To address the cracking and leaking of concrete in frozen shaft linings in deep and thick topsoil layers in coal mines, hybrid-fiber-reinforced concrete (HFRC) was developed. First, the composition of the reference concrete was obtained by investigating high-strength concrete commonly used in shaft linings, and two dosages of polyvinyl alcohol fiber (PVAF) and polypropylene plastic steel fiber (PPSF) were obtained by the mixing test. Then, tests of early cracks of concrete were conducted; results showed that HFRC could almost avoid early cracks, exhibiting an advantage in early crack resistance. Thus, HFRC can play a significant role in improving the durability of frozen shaft linings in complex underground environments. Furthermore, a series of mechanical property tests were carried out. The results showed that the compressive strength of HFRC was similar to that of the reference concrete, but the tensile and flexural strength of HFRC was 42.7% and 35.1% higher than that of the reference concrete, respectively. Finally, an analog simulation model test of shaft linings was conducted. The new type of shaft lining structure containing hybrid fibers (HFs) exhibited plastic deformation characteristics under load, and the maximum hoop strain was −3562 με. It addressed the problem of high brittleness of frozen shaft lining structures of ordinary high-strength concrete and improved the toughness and crack resistance. HFRC is an ideal material for frozen shaft lining structures in deep and thick topsoil. When mining underground coal resources, it is necessary to construct tunnels from the ground surface for the transportation of coal, waste, personnel, materials, and equipment, and for ventilation and drainage. This tunnel is called a vertical shaft. A concrete wall is built to maintain its stability, which is termed the shaft lining structure.The artificial freeze method is usually used to construct vertical shafts passing through deep and thick topsoil layers. Through artificial refrigeration, strata around the shaft is turned into frozen soil to form a frozen wall. With the protection of the frozen wall, the shaft can be excavated, and concrete is poured to construct the lining structure for support. When the shaft lining is constructed, artificial refrigeration is stopped, and the frozen wall thaws until it is completely melted. Subsequently, the shaft lining structure of the reinforced concrete protects the shaft from external pressure from water and soil . AccordiOwing to the strong external pressure from soil and water in deep and thick soil layers, frozen shaft lining structures have been designed with high-strength concrete, and the thickness is generally over 1.0 m ,4. HowevThe results of the present study ,18 show A test method for early cracking of concrete, mechanical property tests of concrete, and an analog simulation model test of shaft lining were designed to compare the material and structural mechanical properties of different concretes, so as to verify the superiority of HFRC shaft lining structures in deep and thick topsoil layers. According to the investigation of several shaft lining structures in thick and deep topsoil layers, concrete commonly used in the shaft lining had a compressive strength of over 70 MPa and excellent fluidity .The Dingji coal mine, which belonged to the Huainan Mining Industry (Group) Co. Ltd., planned to construct a new auxiliary shaft to achieve safe and efficient production in the future. The designed inner and outer diameter of the lower section of the new auxiliary shaft were 8.6 m and 9.75 m, respectively (the thickness was 1.15 m). The designed compressive strength of the high-strength concrete of the shaft lining was 70 MPa. The topsoil layer around the shaft was 535 m thick. The materials of the high-strength concrete were selected as the raw materials in the following tests. The mix proportion of the high-strength concrete was selected as the reference mix proportion of concrete in this study. The size and parameters of the shaft lining were selected as the prototype size for the analog simulation model test.P.O 52.5R Portland cement ) was selected, and its main performance indexes are listed in 2/g and a density of 2.89 g/cm3. The silicon powder had a specific surface area of 250,000–350,000 cm2/g. The contents of the three admixtures are listed in Fly ash was obtained from the Pingwei Thermal Power Plant  and was processed into Grade I before addition. Its water demand ratio was 89%, loss on ignition was 0.95%, and fineness was 6.4%. Ultrafine slag was obtained from Sinosteel Building Materials Co., Ltd. . Its powder had a specific surface area of 8000 cmThe composition of reference concrete are listed in Polyvinyl alcohol fibers (PVAF) and polypropylene plastic steel fibers (PPSF), obtained from Jianqing Fiber Company , were used. Their appearances are shown in A plate steel mold was used in the test for the early cracking of concrete. Its size was 600 mm × 600 mm × 63 mm. The four sides of the mold were welded from box irons and fixed with a steel bottom plate by bolts. Double row bolts with a diameter of 6 mm and spacing of 60 mm were arranged in the frame. The long and short bolt lengths were 100 mm and 50 mm, respectively. The surface of the bottom plate was laid with polyethylene film as the isolation layer.An electric fan with a maximum wind speed of 5 m/s was used to blow the poured concrete. An anemometer with an accuracy of ±0.5 m/s was used to monitor wind speed. A thermometer with an accuracy of ±0.5 °C was used to monitor the temperature of the environment. A relative hygrometer with an accuracy of ±1% was used to monitor the relative humidity. A DJCK-2 crack width gauge with a scale of 0.01mm was used to measure the crack width.A Quanta250 scanning electron microscope  was used to observe the internal structures of both the reference concrete and the HFRC.A 2000-kN universal testing machine  and Test Master software  were used for the compressive test and the splitting tensile test.A 1000-kN universal testing machine testing machine  and Test Master software were used for the flexural test.A sidewall confining pressure loading device  was used for the analog simulation model test. The device had six holes for monitor wires to pass through. It was controlled by oil pressure. 32 strain gauges were prepared to paste on the surface of the model and monitor the axial strain and hoop strain during loading. The distribution of the strain gauges is shown in A set of orthogonal tests were carried out to select the optimal contents of the two fibers. The collapse degree of the concrete slurry, the 28-day compressive strength and splitting tensile strength test results are listed in To obtain the optimum combination of the two types of fibers, the variance method was adopted to analyze the results. The results show that the slump and the compressive and tensile strength of HFRC were affected, firstly by the amount of PVAF, and secondly by the amount of PPSF. Finally, the optimum fiber combination was determined as 0.9 kg PVAF and 5.0 kg PPSF in reference concrete of one cubic meter.The tests were carried out in accordance with “Standards for Testing Methods of Long-term Performance and Durability of Ordinary Concrete (GB/T 50082-2009)” .First, the environment temperature was adjusted to 20 ± 5 °C and the relative humidity to 55%–60%. Concrete slurry was then poured into the plate steel mold a and wasAfter forming the specimen for 30 min, the wind speed of the fan was turned to 4–5 m/s at 100 mm above the center of the specimen surface, and the wind direction was adjusted to be parallel to the specimen surface and the crack inducers. Two specimens of reference concrete and HFRC were prepared for the tests. After 24 h (the timer started by adding water for stirring concrete), the cracks were measured. Crack length was measured with a steel ruler and crack width with a DJCK-2 crack width gauge b.A compressive, splitting tensile test and a flexural test were carried out in accordance with “Standard for Test Method of Mechanical Properties in Ordinary Concrete” . The speAccording to the analog simulation theory and the size of the test equipment, the size of the shaft lining model was determined to be 5/59 of the prototype, and raw materials, mix proportion, strength, reinforcement ratio, and other parameters were the same as the prototype.Two 28-d reference concrete and two 28-d HFRC specimens of shaft lining were made. They were then sent to a workshop and their upper and lower annular planes were polished for smoothness. The model specimen is shown in To obtain the stress and strain data for the model specimen during the loading process, strain gauges were placed in corresponding locations on both the internal and the external surfaces of the specimen. The inside surface of the model specimen was divided into a high and a low level, and four equidistant measurement points were marked on the inside surface at each level. Four such points were also marked on the outer surface at each level in the same way. Finally, one vertical and one horizontal gauge was placed at each measurement point to measure axial and hoop strain, respectively. Strain gauges were connected to a computer by monitoring wires. The monitoring wires attached to the strain gauges pasted on the outer surface of the model were drawn from the inside of the loading device and out through the wire holes. The load was measured using a hydraulic sensor attached to the loading device. Before loading onto the device, two rubber rings were placed on the upper and lower end faces of the model specimen. The two rubber rings could be deformed with the specimen in the axial direction when it was being loaded. Rigid bolts were used to constrain the plates to ensure that the model specimens remained in a plane strain state during the loading process. The confining pressure from the soil was simulated by high-pressure oil from a pump. The load was applied stably and step by step. The loading device is shown in crA is the nominal fracture area, i,maxω is the maximum width of crack, and i, il is the length of crack i.In the test method for early cracking of the reference and the HFRC, the nominal total area of cracks in the specimen were calculated using Equation (1), below :(1)Acr=∑crη is the crack reduction coefficient, mcrA is the nominal fracture area of the reference concrete, and fcrA is the nominal fracture area of the HFRC.The crack reduction coefficient is calculated according to Equation (2), as follows:The calculation results of Equations (1) and (2) are listed in The SEM images of 28-d reference concrete and HFRC are shown in The compressive strength and splitting tensile strength of the reference concrete and the HFRC were obtained from the mixing test. ff is the flexural strength (MPa), F is the load(kN), l is the span between supports (mm), b is the thickness of the specimens (mm), and h is the height of the specimens (mm).The flexural strength was calculated using Equation (3), below . The coeAll the results of the three mechanical properties tests are listed in From In addition, as shown in The reference concrete specimens fault seriously and showed obvious failure characteristics under uniaxial compression. However, the HFRC specimens were intact during fracturing, although several vertical cracks appeared, which is similar to the form of fracturing concrete under a multi-axial state. This suggests that the internal fibers of the HFRC bonded well with the concrete matrix, and the deformation of the specimens was effectively restrained. The fibers clearly improved the stress state of the concrete and prevented specimens from failure.As shown in PVAF and the PPSF played an anti-cracking role in the early and late stages of concrete failure. Mixed fibers in the concrete showed a positive hybrid effect. The failure forms of shaft lining model specimens are shown in The parameters and capacities of the shaft lining model are listed in After sorting through the data, the curves of the relationship between the hoop strains and confining loads of the specimens were obtained, as shown in The hoop stress of concrete was calculated using the damage constitutive model, according to “Code for Design of Concrete Structures” . Figure The early crack resistance test showed that there were 26 cracks on the reference concrete specimen but only one short and narrow crack on the HFRC specimen. The fracture reduction coefficient of the HFRC reached 99.74%, compared with the reference concrete, almost completely avoiding the generation of early cracks. It improved the durability of the frozen shaft lining in the underground complex environment in a remarkable manner.The results of the mechanical properties tests show that the HFRC and the reference concrete have similar values of compressive strength. However, the HFRC has a tensile strength and a flexural strength that were 42.7% and 35.1% higher than the reference concrete, respectively, which would be quite beneficial to crack prevention and the seepage resistance of the shaft lining in frozen, deep, and thick topsoil.When the HFRC shaft lining model was damaged, micro-cracks appeared in a limited area; macro-cracks did not appear in a large area as is common in ordinary brittle concrete. Furthermore, these micro-cracks developed slowly from the connections between fibers and the concrete matrix. Therefore, in a practical engineering application, a shaft lining structure made of HFRC can modify the brittleness of high-strength concrete and improve the crack resistance, impermeability, and durability of frozen shaft lining structures.The model test showed that the shaft lining of the HFRC had an obvious plastic flow deformation stage during the loading, with the maximum hoop strain up to −3562 με. The results indicate that the HFRC is clearly tough and can improve the structure of the shaft lining.In this study, large temperature differences between the inner and outer surfaces of the concrete were considered qualitatively, but not quantitatively. In the future, the applicability of the shaft lining structure of HFRC under the condition of large temperature differences should be studied quantitatively to improve the research of this paper.


We identified pdm3m7, a new allele of the pdm3 gene encoding a POU family transcription factor, in a lethality-based genetic screen for modifiers of Wingless (Wg) signaling in Drosophila. Interestingly, pdm3m7 larvae showed slow locomotion, implying neuromuscular defects. Analysis of larval neuromuscular junctions (NMJs) revealed decreased bouton number with enlarged bouton in pdm3 mutants. pdm3 NMJs also had fewer branches at axon terminals than wild-type NMJs. Consistent with pdm3m7 being a candidate wg modifier, NMJ phenotypes in pdm3 mutants were similar to those of wg mutants, implying a functional link between these two genes. Indeed, lethality caused by Pdm3 overexpression in motor neurons was completely rescued by knockdown of wg, indicating that Pdm3 acts upstream to Wg. Furthermore, transient expression of Pdm3 induced ectopic expression of wg-LacZ reporter and Wg effector proteins in wing discs. We propose that Pdm3 expressed in presynaptic NMJ neurons regulates wg transcription for growth and development of both presynaptic neurons and postsynaptic muscles.Wnt is a conserved family of secreted proteins that play diverse roles in tissue growth and differentiation. Identification of transcription factors that regulate  Pituitary-specific positive transcription factor 1, Octamer transcription factor-1, Uncoordinated-86 domain (POU) transcription factors1 belong to a subfamily of homeodomain transcription factors, and are highly conserved in all metazoans5. POU domain consists of two DNA binding domains, POU homeodomain and POU specific domain, and these two domains are linked by a flexible linker6. Based on sequence homology of the POU domain and the linker, POU proteins are grouped into six classes5. POU proteins are often expressed in spatiotemporally restricted patterns during development, implying that they may be specialized for differentiation of specific cells or tissues by activating required signal transduction pathways5.Transcription factors play essential roles by inducing genes during the formation of body plans, organ development, tissue specificity, and generation of diverse cell types. Numerous transcription factors are grouped based on similarity in their sequences and domain structures. Drosophila POU domain motif 3 (Pdm3) protein is reported to function in olfactory receptor neurons (ORNs) by regulating olfactory receptor gene expression and axon targeting, and in ring (R) neurons by regulating the development of ellipsoid body (EB) and axon targeting to EB in the central brain8. Pdm3 is also important for the axon targeting of a type of tracheal dendrite (td) neurons9. In particular, td neurons that normally form synapse in the nerve cord change their target to the central brain by ectopic expression of Pdm3. Besides the neuronal functions of Pdm3, Pdm3 also acts as a repressor of abdominal pigmentation in D. melanogaster10, and plays a role in female-limited color dimorphism in abdomen of D. montium11. Despite these studies, it is still unknown how Pdm3 performs these neuronal and non-neuronal functions.The class VI pdm3f00828 and pdm31 homozygotes exhibit defects in axon targeting, odor perception, and locomotion8. pdm3f00828 allele has insertion of a piggyback element in an intron near the 3′ end of the pdm3 gene, and pdm31 has a premature stop codon in the middle of the coding region that results in the deletion of the POU domain8. We identified a new pdm3 allele, pdm3m7, as a suppressor of lethality induced by Sol narae (Sona) overexpression in a genetic screen. Sona is a fly ADAMTS  whose family members are secreted metalloproteases important for cell proliferation, cell survival and development14. We have shown that Sona positively regulates Wingless (Wg) signaling and is essential for fly development, cell survival, and Wg processing14. Wg is a prototype of Wnt family that initiates signal transduction cascade as extracellular signaling proteins, and activation of Wnt signaling leads to transcriptional induction of multiple genes for regulation of cell proliferation, cell survival, cell fate decision, and cell migration17. Wg is important for the development of all appendages, and the wing imaginal disc has been a great tool to study Wg signaling because Wg secreted from its dorsal-ventral midline is crucial for growth and development of wings18.19. Among multiple types of boutons such as type Ib, Is, II, and III, Wg is secreted at a high level from the glutamatergic type Ib bouton known as the main localization site of Wg protein and Wg signaling components, and is absent or at very low levels in other types of boutons20. Type Ib boutons also have more extensive SSR compared to other bouton types, so are easily detected by the high level of Discs-Large (Dlg) as a postsynaptic marker. Type Ib boutons in NMJs of wg mutants show reduction in bouton number but increase in bouton size24. Components in Wg signaling such as Arrow (Arr) that positively regulates Wg signaling as a coreceptor of Wg also shows its mutant phenotype similar to wg, but Shaggy (Sgg)/GSK3β that negatively regulates Wg signaling as a kinase shows opposite phenotype to wg25. Thus, dynamic regulation of Wg signaling is essential for the development of NMJ.Wg also plays an essential role in the development of NMJ. During larval development, NMJs continue to form synaptic boutons that are specialized structures with axon terminals of motor neurons surrounded by reticular subsynaptical reticulum (SSR) formed by the plasma membrane of postsynaptic muscle26. Futsch is a homolog of mammalian MAP1B, and both Futsch and MAP1B are phosphorylated at a conserved site by Sgg/GSK3β27. The phosphorylated MAP1B does not bind microtubules, which results in reduced stability of microtubules28. Therefore, localization of Futsch at NMJ faithfully reflects the stability of microtubules that is dynamically regulated by Wg signaling. Loss of futsch phenotype is similar to the loss of wg phenotype in NMJ26.Secreted Wg also signals to the presynaptic motor neuron to regulate Futsch, one of the microtubule-associated proteins (MAPs)pdm3 is identified as a suppressor of Sona-induced lethality. Based on the involvement of Sona in Wg signaling and the neuronal role of Pdm3, we specifically studied the roles of Pdm3 in NMJ. Similar to loss of wg, loss of pdm3 in NMJ caused decrease in number but increase in size of boutons. Lethality induced by overexpressed Pdm3 was completely rescued by the knockdown of wg in motor neurons but not vice versa. This indicated that Pdm3 functions upstream to Wg, and prompted us to test whether Pdm3 can induce wg transcription. Indeed, transient expression of Pdm3 in wing discs induced wg transcription and Wg effector proteins. Based on these data, we propose that one of the main functions of Pdm3 in NMJ is to induce wg transcription.We report here that sona, we carried out a lethality-based genetic screen using ethyl methanesulfonate (EMS) as a mutagen based on the late-pupal lethality induced by Sona driven by 30A-Gal4  that contains multiple morphological markers. The meiotic mapping revealed that the lethal region in m7 is located in between the cinnabar (cn) and curved (c)  genes on the right arm of the second chromosome  and their progeny was checked by complementation test with pdm3f00828 and arr2 mutants in order to find flies with a single mutation, pdm3m7 and arrm7.To map the position of the lethal site in suppressor (c) Fig. . Subsequome Fig. . Pdm3 ispdm3m7 has a defective hobo element in the first exon of the pdm3 gene that is upstream of the initiation codon . None of the other suppressors had the hobo element in the pdm3 gene, indicating that insertion of the hobo element is unique to the m7 suppressor, and occurred subsequent to the point mutation in the arr gene. We found that the level of Pdm3 is extremely low in pdm3m7 wing discs, establishing that insertion of the hobo element negatively affects the expression of Pdm3 BSC267 and Df(2 R)Exel6058, were missing only the pdm3 gene in the entire genome, so Df(2 R)BSC267/Df(2 R)Exel6058 flies were used as a deletion mutant of pdm3 in this study . Thus, pdm3m7 is an authentic sona suppressor, and pdm3 shows a positive genetic interaction with sona.We found that not only sona suppressors revealed that sona itself and most suppressors are linked to Wg signaling13, which raised an interesting possibility that Pdm3 is also involved in Wg signaling. We noticed that pdm3 mutant larvae are slow in locomotion  and a postsynaptic marker Dlg to detect Type Ib boutons at muscles 6 and 7 in the 2nd abdominal (A2) and the 3rd abdominal (A3) segments20.Further analysis of pdm31 NMJ was reduced by 40% and 16% compared to the wild-type counterparts in A2 and A3 segments, respectively . Only 3% of wild-type axon branches was fused based on this definition (8 out of 282).Some ons Fig. , which ipdm3 boutons was an abnormally high level of Dlg in 42.9% of NMJs examined  had high level of Dlg. This suggests that SSR is not properly developed in pdm31 NMJs. We also checked the localized pattern of a glutamate receptor GluRIIA in pdm3 and wg boutons. It has been shown that the GluRIIA pattern in wild-type bouton is cluster-like but that in wg boutons is diffused without any clusters20. Consistent with this report, cluster-like pattern of GluRIIA was found in CS, +/wgCX4, and +/wgts control boutons but wgts/wgCX4 boutons showed diffused pattern  axis. We found that Pdm3 is expressed more in the anterior part than posterior part of ventral ganglion, which is consistent with severer pdm3 phenotype in A2 than A3  had other defects such as wing drooping . 100% of ing Fig. , planar ing Fig. , and incpatched (ptc) is expressed /POU class 6 homeobox 1 (POU6F1) mainly expressed in brain and spinal cord. Brn-5 is heavily expressed in embryonic brain but also expressed in adult brain and multiple adult organs such as kidney, lung, testis, and anterior pituitary9. Ectopic expression of Pdm3 induced lethality without exception, indicating that expression of Pdm3 in fly tissues is generally repressed in vivo in order to express Wg under the strict spatiotemporal control. An important question is whether Pdm3 directly transcribe wg. We found that wg transcription is induced only after 36 hours of transient overexpression of Pdm3. It is possible that the level of Pdm3 needs to be over a threshold to induce wg transcription. Alternatively, Pdm3 may need to turn on other components to indirectly induce wg transcription. DNA sequence of Brn-5 binding site has been reported41, so analysis on wg and wnt regulatory regions will help understand the mechanism of wnt induction by Pdm3 and Brn-5.Most of Pdm3 functions identified so far are related to the maturation of neurons such as olfactory neurons, R neurons and td neurons as well as their postsynaptic partnerspdm3 and wg mutants. Therefore, pdm3 and wg may play more prominent roles in the A2 than the A3 segment. In fact, the level of Pdm3 was higher in the anterior region than the posterior region of ventral ganglion, which suggests that more Wg may be present in the NMJs of anterior abdominal segments. Consistent with this idea, the number of type Ib boutons in the A2 segment was 1.8 times more than A3 segment. One difference between pdm3 and wg mutants is the lack of certain phenotypes in the A3 segment of pdm3 NMJs: the size of boutons and the number of axon terminals in A3 were not affected in pdm3 mutant. It is possible that Pdm3 turns on both common and segment-specific genes besides wg, and A3 segment-specific components may alleviate the loss of wg phenotype in the A3 segment. Similarly, other proteins induced by Pdm3 may also play important roles in NMJ growth, differentiation and maintenance. In fact, multiple signaling pathways including Glass-bottom-boat (Gbb) pathway also play roles in NMJ development43. Gbb is secreted from muscles and induces development of both pre- and post-synaptic structures, similar to Wg signaling.We consistently found more significant NMJ phenotypes in A2 than A3 in both hobo element in the pdm3m7 allele. The hobo element belongs to Ac family found in maize and has short inverted terminal repeats44. Laboratory and wild strains of D. melanogaster have average 28 and 22 copies of hobo elements in the genome that are either full-length or defective, respectively46. Because other suppressors identified in the genetic screen using Sona overexpression did not have hobo element in the pdm3 gene, the transposition of the hobo element to the pdm3 gene may have occurred subsequent to the generation of a point mutation in the arr gene by EMS. Since both arr and pdm3 are positively involved in Wg signaling, this hobo insertion may have helped the original arrm7 mutation to further decrease the activity of Wg signaling under the condition of Sona overexpression.We identified a defective pdm3 mutants show minor but consistent defects in planar cell polarity in a restricted region of the wing as well as adhesion between the dorsal and ventral wing blades. Other phenotypes such as wing drooping and premature death were also observed in all pdm3 mutants, but these may be due to malformation of synaptic structures. Pdm3 also plays a role in female-limited color dimorphism in abdomen of D. montium11. The authors found in sexually dimorphic females that the first intron of the pdm3 gene has four tandem sets with predicted binding sites for the HOX gene Abdominal-B (Abd-B) and the sex determination gene doublesex (dsx). Interestingly, it has been shown that Wg expression is repressed by the combinatory work of Abd-B and Dsx proteins47. Taken together, it is possible that transcription of wg and pdm3 is co-repressed by Abd-B and Dsx. Such co-repression of wg and pdm3 transcription may be also required for synaptic growth and differentiation in neurons. Further studies on Pdm3 will help understand how this understudied transcription factor is involved in the final differentiation of various cell types.Besides the neuronal roles of Pdm3, all Ok6-Gal4, BG57-Gal4 and repo-Gal448 were obtained from S.-B. Lee’s lab, and UAS-pdm38, UAS-GFP:wg49, pdm3f°°828 and pdm31 8 were obtained from the labs that produced them. UAS-sona12 is produced in our lab. UAS-wg RNAi (#4889R-3) was obtained from Fly Stocks of National Institute of Genetics. All other lines such as UAS-GFP (#1533), UAS-GFP:CD8 (#5130), UAS-pdm3 RNAi (#26749), Df(2 R)BSC267, DF(2 R)Exel6058, were obtained from Bloomington Drosophila Stock Center.12, and that of NMJs was performed as described with slight modifications51. To obtain larvae for NMJ analysis, flies were cultured at 25 °C except wg mutants at 18 °C. To control the population size of larvae in each vial, eggs were harvested on grape plates and incubated at 25 °C for 24 hours. Then, 30 larvae were transferred to a food vial, and were cultured until the wandering larval stage for dissection. In case of wg larvae, 30 larvae were cultured at 18 °C until dissection. For NMJ staining, larvae were dissected with HL3.1 solution on sylgard plates and fixed for 20 minutes in 5% formaldehyde/HL3.1 solution52 or Bouin’s solution50. Fixed samples were rinsed 3 to 4 times with PBS or HL3.1 and then were blocked in 5% BSA in PBS before antibody treatment. The following antibodies were used: anti-Pdm3 7, anti-β-Gal , anti-Dlg 53, anti-HRP-Cy3 , anti-Wg , anti-Futsch , anti-α-Tubulin , anti-Glutamate receptor IIA ). For GluRIIA staining, the samples were fixed by Bouin’s solution.Immunohistochemical staining of larval wing discs was performed as described19. Therefore, type Ib boutons are defined as round-shaped structures in NMJ branches that are stained with HRP and have high level of Dlg, and only ones that were qualified to this definition were counted as boutons. To obtain images containing boutons, type Ib boutons visualized with HRP and Dlg were taken with 1 μm interval for 7–10 Z stacks at 400X magnification by a confocal laser microscope of Carl Zeiss (NFEC-2010-09-141569) with Zen 2009 program. To manually count number of boutons, all Z stack images were then merged and type Ib boutons at muscles 6 and 7 in A2 or A3 segments in a given image were counted. The number of images used for counting for each genotype was 17–52. To measure size of boutons, images of terminal boutons were captured at 2,000X magnifications and then area of boutons in merged images was measured by Zen 2009 program.We stained the late 3rd instar larvae for a presynaptic marker HRP and a postsynaptic marker Dlg to detect NMJs. Type Ib boutons have more extensive SSR compared to other bouton types , so are easily detected by the high level of Dlg54. Flies were put into a chamber with a grape juice-containing agar plate that has yeast paste at the center. After 4 hours of egg-laying, plates were incubated for 20 hours at 25 °C. Larvae were transferred to distilled water on cover glass, and washed again with distilled water. Water was then removed and the 1:1 mixture of lactic acid and Hoyer’s mount solution was applied. After waiting for about 1 minute, the sample on a cover glass was placed on the slide glass, and then incubated for overnight at 65 °C.The cuticle preparation was performed as described with slight modificationsStatistical analysis was performed using ANOVA to compare different genotypes to a wild-type control within experimental groups. Data are presented as mean ± SEM. To determine statistical significance, t-test and one-way ANOVA of Microsoft Excel 2019 were used.Movie 1.Supplementary Information.


In the present article, the research results of concretes modified with a polymer dispersion of vinyl-benzene and acrylic (PC1) and with a styrene–butadiene dispersion (PC2) are discussed. Concretes were exposed to diesel, non-ethylated fuel, and the standard mixture of light liquids for 1000 h. Concretes modified with polymer dispersions, especially with the styrene–butadiene dispersion (PC2), indicated a smaller degree of liquid penetration into the depth of samples compared to the control concrete. The compressive strength for tested concretes, determined after 1000 h of storage in crude-oil products, in comparison to the strength of samples stored in de-mineralized water was significantly decreased, with the maximum differences equal to 12% for PC2 concrete. Unfortunately, these substances pollute our surrounding environment [Currently, concrete constitutes the most frequently applied structural material. It is used in various conditions, including in environments subjected to the action of crude-oil products as well as so-called “light liquids”. Light liquids are generally organic compounds derived from the processing of crude oil. These substances are used in various areas of the economy, including as fuels and lubricants. According to the definition given in PN-EN 858-1, a light liquid means a liquid with a density that does not exceed 0.95 g/cmironment . The maiironment , floors ironment , as wellironment . The tasConcrete and reinforced concrete structures working in such aggressive environmental conditions must have adequate durability, reaching at least 50 years. Requirements concerning the durability of concrete are formulated in PN-EN 206, while those for reinforced concrete structures are specified in PN-EN 1992-1-1.Long-term exposure to crude-oil products destructively influences the properties of concrete ,10,11,12Tests conducted so far have indicated that crude-oil products affect the concrete physically and chemically, causing the loosening of the contact zone between the aggregate and paste as well as slowing down or even stopping the cement hydration processes ,25. The Due to the destructive impact that crude oil has on concrete and the natural environment that surrounds us, a program of testing concrete modified with polymers has been prepared and carried out. The aim of the present research was to determine the influence of the action of crude-oil products  as well as light liquids on the mechanical (compressive strength) and physical properties  of concretes modified with polymers. An admixture of polymers causes the formation of a non-continuous membrane on cement grains ,29, whicControl concrete (CC);Concrete modified with a polymer dispersion of vinyl-benzene and acrylic (PC1); andConcrete modified with styrene–butadiene dispersion (PC2).The testing program included preparation of three types of concrete: 2Oeq) and sulfate resistance . The chemical composition and physical properties of the cement are shown in Initially, a concrete strength class of C35/45 and a water-tightness degree of W8 were assumed, as is typical for surface concrete used for petrol stations. For practical reasons, a concrete mix with a consistency of S3–S4 was applied. For this purpose, super-plasticizer based on poli-carbo-xylan was added to the mix of the control concrete (CC) and the PC1 mix. No super-plasticizer was applied for the PC2 mix, as the dispersion that was used indicated the fluxing action. All concretes were prepared following Portland cement CEM I 42,5 N-SR3/NA guidelines destined for structures subjected to aggressive environments, in which the concrete is characterized by a low alkali content . The water absorption and water permeability tests were determined in accordance with the procedures set out in the Polish standard PN-B 06250.The water absorption test was carried out on three samples of regular shapes for each type of concrete. The samples were gradually saturated with water up to a constant weight, then they were dried in a drier at 105 °C to a constant weight, and their water absorption was calculated. In accordance with the requirements given in this standard, the absorption of concrete should not be greater than 5% in the case of concrete being directly exposed to external weathering.Furthermore, tests of water permeability through concrete were carried out on six cubic or cylindrical samples. The surface of hardened cement grout was roughened on a 100 mm diameter circle on the wall which was to be affected by water. The remaining surfaces were covered with a waterproof coating. The samples were fixed in the apparatus, loaded with water at a pressure of 200 kPa, and the pressure was increased by another 200 kPa every 24 h. The test ended when three of six samples showed signs of leakage or when the required degree of water resistance was achieved. For example, a W8 degree of water resistance means that the samples were exposed to water at a pressure of 800 kPa for a minimum of 24 kPa, and the samples showed no signs of leakage at that time. The samples were split to determine the depth of water penetration into the concrete after testing.Other tests were carried out in accordance with the requirements of the European standards listed above.Test results for the consistency and density of individual concrete mixes are presented in 3  was equal to 62 MPa. Both polymer-modified concretes indicated slightly lower strengths: 53.5 MPa for concrete PC1 and 55.5 MPa for concrete PC2. All concretes were designed and executed with the same w/c ratio; however, modification of cement composite structures with polymers may cause a slight (up to a few percent) reduction in the compressive strength because of the difficult access of water to cement grains ,23. ThisMoreover, measurements of water absorption and permeability were made according to PN-B 06250. Water absorption was determined for three cubic samples with dimensions equal to 150 mm, while permeability was determined on six samples for each concrete type. The water permeability test was stopped after reaching the water pressure corresponding to the W8 water-tightness class. Then, the samples were split, and the depth of penetration of water into the sample was measured. Obtained results for water absorption and permeability are summarized in All concretes had very low water absorption values: 4.0% for control concrete, 2.5% for the concrete modified with styrene–butadiene polymer, and 3.3% for the concrete modified with vinyl-benzene and acrylic polymer. The obtained values were less than the value of 5% required by PN-B 06250 for concretes exposed to external factors.The depths of penetration for water acting under a pressure of 800 kPa (required for the assumed W8 water resistance level) were 15 mm and 30 mm, respectively, for the styrene–butadiene polymer and vinyl-benzene and acrylic polymer concretes. These values correspond to a high level of water-tightness for concretes exposed to an aggressive environment.90% (m/m) de-mineralized water;0.75% (m/m) sodium hydroxide;3.75% (m/m) sodium orthophosphate;0.50% (m/m) (meta) sodium silicate;3.25% (m/m) sodium carbonate; and1.75% (m/m) sodium metaphosphate.Later in the research program, samples of individual concrete were subjected to resistance tests with crude-oil products and light liquids according to the procedure included in the code PN-EN 858-1. In compliance with this, concrete samples were stored for 1000 h in diesel in accordance with PN-ISO 8217 at a temperature of 23 ± 2 °C, non-ethylated fuel according to PN-EN 228 also at the temperature 23 ± 2 °C, as well as in a liquid mixture in accordance with point 8.1.4.1 of the code PN-EN 858-1 at a temperature of 40 ± 2 °C. The liquid mixture consisted of:In pursuance of specifying this procedure, a part of each sample was stored as a control in de-mineralized water at a temperature of 40 ± 2 °C. After this, the samples were taken out of the water and rinsed and dried in air for 24 h at a temperature equal to 20 °C. Then, the compressive strengths for individual samples were determined. The samples stored in water are shown in As the results show, after 1000 h of storage in de-mineralized water at a temperature of 40 °C, samples indicated further increases in strength for each concrete in comparison to the 28-day strength.From analyzing the results obtained for the different concretes, it may be stated that the strengths of the concretes after storage in crude-oil products and those in the standard mixture of liquids had comparable 28-day strengths. The only exception was for the PC1 concrete, which, after storage in non-ethylated fuel, indicated about a 10% decrease in strength compared to the 28-day value.In comparing the strengths of the tested concretes after storage in crude-oil products and in de-mineralized water, there was a reduction in the compressive strength for the control concrete (CC) and for the concrete modified with styrene–butadiene dispersion (PC2). For the control concrete, the average value of this decrease was about 4%, while for PC2 concrete it was 12%. Such a significant strength reduction for PC2 concrete is connected with the meaningful increase in strength for the samples stored in de-mineralized water. Presumably, cement hydration reactions slowed down during the stage of maturation, and the following elongation may be caused by the presence of polymer membranes that penetrated the composite structure and hindered water migration.However, the differences in the compressive strength levels for particular concretes stored for a standard time, equal to 1000 h, in crude-oil products were so slight that they may be ascribed to measurement variations. Presumably, the storage time for the tight concretes tested was too short to determine any further negative influence of crude-oil products in the environment on the strength parameters of these concretes.Additional samples stored in non-ethylated fuel were split in order to determine the water penetration depth. The greatest depth for non-ethylated fuel, equal to 28 mm, was obtained for the control concrete (CC). The concrete modified with the vinyl-benzene and acrylic (PC1) polymer dispersion indicated a slightly lower depth equal to 20 mm. Non-ethylated fuel penetrated, to the smallest extent, the concrete modified with the styrene–butadiene dispersion (PC2), up to a depth equal to 5 mm. In Tests for 28-day strength indicated that concretes modified with polymer dispersions are characterized with a lower compressive strength than a control concrete (CC). For the PC1 modified concrete, the difference was almost 14%, while for PC2 it was 11%;Concretes modified with polymer dispersions, especially with the styrene–butadiene dispersion (PC2), indicated a smaller degree of liquid penetration into the samples. The penetration depth for non-ethylated fuel for PC2 concrete, after 1000 h of storage, was only 5 mm, while for the control concrete (CC) it was 28 mm. The water penetration depth determined after 28 days of concrete maturation was 60 mm for the control concrete and 15 mm for the PC2 concrete. There was a consistent general trend for the penetration depths for both liquids in the concrete, but the water penetration depths were greater, because pressure was applied on these samples;The compressive strength for the tested concretes determined after 1000 h of storage in crude-oil products was close to the 28-day strength. However, compared to the strength of the samples stored in de-mineralized water, there was a significant decrease in results, with the maximum difference equal to 12% for PC2 concrete.Based on the conducted tests, the following conclusions may be drawn:The performed tests clearly demonstrate the beneficial effect of polymer dispersion on the increase of concrete durability and strength. Polymer films penetrate the cement composite structure, sealing the matrix as well as the contact zone: aggregate–cement paste.


The properties of the hydrogel were thoroughly investigated and the hydrogel was presented as multifunctional. The introduction of Ca2+-TA@SL composites endowed the hydrogel with excellent conductivity, adhesion and ultraviolet (UV) resistance, and improved antioxidant and antibacterial properties. More importantly, the Ca2+-TA@SL-PAM hydrogel electrode could accurately detect physiological signals of human , electromyography (EMG).Inspired by mussel adhesion chemistry and ion electronics, a novel Ca Recently, conductive hydrogels (CHs) have captured the interest of scholars in the field of wearable electronics for emerging human–machine interfaces . CHs int2+ (CaCl2) was introduced into the composite through chelation. As far as we know, TA with a large amount of catechol structures can adsorb biocompatible Ca2+.Here, inspired by mussel adhesion chemistry and ion electronics, we tried to use tannic acid (TA), a dopamine-like substance, to be doped with sulfonated lignin (SL) to construct composites base with adhesive properties. To further enhance the conductivity, Ca2+-TA@SL composites were incorporated into the polyacrylamide (PAM) hydrogel network, which can endow the hydrogel with great adhesion and conductivity. Meanwhile, TA and SL can impart the hydrogel with anti-UV and mild anti-bacterial ability. In addition, the TA can enhance the antioxidant properties of hydrogel. Importantly, the Ca2+-TA@SL-PAM CHs, as a non-invasive adhesive electrode, can achieve the accurate capture of human ECG and EMG signals. We believe that biomass-based CHs can provide more application ideas in the field of human-machine interfaces.Subsequently, the above Ca2+ was absorbed onto the TA@SL composite. Next, acrylamide (AM), ammonium persulfate (APS), N, N’-Methylene bisacrylamide (BIS), and tetramethylethylenediamine (TMEDA) were added to the Ca2+-TA@SL suspension to form Ca2+-TA@SL-PAM hydrogel. The detailed preparation procedure and composition of the Ca2+-TA@SL-PAM hydrogels were listed in Lignin was separated from the black liquor of radiated pine Kraft pulping; SL was prepared via lignin sulfonation . TA@SL c2+-TA@SL-PAM hydrogel were observed by field-emission scanning electron microscopy , the element distribution of TA@SL was observed by Octane Elect EDS . The visible (550 nm) and UV (365 nm) transmittance of the PAM and Ca2+-TA@SL-PAM hydrogels with 3 mm thicknesses were measured using an optical transmittance meter . The hydrogel conductivity was measured by an LCR meter. The conductivity (σ) is calculated by σ = d/RS, in which R is the resistance, and d and S are the hydrogel’ geometry factors corresponding to the length and cross-sectional area, respectively. Detailed antibacterial and antioxidant assay of Ca2+-TA@SL-PAM hydrogel can be found in the The morphology of the SL, TA@SL and Ca visible 50 nm andThe PAM-5 hydrogel was cut into cubes and then adhered to specific skin locations. The ECG (electrocardiograph) and EMG (electromyography) signals of volunteer were collected using a multi-channel bio-signal acquisition and process system .2+-TA@SL-PAM hydrogel. 2+ was successfully absorbed onto the TA@SL composite through complexation with catechol groups on TA@SL composites [2+-TA@SL-PAM hydrogel. mposites . Finally2+-TA@SL-PAM hydrogel, making the resultant hydrogel have enhanced conductivity, adhesion, transparency, antioxidant, antibacterial, and anti-UV properties. As shown in 2+ to achieve the circuit connectivity (0.91 S/m). The interaction of catechol groups on the TA and Ca2+ inspired by mussel adhesion chemistry can impart adhesion to the hydrogel [S. aureus), while the SL based PAM hydrogel and Ca2+-TA@SL-PAM hydrogel can scavenge some of the S. aureus. Notably, PAM-5 hydrogel can even achieve a 98% S. aureus killing rate. 2+-TA@SL-PAM hydrogel with different TA@SL content. Due to the presence of odd electrons, the DPPH radical generates a strong absorption peak at 517 nm [2+-TA@SL-PAM hydrogel. Compared with the control group, the DPPH scavenging rate even reached 75% when the TA@SL content reached 16 mg/mL (2+-TA@SL-PAM hydrogel has effective antioxidant properties. Generally, the antioxidant ability of the hydrogel is derived from the action of TA and lignin; the phenolic hydroxyl group on TA and SL can reduce the free DPPH radical to diphenyl-picrylhydrazine [hydrogel . As pres16 mg/mL g. These tivity 0.1 S/m. Th2+ have a greater advantage in the preparation of non-invasive electronics [2+-TA@SL-PAM hydrogel electrodes was evaluated in detail. As presented in Biocompatible Cactronics . Herein,2+ introduction imparted the hydrogel with electrical conductivity. The fabricated hydrogel electrode could accurately collect the physiological signals from the human, including EMG and ECG. Considering the prominent properties and quick feedback of the human motion, the Ca2+-TA@SL-PAM hydrogel electrode has great potential in visual medical electronics.A novel PAM based ionic conductive hydrogel electrode was successfully developed—inspired by mussel and ion electronics. The TA@SL composite application endowed the hydrogel with excellent adhesion, UV resistance, antioxidant, and moderate antibacterial properties, but without the destruction of transparency. The Ca


We held a multidisciplinary discussion (MDD) about a 61‐year‐old woman who had an interstitial lung disease (ILD) without extrathoracic lesions that met the classification criteria for interstitial pneumonia with autoimmune features (IPAF) and the proposed diagnostic criteria for immunoglobulin G4 (IgG4)‐related respiratory disease (IgG4‐RRD). Clinically, the marked progression of lung‐limited diffuse lesions was consistent with IPAF. Serum IgG4 and rheumatoid factor levels simultaneously increased and did not contribute to a diagnosis. Pathologically, the significant hyperplasia of lymphoid follicles was consistent with rheumatoid arthritis (RA)‐associated ILD. Pulmonary venous occlusions by intimal fibrosis and intimal thickening were not important because these occlusions are found in IgG4‐related lung disease (IgG4‐RLD) and also in IPAF or ILDs related to connective tissue diseases (CTDs). Radiologically, fibrosing shadows that remained in the lung periphery after treatment were compatible with RA‐associated chronic ILD. We concluded that the present case was IPAF that met the proposed diagnostic criteria for IgG4‐RRD. We held a multidisciplinary discussion on a case of interstitial lung disease without extrathoracic lesions that met the classification criteria for interstitial pneumonia with autoimmune features (IPAF) and the proposed diagnostic criteria for immunoglobulin G4 (IgG4)‐related respiratory disease (IgG4‐RRD). We concluded that the present case was IPAF that met the proposed diagnostic criteria for IgG4‐RRD. Interstitial pneumonia with autoimmune features (IPAF) is the term that was proposed in 2015 to describe idiopathic interstitial pneumonias (IIPs) with clinical features that imply some autoimmune background, but do not meet established criteria for connective tissue disease (CTD) Immunoglobulin G4‐related disease (IgG4‐RD) is a systemic fibroinflammatory disorder that is characterized by tumefactive lesions, the intensive infiltration of IgG4‐positive plasma cells, and storiform fibrosis and is generally accompanied by elevated serum IgG4 levels We herein present a case of interstitial lung disease (ILD) diagnosed as IPAF that met the proposed IgG4‐RRD criteria after a detailed multidisciplinary discussion (MDD).A 61‐year‐old woman developed a wet cough with dyspnoea in February 2018. She had a history of type 1 diabetes mellitus (DM) diagnosed in adolescence. She was not taking any medication other than insulin. She was a current smoker (20 cigarettes daily for 41 years). She had no history of alcohol consumption. She had worked as a cook in a restaurant with no history of dust inhalation. Her house was built 17 years earlier and received adequate sunshine. She had an ibuprofen allergy. Her brother had rheumatoid arthritis (RA) and her son had a history of neurosarcoidosis.Although her wet cough attenuated over the course of one month, dyspnoea progressed. In April 2018, she visited a nearby hospital and an interstitial shadow was detected using chest computed tomography (CT). She was subsequently referred to our hospital.Chest CT on referral Fig. A revealeOn admission, her height was 151 cm and weight was 70 kg. She had no fever and her resting respiratory rate was 16/min; however, she had distinct dyspnoea on effort. Her fingers were clubbed, and fine crackles were audible in the lower chest. There were no palpable surface lymph nodes. She had neither arthralgia nor skin lesions that suggested CTDs.SLB was performed by video‐assisted thoracoscopic surgery (VATS). Specimens were taken from a radiologically less affected part and an intensively affected part; the right upper lung S2) and the right lower lung S10), respectively  Fig. G. Some pSince it took four months from her referral to the initiation of treatment, her interstitial shadows and dyspnoea had progressed Fig. B. HoweveWe herein report a case of ILD without extrathoracic lesions that met both the IPAF criteria and the proposed IgG4‐RRD criteria. As difficulties were associated with differentiating between these two conditions, we held an MDD.Clinically, IPAF was more likely than IgG4‐RLD without extrathoracic lesions because it was unusual to detect no extrathoracic IgG4‐related lesions despite the marked progression of extensively diffuse pulmonary lesions. This progression of lung‐limited diffuse lesions was acceptable for IPAF. IgG4‐RLD without extrathoracic lesions is very rare Serologically, difficulties were associated with distinguishing IPAF from IgG4‐RLD without extrathoracic lesions because elevated serum IgG4 levels are not specific to IgG4‐RD; they are also often elevated in CTDs, particularly in RA and Churg–Strauss syndrome (CSS) Pathologically, IPAF was more likely than IgG4‐RLD without extrathoracic lesions because the significant hyperplasia of lymphoid follicles in the present case is sometimes detected in RA‐related ILD Radiologically, it was more reasonable to diagnose the case as IPAF than IgG4‐RLD without extrathoracic lesions. Chest CT before steroid therapy Fig. A, B reveWe concluded that the present case was IPAF that met the proposed IgG4‐RRD criteria. The present case suggests that the concept of IgG4‐RRD needs more refinements. We should proceed with investigation into pathogenesis of the disease. The present case also reveals that we need further study to clarify the relationship between IPAF and IgG4.Appropriate written informed consent was obtained for publication of this case report and accompanying images.This case was presented and discussed at the 20th Tokyo Diffuse Lung Diseases Study Conference held in Tokyo on 26 October 2019 under the joint sponsorship of the Tokyo Diffuse Lung Diseases Study Group and Shionogi Pharmaceutical Co., Ltd. I, Nobuhito Arakawa, declare on behalf of my co‐authors and myself that we have no financial relationship with any commercial entity having an interest in the subject of this manuscript.


Clostridium difficile and antibiotic resistance. Our institution reduced the duration of piperacillin-tazobactam prophylaxis following ILE from 4 to 1 day or less in January 2015. We evaluated short-term outcomes in ILE patients before and after this institutional change. METHODS/STUDY POPULATION: Retrospective cohort study of all ILE patients from 2012 to 2016. We confirmed antibiotic duration directly from nursing medication administration records. The primary outcomes of this study were rates of C. difficile and postoperative pneumonia. Secondary outcomes include other infection, length of hospital stay, and readmission within 30 days. We used logistic regression to analyze impact of days of antibiotics and χ2 or Fischer exact tests for categorical variables. RESULTS/ANTICIPATED RESULTS: Of 104 ILE patients, 40.4% (n=42) were after January 2015, 11.5% developed pneumonia and 5.8% developed C. difficile colitis. ILE patients received more days of antibiotics before the institutional change compared with after . For a 1-day increase in antibiotic duration, the odds of acquiring C. difficile increased significantly by 1.2 (p=0.03). Before compared with after the institutional change, rates of C. difficile were 8.1% Versus 2.4% (p>0.2), rates of pneumonia were 11.3% Versus 11.9% (p>0.2), and length of stay was 10.9 Versus 10.5 days (p>0.2), respectively. DISCUSSION/SIGNIFICANCE OF IMPACT: Institutional policy can have an impact on patient outcomes. Antibiotic stewardship is associated with reduced rates of inpatient C. difficile. Our study suggests reduced antibiotics are not associated with pneumonia, although larger studies are necessary to confirm this finding. Surgeons should consider the benefit of decreased rates of C. difficile before administering prolonged antibiotic prophylaxis following esophagectomies.OBJECTIVES/SPECIFIC AIMS: Ivor-Lewis esophagectomy (ILE) is an invasive surgical procedure with a high incidence of postoperative pneumonia. Antibiotic prophylaxis could reduce respiratory infections but increase


Nitrofurantoin is a commonly used treatment for urinary tract infections with a risk for pulmonary toxicity. We report a case of a 48-year-old woman on a prophylactic regimen of nitrofurantoin who exhibited classic signs of bacterial sepsis including elevated procalcitonin (PCL) and C-reactive protein (CRP) levels two days post-nephrolithotripsy. The microbial analysis did not reveal an infectious cause for the initial symptoms and, subsequently, the patient developed a dry cough, fever, chills, and transient hypoxemia requiring supplemental oxygen. Pulmonary imaging revealed significant abnormal features inconsistent with the patient’s symptoms which indicated an inflammatory/immune reaction to nitrofurantoin. Treatment discontinuation improved the patient’s symptoms and reduced PCL and CRP levels to within normal limits. A high index of suspicion for nitrofurantoin-associated pulmonary toxicity is warranted for patients on a regimen of nitrofurantoin who exhibit severe pulmonary symptoms and elevated PCL and CRP levels with no corresponding infection. Nitrofurantoin is used to treat uncomplicated urinary tract infection, and its use can lead to acute, subacute, or chronic pulmonary toxicity . NitrofuA 48-year-old female with history of hypertension, obesity, polycystic ovarian syndrome, and right-sided nephrolithiasis experienced right-flank severe colicky pain with nausea and non-bloody vomiting within 24 hours post-urology intervention of right ureteroscopy with laser lithotripsy, stone extraction, and double-J stent placement. The patient’s symptoms were secondary to the displacement of a ureteral stent that resulted in spontaneous expulsion of the stent with subsequent mild improvement of symptoms. However, the patient then developed shaking, chills, and a fever of 39.1°C (102.4°F) associated with lethargy and weakness. The patient denied urinary, respiratory, or other symptoms, and a chart review indicated the patient had been discharged home post-surgery on a prophylactic regimen of 100 mg nitrofurantoin administered twice daily.On examination, the patient appeared lethargic and had dry oral mucous membranes. Abdominal exam revealed a soft, non-distended abdomen with mild right flank discomfort. Chest auscultation was clear except for bibasilar mild crackles. Vitals were normal except for mild tachycardia. Lab work showed leukocytosis of 19,000 cells/µL  with elevated lactic acid of 2.5 mmol/L , serum creatinine (SCr) of 1.3 mg/dL (baseline SCr: 1.0 mg/dL), and elevated blood urea nitrogen (BUN) of 26 mg/dL. In addition, PCL and CRP levels were elevated to 25.5 ng/mL and 39.3 mg/dL, respectively, indicating a bacterial infection. Urinalysis was unremarkable for infection or hematuria. To evaluate flank pain, a computed tomography (CT scan) of the abdomen and pelvis without contrast was performed which revealed multiple small calculi in both kidneys with mild right hydronephrosis, ureteral edema, and stranding of the right kidney and collecting system suggesting reactionary changes due to recent instrumentation versus possible infection , and chills associated with transient hypoxia with a pulse oximetry desaturation to 86% requiring 1 L nasal cannula oxygen to maintain normal oxygenation. A chest radiograph revealed an asymmetric left suprahilar density and right infrahilar density consistent with atelectasis, aspiration, or multifocal pneumonia (Figure Comprehensive respiratory viral panel, urine Legionella, and streptococcal pneumonia antigen were negative. Sputum cultures could not be taken because of nonproductive cough. Preliminary urine cultures and blood cultures revealed no growth. Due to atypical presentation of pulmonary symptoms, follow-up CT angiography of the chest revealed no evidence of pulmonary emboli. The ground glass opacities were present throughout both lungs with small bilateral pleural effusions and adjacent bibasilar atelectasis Figure .Pulmonary imaging findings were out of proportion to the clinical condition of the patient. Urology and infectious disease physicians evaluated the patient, and it was considered that nitrofurantoin use was contributing to acute onset of bilateral multifocal pulmonary infiltrates leading to mild transient cough and transient pulse oximetry desaturation associated with physiological systemic inflammatory stress response as evidenced by fever, leukocytosis, and significantly elevated PCL and CRP. After the third day of hospitalization, the piperacillin-tazobactam regimen was discontinued, and the patient was observed off all antibiotics. The patient continued to improve without antibiotics and did not require supplemental oxygen with resolution of symptoms within the next three days. We determined that inflammatory findings present in imaging of the right kidney and collecting system were likely secondary to recent urological instrumentation with laser lithotripsy while the transient right-sided flank pain was probably secondary to displacement of the ureteral stent leading to ureteral colic. Serum inflammatory markers continued to improve without antibiotics after cessation of nitrofurantoin use with decreases in serum PCL and CRP levels from admission to discharge day specifically 25.5 ng/mL to 2.1 ng/mL and 39.3 mg/dL to 14.5 mg/dL, respectively. The patient was discharged after six days of hospitalization, and it was strongly recommended the patient avoid use of nitrofurantoin in the future to prevent recurrence of pulmonary toxicity. The patient continues to have a stable clinical condition after discharge and no complications have been reported since then.Nitrofurantoin is used to treat acute uncomplicated cystitis and as a prophylaxis for recurrent uncomplicated urinary tract infections . Beers cNitrofurantoin pulmonary toxicity is more commonly observed in women compared to men though the high prevalence observed in women is probably secondary to a greater susceptibility of women to recurrent urinary tract infections requiring antibiotic therapy ,3,5,6,11Patients with nitrofurantoin-induced pulmonary toxicity usually present with fever, cough, dyspnea, eosinophilia in blood, and bilateral lung infiltrates seen on chest images ,4-7. TheA majority of patients with nitrofurantoin-associated pulmonary toxicity show rapid improvement with the cessation of nitrofurantoin . HoweverThe pulmonary biopsy is usually not needed in patients with nitrofurantoin-induced toxicity though it may be considered when the patient does not show improvement after discontinuation of nitrofurantoin, and there is a high index of suspicion for alternative diagnoses. In an acute reaction, a pulmonary biopsy usually reveals an acute reaction with eosinophils, interstitial inflammation, vasculitis, and alveolar exudates . HoweverMost patients with acute, subacute, and chronic lung injury respond well to cessation of nitrofurantoin therapy resulting in spontaneous clinical recovery; however, those with severe pulmonary toxicity may need systemic corticosteroid therapy ,5,7,13. Despite a generally good prognosis, fatalities have been reported in the literature. According to the medicine and healthcare products regulatory authority report, from 1963 until 2010, 11 out of 392 patients affected with nitrofurantoin-associated lung injury were reported to have a fatal outcome [Our patient’s presentation of nitrofurantoin-induced toxicity was unique in that it mimicked severe bacterial sepsis including tachycardia, lethargy, high-grade fever, chills, mild nonproductive cough with dyspnea, and hypoxia associated with elevated PCL and CRP levels in addition to leukocytosis without eosinophilia. PCL is considered a more specific marker for bacterial infection compared to other inflammatory markers such as serum leukocyte count, CRP, and erythrocyte sedimentation rate though recent work indicates comparable diagnostic performance with PCL and CRP as sepsis markers ,9. HowevNitrofurantoin-induced pulmonary toxicity can occur as early as within 24 to 48 hours of initiation of drug exposure, and recent medications use by the patient should be explored at the time of presentation for all patients exhibiting respiratory symptoms and sepsis with no identifiable infectious cause. Though elevated PCL is a marker for bacterial sepsis, it can be elevated in nitrofurantoin-pulmonary toxicity without bacterial infection. Early cessation of nitrofurantoin therapy in patients with hyperacute pulmonary toxicity carries a good prognosis and results in early resolution of symptoms without the need for systemic steroids; patients should be recommended to not use nitrofurantoin in the future to avoid toxicity recurrence.Due to the higher prevalence of nitrofurantoin-induced pulmonary toxicity, physicians should be cautious in prescribing nitrofurantoin, and patients should be informed about the possibility of pulmonary toxicity along with other adverse effects at the time of prescription. Providers should also continue long-term surveillance of patients on a long-term nitrofurantoin regimen to ensure early detection of toxicity and to discontinue nitrofurantoin promptly.


Ever-growing globalization and industrialization put forward impending requirements for green and sustainable logistics (G&SL). Over the past decades, G&SL initiatives triggered worldwide deliberations, aiming at easing negative transport externalities and improving supply chain performance. This review-based paper attempts to offer a joint quantitative and qualitative understanding for the overall evolutionary trend, knowledge structure, and literature gaps of the G&SL research field. Employing the science mapping approach, a total of 306 major paper published from 1999 to 2019 were retrieved, elaborated on, and synthesized. Visualized statistics regarding publication years, journal allocation/co-citation, inter-country/institution collaboration, influential articles, co-occurred keywords, and time view clusters of research themes were analyzed bibliographically. On this basis, a total of 50 sub-branches of G&SL knowledge were classified and thematically discussed based on five alignments, namely (i) social-environmental-economic research, (ii) planning, policy and management, (iii) application and practice, (iv) technology, and (v) operations research. Finally, the current knowledge obstacles and the future research opportunities were suggested. The findings contribute to portray a systematic intellectual prospect for the state quo, hotspots, and academic frontiers of G&SL research. Moreover, it provides researchers and practitioners with heuristic thoughts to govern transportation ecology and logistics service quality. Sustainable development has inspired many green and sustainable logistics (G&SL) activities to reduce the negative effects of freight transportation  and imprThe operation management of physical distribution is one of the most significant and challenging sub-issues of the macro supply chain management (SCM) , becausePrevious studies reviewed G&SL from different perspectives. By reviewing 115 papers, Zhang et al.  analyzedHowever, based on the time of publication and the number of papers contained, the existing studies are outdated and incomplete, unable to provide a comprehensive analysis of the booming G&SL research in the past two years. Also, it is more difficult to integrate the multitudinous research directions to build a complete knowledge structure for G&SL. Therefore, it is of great theoretical and practical significance to objectively and quantitatively investigate the overall progress of G&SL.This study aims to conduct a comprehensive review of the global G&SL literature, so as to explore the state-of-the-art, hotspots and research trend, as well as to build the G&SL knowledge classification system. Specifically, first, tracking and analyzing the evolution of the G&SL research field from (i) publication year and journals; (ii) countries, regions, and organizations; (iii) influential documents; (iv) keywords clustering and research themes. Second, establishing the knowledge taxonomy based on the scientometric results. Third, identifying the research gaps and the future research opportunities.The novelty of this study lies in two aspects. One is to integrate the science mapping approach into the systematic literature review process to visualize the relationships among the G&SL literature. Science mapping approach is composed of data mining and bibliographic analysis, which can minimize subjective arbitrariness and grasp useful information to facilitate in-depth thematic analysis. Another is that this study further extends the bibliography to illuminate the emerging knowledge branches, gaps, and agendas in G&SL research, which will contribute to the improvement of G&SL practice and research innovation. The findings are expected to provide researchers and practitioners with a panoramic description and in-depth understanding of G&SL research. Additionally, the proposed knowledge structure can also be used as a handbook-like tool to further collect, analyze, and expand knowledge in the G&SL field and to provide references for other innovative logistics initiatives.The rest is organized as follows. In This review-based study conducted a systematic investigation on the academic development of global G&SL research with the aids of science mapping. Science mapping is a quantitative analysis approach that uses mathematical statistics and visualization techniques to study bibliographic networks  in a specific field . This apIn step 1, the statistics was obtained after a comprehensive retrieval from two electronic databases, Web of Science (WoS) core and Scopus. Two rounds of selection were then performed to refine, classify, and encode the documents. The year publication trend, journal allocation and the most cited articles were described.Journal co-citation analysis: to identify the most cited journals and the research domains they belong to. This analysis helps to reveal the distribution of published journals and cited journals of the reviewed documents, so as to identify popular journals in G&SL research domain. (ii) Countries/organizations collaboration analysis: to visualize the collaborative research network of G&SL among countries and organizations, so that the readers can quickly understand the partnerships between major research communities and institutions around the world. (iii) Document co-citation analysis: to highlight the influential G&SL articles and the corresponding reference relationships. By analysis of the papers with high citation, the emerging trend of scholars’ research interest to G&SL is easier to grasp. (iv) Keywords co-occurrence analysis: to map out the co-occurred time zone of the hotspots G&SL keywords and cluster them into several research themes. Network analysis of co-occurred keywords is used to clarify the knowledge structure of G&SL as well as to present the research hotspots and potential research opportunities in the future.Four scientometric tests were carried out in step 2, namely (i) In step 3, the hierarchical knowledge structure of G&SL was proposed for thematic discussion.ijc is the co-occurred or co-cited times of item i and item j, iW and jW denote the node sizes of item i and item j respectively [ix and jx are the positions of the nodes.The text mining software VOSviewer was adopted for science mapping, combining with another software CiteSpace to portray the time view of the clustered keywords based on the same data. VOSviewer, developed by van Eck and Waltman , is a coectively . The stoectively , which cFor a detailed operation manual of bibliographical experiments using science mapping approach, readers are advised to refer Jin et al.  and Hu eThe advanced retrieval function in Scopus and WoS core collection database was used to retrieve the G&SL related papers published during 1999 to August 2019 see . To ensuSustainability ranks first , followed by Journal of Cleaner Production , Transportation Research Part D: Transport and Environment  and International Journal of Production Economics . Among the top 15 journals, eight are from UK, four from The Netherlands, two from Switzerland, and one from Germany. The papers are mainly distributed in the three academic fields of environment, traffic engineering and operations management, but they obviously account for a larger proportion in the environmental science and sustainable field, which is in line with the connotation of G&SL.All 306 documents were found in 81 different journals. As shown in European Journal of Operational Research , International Journal of Production Research , Expert Systems with Applications , Omega  and Computers & Operations Research , which can offer quantitative methods for the decision-making and optimization issues related to G&SL. The second cluster is transportation research (TR), such as Transportation Research Part A , Part D , Part E , and Journal of Transport Geography , which accumulates enormous knowledge towards transportation planning, technology and operations that can enlighten G&SL research from real-life transport demand and practice. The third cluster, including Supply Chain Management , International Journal of Physical Distribution & Logistics Management  and Journal of Business Logistics , etc., reveals that a large amount of G&SL research was conducted based on the research foundation of logistics and supply chain management (SCM). Among all the publications, Journal of Cleaner Production  and International Journal of Production Economics  are the two most co-cited journals. They often act as hubs, integrating the results of OR, TR and SCM with social, environment or economic implications to provide cross-domain knowledge crucial to the diverse development of G&SL.As shown in According to Two evidence can be observed from Second, the international collaboration is not significant. Taking mainland China for instance, about 70 percent of 49 publications are completed entirely by domestic institutions. The Swedish publications do not have any co-authors from other countries or regions. This phenomenon may be due to the large differences in the background and model of G&SL development in different countries . MoreoveAmong the 402 organizations that contributed to G&SL research, those with more than five documents and over 30 citations were built into a network of 22 items and 22 links, as shown in Through the document co-citation test of the portfolio, the most influential G&SL publications in the past two decades were analyzed and the co-citation network was constructed. In VOSviewer, the minimum number of citations was set to 30 to build a co-cited visual network map of 83 items and 350, as shown in The top 15 most cited papers are presented in The keywords co-occurrence analysis was conducted to describe the internal composition and structure of G&SL and to reveal the frontiers . The optKeyword co-occurrence network is a static expression of a particular area that does not take into account changes over time in the manner that the terms are used . Figure Through the aforementioned analysis, the research progress, evolutionary trend, and hot-discussed topics of global G&SL are clarified. However, the generic scientometric results cannot accurately reflect the explicit division of the multifarious knowledge of a domain . Based oNearly a quarter of the literature (71 out of 306 papers) focused on evaluating and quantifying how the potential green logistics initiatives improve the “triple bottom line”  of existing freight activities. The subjects of these studies were basically originated from four aspects: carbon emission, energy consumption, social sustainability, and external cost-and-benefit. Mattila and Antikainen  providedAnother part of emphasis was given to SEE performance of G&SL based on logistics operations and business. Guo and Ma  evaluateExcept for the three-dimensional evaluation system, some scholars also analyzed the critical success factors and barriers for G&SL initiative implementation from the SEE perspectives. For instance, Arslan and Sar  found thThis knowledge branch focuses on two basic G&SL topics, (i) the planning, development, and policymaking from industrial level, and (ii) the collaboration strategy and management from project level. For the former, Lindholm and Blinge  indicateFor the latter, the collaboration and game among logistics service providers (LSP), government, shippers, and enterprises are paid more attention. Commonly, a positive cooperation strategy of stakeholders will significantly improve the operational performance of G&SL  and evenFurthermore, several novel business and operational modes of logistics aiming at improving the sustainability in transportation process were proposed, e.g., freight consolidation , smart lOver the past decade, research on the G&SL practices were carried out over a broad range, including SCM, reverse logistics (RL), e-commerce, urban distribution, multimodal transport, and other dedicated logistics such as food  and manuReverse logistics is convincingly one of the most efficient solutions to reduce environmental pollution and waste of resources by capturing and recovering the values of the used products . LegislaThe unsustainability of urban logistics makes it the most urgent goal of greening. Huge logistics demand, such as rapid business-to-business and business-to-customer logistics activities, make freight transportation in big cities face the dilemma of air pollution, poor accessibility, and livability . The praCompared with G&SL in urban domain, the sustainability issues regarding inter-city or regional logistics are more emphasized on the intermodal application. The shift of road-based modal to other transportation system, such as rail and water has the potentials of ensuring environmental sustainability, flexibility, and cost reduction . HoweverDeveloping advanced facilities and technologies is a sustainable and forward-looking solution to meet the challenge of freight transport. Many emerging logistics systems were proposed in recent years. Such as urban consolidation center , electriElectric vehicles (EVs) technology, which has been widely applied in passenger transport, is also waving a revolution in the field of G&SL. Current research on freight EVs mostly focuses on energy efficiency , fleet oFor reducing the negative externalities such as traffic congestion and disturbance, another interesting concept, i.e., transferring the ground logistics process to underground space, namely the Underground Logistics System (ULS), has aroused increasing attention. ULS refers to using a group of hierarchical underground nodes, pipelines, and tunnels to distribute cargo flows in and between cities with 24-h automated operations . ULS canThe operations research (OR) of G&SL issues that are originated from real-world applications is always being a well-concerned topic because it is directly related to the quality of some critical decision-making in logistics operation. The OR method applied for G&SL is defined as a better of science to identify the trade-offs between environmental aspects and costs, so that the corresponding decisions such as location, transportation, warehousing, and inventory can be optimized and the limited resources can be reasonably assigned . Dekker Through the above scientometric analysis and thematic discussion, the comprehensive research trend, mainstream academic topics, and knowledge taxonomy of G&SL domain were revealed. Although researchers and practitioners achieved substantial results in promoting G&SL theory and practice, there are still some shortcomings that need to be elaborated in future studies.In terms of research model, international cooperation is still lacking. The broad applicability of most G&SL knowledge based on local cases deserves further discussion, such as planning methods and evaluation systems. European countries made great efforts in rebuilding the integration of green logistics. However, the lack of international cooperation and universal solutions hinders the dissemination and deepening of knowledge, and the current achievements are far from enough to promote the globalization of G&SL, which is reflected in the imbalance of global G&SL practice.To fill this gap, although it is recognized that logistics policy has a strong regional character, cross-institutional and cross-national collaborative research on market operation, industrial metrics, technology innovation and macro development strategies should be strengthened under the trend of supply chain globalization. For example, more attention can be paid to the horizontal comparison of green logistics mode, scheme and performance under different case backgrounds. Additionally, more empirical studies are needed to be carried out in some developing countries in Asia and elsewhere in the world, considering they are the fast growing economies with higher population and logistics demand.Although the knowledge branch of research is flourishing, it is acknowledged that there is still a need to supplement the overall or holistic research to improve the knowledge system of G&SL. Research on sustainability and green has always been complex and multi-variable, interactive, with far-reaching implications. Besides, sustainability and green are public and social issues. Current theoretical applications are limited to the analysis of local or one-way relationships, such as LSP/retailer/carrier responses to green policies, planning and performance evaluation of green and sustainable initiatives.The operation and decision-making of G&SL involve many stakeholders, such as local authorities, manufactures, LSP, carriers, customers, and even the sharers of transportation resources. The impact of G&SL should also be long-term and dynamic. Thus, the whole picture includes multiple perspectives, such as the dynamic evaluation of the whole life-cycle of green logistics practice, the decision interaction among multiple stakeholders, and the follow-up research and report on a new green technology or practice.Without green innovation technologies, the effect of implementing G&SL from a management perspective alone is minimal. However, it takes a lot of time for some innovative technologies that can fundamentally improve the negative effects of logistics to move from laboratory to application. Applications such as the EV took decades to implement . AlthougThe introduction of a new thing does require a long period of demonstration, such as the reliability of the technology, the acceptability of the market and the ambiguity of the real benefits. However, the problem is often the gap and lag in the research of application management in the transition from technical problems to market application and practice management. Therefore, building effective platforms based on multidisciplinary, cross-organizational collaboration to accelerate the research and application of innovative technologies is particularly important for G&SL practices, such as ULS, RL, and CS. Such calls are all the more urgent in their own research.The concept of green and sustainable logistics has received increasing attention and consideration government sectors, scholars, practitioners, and international organizations. A large amount of practical achievement was made at both the industrial and theoretical levels. This study reviewed 306 valuable contributions regarding G&LS over the past two decades through a three-step review program. They were described in year publication, journal allocation and citation counts. Then, the bibliographic networks of countries, organizations, journal and document co-citations, keyword co-occurrence and timezone clusters of research themes were visualized to help understand the overall research status and academic progress worldwide. Grounded in the scientometric analysis, an integrated knowledge taxonomy of the G&SL field was presented, including five major alignments and 50 sub-branches.Sustainability, Journal of Cleaner Production, Transportation Research Part D: Transport Environment and International Journal of Production Economy are the top four journals, which contributed over a quarter of all G&SL papers since 1999. The maps of journal allocation and co-cited journals show that the current research is most relevant to the environmental science and transportation science. In terms of countries, China, the United States, the UK, Sweden, and India are the major territories of G&SL research. The network across co-authored organizations and countries revealed that the collaboration among different research communities is not strong. Hence an active and robust global collaboration atmosphere has not formed yet.Results indicate that the chronological publication of G&SL shows a trend of rapid increase. The quantity of literature published in 2018 is fifteen times more than that of 10 years ago. The map of co-occurred keywords showed that the most frequently discussed G&SL themes in each cluster were sustainability and management (cluster #1), freight transportation and carbon emission (cluster #2), model and reverse logistics (cluster #3), and green supply chain and green logistics (cluster #4). The timezone view of keywords showed that articles related to collaboration, transportation planning, modal shift and stakeholder were largely published during the recent years. On this basis, the knowledge taxonomy of G&SL was manually synthesized from five aspects: (i) evaluation on SEE impacts of G&SL initiatives; (ii) planning, policy, and management research; (iii) real-world application areas and practices; (iv) emerging technologies and (v) operations research and optimization methods for G&SL decision-making.Finally, the potential roadmap for filling current research gaps was recommended, which were divided into three streams: (i) more global research collaboration should be advocated to jointly develop and supplement the comprehensive evaluation framework of G&SL performance; (ii) future research efforts could focus on the interactive and dynamic relationships among sustainable development goals, green policies and the decision-making of multiple stakeholders; (iii) the application-oriented platforms and management research for some most advanced green logistics initiatives would be highly beneficial in promoting G&SL innovation.However, it should be noted that the data used in this study was confined to those research articles and review articles that were published in the peer-reviewed journals, and they were retrieved only from the two mainstream databases considering the applicability of software. Although the indexed documents could represent most of the convictive viewpoints of G&SL research, some valuable articles that were published in other forms or included in other databases might be overlooked inevitably. To sum up, this review has great room for improvement in terms of material selection. A systematic investigation incorporating valuable conference proceedings, reports, and books in the field of green logistics or green supply chain is expected to portray a more comprehensive knowledge map for future research. Additionally, the in-depth review of the hotspot themes in G&SL domain e.g., OR application and SCM, may also contribute to multidisciplinary integration and interaction.