Individual Compartments Research Articles

Passive Mechanical Properties of Human Medial Gastrocnemius and Soleus Musculotendinous Unit.

The in vivo characterization of the passive mechanical properties of the human triceps surae musculotendinous unit is important for gaining a deeper understanding of the interactive responses of the tendon and muscle tissues to loading during passive stretching. This study sought to quantify a comprehensive set of passive muscle-tendon properties such as slack length, stiffness, and the stress-strain relationship using a combination of ultrasound imaging and a three-dimensional motion capture system in healthy adults. By measuring tendon length, the cross-section areas of the Achilles tendon subcompartments (i.e., medial gastrocnemius and soleus aspects), and the ankle torque simultaneously, the mechanical properties of each individual compartment can be specifically identified. We found that the medial gastrocnemius (GM) and soleus (SOL) aspects of the Achilles tendon have similar mechanical properties in terms of slack angle (GM: −10.96° ± 3.48°; SOL: −8.50° ± 4.03°), moment arm at 0° of ankle angle (GM: 30.35 ± 6.42 mm; SOL: 31.39 ± 6.42 mm), and stiffness (GM: 23.18 ± 13.46 Nmm−1; SOL: 31.57 ± 13.26 Nmm−1). However, maximal tendon stress in the GM was significantly less than that in SOL (GM: 2.96 ± 1.50 MPa; SOL: 4.90 ± 1.88 MPa, p = 0.024), largely due to the higher passive force observed in the soleus compartment (GM: 99.89 ± 39.50 N; SOL: 174.59 ± 79.54 N, p = 0.020). Moreover, the tendon contributed to more than half of the total muscle-tendon unit lengthening during the passive stretch. This unequal passive stress between the medial gastrocnemius and the soleus tendon might contribute to the asymmetrical loading and deformation of the Achilles tendon during motion reported in the literature. Such information is relevant to understanding the Achilles tendon function and loading profile in pathological populations in the future.

Liposome-Assisted Delivery of Biomembrane-Impermeable Cargo to Primitive Cells

We report on a platform for targeted delivery, and incorporation of membrane-impermeable compounds into cell-like unilamellar lipid bilayer microcompartments. We perform the delivery by means of an open-space microfluidic device, which perfuses individual surface-adhered compartments (primitive model cells) with cargo-loaded small unilamellar liposomes, composed of cationic, i.e. fusogenic, lipids. We show specifically that fluorescently labelled RNA and DNA oligomers can be successfully internalized in this way.

Plasticity of the Cuticular Transpiration Barrier in Response to Water Shortage and Resupply in Camellia sinensis: A Role of Cuticular Waxes.

The cuticle is regarded as a non-living tissue; it remains unknown whether the cuticle could be reversibly modified and what are the potential mechanisms. In this study, three tea germplasms (Wuniuzao, 0202-10, and 0306A) were subjected to water deprivation followed by rehydration. The epicuticular waxes and intracuticular waxes from both leaf surfaces were quantified from the mature 5th leaf. Cuticular transpiration rates were then measured from leaf drying curves, and the correlations between cuticular transpiration rates and cuticular wax coverage were analyzed. We found that the cuticular transpiration barriers were reinforced by drought and reversed by rehydration treatment; the initial weak cuticular transpiration barriers were preferentially reinforced by drought stress, while the original major cuticular transpiration barriers were either strengthened or unaltered. Correlation analysis suggests that cuticle modifications could be realized by selective deposition of specific wax compounds into individual cuticular compartments through multiple mechanisms, including in vivo wax synthesis or transport, dynamic phase separation between epicuticular waxes and the intracuticular waxes, in vitro polymerization, and retro transportation into epidermal cell wall or protoplast for further transformation. Our data suggest that modifications of a limited set of specific wax components from individual cuticular compartments are sufficient to alter cuticular transpiration barrier properties.

Environmental parameters and microbial community profiles as indication towards microbial activities and diversity in aquaponic system compartments

BackgroundAn aquaponic system couples cultivation of plants and fish in the same aqueous medium. The system consists of interconnected compartments for fish rearing and plant production, as well as for water filtration, with all compartments hosting diverse microbial communities, which interact within the system. Due to the design, function and operation mode of the individual compartments, each of them exhibits unique biotic and abiotic conditions. Elucidating how these conditions shape microbial communities is useful in understanding how these compartments may affect the quality of the water, in which plants and fish are cultured.ResultsWe investigated the possible relationships between microbial communities from biofilms and water quality parameters in different compartments of the aquaponic system. Biofilm samples were analyzed by total community profiling for bacterial and archaeal communities. The results implied that the oxygen levels could largely explain the main differences in abiotic parameters and microbial communities in each compartment of the system. Aerobic system compartments are highly biodiverse and work mostly as a nitrifying biofilter, whereas biofilms in the anaerobic compartments contain a less diverse community. Finally, the part of the system connecting the aerobic and anaerobic processes showed common conditions where both aerobic and anaerobic processes were observed.ConclusionDifferent predicted microbial activities for each compartment were found to be supported by the abiotic parameters, of which the oxygen saturation, total organic carbon and total nitrogen differentiated clearly between samples from the main aerobic loop and the anaerobic compartments. The latter was also confirmed using microbial community profile analysis.

Multi-Compartment Spatially-Derived Radiomics From Optical Coherence Tomography Predict Anti-VEGF Treatment Durability in Macular Edema Secondary to Retinal Vascular Disease: Preliminary Findings.

Objective: Diabetic macular edema (DME) and retinal vein occlusion (RVO) are the leading causes of visual impairments across the world. Vascular endothelial growth factor (VEGF) stimulates breakdown of blood-retinal barrier that causes accumulation of fluid within macula. Anti-VEGF therapy is the first-line treatment for both the diseases; however, the degree of response varies for individual patients. The main objective of this work was to identify the (i) texture-based radiomics features within individual fluid and retinal tissue compartments of baseline spectral-domain optical coherence tomography (SD-OCT) images and (ii) the specific spatial compartments that contribute most pertinent features for predicting therapeutic response. Methods: A total of 962 texture-based radiomics features were extracted from each of the fluid and retinal tissue compartments of OCT images, obtained from the PERMEATE study. Top-performing features selected from the consensus of different feature selection methods were evaluated in conjunction with four different machine learning classifiers: Linear Discriminant Analysis (LDA), Quadratic Discriminant Analysis (QDA), Random Forest (RF), and Support Vector Machine (SVM) in a cross-validated approach to distinguish eyes tolerating extended interval dosing (non-rebounders) and those requiring more frequent dosing (rebounders). Results: Combination of fluid and retinal tissue features yielded a cross-validated area under receiver operating characteristic curve (AUC) of 0.78±0.08 in distinguishing rebounders from non-rebounders. Conclusions: This study revealed that the texture-based radiomics features pertaining to IRF subcompartment were most discriminating between rebounders and non-rebounders to anti-VEGF therapy. Clinical Impact: With further validation, OCT-based imaging biomarkers could be used for treatment management of DME patients.

Expression of placental glucose transporter proteins in pregnancies complicated by fetal growth disorders.

During pregnancy fetal growth disorders, including fetal macrosomia and fetal growth restriction (FGR) are associated with numerous maternal-fetal complications, as well as due to the adverse effect of the intrauterine environment lead to an increased morbidity in adult life. Accumulating evidence suggests that occurrence of fetal macrosomia or FGR, may be associated with alterations in the transfer of nutrients across the placenta, in particular of glucose. The placental expression and activity of specific GLUT transporters are the main regulatory factors in the process of maternal-fetal glucose exchange. This review article summarizes the results of previous studies on the expression of GLUT transporters in the placenta, concentrating on human pregnancies complicated by intrauterine fetal growth disorders. Characteristics of each transporter protein found in the placenta is presented, alterations in the location and expression of GLUT isoforms observed in individual placental compartments are described, and the factors regulating the expression of selected GLUT proteins are examined. Based on the above data, the potential function of each GLUT isoform in the maternal-fetal glucose transfer is determined. Further on, a detailed analysis of changes in the expression of glucose transporters in pregnancies complicated by fetal growth disorders is given, and significance of these modifications for the pathogenesis of fetal macrosomia and FGR is discussed. In the final part novel interventional approaches that might reduce the risk associated with abnormalities of intrauterine fetal growth through modifications of placental GLUT-mediated glucose transfer are explored.

Reproducibility of fluid status measured by bioelectrical impedance analysis in healthy volunteers: akey requirement to monitor fluid status in the intensive care unit.

Little is known about the use of bioelectrical impedance analysis (BIA) in critically ill patients. The objective of this study was to evaluate the reproducibility of BIA measurements by comparing non-dominant versus dominant body-side measurements at 2 separate time points in healthy volunteers in order to extrapolate key elements that may be of relevance in critically ill patients. Aprospective observational validation experiment was carried out in healthy volunteers. Full-body and segmental multiple frequency BIA measurements were carried out at the non-dominant and the dominant body side, consecutively, and on 2 separate occasions within 1 week. Parameters of interest were both raw data (impedance and phase angle) at the individual frequencies (5-50-100-200 kHz) and body fluid compartment volume estimations (total body water, extracellular water volume, intracellular water volume, volume excess). Atotal of 42 measurements were performed in 22 volunteers. Median (interquartile range) age and time between measurements was 26 years (24; 35) and 2.07 days (1.00; 2.99), respectively. The intraclass-correlation coefficients (ICCs) for body fluid compartment volumes estimated by full-body BIA, were greatly above 90%, showing excellent agreement, except for volume excess which showed moderate agreement. Full-body raw impedance and phase angle measurements showed highly variable and much lower ICCs. For both estimated body fluid compartment volumes and raw measurements, segmental BIA showed also highly variable and low ICCs. Overall this study showed that in healthy volunteers, BIA-derived fluid parameters are reproducible, and differences can be attributed to the changes in clinical status.

Growth performance and nutrient digestibility of weaner rabbits fed African locust bean (Parkia biglobosa) fruit pulp

The need to seek for alternative feed materials to replace maize as source of energy to support growth in weaner rabbits necessitated this study. A feeding trial of 84 days using unsexed rabbit crosses (New Zealand white and Chinchilla) was conducted to assess the replacement value of African Locust bean fruit pulp for maize in weaner rabbits diet. Five diets were formulated, such that 100% maize and 0.00% fruit pulp of African fruit pulp at (T1), 75% maize and 25% inclusion of African locust bean at (T2), 50% maize and 50% African locust bean pulp as (T3), 25% maize and 75% African fruit pulp at (T4) while (T5) contain 100% fruit pulp of African locust bean and 0.00% maize, respectively. Seventy-five (75) weaner rabbits with average initial weight between 610.67g - 622.60g were randomly allocated in a complete randomized design (CRD) experiment to the five diets with fifteen (15) weaner rabbits in each treatment which were subdivided into three (3) replicates having five (5) weaner rabbits in each replicate. The weaner rabbits were housed in individual cage compartment. Data collected were subjected to one-way analysis of variance (ANOVA). Proximate and Phytochemical composition of sun dried fruit pulp of African locust bean used in the diets showed dry matter content of 91.38 %, crude protein 6.70 %, crude fiber 11.21 % Ether extract 5.04 %, ash 5.02 and nitrogen free extract 63.41 %. The metabolizable energy contained in the ingredient was 3079.41 ME/Kcal/Kg. Proximate composition of the experimental diets fed to weaner rabbits carried out indicated that T1 had dry matter content of 93.28 %, crude protein of 19.80 % and 12.41 % of crude fiber. Ether extract was higher in T5 with 8.48 %, while nitrogen free extract contained in T3 had the highest value of 51.17 %. However, ash content in T5 has the highest value of 7.83 %. The metabolizable energy in all the dietary treatment shows that T5 had the highest value of 3013.21 ME/Kcal/Kg. Nutrient digestibility in weaner rabbits showed crude protein digested during the experimental work in the treatment ranged from 69.46 % to 73.73 %. Crude fiber digested was between the ranges of 42.20 % to 58.12 % in all the treatments. Ash content ranged from 50.18 % to 56.35. T5 has the highest percent nutrient of ether extract digested with 83.36 % while the lowest value was obtained in T1 with 58.99 respectively. Values in nitrogen free extract ranged from 65.20 % to 72.43 % and showed a significance difference (P< 0.05) in all the treatment. Data were collected on daily feed intake and weekly weight. Feed conversion ratio, protein efficiency ratio, energy efficiency ratio, nutrient digestibility and feed cost benefit N (kg) were calculated. Weaner rabbits fed 75% maize and 25% African fruit pulp (T2) consumed more feed and significantly gained more weight (P < 0.05) than those fed the other diets. Feed conversion, protein efficiency ratio and energy efficiency ratio were not significantly (P>.0.05) influenced by the treatment diets while nutrient digestibility showed a significance difference (P < 0.05) in all the treatments. Average feed cost/weight gain of production with maize was estimated at N311.25 in T1 while the African locust bean fruit pulp was N161.81 in T5, a cost difference of N149.44, this resulted to a progressive reduction in the cost treatment of feed with increase in the level of fruit pulp of African locust bean meal. The results of this study indicated that when 25 % fruit pulp of African locust replaced maize, the rabbits performed better than the control diet and also of more economical option.

Fully automated 3D segmentation of MR-imaged calf muscle compartments: Neighborhood relationship enhanced fully convolutional network

Automated segmentation of individual calf muscle compartments from 3D magnetic resonance (MR) images is essential for developing quantitative biomarkers for muscular disease progression and its prediction. Achieving clinically acceptable results is a challenging task due to large variations in muscle shape and MR appearance. In this paper, we present a novel fully convolutional network (FCN) that utilizes contextual information in a large neighborhood and embeds edge-aware constraints for individual calf muscle compartment segmentations. An encoder–decoder architecture is used to systematically enlarge convolution receptive field and preserve information at all resolutions. Edge positions derived from the FCN output muscle probability maps are explicitly regularized using kernel-based edge detection in an end-to-end optimization framework. Our method was evaluated on 40 T1-weighted MR images of 10 healthy and 30 diseased subjects by fourfold cross-validation. Mean DICE coefficients of 88.00–91.29% and mean absolute surface positioning errors of 1.04–1.66 mm were achieved for the five 3D muscle compartments.

Heterogeneity of Sensory-Induced Astrocytic Ca2+ Dynamics During Functional Hyperemia.

Astrocytic Ca2+ fluctuations associated with functional hyperemia have typically been measured from large cellular compartments such as the soma, the whole arbor and the endfoot. The most prominent Ca2+ event is a large magnitude, delayed signal that follows vasodilation. However, previous work has provided little information about the spatio-temporal properties of such Ca2+ transients or their heterogeneity. Here, using an awake, in vivo two-photon fluorescence-imaging model, we performed detailed profiling of delayed astrocytic Ca2+ signals across astrocytes or within individual astrocyte compartments using small regions of interest next to penetrating arterioles and capillaries along with vasomotor responses to vibrissae stimulation. We demonstrated that while a 5-s air puff that stimulates all whiskers predominantly generated reproducible functional hyperemia in the presence or absence of astrocytic Ca2+ changes, whisker stimulation inconsistently produced astrocytic Ca2+ responses. More importantly, these Ca2+ responses were heterogeneous among subcellular structures of the astrocyte and across different astrocytes that resided within the same field of view. Furthermore, we found that whisker stimulation induced discrete Ca2+ “hot spots” that spread regionally within the endfoot. These data reveal that astrocytic Ca2+ dynamics associated with the microvasculature are more complex than previously thought, and highlight the importance of considering the heterogeneity of astrocytic Ca2+ activity to fully understanding neurovascular coupling.

Standard trees versus underwood: Historical patterns of tree taxon occurrence in coppice forests

AbstractQuestionsCoppice woods were once widespread in Europe. It is usually assumed that underwood tree taxon composition was not directly influenced by people, whereas especially Quercus was promoted among standard trees. However, no work has quantitatively tested these assumptions. Our main question was whether there were any patterns in our data to suggest that certain trees occurred more frequently as standards than as underwood, possibly as a result of management decisions.LocationCzech Republic.MethodsWe constructed a database covering 12,000 hectares of forests around AD 1900. We recorded the occurrence of tree taxa for standards and underwood in 1,507 compartments in 21 forest units. We visualised patterns in these data by bar graphs and chord diagrams. We analysed correlation patterns through association analysis and co‐correspondence analysis.ResultsThere was a statistically significant association between the occurrences of standards and underwood in individual compartments for Quercus, Alnus, Carpinus, Fagus, Larix, Populus tremula and Tilia. Only Quercus standards and underwood showed a positive association. The taxon composition of standards and underwood in forest compartments was statistically significantly correlated.ConclusionsNo simple answer can be given to the question whether or not the taxon composition of standards and underwood in coppice woods differed as a result of management. At the level of individual tree taxa, some trees were more common as standards while others, as underwood. However, the prevalence of Quercus among standards was not confirmed because in our study area Quercus was a common underwood tree as well. At the same time, no significant differences were apparent in the taxon composition of standards and underwood in individual forest compartments. It seems difficult to differentiate between the effects of human selection and indirect selection through natural properties and environmental factors. Our results also highlight the need to carefully consider the properties of written historical sources.

Computed Tomographic Assessment of Individual Paranasal Sinus Compartment and Nasal Conchal Bulla Involvement in 300 Cases of Equine Sinonasal Disease.

Background: Computed tomographic (CT) imaging has allowed new anatomical studies and detailed clinical imaging of the complex, overlapping equine sinonasal structures. Despite the widespread use of CT, no study has specifically identified which compartments are most commonly affected with sinus disorders. CT has also shown the presence of intercurrent, ipsilateral nasal disorders, especially infection of the nasal conchal bullae (NCB) in many cases of sinus disease, but the frequency of intercurrent NCB infections has not been reported.Objectives: To identify which sinus compartments are most commonly affected in horses with clinical sinus disorders and to record the prevalence of NCB involvement in such cases.Study Design: Retrospective examination of CT images of horses with confirmed unilateral sinus disease.Methods: The CT images of 300 horses, from three different equine hospitals with clinically confirmed sinus disease [mainly dental (53%) and primary sinusitis (25.7%)] were retrospectively examined to determine which sinus compartments and NCBs were affected.Results: The rostral, more dependent sinus compartments were most commonly involved, i.e., the rostral maxillary sinus in 284/300 (94.7% affected) and the ventral conchal sinus (87% affected). The caudal maxillary sinus (65.3%), dorsal conchal sinus (52.7%), frontal sinus (26%), ethmoidal sinus (32%) and sphenopalatine sinus (28.7%) were less commonly affected. There was infection or destruction of the ipsilateral NCBs in 56% of horses with sinus disorders, including the ventral NCBs in 42.3%, dorsal NCBs in 29% and both NCBs in 18% of cases.Main Limitations: The horses with sinonasal disease that underwent head CT imaging include more problematic cases and horses of high value, rather than the general horse population.Conclusions: The more dependant (i.e., the RMS and VCS) sinus compartments are most commonly involved in sinus disorders, with the RMS involved in nearly every case. The more dorsally located sinuses (i.e., caudal group) are less commonly involved. Many horses with sinus disease also have disorders of their nasal conchal bullae and so the term sinonasal disease seems appropriate for these disorders.

Isolated and combined effects of thermal stress and copper exposure on the trophic behavior and oxidative status of the reef-building coral Mussismilia harttii.

Global warming and local disturbances such as pollution cause several impacts on coral reefs. Among them is the breakdown of the symbiosis between host corals and photosynthetic symbionts, which is often a consequence of oxidative stress. Therefore, we investigated if the combined effects of thermal stress and copper (Cu) exposure change the trophic behavior and oxidative status of the reef-building coral Mussismilia harttii. Coral fragments were exposed in a mesocosm system to three temperatures (25.0, 26.6 and 27.3°C) and three Cu concentrations (2.9, 5.4 and 8.6μgL-1). Samples were collected after 4 and 12 days of exposure. We then (i) performed fatty acid analysis by gas chromatography-mass spectrometry to quantify changes in stearidonic acid and docosapentaenoic acid (autotrophy markers) and cis-gondoic acid (heterotrophy marker), and (ii) assessed the oxidative status of both host and symbiont through analyses of lipid peroxidation (LPO) and total antioxidant capacity (TAC). Our findings show that trophic behavior was predominantly autotrophic and remained unchanged under individual and combined stressors for both 4- and 12-day experiments; for the latter, however, there was an increase in the heterotrophy marker. Results also show that 4 days was not enough to trigger changes in LPO or TAC for both coral and symbiont. However, the 12-day experiment showed a reduction in symbiont LPO associated with thermal stress alone, and the combination of stressors increased their TAC. For the coral, the isolated effects of increase in Cu and temperature led to an increase in LPO. The effects of combined stressors on trophic behavior and oxidative status were not much different than those from the isolated effects of each stressor. These findings highlight that host and symbionts respond differently to stress and are relevant as they show the physiological response of individual holobiont compartments to both global and local stressors.

Controllable synthesis of patchy particles with tunable geometry and orthogonal chemistry

HypothesisSelf-assembly using anisotropic colloidal building blocks may lead to superstructures similar to those found in molecular systems yet can have unique optical, electronic, and structural properties. To widen the spectrum of achievable superstructures and related properties, significant effort was devoted to the synthesis of new types of colloidal particles. Despite these efforts, the preparation of anisotropic colloids carrying chemically orthogonal anchor groups on distinct surface patches remains an elusive challenge. ExperimentsWe report a simple yet effective method for synthesizing patchy particles via seed-mediated heterogeneous nucleation. Key to this procedure is the use of 3-(trimethoxysilyl)propyl methacrylate (TPM) or 3-(trimethoxysilyl)propyl acrylate (TMSPA), which can form patches on a variety of functional polymer seeds via a nucleation and growth mechanism. FindingsA family of anisotropic colloids with tunable numbers of patches and patch arrangements were prepared. By continuously feeding TPM or TMSPA the geometry of the colloids could be adjusted accurately. Furthermore, the patches could be reshaped by selectively polymerizing and/or solvating the individual colloidal compartments. Relying on the chemically distinct properties of the TPM/TMSPA and seed-derived domains, both types of patches could be functionalized independently. Combining detailed control over the patch chemistry and geometry opens new avenues for colloidal self-assembly.

Microdissected pancreatic cancer proteomes reveal tumor heterogeneity and therapeutic targets.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a relative paucity of cancer cells that are surrounded by an abundance of nontumor cells and extracellular matrix, known as stroma. The interaction between stroma and cancer cells contributes to poor outcome, but how proteins from these individual compartments drive aggressive tumor behavior is not known. Here, we report the proteomic analysis of laser-capture microdissected (LCM) PDAC samples. We isolated stroma, tumor, and bulk samples from a cohort with long- and short-term survivors. Compartment-specific proteins were measured by mass spectrometry, yielding what we believe to be the largest PDAC proteome landscape to date. These analyses revealed that, in bulk analysis, tumor-derived proteins were typically masked and that LCM was required to reveal biology and prognostic markers. We validated tumor CALB2 and stromal COL11A1 expression as compartment-specific prognostic markers. We identified and functionally addressed the contributions of the tumor cell receptor EPHA2 to tumor cell viability and motility, underscoring the value of compartment-specific protein analysis in PDAC.

Emerging heterogeneous compartments by viruses in single bacterial cells

Spatial organization of biological processes allows for variability in molecular outcomes and coordinated development. Here, we investigate how organization underpins phage lambda development and decision-making by characterizing viral components and processes in subcellular space. We use live-cell and in situ fluorescence imaging at the single-molecule level to examine lambda DNA replication, transcription, virion assembly, and resource recruitment in single-cell infections, uniting key processes of the infection cycle into a coherent model of phage development encompassing space and time. We find that different viral DNAs establish separate subcellular compartments within cells, which sustains heterogeneous viral development in single cells. These individual phage compartments are physically separated by the E. coli nucleoid. Our results provide mechanistic details describing how separate viruses develop heterogeneously to resemble single-cell phenotypes.

Cooling Performance Analysis of the Lab-Scale Hybrid Oyster Refrigeration System

Compared with the waste-to-heat and electricity-based hybrid refrigeration system, the innovative lab-scale refrigeration system integrated with the DC and AC cooling units that able to use solar and electricity as energy resources. Previous studies found that temperature control and uniform temperature distribution in refrigeration systems are both critical factors reducing vibrio growth on raw oysters and saving energy consumption. Therefore, this refrigeration system also equipped a specially designed divider and was used to test various air circulation strategies to achieve uniform temperature distribution in six individual compartments. The objective is to investigate and evaluate the effects of air circulation strategies and operating conditions on the cooling performance, including temperature distribution, standard deviation of compartment temperatures, and cooling time using a factorial design method. Results indicated the maximum temperature difference between the compartments was 8.9 ± 2.0 °C, 6.7 ± 2.0 °C, and 4.8 ± 2.0 °C in the scenarios of no air circulation, natural air circulation, and combined natural and forced air circulation, respectively. The interaction of fan location and fan direction showed a significant effect on the compartment temperatures while there was no significant effect on cooling time. A circulation fan on the lower part of the 12-volt section with an air supply from the 12- to 110-volt section was determined as the optimal condition to achieve relatively uniform temperature distribution. Refrigeration system also achieved a cooling temperature of 7.2 °C within 150 min to meet regulations. To that end, the innovative hybrid oyster refrigeration system will benefit oyster industries, as well as the aquaculture farmers in terms of complying with regulations and energy savings.

First Report of Collar and Stem Rot Caused by Rhizoctonia solani AG1-IA on Sesbania sesban in India.

Sesbania sesban (L.) Merr., (family Fabaceae) commonly called as "dhaincha" in India, is a multi-purpose crop used as a cover crop, as green manure, in the paper industry as well as animal fodder. The leaves of Sesbania contain high amounts of pinitol, which acts as an anti-diabetic agent (Misra and Siddiqi 2004). During July/August from 2017 to 2019, Sesbania plants exhibiting typical Rhizoctonia-like symptoms, including collar rot, wilting, and necrotic lesions on stems were regularly observed at ICAR-Central Potato Research Institute Regional Station, Meerut, Uttar Pradesh. The disease incidence ranged between 5 and 10% in a Sesbania crop being grown on 25 ha in Sesbania-potato rotation. Ten diseased plants were collected from different fields and brought to the laboratory for diagnosis. Affected stem pieces approximately 5 mm in size were surface sterilized with 2% sodium hypochlorite, washed twice in sterilized water and air dried. Four diseased pieces per plate were inoculated on 2% water agar amended with 2% streptomycin sulfate and incubated at 28±1℃ in the dark. All four affected pieces began to produce Rhizoctonia-like colonies after 48 h of incubation and in total eight isolates were purified and stored at 4℃ for further use. The colonies of eight isolates were evaluated and all were whitish during early growth and became light brown after 72 h. Dark-brown sclerotia appeared in the random pattern on PDA after 120 h. Microscopic observations showed that all isolates had hyphal branching at right angles with slight constriction at the base of the branch, presence of dolipore septum near the branching and multinucleate individual hyphae compartments (Sneh 1991). Based on these morphological characteristics, the fungus was identified as Rhizoctonia solani. All isolates were further characterized to determine anastomosis group (AG) by pairing with a known AG tester of R. solani AG-1-IA (ITCC 7650), AG-1-IB (ITCC 5650), and AG-3 (RS-20) procured from Indian Type Culture Collection, Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi and ICAR-Central Potato Research Institute, Shimla, Himachal Pradesh, India, respectively. All eight isolates showed positive anastomosis with a known AG-1-IA tester isolate while no anastomosis was observed with other known tester isolates (Carling 1996). Furthermore, a single ~265 bp amplicon was amplified with AG-specific primer, which was specific to R. solani AG1-IA group; confirms the AG-specific identity of the isolates (Matsumoto 2002). Amplification was not observed with AG1-IB, AG3 and AG2 specific primers (Khodayari et al. 2009). The selected four isolates were molecularly characterized by amplifying the internal transcribed spacer (ITS) and ribosomal DNA (rDNA) 5.8s regions by polymerase chain reaction using ITS1 and ITS4 primer pairs (White et al. 1990). The nucleotide BLAST (BLASTn) analysis of the resulting four sequences i.e. GenBank acc. no. MT105386, MT105387, MT105388, and MT105389 supported the identification of the isolates as AG-1-IA sub-group and showed 95.12%, 98.93%, 96.79%, and 98.04%, respectively, sequence homology with known cultures of R. solani AG1-IA isolated from rice in China (KC285893), and India (MK481078). To confirm pathogenicity, Sesbania plants were grown in pots and maintained in the greenhouse at 25℃ with a 12-h-light/dark photoperiod. After 35 to 40 days of growth, the stems of ten Sesbania plant were artificially inoculated with PDA plugs containing R. solani mycelia (Jia et al. 2007) and covered with aluminium foil. Plants inoculated with noncolonized agar plugs served as control. After 96 h of incubation, all the plants inoculated presented the typical collar and stem rot symptoms. No symptoms were observed in the control plants. R. solani was re-isolated cent percent from these ten infected plants fulfilling Koch's postulates. R. solani AG1-IA has been reported to cause sheath blight and banded leaf and sheath blight diseases of rice and maize, respectively (Ogoshi 1987). To our knowledge, this is the first report of Sesbania sesban infected by R. solani AG1-IA, and serve as a host for the pathogen. The result from our findings will be helpful for planning of crop rotations in an agro-ecosystem.

Digestion of Whole Mouse Eyes for Multi-Parameter Flow Cytometric Analysis of Mononuclear Phagocytes.

The innate immune system plays important roles in ocular pathophysiology including uveitis, diabetic retinopathy, and age-related macular degeneration. Innate immune cells, specifically mononuclear phagocytes, express overlapping cell surface markers, which makes identifying these populations a challenge. Multi-parameter flow cytometry allows for the simultaneous, quantitative analysis of multiple cell surface markers in order to differentiate monocytes, macrophages, microglia, and dendritic cells in mouse eyes. This protocol describes the enucleation of whole mouse eyes, ocular dissection, digestion into a single cell suspension, and staining of the single cell suspension for myeloid cell markers. Additionally, we explain the proper methods for determining voltages using single color controls and for delineating positive gates using fluorescence minus one controls. The major limitation of multi-parameter flow cytometry is the absence of tissue architecture. This limitation can be overcome by multi-parameter flow cytometry of individual ocular compartments or complimentary immunofluorescence staining. However, immunofluorescence is limited by its lack of quantitative analysis and reduced number of fluorophores on most microscopes. We describe the use of multi-parametric flow cytometry to provide highly quantitative analysis of mononuclear phagocytes in laser-induced choroidal neovascularization. Additionally, multi-parameter flow cytometry can be used for the identification of macrophage subsets, fate mapping, and cell sorting for transcriptomic or proteomic studies.

Coupling Structural and Adsorption Properties of Metal-Organic Frameworks: From Pore Size Distribution to Pore Type Distribution.

Metal-organic frameworks (MOFs) attract a rapidly growing attention across the disciplines due to their multifarious pore structures and unique ability to selectively adsorb, store, and release various guest molecules. Pore structure characterization and coupling of adsorption and structural properties are imperative for rational design of advanced MOF materials and their applications. The pore structure of MOFs represents a three-dimensional network comprised of several types of pore compartments: interconnected cages and channels distinguished by their size, shape, and chemistry. Here, we propose a novel methodology for pore structure characterization of MOF materials based on matching of the experimental adsorption isotherms to in silico-generated fingerprint isotherms of adsorption in individual pore compartments of the ideal crystal. The proposed approach couples structural and adsorption properties, determines the contributions of different types of pores into the total adsorption, and estimates to what extent the pore structure of the sample under investigation is different from the ideal crystal. The MOF pore structure is characterized by the pore type distribution (PTD), which is more informative than the traditional pore size distribution that is based on oversimplistic pore models. The method is illustrated on the example of Ar adsorption at 87 K on hydrated and dehydrated structures of Cu-BTC, one of the most well-known MOF materials. The PTD determined from the experimental isotherm provides an estimate of the crystal fraction in the sample and the accessibility and degree of hydration of different types of pore compartments. In addition, the PTD determined from the experimental adsorption isotherm is used to predict the isosteric heat of adsorption that provides important information on the specifics of adsorption interactions. The results are found to be in excellent agreement with experimental data. Such detailed information about the pore structure and adsorption properties of practical MOF samples cannot be obtained with currently available methods of adsorption characterization.