlation was observed between students' interest in nursing and their perceived ability with nursing role perception (r = .282, P < .01) and all its dimensions. CONCLUSION: Overall, nursing students demonstrated a favorable perception of nursing role function. However, their perception of mental and spiritual care was relatively weak. These findings highlight the need to review nursing education programs and incorporate the spiritual care dimension to enhance students' understanding of and preparation for their role as nurses.

Nanohybrid Based on (Mn, Zn) Ferrite Nanoparticles Functionalized With Chitosan and Sodium Alginate for Loading of Curcumin Against Human Breast Cancer Cells.

This study reports on the synthesis of Mn1 - xZnxFe2O4 (Mn, Zn ferrite) magnetic nanoparticles (MNPs) as drug delivery carriers for effective therapeutic outcomes. The MNPs were prepared using the coprecipitation method, and their magnetic properties were investigated based on their composition. Among the compositions tested, Mn0.8Zn0.2Fe2O4 MNPs exhibited superparamagnetic properties with a saturation magnetization moment of 34.6 emu/g at room temperature (25°C). To enhance the water solubility of curcumin (Cur), known for its hydrophobic nature, it was successfully loaded onto alginate (Alg)/chitosan (Chit)@Mn0.8Zn0.2Fe2O4 nanoparticles (NPs). The nanocomposite was characterized by field emission scanning electron microscopy (FE-SEM) which revealed a particle size of approximately 20 nm. The crystalline structure of the NPs was analyzed using X-ray diffraction, while Fourier-transform infrared (FTIR), energy-dispersive X-ray, and map analysis techniques were employed for further characterization. In terms of drug release, there was an initial burst release of Cur (around 18%) within the first hour, followed by a slower release (approximately 61%) over the next 36 h. The anti-tumor properties of the Cur-loaded NPs were evaluated using the Methyl Thiazol Tetrazolium (MTT) assay and quantitative real-time polymerase chain reaction. The MTT assay confirmed a higher cytotoxic effect of Cur-loaded Alg/Chit@Mn0.8Zn0.2Fe2O4 NPs on the MCF-7 breast cancer cell line compared to free Cur, highlighting the significance of incorporating Cur into nano-sized carrier systems.

Higher-Dose Gabapentinoids and the Risk of Adverse Events in Older Adults With CKD: A Population-Based Cohort Study.

RATIONALE & OBJECTIVE: Gabapentinoids are opioid substitutes whose elimination by the kidneys is reduced as kidney function declines. To inform their safe prescribing in older adults with chronic kidney disease (CKD), we examined the 30-day risk of serious adverse events according to the prescribed starting dose. STUDY DESIGN: Population-based cohort study. SETTING & PARTICIPANTS: 74,084 older adults (64% women; median age, 79 [interquartile range, 73-85] years) with CKD (defined for this study as an estimated glomerular filtration rate <60 mL/min/1.73 m2 and excluding those receiving dialysis) and a newly prescribed gabapentinoid between 2008 and 2020 in Ontario, Canada. EXPOSURE: Higher-dose gabapentinoids (gabapentin >300 mg/d or pregabalin >75 mg/d) versus lower-dose gabapentinoids (gabapentin ≤300 mg/d or pregabalin ≤75 mg/d). OUTCOMES: The primary composite outcome was the 30-day risk of a hospital visit with encephalopathy, a fall, or a fracture or a hospitalization with respiratory depression. ANALYTICAL APPROACH: Comparison groups were balanced on indicators of baseline health using inverse probability of treatment weighting using propensity score analysis that generated a pseudosample for the reference group with a distribution of measured covariates similar to the exposed group. Weighted risk ratios were estimated using modified Poisson regression, and weighted risk differences were estimated using binomial regression. Prespecified subgroup analyses were conducted by estimated glomerular filtration rate category and type of gabapentinoid. RESULTS: Among 74,084 patients identified with CKD and a new prescription for gabapentin or pregabalin, 41% started at >300 or >75 mg/d, respectively. From this set of patients, a weighted study population with a size of 61,367 was generated. Patients who started at a higher dose had a higher 30-day risk of the primary outcome than patients who started at lower dose. Within the weighted population, the numbers of events for higher versus lower dose were 585 of 30,660 (1.9%) versus 462 of 30,707 (1.5%), respectively. The weighted risk ratio was 1.27 (95% CI, 1.13-1.42), and the weighted risk difference was 0.40% (95% CI, 0.21%-0.60%). In subgroup analyses, neither multiplicative nor additive interactions were statistically significant. LIMITATIONS: Residual confounding. CONCLUSIONS: In this population-based study, starting a gabapentinoid at a higher versus a lower dose was associated with a slightly higher risk of a hospital visit with encephalopathy, a fall, or a fracture or hospitalization with respiratory depression. If verified, these risks should be balanced against the benefits of using a higher-dose gabapentinoid.

Identification and characterization of murine glycoprotein 2-expressing intestinal dendritic cells.

The intestinal lamina propria (LP) contains distinct subsets of classical dendritic cells (cDC), each playing key non-redundant roles in intestinal immune homeostasis. Here, we show that glycoprotein 2 (GP2), a GPI-anchored protein and receptor for bacterial type-I fimbriae, is selectively expressed by CD103+CD11b+ cDC in the murine small intestine (SI). GP2 expression was induced on CD103+CD11b+ cDC within the SI-LP and was regulated by IRF4, TGFßR1- and retinoic acid signalling. Mice selectively lacking Gp2 on CD103+CD11b+ cDC (huLang-Cre.gp2fl/fl mice) had normal numbers and proportions of innate and adaptive immune cells in the SI-LP suggesting that GP2 expression by CD103+CD11b+ cDC is not required for intestinal immune homoeostasis.

Postpartum depression and its correlates: a cross-sectional study in southeast Iran.

BACKGROUND: Postpartum depression has a negative impact on both infants and women. This study aimed to determine the correlates of postpartum depression in women in southern Iran. METHODS: This cross-sectional study was performed on 186 mothers who had recently given birth to a baby. Data were collected using the demographic form, Quality of Prenatal Care Questionnaire, Edinburgh Postnatal Depression Scale (EPDS), Depression, Anxiety and Stress Scale-21 items (DASS-21) 3 days after delivery and EPDS and DASS-21 6 months after childbirth. RESULTS: Postpartum depression (PPD) was 24.2% and 3.2% 3 days and 6 months after delivery, respectively. Anxiety, Prenatal Care Quality and educational level predicted 34.0% of the variance of PPD 3 days after delivery (R2 = 34.0%). Anxiety, type of delivery, and stress predicted 24% of the variance of PPD 6 months after delivery (R2 = 24.0%). CONCLUSIONS: With an increase in stress and anxiety and a reduction in the quality of prenatal care, the risk of postpartum depression increases. Therefore, attention to the quality of prenatal care and postpartum stress and anxiety should be carefully evaluated to prevent PPD. Psychological support and interventions are recommended to promote the mental health of women before and after childbirth.

Solid lipid nanoparticles and nanostructured lipid carriers: a review of the methods of manufacture and routes of administration.

The bioavailability of drugs is dependent on several factors such as solubility and the administration route. A drug with poor aqueous solubility, therefore, poses challenges with regards to its pharmaceutical advance and ultimately its biological usage. Lipid nanoparticles have been used in pharmaceutical science due to their importance in green chemistry. Their biochemical properties as 'green' materials and biochemical processes as 'green' processes mean they can be environmentally sustainable. Generally, lipid nanoparticles can be employed as carriers for both lipophilic and hydrophilic drugs. The proposed administration route for nanoparticles can present advantages and disadvantages which should be considered by a formulator. Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) are attractive delivery systems because of their ease of manufacture, biocompatibility, biodegradability, and scale-up capacity of formulation constituents. The easy and simple scalability of novel SLNs and nano lipid carriers, along with their various processing procedures, recent developments, limitation and toxicity, formulation optimization and approaches for the manufacture of lipid nanoparticles, lyophilization and drug release are comprehensively discussed in this review. This review also summarizes the research data related to the various preparation methods and excipients used for SLNs and NLCs in recent years.

Assessment of Mortality Rate, Need for ICU Admission and Ventilation in COVID-19 Patients with Diabetes Mellitus.

INTRODUCTION: Coronavirus disease 2019 (COVID-19) has become a threat to public health. People with chronic diseases, such as diabetes, are at a greater risk of severe diseases and death upon contracting this new disease. Due to the novelty of COVID-19, no specific information is available about the degree of its mortality and risk factors among diabetic patients. Therefore, this study aims to compare diabetic and nondiabetic COVID-19 patients regarding mortality rate, the need for intensive care unit (ICU) admission, invasive and noninvasive ventilation, and the associated risk factors. METHODS: This was a cross-sectional study performed on the medical records of 650 adult COVID-19 patients (325 diabetics and 325 nondiabetics) admitted to Razi Hospital in Ahvaz from March 2020 to September 2020. RESULTS: The mean age of the patients was 61.3 years in the diabetic group and 52.3 years in the nondiabetic group. Men comprised 48.3% of the diabetic group and 59.7% of the nondiabetic group. Diabetic patients suffered from significantly more underlying diseases, such as ischemic heart disease (IHD), hypertension (HTN), chronic kidney disease (CKD), and acute renal failure (ARF) compared to the nondiabetic group (p < 0.0001). Also, when compared with the nondiabetic group, the diabetic group had a significantly higher mortality rate (17.5% vs. 12%; p = 0.047, respectively), more ICU admissions (35.4% vs. 27.7%; p = 0.035, respectively), and a greater need for invasive ventilation (17.5% vs. 11.4%; p = 0.026, respectively). CONCLUSION: In diabetic patients, the mortality rate, need for ICU admission, and need for invasive ventilation were significantly higher than nondiabetic patients. Our logistic regression analysis in diabetic patients with COVID-19 showed that age, CKD, and ARF were the risk factors affecting mortality. In contrast, age and CKD were the risk factors affecting the rate of ICU admission, and CKD and ARF were the risk factors affecting the need for invasive ventilation.

Induction of diapause by clock proteins period and timeless via changes in PTTH and ecdysteroid titer in the sugar beet moth, Scrobipalpa ocellatella (Lepidoptera: Gelechiidae).

The sugar beet moth, Scrobipalpa ocellatella (Boyd), one of the most severe sugar beet pests, causes quantitative and qualitative yield losses late in the autumn. Previously, it was shown that low temperature and short-day photoperiod together cause diapause induction in pupae. Here, the interaction of the critical elements of the diapause induction, including the period (PER), timeless (TIM), prothoracicotropic hormone (PTTH), and ecdysteroid titer, were investigated. Immunohistochemistry results showed that the number of period immunoreactivity (PER-ir) and TIM-ir cells in nondiapause pupae (NDP) was lower than in the brain of the diapause pupae (DP). Moreover, the number of PER-ir and TIM-ir cells in the protocerebrum and optic lobe (OL) of NDP was lower than DP. Moreover, lower PTTH content in the brain and hemolymph of DP was confirmed by competitive enzyme-linked immunosorbent assay. Enzyme immunoassay showed a lower 20-hydroxyecdysone (20E) titer in the hemolymph of the DP compared with the NDP. Within a short-day condition, PER and TIM titers increased in the brain leading to decreasing PTTH titers in the brain and hemolymph that caused decreasing 20E titer in the hemolymph, leading to the induction of diapause. This study suggests that PER and TIM could be one of the brain factors that play an essential role in regulating diapause in S. ocellatella.

One Cell, One Drop, One Click: Hybrid Microfluidics for Mammalian Single Cell Isolation.

Generating a stable knockout cell line is a complex process that can take several months to complete. In this work, a microfluidic method that is capable of isolating single cells in droplets, selecting successful edited clones, and expansion of these isoclones is introduced. Using a hybrid microfluidics method, droplets in channels can be individually addressed using a co-planar electrode system. In the hybrid microfluidics device, it is shown that single cells can be trapped and subsequently encapsulate them on demand into pL-sized droplets. Furthermore, droplets containing single cells are either released, kept in the traps, or merged with other droplets by the application of an electric potential to the electrodes that is actuated through an in-house user interface. This high precision control is used to successfully sort and recover single isoclones to establish monoclonal cell lines, which is demonstrated with a heterozygous NCI-H1299 lung squamous cell population resulting from loss-of-function eGFP and RAF1 gene knockout transfections.