Khalas, Khinaizi, Sukkary, Shaham, Zahidi and Fardh were prepared. Two types of flour were used (white flour and whole wheat) at two different baking temperatures: 180 and 200 °C. The incorporation of date seed had no or slight effect on moisture, ash, fat and protein content of the baked cookies. On the other hand, incorporation significantly affected the lightness and hardness of cookies; the higher level of addition, the darker and crispier the resulting cookies. The sensory analysis indicated that the produced cookies were acceptable in terms of smell, taste, texture and overall acceptability. The results indicate that the most acceptable cookies across all evaluated parameters were produced using whole wheat flour with 7.5% levels of date seed powder using Khalas and Zahidi varieties. Overall, the analysis indicated that cookies with acceptable physical characteristics and an improved nutritional profile could be produced with partial replacement of the white/whole wheat flour by date seed powder.Date seeds are a major waste product that can be utilised as a valuable and nutritional material in the food industry. The aim of the present study was to improve cookies quality in terms of functional and textural value and assess the effect of date seed powder flour substitution on the physical and chemical characteristics of cookies. Three substitution levels  of flour by fine date seed powder from six varieties locally named  The arid and semiarid regions of countries in North Africa and the Middle East are good habitats for the date palm plant ,2,3,4. DCookies hold an important position as snack foods due to the variety of taste, texture and digestibility ,10,11,12In most of the date-processing plants, including date confectionery and date syrup, the seed is considered as the major waste product which amounts to approximately 10% of the fruit mass. Thus, the utilisation of date waste is important for date cultivation and to increase the income in date-processing units. Currently, seeds are used mainly for animal feed for various livestock and sometimes as a soil additive ,21. The At the same time, baked food items such as cookies, biscuits, muffins and cakes are very popular among consumers due to their taste and availability. When considering the health aspects, they are usually high in sugar, whereas they are low in antioxidants, minerals and fibre content . Hence, Therefore, the purpose of this study was to determine the physicochemical and sensory attributes of date seed powder (DSP)-substituted cookies in order to enable the development and production of better cookies in terms of quality and nutritional value while reducing waste.Khalas, Khinaizi, Sukkari, Shaham, Zahidi and Fardh were used. The seeds were collected from local farms located in Al Ain, United Arab Emirates. Flour , sugar, salt, palm oil, food-grade ammonium bicarbonate (NH4HCO3) and food-grade sodium bicarbonate (NaHCO3) were purchased from a local market .Date seed powder of six fully ripe Emirati date varieties locally known as The received seeds were cleaned, washed with water, air-dried, ground into powder according to  and siev3, 1.0 g salt, 0.4 g NH4HCO3 and 17.6 g water. The flour was replaced with preparations of date seed powder to a level of 2.5%, 5.0% and 7.5% in the cookie recipe, and cookies with no substitution were produced as control samples. Two types of flour—white (WF) and whole wheat flour (WW)—and two baking temperatures of 180 °C and 200 °C  were used, making twelve different combinations for each variety of date seeds. The dough was kneaded and sheeted to a uniform thickness using a pasta machine set at 3 mm. Then, the dough was cut with a stainless-steel cutter into circular shapes of a diameter of 4.5 cm. The cookies were baked in a wall oven  and were packed in sealed plastic bags and stored at −20 °C until further analysis.The recipe of cookies described in the American Association of Cereal Chemists (AACC) method 1054  was usedMoisture percentage was determined by the oven dry method  method 934.01)  using anThe colour of the date seed powder of six varieties and cookie samples was determined using a Hunter Colourimeter fitted with an optical sensor  on the basis of the L*, a*, b* colour system. L* is the lightness component that measures black (0) to white (100), a* parameter goes from green to red, and b* parameter from blue to yellow. Throughout this manuscript, the colour analysis was performed only on the basis of lightness (L*). Six cookies were evaluated for each formulation.Measuring hardness was determined using Texture Analyzer  equipped with a 3-Point Bending Rig in a compression mode with a sharp blade-cutting probe (TA7). Pre-test and test speeds were 1 and 0.5 mm/s, respectively, at a trigger load of 3.0 g. Hardness (maximum peak force) was measured with at least 3 cookies for each sample. The peak force, when the cookie began to break, was reported in Newton (N). Cookies were subjected to a sensory evaluation using 30 untrained panellists recruited from within the university community. Sensory properties  were evaluated. The ratings were on a 9-point hedonic scale ranging from 9 (like extremely) to 1 (dislike extremely).p < 0.05 to determine the level of significance.The data were shown as mean ± standard deviation (SD). Data were assessed by Tukey test using  statistical software package at Sukkari seed powder was the least moist, followed by Zahidi Fardh, Khinaizi, Shaham and Khalas seeds. Differences in cultivars, as well as cultivation under divergent climatic conditions, make it possible to have discrete moisture levels. Previous studies on date seed flour also demonstrated different moisture contents in different varieties [Sukkari seed powder had the lowest, while seeds of the Fardh variety had the highest. In terms of crude fat, Zahidi seeds had a high amount of fat, opposite to the Khinaizi seed variety. No significant difference was observed in protein content between the varieties used. The lowest carbohydrate content was in Khalas powder; it was significantly different from Sukkari, Shaham and Zahidi varieties. In previous findings for Khalas, Sukkari and Fardh, the results are quite different from this study [Khinaizi, Shaham and Zahidi—could not be found.The chemical composition for DSP varieties used in making cookies is shown in arieties ,31. For is study ,31,32. Tis study . Data abThe percentage of moisture, ash content and crude fat content of different cookie samples are shown in Khalas cookies at 2.5% addition, where the WW cookies had higher moisture than the WF ones. The reasons for this difference are not clear.In Shaham cookies that exhibited no significant differences between the two baking temperatures.When considering the differences between baking temperatures (for the same flour type) overall, as expected, the cookies baked at 200 °C had lower moisture content than those baked at 180 °C. This observation was true for the controls as well as the composite cookies. The exceptions were the Shaham had the highest values, while at 7.5%, the highest was Khalas. For the 180 °C WW cookies, at all levels of addition, the Khalas variety cookies exhibited the highest values. For WF cookies baked at 200 °C, there was no clear pattern emerging on the basis of date seed variety. For WW cookies baked at 200 °C, Khalas showed the highest moisture values at all addition levels.When considering the differences among varieties, for the 180 °C WF cookies, at 2.5 and 5% addition, In previous studies, improved water holding capacities resulted in bread formulated with defatted date seed powder  and extrZahidi cookies had the highest values at all levels of addition. For WW cookies at 180 °C, again, the Zahidi variety had the highest values at the higher levels of addition (5% and 7.5%). For WF cookies prepared at 200 °C, the Zahidi variety had significantly higher values than the rest, although no significant differences were observed between levels of addition. For WW cookies baked at 200 °C, Fardh cookies had the highest values at all levels of addition. Overall, the results regarding the ash content are somewhat surprising as they did not correspond to the ash content of the DSP.Ash content differences between cookies are listed in Fardh. This may be due to the addition of lower amounts of date seed powder in composite cookies, leading to no significant difference in ash content between the control and composite samples. High ash content is an indication of high mineral content. Date seeds are found to be rich in minerals [There was no significant improvement in ash content of some of the composite cookies compared to control samples, although a considerable amount of ash was observed in date seed powder, especially minerals ,36, and minerals . TherefoThe fat content results in The different stages of cookie preparation change the physical and chemical properties of the various flour constituents, including fat, present in the cookie dough formula . MoreoveThe incorporation of date pits in the form of powder at different levels into cookies resulted in similar fat content, and this can be attributed to the high amount of fat in the oil and flour used  that masks the fat content coming from date seed powder. The same explains the insignificant difference in the protein content of the cookies, where the protein content came mainly from the high amount of protein in whole wheat and white wheat flour. However, using even higher date seed powder addition levels, as shown by Platat et al. , may makIn terms of protein content, the incorporation of date seed powder of any variety in making cookies did not affect the protein content when compared to cookies with zero incorporation, which agrees with Platat et al. . The datSukkari date seed powder was the lightest in colour, and Khalas was the darkest; however, the differences in the level of lightness between the date seed powder did not affect the final colour of the cookies. White flour cookies with no addition of date seed powder were obviously lighter than cookies of whole wheat flour, and the lightness of the composite cookies displayed a decreasing trend with the increasing level of addition; the higher level of addition, the darker the cookie. These results are in accordance with Gómez and Martinez [The colour measurements of date seed powder varieties and the cookies substituted with various date seed powder are listed in Martinez , AksoyluMartinez  and AshoMartinez , who repShaham date seed powder was not affected by the addition level and was almost similar to the texture of Zahidi cookies, which were softer when prepared with white flour at 200 °C at an addition level of 7.5%. The effect of the addition level of either Fardh or Sukkari seed powder on the texture of cookies was different; the more powder added, the softer the cookies. Moreover, the case was the same in Khalas cookies, except for whole wheat flour Khalas cookies baked at 180 °C, where softer cookies were obtained. While in whole wheat flour Khinaizi cookies at any baking temperature, the softness was not affected, differing from white flour Khinaizi cookies, where a decrease in hardness with the addition level was recognisable.Cookies’ hardness assessments are listed in Shahm and Zahidi. In the case of whole wheat flour, a significant difference was noticeable in Khalas and Sukkari at higher addition levels and the 7.5% addition level in Fardh at 200 °C, whereas at 180 °C, it was only in Fardh and 7.5% addition level in Sukkari.With the addition of date seed flour, cookies become softer when the control is white flour, except for Sukkari cookie samples at 200 °C, which showed a prominent significant difference from the control. However, it was not significantly different from Khinaizi and Fardh at the same treatment conditions. In comparison among varieties, the increase in the softness was quite underlined in Fardh cookies in all the treatment conditions at 7.5% inclusion level except white flour Fardh cookies at 200 °C. In that treatment condition, all the inclusion levels of 2.5%, 5% and 7.5% were also significantly different from the control, although it did not show significant differences between the addition levels. The most pronounced decrease in the hardness of the obtained composite cookies was observed for the 7.5% addition level of white flour Composite cookies were softer in accordance with similar studies on seed incorporation, such as amaranth flour composite cookies ,40. AccoThe evaluation of sensory properties of cookies is illustrated in Khalas and Zahidi. It is noteworthy that in some instances, date seed composite cookies with high substitutions are the ones which are more preferred compared to zero or low-level samples. When the texture is considered, consumer acceptability is high in the samples with high date seed flour substitutions. This may be due to the softness of biscuits. According to the results of hardness, date seed composite cookies have lower hardness compared to the control samples. Hardness is a textural property that plays a major role in the evaluation of baked goods, as it is associated with the human perception of freshness—the lower this parameter is, the more desirable the product [The consumers’ evaluation in terms of colour, texture and taste showed that the produced cookies were highly acceptable. A darker cookie colour correlates with other physical, chemical and sensorial indicators of product quality. Colour is considered a fundamental physical property of foods and agricultural products, and it affects the assessment of external quality in both the food industry and food engineering research ,45. When product . HoweverKhalas’ moisture was the highest. It appears that the moisture content level is primarily affected by the baking temperature and the type of flour, while varietal differences seem to be less pronounced. When considering the ash content, Zahidi composite cookies at the 7.5% addition level in both flour types had the highest values. The colour of cookies was significantly affected by the incorporation of date seed flour, making them darker. This may be the reason for the somewhat reduced score for the colour attribute in sensory, even though some addition levels show higher consumer acceptance. However, the sensory analysis results showed that the overall acceptance of the composite cookies was higher than the others with whole wheat flour at both temperature levels. Therefore, based on the sensory analysis, Khalas and Zahidi composite cookies at 7.5% addition level with whole wheat flour can be recommended to obtain the most preferred final product. As expected, an increased softness was observed in date seed composite cookies, except for the Shaham and Zahidi varieties (where the increment was not significant), leading the path to formulate high-quality cookies in terms of texture.This study concluded that the by-products or the waste of date-processing units, such as date seeds, can be used to enhance the quality of cookies in terms of their physical, chemical and textural properties. There were no significant differences in the protein content of cookies with date seed flour incorporated since the composite cookies had a protein content similar to the control samples. For cookies baked at 180 °C, whole wheat flour formulations had lower moisture content than those made with white flour. A similar trend was observed at 200 °C, with whole wheat flour formulations having lower moisture content than white flour for each date variety. Different varieties with different addition levels showed a variation in moisture levels, but at all addition levels,


Cyclic diguanylate (c-di-GMP) signal transduction systems provide bacteria with the ability to sense changing cell status or environmental conditions and then execute suitable physiological and social behaviors in response. In this review, we provide a comprehensive census of the stimuli and receptors that are linked to the modulation of intracellular c-di-GMP. Emerging evidence indicates that c-di-GMP networks sense light, surfaces, energy, redox potential, respiratory electron acceptors, temperature, and structurally diverse biotic and abiotic chemicals. Bioinformatic analysis of sensory domains in diguanylate cyclases and c-di-GMP-specific phosphodiesterases as well as the receptor complexes associated with them reveals that these functions are linked to a diverse repertoire of protein domain families. We describe the principles of stimulus perception learned from studying these modular sensory devices, illustrate how they are assembled in varied combinations with output domains, and summarize a system for classifying these sensor proteins based on their complexity. Biological information processing via c-di-GMP signal transduction not only is fundamental to bacterial survival in dynamic environments but also is being used to engineer gene expression circuitry and synthetic proteins with à la carte biochemical functionalities. The ability of bacteria to monitor their cell status and surroundings is required for their survival in dynamic environments –3. Bacte–9–Here, we use the term sensory perception to refer to the mechanisms responsible for converting an input stimulus into a biochemical output and the term stimulus to refer to any agent or change inside or outside the cell. Catalytic mechanisms for c-di-GMP synthesis and degradation are understood in detail –16, and –17–c-di-GMP is synthesized by diguanylate cyclases (DGCs) that possess a GGDEF domain, and it is degraded by c-di-GMP-specific phosphodiesterases (PDEs) that possess an EAL or HD-GYP domain . Product21–25–We analyzed sensory inputs and domain architectures of proteins containing GGDEF, EAL, and/or HD-GYP domains using the Pfam database  , the NatA key observation is that GGDEF, EAL, and HD-GYP domains are found in thousands of unique protein domain architectures with putative sensor and receiver domains (Tables S1 and S2). There are many ways of classifying signal transduction proteins. However, one practical system for grouping the diverse GGDEF, EAL, and HD-GYP domain-containing architectures is the complexity scheme –33, whic–One-component systems (OCSs) combine sensory and regulatory functions in a single protein , althougIn contrast, two-component systems (TCSs), which are mostly involved in sensing extracellular stimuli, are minimally comprised of a sensor histidine kinase and a cognate response regulator . Some TCEscherichia coli McpA, CheA, CheW, CheR, CheB, and CheY proteins, which correspond to the methylating chemotaxis protein, histidine kinase, coupling protein, methyltransferase, methylesterase, and response regulator of the chemosensory pathways regulating flagellar motility, respectively , 36Escheectively  or type Per-Arnt-SIM (PAS); cGMP-phosphodiesterase, Anabaenaadenylyl cyclases, and E. coliFhlA (GAF); and calcium channels and chemotaxis receptors (CACHE) domain clans—are evidently prevalent among these unique conserved architectures  different databases define protein domains using algorithms that group similar domains differently, (ii) different algorithms are varied in their ability to detect protein domains, and (iii) nomenclature can vary between databases and is not always consistent with relationships deduced from sequence alignments or statistical models. We highlight the relevant differences in nomenclature between databases, as well as the trends and knowledge gaps in c-di-GMP signaling that emerge from this assessment. We also summarize basic principles from biochemistry explaining how these modular input devices regulate the activity of output domains. In the sections that follow, sensory domains with known functions are presented first and in their order of overall abundance in GDDEF, EAL, and HD_5 (HD-GYP)-domain containing proteins (Table S1); putative phosphoreceiver domains are discussed afterward. Regardless of their abundance, putative sensory domains with unknown functions are discussed together in a final section.2) elicit rearrangements in the core PAS fold β-sheet structure that are propagated to the output domain via torque of a linker region situated at the C terminus of the PAS domain -binding domain that facilitates hypochlorite and/or Zn sensing in some DGCs , 70. Thea linker , 70.2-carrier proteins, hemerythrin domains are sometimes denoted bacterial hemerythrin-like (Bhr), particularly in the context of c-di-GMP signaling. Hemerythrin domains are defined by a binuclear nonheme iron center in which 7 amino acids coordinate 2 iron (Fe) ions (2 to this di-iron center causes it to oxidize from the diferrous (Fe2+) to diferric (Fe3+) state  ions . The bin+) state , 72. Whi+) state , 73.2 sensing. Based on in vitro reactivity, however, it is possible that GCSs could function in CO (2 in the binding pocket by Fe(II)-heme and distal threonine and tyrosine residues   that hav75–on in CO , 78, 79 on in CO , 78, 79,residues  causes tresidues , and thiresidues .Clostridium spp.  domain-containing proteins at all . The BLU93–cNMP_binding domains, encompassed within the CAP_ED domains in CDD (Table S2), are intracellular sensory domains that bind cGMP or cAMP , 98. Fir2 and redox sensing.This domain is always found at the N terminus of an EAL domain. Although its structure is unknown, the CSS motif domain is comprised of a periplasmic region flanked by putative TM helices . The perreceiver (REC) domain by the CDD  domain, named after its conserved methionine-histidine-tyrosine-threonine amino acid pattern , is an iThe FIST-FIST_C domain architecture is also present in some proteins containing GGDEF, EAL, and HD-GYP domains . It has The Pfam v34.0 aspartate chemoreceptor signal-transduction ligand-binding clan (4HB_MCP) contains both the chemoreceptor 4HB_MCP_1 family (which overlaps with the Tar_Tsr_LBD domain from CDD)  and the 7 transmembrane receptors with diverse intracellular signaling modules extracellular domain 2 (7TMR-DISMED2) is a TM domain predicted to sense carbohydrates via a conserved jellyroll fold (cystathionine beta-synthase (CBS) domains are thought to be energy-sensing modules that may bind AMP, ATP, and S-adenosylmethionine (diguanylate cyclases and phosphodiesterases and two-component systems (DICT), the membrane-associated sensor 1 , and the cyclases/histidine kinases associated sensory extracellular 6C (CHASE6_C) domain  domain , which iin vitro and in vivo, and highlight some of the important limitations and questions that remain.Here, we categorize the sensory functions of c-di-GMP signaling networks, describe the physiological relevance of several signal transduction mechanisms that have been substantiated The literature is rife with descriptions of genetic linkages between stimuli and DGCs or PDEs causing changes in intracellular c-di-GMP. While these observations are discussed and summarized in this review , we caution that mechanisms of sensory perception can be difficult to interpret from genetic linkages without additional data. For instance, deletion of a housekeeping PDE gene might have a dominant gain-of-function phenotype (elevated cellular c-di-GMP levels) that may confound interpretations of the genetic linkage. Similarly, a pitfall may come from deletion of a DGC gene that is highly expressed under the conditions tested, thereby producing a dominant loss-of-function phenotype.Additionally, a small number of reports make putative stimuli-receptor linkages relying solely on c-di-GMP-dependent transcriptional reporter measurements as a proxy for direct, quantitative c-di-GMP measurements. Although c-di-GMP bioreporters are easy, quick, and inexpensive compared with more laborious analytical chemistry protocols relying on mass spectrometry infrastructure for quantifying c-di-GMP, reports relying solely on bioreporters have been excluded from this review because without direct measures of c-di-GMP levels, we cannot rule out that gene transcription might be controlled by other factors that are c-di-GMP -independent.Pseudomonas aeruginosa (Xanthomonas spp. (Thermosynechococcus elongatus (126–in vitro for DGCs and PDEs from many other species (Light sensing is one of the best understood sensory functions of c-di-GMP networks. It is facilitated by photosensitive domains in (i) GGDEF and EAL domain-containing proteins or (ii) short photoreceptor proteins that modulate the activity of EAL domain-containing proteins through protein-protein interactions. Photoreceptors identified in c-di-GMP networks use light-sensitive BLUF, GAF, or LOV (PAS9) domains . Compellruginosa , Xanthomnas spp. , 125, anlongatus , 126–128 species , and in Klebsiella pneumoniae, which consists of a BLUF domain linked to an EAL domain. Light increases the PDE activity of this protein 4-fold  . The GAFZ/E isomerization at a carbon double bond that causes the whole chromophore to rotate in its binding pocket  molecule in the LOV domain forms the canonical thioester bond between the flavin moiety and the key cysteine residue. The protein also creates electrostatic interactions between two arginine residues and the phosphate group of the FMN molecule  or malate (CtpM) retain surface-sensing functions -dependent surface recognition has also been linked to c-di-GMP signaling in several bacterial species  149–153149–149–153. C. crescentus has tight adherence (tad) type IV pili and a single flagellum located at one of its cell poles. It uses these appendages to orchestrate the deployment of an exopolysaccharide holdfast within seconds of encountering a surface , suggesting that this organelle does not simply act as a tether  or PAS domain at the N terminus to regulate enzymatic activity in an O2-dependent manner  . DgcO and PdeO assemble into a functional complex in vitro and, under certain conditions , are also thought to comprise part of the E. coli RNA degradosome via interactions with a scaffolding protein, RNase E  , 167. O2 RNase E , 169. An RNase E . Togethea PNPase .Ferrovum sp. strain PN-J185  form and not the oxidized (ferric) form. These two states were reversibly interconverted in the purified protein, suggesting that the domain is used for redox sensing. Although exposure to air oxidized the protein, absorption spectra did not show evidence of O2 binding, indicating that an O2 adduct does not form (in contrast to what is observed for invertebrate hemerythrins)  , 170. Inythrins) . The rolP. aeruginosa protein RmcA has also been implicated in the response of P. aeruginosa to its own redox-active products, a class of small molecules called phenazines. These compounds contribute to redox homeostasis by coupling the oxidation of NADH to the reduction of O2. Accordingly, a phenazine-null mutant shows an increased NADH/NAD+ ratio  , the D+ ratio . Like th parent) . RmcA is parent) . RmcA PAosa RmcA . Interesosa RmcA . The uniosa RmcA .Bacillus cereus group is a cohort of Bacillus strains that are closely related phylogenetically  but that have been historically treated as separate species due to other differences, such as the presence of unique plasmids  spectrum of recombinant, purified CdgF suggests that it binds a flavin molecule. Shifting the protein from the oxidized to the reduced state correlated with a switch from DGC to PDE activity. The authors of this study thus speculated that CdgF acts to promote biofilm formation in oxygenated environments  interact with redox-sensitive extracytoplasmic proteins or (ii) themselves contain a redox-sensitive cysteine pair that forms a disulfide bond.Diverse Gram-negative bacteria have protein systems that sense the redox state of the periplasm and transduce this information into modulation of c-di-GMP levels in the cytoplasm. Compared with the conditions of the cytoplasm, those of the periplasm are more sensitive to the external environment. While the cytoplasm is typically maintained in a reducing state, the periplasm is oxidizing and supports the generation of disulfide bonds, which form between two cysteines in proteins or small molecules. Periplasmic disulfide bond formation can be an important component of protein folding or regulation of activity and is usually mediated by well-characterized Dsb proteins, which facilitate electron transfer between target proteins/small molecules and the quinone pool of the electron transport chain . For sevY. pestis is the causative agent of plague and is carried by fleas. The formation of robust biofilms in the flea gut promotes the transmission of Y. pestis to mammalian hosts. Biofilm formation is stimulated by reducing conditions and is positively regulated by c-di-GMP. A recent study implicated the HmsCD proteins in this effect  one in which PdeC forms the disulfide bond in the CSS domain, which results in low PDE activity and therefore relatively high concentrations of c-di-GMP; (ii) one in which the cysteines in the CSS domain remain in the free thiol form, allowing them to be used for dimerization with another PdeC molecule, which increases PDE activity and therefore decreases c-di-GMP concentration; and (iii) one in which the region that contains the free thiols is processed by periplasmic proteases to yield a shorter protein that is still able to dimerize and degrade c-di-GMP and therefore decrease the c-di-GMP concentration. The formation of the first state is possible when cells are grown under aerobic conditions. In this case, the DsbA/DsbB system and the oxidizing environment of the periplasm promote disulfide bond formation. The second and third states occur in reducing environments, which favor the formation of free thiols as opposed to disulfide bonds. After the third state has been formed, it is slowly degraded by further proteolysis. Together with in vivo results, these observations indicate that PdeC functions to modulate matrix production in poorly oxygenated subzones of E. coli biofilms. The fact that PdeC is just one of five similar CSS-domain PDEs in E. coli suggests that periplasmic conditions exert a substantial influence on c-di-GMP-dependent processes in this organism and exemplify the potential for elaborate linkages between environmental sensing and multicellular behavior.SS motif . The cyt of hmsT mRNA abolishes this phenotype . Functional homologs of TdcA were also identified in distantly related proteobacteria . H. HShewancholerae , is an acholerae , 89. In cholerae , 89. Whecholerae . The actcholerae .An analogous TCS is the NosP-NahK-NarR system, but in contrast to HnoXKBD system, NO promotes PDE activity . Here, NDinoroseobacter shibae is inhibited by an HNOB domain-containing partner protein called H-NOX when it binds NO  . One mecruginosa . The PDEruginosa . Anotherriovorus . In Bdelehaviors . The Bd1ehaviors . Bdellovcyclases  that proLeptospira interrogans, activating the adjacent GGDEF domain and c-di-GMP synthesis , while others decrease them significantly . Moreover, micromolar concentrations of these compounds caused corresponding changes in cellulose levels. l-Arginine was exceptional among the identified environmental nutrients because S. Typhimurium cells responded to it at very low concentrations . Genetic linkage analysis identified that a Δstm1987 mutant lost its ability to respond to l-arginine, although the authors noted three other genes that might encode secondary, downstream c-di-GMP-metabolizing enzymes that might contribute to the phenotype, albeit to a lesser extent than stm1987 -based c-di-GMP biosensor , 199 in Because quorum sensing regulates a constellation of genes , 201, dicis-2 double bond, are an important group of quorum-sensing autoinducers. Structural biology has provided evidence indicating that the Burkholderia diffusible signaling factor (BDSF) interacts with the PAS9 domain of the c-di-GMP specific PDE RpfR . Membrane-associated DGC rpHK1S-Z16 from orimetry . Also, mPseudomonas fluorescens GcbC likely binds to citrate via its dCACHE_1 domain, as evidenced by a homology model of the dCACHE_1 domain, which was based on a crystal structure from rpHK1S-Z16 from R. palustris, a homolog of GcbC. A mutational analysis was used to confirm that the critical residues for binding citrate in rpHK1S-Z16 were operational in citrate sensing for P. fluorescens GcbC (Accumulating studies reveal additional, diverse chemosensory functions for c-di-GMP networks, including the sensing of tricarboxylic acid intermediates, host factors, and metal ions. For instance, ens GcbC . Furtherens GcbC . When Gcens GcbC , citrateens GcbC . Howeverens GcbC .V. cholerae, CdpA may rely on FIST-FIST_C domains to detect heme. Heme binding inhibits the EAL domain of CpdA (V. cholerae.In  of CpdA , and theE. coli DgcZ, which responds to femtomolar concentrations of Zn2+ via its CZB domain (2+ concentrations in vitro led to the strong inhibition of DGC activity (2+, indicating that cellular oxidants, which are also affected by metal toxicity (Finally, numerous studies have linked bacterial exposure to metal or metalloid ions to modulation of c-di-GMP levels –208. Mec–B domain , 70. Incactivity , 70. Inttoxicity , 210, catoxicity .Rhodobacter sphaeroides BphG protein (Synechocystis spp. Slr1143 protein (in vivo experiments (K. pneumoniae protein (An understanding of sensory perception in c-di-GMP networks is providing an opportunity to use these systems for biotechnology. The most notable application so far has been optogenetics, which relies on the heterologous expression of genes coding for light-sensitive proteins and illumination to alter cellular behavior . Light i protein  and the  protein . BphS reeriments . BphS caeriments , or BlrP protein —to creat protein , 214. Th protein . The fla protein .To date, sensory perception in c-di-GMP networks has been studied in at least 40 bacterial species from 35 genera (Tables 1 to Despite recent advances, we propose that most sensory functions of c-di-GMP networks remain undiscovered. This idea is evidenced by the diverse repertoire of putative modular sensory and receiver domains predicted to be in c-di-GMP signaling proteins  or lipidMany gaps in our knowledge of well-studied sensor systems remain too. For example, the molecular basis for single-cell heterogeneity in c-di-GMP-dependent chemosensing  or surfa2, and metal ions and to control bioprocesses in a variety of research and industrial contexts. It is also possible that sensory perception may be exploited to devise new therapeutics to treat infections with biofilm etiology.Finally, investigations of sensory perception in c-di-GMP networks are leading to innovations in optogenetics and may provide other biosensors for use in synthetic biology. Such modular sensory devices could be used to sense temperature, O


Legionella pneumophila is a facultative intracellular pathogen that uses the Dot/Icm Type IV secretion system (T4SS) to translocate many effectors into its host and establish a safe, replicative lifestyle. The bacteria, once phagocytosed, reside in a vacuolar structure known as the Legionella-containing vacuole (LCV) within the host cells and rapidly subvert organelle trafficking events, block inflammatory responses, hijack the host ubiquitination system, and abolish apoptotic signaling. This arsenal of translocated effectors can manipulate the host factors in a multitude of different ways. These proteins also contribute to bacterial virulence by positively or negatively regulating the activity of one another. Such effector–effector interactions, direct and indirect, provide the delicate balance required to maintain cellular homeostasis while establishing itself within the host. This review summarizes the recent progress in our knowledge of the structure–function relationship and biochemical mechanisms of select effector pairs from Legionella that work in opposition to one another, while highlighting the diversity of biochemical means adopted by this intracellular pathogen to establish a replicative niche within host cells. Legionella pneumophila exploit eukaryotic cell functions and influence multiple signaling events by translocating over 330 effectors via its Type IV secretion system, also known as the Icm/Dot transporter  have been described in Gram-negative and Gram-positive bacteria. Specialized systems, such as type III and type IV secretion apparatus, have been central to the evolution of many intracellular pathogenic Gram-negative bacteria , 2, 3. Tnsporter , 10, 11.Salmonella typhimurium functions as a guanine-nucleotide exchange factor (GEF) targeting the Rho-family of GTPases  , 15. PTMLegionella represents one of the most elaborate cases of cross talk between host cellular processes and the effectors it translocates. This pathogen has become a popular model for understanding both the role of the effectors in infection and the affected host signaling mechanisms, examples of which have been covered extensively in several past reviews  to the ε-amino group of lysine residues of substrates through the sequential actions of a trio of enzymes, an ATP-dependent Ub-activating E1 enzyme, a Ub-conjugating E2 enzyme, and an E3 Ub ligase  to ubiquitinate serine residues of host targets via a phospho-ribose (PR) linker connecting the hydroxyl group with Arg42 of Ub . These effectors were first described as cysteine-dependent deamidases based on their structural similarity to a group of bacterial deamidases of the Cif family, such as CHBP. They catalyze the conversion of Gln40 of Ub or the Ubl modifier NEDD8 to Glu40 , ubiquitinating it  (B). Formation of this intermediate is accompanied by release of ammonia, aided by His231MavC acting as a proton donor to the leaving group. This intermediate is subsequently attacked by the ε−amino group of Lys92Ube2N resulting in an isopeptide cross-link between Ub and the E2 target. The same intermediate is also prone to a nucleophilic attack by water, especially in the absence of the amine nucleophile, resulting in the deamidation of the glutamine side chain  B. Formatde chain . Recentlde chain , 46, 47.via the active-site thioester linkage in Ube2N∼Ub , MvcA catalyzes the removal of Ub from the ubiquitinated Ube2N (Ub-Ube2N), the product of the MavC-catalyzed modification. Cleavage of the isopeptide cross-link by MvcA leads to regeneration of native Ube2N in a reaction akin to cysteine-dependent deubiquitinase activity of eukaryotic DUBs (C). Hydrolysis of this intermediate releases Ub as the Q40E derivative. Thus, MvcA and MavC function similarly during the first step of their catalysis, forming the thioester intermediate with Ub accompanied by the departure of an amine group (ammonia in the case of MavC and Ube2N in the case of MvcA). The difference lies in the second step, wherein the MvcA catalysis involves water as the nucleophile in contrast to the Lys92Ube2N amine nucleophile in the MavC reaction (B and C). In addition to the same catalytic triad, contact regions between MvcA and its substrates also mimic those in the MavC-substrate complexes (D), pointing to an evolutionary adaptation that enables two enzymes sharing a common fold and mechanistic features to catalyze opposite reactions.Remarkably, despite striking structural similarity with MavC, including identical catalytic residues and a similar INS domain , A and Ctic DUBs , 47, 54.tic DUBs C. Hydrolreaction , B and Complexes D, pointiLegionella necessitated by a specific requirement for Ub attachment and removal. In the initial stages of infection, MavC ubiquitinates Ube2N and dampens NF-κB signaling  . Thus, unlike MavC and MvcA, which have evolved to specifically inhibit Ube2N and restore it, respectively, at different time points during Legionella infection , s, smavC agulation , 57. Surgulation , 46. Thegulation D. Thus, nfection , 44, lpgnfection . Gan et The Rab family of small GTPases are critical mediators of eukaryotic endocytic and secretory vesicular trafficking events , 59, 60.2+ coordination), and Switch II (consists of the DXXG motif that links binding of Mg2+ and the γ-phosphate of GTP). The inactive and active states of these GTPases are distinguished by the conformation of the switch loops . Neunuebel et al. showed that translocation of SidM, SidD, and LepB is temporally regulated. Levels of SidM are high immediately following infection, commensurate with the recruitment of Rab1 to the LCV. Two hours postinfection, the levels of SidM start to decline as the levels of SidD and LepB rise, both of which are required to release Rab1 from the maturing LCV back to the host cytosol : an N-terminal adenylyl-transferase domain (ATase), a C-terminal lipid phosphatidylinositol-4-phosphate binding domain of SidM (P4M), and a central GEF domain . As far as GEF activity is concerned, Rab1 substrate specificity for SidM stems from interactions with residues in the Switch I loop (Asp34 to Ile41). Crystal structures show substantial conformational reorganization in the Rab1 switch regions upon SidM binding (A and B). Although the mode of Rab1 activation is similar to that observed in eukaryotic GEFs, the GEF domain of SidM is structurally distinct from the eukaryotic GEFs (C). Conformational changes during eukaryotic GEF-catalyzed nucleotide exchange involve structural rearrangements within the switch regions, with a more pronounced change in Switch I (A). When a GEF binds to the Switch I loop of the Rab GTPase, it destabilizes the interaction of the GTPase with the phosphate and the Mg2+ ion, pulling Switch I into an open conformation (A and B). This destabilization also displaces the conserved Tyr36 (or Phe in some GTPases) from its interaction with the guanine nucleobase while causing the P-loop to lose its interactions with the phosphate groups of the nucleotide, thus lowering the affinity for GDP even further (A and B). SidM binding also affects the Switch II region, stabilizing it in a conformation reminiscent of a GTP-bound Rab1 , features three distinct functional domains A: an N-tF domain . Once trF domain , where iF domain , 82, 83.F domain B; Step 1 binding , A and Btic GEFs  Fig. 3CCA: an N-Switch I A. When aormation , A and B further , 87, 88. further , 84, 86. further , A and Bund Rab1 .Figure 4B; Step 2) through the covalent attachment of an AMP moiety onto Tyr77 of Switch II (C)  that catalyzes AMPylation of glutamine synthetase for regulating its activity . Two additional aspartates (Asp150SidM and Asp249SidM) contribute to the binding of the Mg2+ ion, with Asp112SidM serving as a general base to promote nucleophilic attack by the phenolic OH group of Tyr77Rab1 . Although the attachment makes little difference to the conformation of Rab1 and hence its GTP affinity, the modification of Tyr77 does prevent the Legionella GAP, LepB, or possibly host GAPs from binding to Rab1 prematurely  while a putative membrane targeting segment in the C-terminal region may assist in the LCV localization (A) . Five catalytic aspartates  and a binuclear bridging water, that acts as the nucleophile, coordinate the two metal ions. Hydrolysis of the adenylyl-O-tyrosyl linkage (AMP-Tyr77) releases AMP and restores Tyr77Rab1 to its unmodified form (A).Overactivation and prolonged LCV retention of Rab1 may cause a drastic effect on host vesicular traffic events that rely on this critical ER-associated GTPase. SidD counteracts SidM-catalyzed AMPylation activity B; Step 3lization A  . The deAtion (A) , 95. The in PPMs A. Five cied form A.Figure B; Step 4), leading to inactivation of Rab1 and their subsequent removal from the LCV. LepB is mechanistically homologous to the eukaryotic Rho/Ras-GAP rather than a Rab-GAP. Traditional Rab-GAPs  and some bacterial GAPs (of the VirA/EspG family) feature a catalytic glutamine finger in addition to the canonical arginine finger (Arg finger) (LepB) instead of the glutamine finger. Glu449LepB occupies a structural position in apo-LepB equivalent to the canonical glutamine in the traditional Rab-GAPs (B). Upon binding to Rab1, Glu449LepB undergoes a pronounced movement that triggers the side chain of Gln70Rab1 to flip toward the γ-phosphate of GTP. As a result, Gln70Rab1 adopts a similar position with respect to the γ-phosphate and a water molecule to what has previously been observed with the catalytic in-trans Gln finger (contributed by the substrate GTPase) in Ras-GAPs (B). The side chain of Gln70Rab1 would orient the water molecule for nucleophilic attack on the γ-phosphate center of GTP to facilitate its hydrolysis much like the catalytic in-cis Gln finger (contributed by the GAP) of TBC-like Rab GAPs (C). Thus, LepB seems to employ the same sort of substrate-assisted catalysis commonly observed in Ras GAP-like catalytic mechanisms  , 97. LepRab-GAPs , 99, 100Rab-GAPs B. Upon bRas-GAPs B. The siRab GAPs , 99, 100Rab GAPs C. Thus, chanisms , 99, 100Legionella to subvert the function of Rab GTPases and facilitate LCV maturation (A). A time-resolved analysis of Legionella effectors that modulate Rab1 function showed that these effectors differ in the specific timing of increase in their levels, which agrees with the role played by them in recruiting Rab1. Allombert et al. demonstrated that AnkX levels begin to rise only after SidM has been translocated and has had a chance to release Rab1 from the Rab1-GDI complex (A) catalyzes phosphocholination of Rab1 using CDP-choline, a modification that is ultimately reversed by Lem3 (A) . . LegioneB) revealed that the Fic domain plays a crucial role in binding the Switch I, II, and C-terminal regions of Rab1 (A and B). Phe107, Ile109, and Asp265 in the active site of AnkX sterically occlude the base of the CMP substrate from binding in the same orientation as seen with nucleotide substrates in other Fic enzymes. Instead, the choline group nestles in this region while the cytidine group stacks against Tyr41 of the CMP-binding domain, forcing a flipped orientation of the bound CMP moiety compared with the nucleotide-binding in AMPylating Fics. Thus, AnkX behaves as a phosphocholine transferase instead of a nucleotide transferase. Upon binding to AnkX, Switch II of Rab1b undergoes a significant structural rearrangement when Phe143AnkX sticks into a hydrophobic pocket formed between Switch II and helix-3 of Rab1b. This encroachment by Phe143AnkX displaces Tyr78Rab1b from its highly conserved position in this hydrophobic cavity (C) . As a result, the otherwise structurally restricted Ser76Rab1 of Switch II can now reach into the active site of AnkX for phosphocholination. The catalytic His229AnkX of the Fic motif acts as a general base in deprotonating the OH group of Ser76Rab1, promoting a nucleophilic attack on the β-P center of the nucleotide (D). The Asp233 residue of the Fic motif positions the catalytic Mg2+, while the Asn235 and Arg237 play a critical role in interactions with the phosphocholine moiety.The different crystal structures of AnkX (in apo and Rab1-bound form) B reveale of Rab1 , 113. Th of Rab1 , A and Bc cavity C 114). . B) revent to it C. As a rcleotide D. The AsLegionella effector Lem3 (A), a phosphodiesterase whose structure is yet to be determined, can reverse the effects of AnkX by removing the phosphocholine group from Ser76 of Rab1 , known so far to modulate Rab1 function, point to the importance of the GTPase in the intracellular lifecycle of the bacteria. Upon activation, Rab1 interacts with other proteins, such as p115 or GM130, in order to guide ER-derived vesicles to fuse with the Golgi apparatus : a deubiquitinase domain (DUB), a phosphodiesterase domain (PDE), a mono-ADP-ribosyltransferase domain (mART), and a C-terminal coiled-coil domain (CC). Biochemical studies have shown that the DUB domain is not essential for the ubiquitinating activity of these proteins  as an intermediate. Subsequently, the PDE domain catalyzes a phospho-transferase-like reaction where the PR-Ub of Ub-ADPR is transferred to the hydroxyl group of a serine residue of the host protein, accompanied by the release of AMP. Several structures of different constructs of these proteins , which facilitates the departure of nicotinamide (Nic) prior to the attack of the Arg side chain on the resulting oxocarbenium center of the Nic-bearing ribose of NAD+  and the second Glu stabilizes the oxocarbenium ion. The conformation of NAD+ observed in SidE mART crystal structures and the placement of critical residues of the RSE motif are consistent with the strain alleviation model of ADP-ribosylation in this enzyme.The mART domain of SdeA, like certain bacterial mART toxins, consists of a helical lobe and a main lobe, together forming the NADg pocket  at theirg pocket B, which  of NAD+ , 127. Thsylation . The consylation B and thePseudomonas aeruginosa (via the formation of a phospho-His-like intermediate (C) (C). Thus, despite the similarity of the PDE domain of SdeA with the bacterial phosphodiesterases, the catalytic motif in the SdeA PDE domain catalyzes a (substituted) phospho-transfer to a serine residue in the ubiquitination reaction instead of water, which would result in hydrolysis of the phosphodiester bond. Incidentally, the PDE domain of the SdeA can also catalyze phospho-transfer to water, ensuing from water attacking the phospho-His intermediate, resulting in the PR-Ub hydrolysis product . H. HPseudoiate (C) . This atiate (C) , 130. Friate (C) C. Thus, Legionella replication in eukaryotes, unchecked activity of these proteins may result in the accumulation of free PR-Ub that would inhibit the host ubiquitination machinery   . Studiesocus (A) , 137.Legionella infection. Inhibition of SidE proteins by SidJ is both temporally regulated and spatially restricted as it requires the host calmodulin (CaM) for its activity , a reaction akin to the acyl-adenylation step of the reaction catalyzed by aminoacyl-tRNA synthetases or activation of Ub by E1. The activated carbonyl of this unstable intermediate is then attacked by the amino group of a free glutamate residue, leading to glutamylation of the SdeA catalytic residue via an isopeptide linkage and the release of AMP (A). This second step likely involves another essential region in SidJ, named the “migrated” nucleotide-binding site . SdeD acts as a hydrolase to catalyze the hydrolysis of PR-ubiquitinated substrates, presumably proceeding via the formation of a histidine-based phosphoramidate intermediate such as the SdeA-PDE mechanism (B). SdeD can remove the AMP moiety from Ub-ADPR, like the hydrolysis reaction catalyzed by the PDE domain of SidE members. However, it cannot transfer the PR-Ub onto target proteins, likely due to the structural differences that alter the conformation and accessibility to the catalytic center in the two effectors. The SdeD loop comprising residues 26 to 48 is missing in SdeA, and conversely, the SdeA loop comprising residues 465 to 513 does not exist in SdeD. It is possible that because of this structural difference, in SdeD, only water can act as the ubiquitin acceptor instead of the hydroxyl group of a serine residue   shares overall structural similarity with the PDE domain of the SidE proteins , 130 aloFig. 7echanism  Fig. 8BBA. SdeD Fig. 8B) . The SdeThe ligase activity of SidE members results in serine ubiquitination of several structurally diverse substrates, the consequences of which affect a variety of cellular processes, ranging from autophagy and vesicular trafficking to tubular ER dynamics and inhibition of protein synthesis. It appears that the seemingly indiscriminate nature of the SidE proteins contributes to the virulence of the pathogen and enables the establishment of the LCV. For example, by PR-ubiquitinating Rab6a and Rab33b, these effectors modulate the Golgi-to-ER retrograde trafficking and prevent the formation of autophagosomes  and all , effectiet al.  inactivation of RavJ by LegL1 (2), polyubiquitination of SidH by LubX, and (3) inhibition of MavQ by SidP.Identification of effector–effector interactions is often the consequence of studying the function of an individual effector. However, recently, studies have systematically explored the interactions between effector pairs by combining a gain-of-function genetic screen in yeast with cellular and biochemical approaches. Urbanus et al.  successfLegionella effector, rearranges the actin cytoskeleton leading to the accumulation of more F-actin on the plasma membrane -containing Legionella effector that binds RavJ . The pret al. (The partnering between a phosphatase (SidP) and a kinase (MavQ) is involved in phosphatidylinositol polyphosphate (PIP) modulation. The amino acid sequence of SidP bears no homology to any known phosphatidylinositol phosphatase (PI phosphatase), while the MavQ sequence is an atypical kinase. Biochemically, SidP is similar to the CX5R-based PI phosphatases that belong to the myotubularin family , 154 in et al.  showed tL. pneumophila allocates about 10% of its protein-coding ability toward functions that require direct interactions with host cellular processes. One of the most remarkable features of Legionella is its capacity to both temporally and spatially regulate the dynamics of its effectors during infection. Three regulatory systems  directly or indirectly regulate the expression of several effector-encoding genes (Legionella triggers the nuclear localization of NF-κB in macrophages to positively upregulate antiapoptotic genes in a Dot/Icm-dependent manner to support intracellular bacterial growth (L. pneumophila is the host amino acid metabolism regulated by mTORC1. As alluded to before, the pathogen frees up host amino acids for its consumption via a concerted albeit temporally regulated action of SidE and Lgt family of effectors (Legionella. On the one hand, the translocated effectors such as MavC and SidE proteins ligate Ub onto host proteins, inactivating or altering their function in the process (ng genes , 161. Thl growth . Anotherffectors . Another process . On the Legionella species, the effector arsenal is quite varied, suggesting that Legionella's genomic flexibility is because of its coevolution with numerous protists species that belong to the phyla Amoebozoa and Percolozoa (Legionella effectors can regulate a myriad of cellular functions (It is not surprising that there is a constant battle between pathogens and their hosts to develop means to adapt and counter-adapt during evolution. In contrast to the high conservation of the secretion system itself among different rcolozoa . Legioneunctions  because unctions . This ununctions . These bunctions , 165. Thunctions .L. pneumophila effectors have been identified using various genetic and biochemical techniques. However, it is still unclear why the pathogen requires so many effectors for its survival within its hosts. Perhaps it is to ensure sustained intracellular replication and is simply an indication of the numerous eukaryotic pathways regulated by the bacteria (Legionella. Despite the diverse nature of the different eukaryotic functions targeted by these effectors, one emerging theme is the existence of a yin and yang type of mechanism. It would not be unreasonable to assume that these principles are co-opted by pathogens in general and not just by bacteria. Future structure–function studies will not only help in evaluating the role of pathogenic effectors but will also aid us in understanding how essential eukaryotic cellular homeostasis is maintained.Many bacteria , making The authors declare that they have no conflict of interest with the contents of this article.


It was found that the PFM amplitude increases with the relative humidity, and that the ferroelectric hysteresis loops at different relative humidity levels show that the coercive bias decreases with the increase in relative humidity. These observed phenomena are attributed to the existence of the water layer between the probe tip and the sample surface in a humid atmosphere, which could affect the effect of the electric field distribution and screening properties at the ferroelectric sample surface. These results provide a better understanding of the water adsorption phenomena at the nanoscale in regard to the fundamental understanding of ferroelectrics’ properties.The effect of relative humidity on the domain structure imaging and polarization switching process of Pb(Zn The superior piezoelectric and ferroelectric properties of ferroelectric materials have made them promising candidates for non-volatile random access memories (NVRAMs), microelectromechanical systems (MEMS), high-performance transducers, sensors, and actuators ,2,3. TheIn the last two decades, thanks to advances in humidity sensor technology ,17,18, hIn this work, two SPM techniques, including PFM and PFS, were used to characterize the domain structure and ferroelectric hysteresis loops of PZN-x%PT single crystals under various relative humidities. The purpose of this work was to study the effect of relative humidity on the domain structure imaging and polarization switching properties of PZN-x%PT single crystals, and to explore the possible mechanisms involved therein.L(010)W(001)T orientations, the samples were cut into dimensions of 4 mm (L) × 4 mm (W) × 0.5 mm (T). Then, the samples were polished to a mirror finish.PZN-x%PT single crystals  grown by an improved flux method were used in this work . After bN) and positive (VP) switching bias could be extracted from the hysteresis loops, and the coercive bias and imprint bias could be calculated by Vc = (VP − VN)/2 and Vi = (VN + VP)/2, respectively [A commercial SPM instrument  was used to carry out the piezoresponse force microscopy (PFM) and piezoresponse force spectroscopy (PFS) measurements for the acquisition of the domain structure and the ferroelectric hysteresis loops, respectively. The PFM and PFS measurements were performed using conductive PtIr-coated silicon tips  . Before each measurement, the probe was calibrated to guarantee data reliability. All the SPM measurements were conducted at room temperature (25 °C), which was much lower than the Curie temperature of PZN-x%PT (160 °C) . In the ectively . The rel2  and in synthetic air  were obtained . In our In addition, the PFM phase images before and after the polarization switching processes in different atmospheres are shown in In this study, the effects of relative humidity on the domain structure imaging and polarization switching properties of PZN-x%PT single crystals were investigated by PFM and PFS techniques. It was found that the PFM amplitude increased with the relative humidity, while the PFM phase had no significant relative humidity dependence. In addition, the ferroelectric hysteresis loops at different relative humidity levels showed that the coercive bias decreased with the relative humidity, while the imprint increased with the relative humidity. The water layer formed between the probe tip and the sample surface due to the condensation of water molecules in the environment was employed to explain the above results. Specifically, the electric field was weakened with the thin water layer at a low relative humidity, while it was enhanced with a thick water layer at a high relative humidity. The study on the polarization switching behaviors in the ambient air and synthetic air revealed that the coercive bias in synthetic air was larger than that in the ambient air, suggesting that the charge species and moisture in the ambient air can facilitate the polarization switching process. These results provided a better understanding of the experimental contribution of the water layer to the domain structure and ferroelectric hysteresis loop measurements of ferroelectric relaxors in SPM studies.


Neural network pruning, an important method to reduce the computational complexity of deep models, can be well applied to devices with limited resources. However, most current methods focus on some kind of information about the filter itself to prune the network, rarely exploring the relationship between the feature maps and the filters. In this paper, two novel pruning methods are proposed. First, a new pruning method is proposed, which reflects the importance of filters by exploring the information in the feature maps. Based on the premise that the more information there is, more important the feature map is, the information entropy of feature maps is used to measure information, which is used to evaluate the importance of each filter in the current layer. Further, normalization is used to realize cross layer comparison. As a result, based on the method mentioned above, the network structure is efficiently pruned while its performance is well reserved. Second, we proposed a parallel pruning method using the combination of our pruning method above and slimming pruning method which has better results in terms of computational cost. Our methods perform better in terms of accuracy, parameters, and FLOPs compared to most advanced methods. On ImageNet, it is achieved 72.02% top1 accuracy for ResNet50 with merely 11.41M parameters and 1.12B FLOPs.For DenseNet40, it is obtained 94.04% accuracy with only 0.38M parameters and 110.72M FLOPs on CIFAR10, and our parallel pruning method makes the parameters and FLOPs are just 0.37M and 100.12M, respectively, with little loss of accuracy. With the development of deep neural networks in recent years, great success has been achieved in computer vision applications ,2,3,4. HThe existing pruning methods are usually classified into two categories based on their compact CNN learning process: (1) Pretraining-dependency pruning, which is based on pretrained filter weights  or dataℓ1-norm , rank ofℓ1-norm  and the ℓ1-norm ,26 of thℓ1-norm ,28,29 anℓ1-norm  during tIn this paper, two novel pruning methods are proposed. First, we pay attention to the information of output feature maps, and propose a novel pruning method: directly calculate the information entropy of feature maps, and the importance of obtaining the corresponding filters , to reduce the redundancy of the network filters. Moreover, we normalize the importance of feature maps of various layers to the same scale to avoid layer-by-layer pruning ratios. Secondly, we propose a new parallel pruning method by combining two methods including our first pruning method based on the entropy of the feature maps and Network Slimming . AlthougWe then compare the proposed methods with other advanced criteria. Experiments demonstrate that our methods can compress various CNN models consisting of VGGNet , DenseNeIn the following, we will first discuss the related work in Weight pruning removes individual neurons in the filter or connections between fully connected layers. Cun et al.  proposedhold. In , pruninghold. In  proposedhold. In .In contrast, generic hardware and software can support filter pruning well, as it removes the entire filter without changing the original convolutional structure. For this purpose, Li et al.  used thePlease note that ,14 invesThe typical pipeline of a conventional pruning algorithm is shown in l-th convolution layer. The shape of filters in l-th convolutional kernel size. l-th layer and its shape is l-th layer is l-th layer and its shape is l-th layer is k-th channel of k-th filter of Assume a pretrained CNN model has L layers, and l-th layer convolution l-th layer Equation   Yil fromWe know that different metrics have different advantages and different pruning effects, resulting in different pruning rates for the same layer of the network. As can be seen in Algorithm 1 A Pruning Algorithm Based on Entropy of Feature Map1:Input: An L-layer Pre-trained CNN model with filter sets 2:Output: The pruned network with filter sets 3:whiledo4:    Compute the feature maps Probability by Equation .5:    Obtain the importance scores for filters Equation .6:    Get the preserved filer set 7:end while8:Get the pruned model without fine-tune.9:whiledo10:    Fine-tune the pruned model.11:end while12:Return the pruned model with filter sets To begin with, the network is trained to be sparse. It is trained to impose sparsity on the scale factor of the BN layer by L1 regularization, and then pruning is performed according to the size of the scale factor. Specifically, the optimization of the objective function is performed as follows.Algorithm 2 A Parallel Pruning AlgorithmRequire: Training set Ensure: The compat network.1:whiledo2:    The model is obtained by sparsity training through Equation 3:end while4:whiledo5:    mask1 = 6:    mask2 = Equation ;7:if then    8:        9:else    10:        11:end if    12:    Pruning the current layer ← mask;13:end while14:Obtain the compat model without fine-tuning.15:whiledo16:    Fine-tune the compat model.17:end while18:Return the compat modelk-th filter of this layer is reserved. (3) The filter retention result mask of this layer is the smallest result in mask1 and mask2. As can be seen from Then, the preserving filters are obtained by comparing the respective thresholds with the importance assessment values of filters. As shown in The experimental results of the parallel pruning method prove the effectiveness of the method, as described in Traditional convolution structure and recent structural variants are two main architectures of the current network structure. The typical former is VGGNet , while tThese networks are pruned by different strategies. For VGGNet, they are all conventional convolutional layers with direct conventional pruning, where the importance of the feature maps obtained from each convolutional layer are evaluated and then pruned according to the importance threshold, as shown in For ResNet, there are some limitations owing to its special structure. For example, in order to complete the summation operation, the channel numbers of each block of the same group need to be consistent. Therefore, it is difficult to directly prune the last convolutional layer of each residual block. Like ResNet164 with three convolution layers per block, most parameters are located in the first two layers. Similarly, each block of ResNet56 has two layers, and most parameters are located in the first layer. Therefore, for each block of ResNet, it is a good choice to keep only the last layer and prune other layers, as shown in DenseNet also has a special structure with certain limitations. Due to the growth rate setting, each dense block generates the same number of feature maps, which are then fused with the previous feature maps. Therefore, it is difficult to directly evaluate the importance of the feature maps generated by the convolutional layers of each dense block. Since each dense block has a BN layer which is not affected by the growth rate, it is a good choice to evaluate the feature maps generated by the BN layer, which is illustrated in To demonstrate the effectiveness and efficiency of our two proposed pruning methods (Algorithms 1 and 2), we conducted extensive experiments on image classification. Representative compact design networks, including VGG16/19 , and ResWe carry out CIFAR experiments on NVIDIA RTX 2060 SUPER GPU and ImageNet experiments on NVIDIA RTX 3090 GPU. All models are implemented and trained using the deep learning framework Pytoch . The effAll networks are trained using stochastic gradient descent (SGD), and we set weight decay and momentum to be As shown in VGG16/19. According to the results of VGG, it can be seen that our algorithm based on the entropy of the feature maps (Algorithm 1) has good results, for example, the compressed VGG16 achieves 93.53% accuracy with only 0.99M parameters and 83.96M FLOPs. using our parallel pruning method (Algorithm 2), the pruned VGG16 has less parameters and less FLOPs with little loss in accuracy. Therefore, our algorithm has the ability to compress the network to a more compact structure.ResNet56/164. On CIFAR10, with similar parameters and FLOPs, our algorithm based on the entropy of the feature maps enables ResNet56 to obtain an accuracy of 93.56% with 0.39M parameters and 69.52M FLOPs, respectively. ResNet164 achieves an accuracy of 94.66% with 0.67M parameters and 111.33M FLOPs, respectively. In addition, using our parallel pruning algorithm is effective in reducing the computation with a slight loss of accuracy. Similarly, we can obtain the same results on CIFAR100. This shows that our algorithm is particularly suitable for pruning residual blocks.DenseNet40. Our algorithm based on the entropy of the feature maps demonstrates that DenseNet40 can obtain 94.04% accuracy on CIFAR10 with only 0.38M parameters and 110.72M FLOPs. Meanwhile, it obtains 74.50% accuracy on CIFAR100 with only 0.40M parameters and 109.55M FLOPs. In addition, our parallel pruning method is able to obtain fewer parameters and computation with little difference in accuracy on both datasets. Overall, our algorithm has better results relative to existing algorithms, so it can work on networks with dense blocks, too.In The results of the comparative experiments on ResNet50 on ImageNet dataset are illustrated in Normalization. From Softmax. To analyze the conventional softmax and the improved softmax, we conducted an analytical comparison. From In this paper, we proposed two novel pruning methods to train compact CNNs. First, our proposed pruning method based on the feature map information entropy acts directly on the feature map, where the accuracy can be well maintained by this information entropy as the filter importance evaluation criterion. Secondly, we further propose a parallel pruning method, which can eliminate the limitations of a single pruning method and significantly reduce the complexity of model. Finally, Our parallel pruning method can be extended by integrating more pruning methods to achieve parallelization and obtain a more compact network model. Numerous experiments have proved the superiority of our filter pruning method over the latest methods.


Propionibacterium acnes and coagulase-negative staphylococci (CoNS) with Staphylococcus epidermidis as a leading species. An as-yet-unanswered question is what enables S. epidermis to colonize skin so successfully. In their recent article, P. D. Fey and his colleagues  have brought us one step closer to answering this question.The stratum corneum is the outermost layer of the epidermis and is thus directly exposed to the environment. It consists mainly of corneocytes, which are keratinocytes in the last stage of differentiation, having neither nuclei nor organelles. However, they retain keratin filaments embedded in filaggrin matrix and possess a lipid envelope which protects the body from desiccation. Despite the desiccated, nutrient-poor, and acidic nature of the skin making it a hostile environment for most microorganisms, this organ is colonized by commensal microbes. Among the classic skin commensals are  Staphylococcus epidermidis the cell wall-anchored protein Aap mediates strong adherence to glycan moieties on corneocytes, making this protein seemingly responsible for the permanent colonization of the skin by coagulase-negative staphylococci (CoNS).Microorganisms that colonize the human skin as commensals must be physiologically equipped to live on this inhospitable habitat over long periods of time and must also be tolerated by the host immune system. Bacteria that stimulate the immune system too much are hardly found as compatible commensals on the skin in the absence of irritation . The firaccumulation-associated protein (Aap). By heterologous expression of Aap on the surface of Lactococcus lactis, Macintosh et al. (S. epidermidis to corneocytes. Evidence for this assumption came from the following observations:)aS. epidermidis strains to corneocytes, suggesting that the target sites were occupied by the A domain, thus preventing S. epidermidis binding; this also ruled out other surface proteins, teichoic acids, or the polysaccharide intercellular adhesin (PIA) (A recombinant A domain was able to block binding of three different in (PIA) , 9 as po)bS. epidermidis adherence.Knocking out PIA and SdrF (Serine-aspartate repeat protein F) expression had no effect on the )cAntiserum raised against the A or B domain of Aap decreased bacterial binding to corneocytes.)dDeglycosylation of the corneocytes’ surface with various deglycosylases decreased adhesion to corneocytes.)eS. epidermidis.Antibody-mediated blocking of the highly expressed corneocyte proteins loricrin and cytokeratin-10 had no effect on the adherence of Aap was first described by the group of G. Peters , 4. The h et al.  demonstrh et al. . The A dh et al. . The strh et al. . TherefoN-glycosidase (PNGase) and O-glycosidase reduced adherence of S. epidermidis by approximately 50%, suggesting that both of these glycan linkages are expressed and serve as binding targets. Pretreatment of corneocytes with glucosidases that remove 5-N-acetylneuraminic acid, fucose, and galactose also decreased adherence while removal of N-acetylglucosamine and mannose residues did not affect adherence. These results suggest that the binding of S. epidermidis to corneocytes is due to the interaction of Aap’s lectin domain with glycoproteins and possibly glycolipids. This is per se not surprising since lectins are carbohydrate binding proteins, i.e., they specifically bind sugar groups that are part of more complex molecules.The deglycosylation experiments were the highlights of this work because they brought us closer to the target structures. The most common sugar tags on human skin surface proteins are N-linked glycosylation at Asn-X-Ser/Thr motifs and O-linked glycosylation at Ser/Thr residues . TreatmeInterestingly, however, Fey and colleagues observed that, although not directly involved in ligand binding, the B domain enhanced binding. They speculate that the B domain may contribute to reduce the electrostatic repulsive forces between the bacterial cell surface and the host surface or increase Aap flexibility. An alternative plausible explanation could be that by omission of the B domain, the distance of the lectin domain from the cell wall is shortened so much that the L-lectin domain becomes partly shielded by the peptidoglycan scaffold of the bacterial cell wall and thus cannot capture the glycan structures on the corneocytes’ surface. A similar situation was described for a cell wall-anchored enzyme (a lipase) wherein a gradual shortening of the cell wall-spanning sequence of the enzyme negatively affected its folding to an active conformation and was correlated with a decreasing activity of the cell-bound lipase .S. epidermidis, the N-terminal region of Aap needs to be processed since it apparently interferes with the Zn2+-dependent dimerization of the B domain   , 14. OnlA (Sep1) . TogetheA (Sep1) , these fS. hominis, S. haemolyticus, S. saprophyticus, and S. capitis. Of note, the Aap-like protein sequences found in S. simulans and S. warneri lacked the A-lectin domain, explaining why these species showed decreased or peripheral adherence to corneocytes . Turning off this cascade contributes to the virulence defect of S. aureus strains and might allow these strains to adapt to skin during colonization where Ebh, SraP, and SasG may act as alternative adhesins (S. aureus and mutants lacking both mgrA and sasG (Bioinformatic analyses revealed that Aap homologous adhesins with a functional lectin-like domain are also present in the other commensal staphylococcal species including neocytes . This isneocytes . The mosadhesins , 18. Theand sasG .S. aureus from S. epidermidis (S. aureus is an aggressive pathogen due to its numerous virulence factors with cell-destructive power. S. epidermidis, on the other hand, is much less virulent, but it is a master of adhesion; there is hardly any implant material to which S. epidermidis has not been found to adhere (S. epidermidis, although not aggressive, is a persistent commensal. The dangerousness of S. aureus is that it can be both: it can be an aggressive and toxin-producing pathogen that causes acute infections, and on the other hand, S. aureus can mutate to commensal nontoxigenic variants with high expression of adherence factors and increased antibiotic resistance which are less distinguishable from S. epidermidis and may cause chronic infections.This brings us to the final point, namely, the old question of what distinguishes pathogenic from nonpathogenic staphylococcal species, or rather, what distinguishes dermidis . There io adhere . In thisS. epidermidis. Aap fulfills two tasks at the same time. The lectin domain mediates binding to glycan structures of skin cells while the truncated isoforms, with their exposed B domain, cause cross-linking between S. epidermidis cells, leading to biofilm and microcolony formation (as illustrated in In summary, the findings by Roy et al. demonstrate the predominant role of Aap in the colonization of the skin and its corneocytes by


IUCrJ (2021). 8, 342–350] report the cryo-EM structure of a Thermotoga maritima encapsulin, a nanocompartment that encapsulates a ferritin-like protein cargo. The 2 Å resolution structure offers insights into the active role of this thermostable encapsulin in regulating iron homeostasis to reduce oxidative stress.Wiryaman & Toor [ Compartmentalization is an essential strategy, widespread throughout all domains of life Archaea, Bacteria and Eukarya. Although the frontiers between different cell types are becoming more and more diffuse, subcellular structures in eukaryotes rely mainly on membrane-bound organelles, whereas protein-based microcompartments are more prevalent in archaeal and prokaryotic cells. Bacterial protein compartments such as ferritins and carboxysomes have critical roles in the vital processes of iron homeostasis and carbon fixation, respectively. Encapsulins make up another protein nanocompartment that contains one or several cargo enzymes  with icosahedral symmetry, with triangulation numbers between 1 and 4, although assembly control does not require auxiliary proteins such as scaffolding proteins or proteases. Although they have little sequence similarity, structural comparison between the encapsulin fold and the capsid protein fold of the Hong Kong 97 (HK97)-like viruses (which have the most successful self-replicating system on Earth) shows high structural similarity  using cryo-electron microscopy (cryo-EM) . Like the fold of the HK97 virus capsid protein, EncTm has three domains: the axial (A) domain, the peripheral (P) domain and the elongated (E) loop. The A domain is responsible for interface interactions at the fivefold symmetry axis, whereas the P domain is at the periphery of pentamers. The E loop is a long β-sheet hairpin with an extended conformation that establishes tight contacts between the twofold symmetry-related subunits from adjacent pentamers; that is, it acts as a molecular hook. An E-loop β-strand forms a β-sheet with a β-strand of the P domain from the other subunit; mostly hydrophobic and aromatic interactions take place at this interface as well as an ionic interaction, all of which contribute to EncTm thermostability.2+ diffusion, iron permeability is probably slowed. The pores at the threefold axis, with an ~6 Å diameter hole, have a ring of Phe residues on the interior side that probably blocks iron transport. Finally, pores in the dimer interface only contain hydrophilic residues, which would allow iron transport.The EncTm nanocontainer encapsulates a ferritin-like protein with ferroxidase activity , and has numerous pores on the five- and threefold axes and around the twofold axis, which regulate iron transport through the protein shell. The pores at the fivefold axis, with an ~5 Å diameter hole, have two rings of His and Tyr residues that point towards the exterior and the interior, respectively; although they allow FeThis study further reports the three tricyclic rings, which correspond to flavin ligands, that surround each of the 20 icosahedral threefold axes on the exterior. The flavin binding site is formed by three different subunits, distinct from the threefold-related subunits Fig. 1. This un


The core objective of current research was determined to nutritional and bioactive profile of maize bran (MB)‐enriched flour in relation to its end‐use product quality. Furthermore, rheological properties of MB‐enriched flour at different levels  were explicated through farinograph and mixograph. Moreover, bread was prepared with the addition of MB‐enriched flour and was characterized for nutritional and textural properties. Results showed that MB‐enriched flour having high water absorption and water retaining potential up to 4%–7% as compared to wheat flour (WF). Moreover, dough height gradually decreased with the addition of MB due to water‐binding ability of bran which causes a decrease in gas retention during fermentation. This resulted in bread volume decrease (4%–7%) as compared to WF. Furthermore, the moisture content and hardness increased with the addition of MB. The water activity of bread slightly increased with the addition of maize bran after 4‐day storage. Conclusively, MB‐enriched flour improved nutritional, textural, and sensorial properties of final product. Maize bran‐enriched flour is rich source of nutritional and bioactive components. Maize bran‐enriched flour bread has high dietary fibers than wheat flour bread. MB‐enriched flour bread explicated high score of sensorial overall acceptability. Cereal brans are generally used in various foods to improve their nutritional profile with special reference to dietary fibers  is a major crop among cereals all over the world. During maize processing, the milling process produces 60–70 g/kg of maize bran , Faisalabad. Maize bran  was procured from Rafhan Maize Product Co Ltd. Faisalabad, Pakistan. Maize bran was commercially micronized (particle size <250 µm) by using milling process.2.1Maize bran (MB) was added with the different proportions of 0% control (WF), 5% (MB5), 10% (MB10), and 15% (MB15) in wheat flour to develop maize bran‐enriched flour.2.2MB‐enriched flour treatment samples were evaluated for proximate composition including moisture content of flour determined by method no. (44–15.02), crude fat content evaluated by method no. (30–10.01), crude protein determined by the percentage of nitrogen in flour method no. (46–19.01), basic ash content was determined by using method no. (08–01.01), crude fiber content was determined by using method no. (32–10.01), described in AACC .2.3Total dietary fiber, soluble, and insoluble dietary fibers of all the treatment groups WF, MB5, MB10, and MB15 were analyzed by the following method no. 32‐05, AACC .2.4Water‐holding capacity (WHC) of wheat flour (WF) and bran‐enriched flour  was measured according to Hemdane et al.  protocol2.5The farinographic characteristics including water absorption capacity, dough development time, dough stability, mixing tolerance index, and dough softness of dough obtained from various blends of wheat flour with maize bran treatments  were analyzed through farinograph by following method no. 54‐21.01 of AACC .2.6Mixographic properties such as mixing time and peak height of the dough obtained from various blends of wheat flour with maize bran treatments  were analyzed through mixograph by following method no. 54‐40.02 of AACC .2.7The ingredients, MB‐enriched flour, milk powder, compressed fresh yeast, sugar, salt, improver, shortening, and water were mixed with mixer and prepared bread with straight dough bread baking method no. 10‐10.03 AACC, . Compari2.8The moisture content of fresh‐baked bread was measured by using a hot air oven at 105 ± 5°C temperature by following the AACC  method n2.9The hardness of the bread was measured by some modification of the bread compression test described by Saeed et al. . In thisThe fracturability of bread was evaluated by a cylindrical ball die that penetrates the bread slices up to 40% followed by method no. 74‐09.02 AACC, .2.10After baking, the bread was cool down for 15 min, and the loaf volume was estimated by the rapeseed replacement method by following the AACC method 10‐05.01 AACC, .2.11The water activity of bread samples was indicated by using the Hygropalm Water Behavior Meter  followed by the method illustrated by Piga et al. .2.12F test value was obtained, and a multiple comparison test was performed on the means, using Duncan's Multiple Range Test at 0.05 levels.Bread treatments WF, MB5, MB10, and MB15 were prepared for sensory evaluation. Sensory acceptance of bread was tested by twenty panelists . Each panelist was presented with four coded samples in a sensory booth and asked to evaluate for appearance, color, taste, flavor, texture, and overall acceptability on a 9‐point hedonic scale, where 1 was disliked extremely and 9 was like extremely. Statistical analysis of data was carried out using SPSS statistics 21. The 2.13F test value was obtained, and a multiple comparison test was performed on the means, using Duncan's Multiple Range Test at 0.05 levels.All experiments were carried out in three replicates. Analysis of variance (ANOVA) was conducted by using the SPSS statistics 21. The 33.1The flour samples were analyzed for moisture content, crude protein, crude fat, crude fiber, NFE, and ash content. The nutritional composition of different flour samples and their mean values are mentioned in Table Moisture content is important aspect to control physiochemical properties of flour. Moisture is important in determining the shelf life of the flour. The mean value of moisture content was 12.48 ± 0.04 in WF (control) sample. The moisture content in MB5, MB10, and MB15 treatments was 12.32%, 12.16%, and 11.69%, respectively. According to Turkish Food Codex (1999) and Codex Alimentarius standards, maximum moisture should be 14.5% and 15.5%, respectively. When the moisture content of flour rises above 14%, it is more susceptible to fungal growth, flavor change, and enzymatic activity , 8.17 ± 0.01% (MB5), 8.32 ± 0.06% (MB10), and 8.54 ± 0.06% (MB15). In all treatments, the average protein content ranges meet the standards (minimum 7.0%) given by Codex Standard for wheat flour  is directly associated with the flour and bran content of the sample. The WHC of wheat flour WF and bran containing composite flour MB5, MB10, and MB15 is shown in Figure m et al.  indicatem et al.  explored3.43.4.1Dough leads to form a wet mass when wheat flour, MB, and water are mixed. The dough is developed during the mixing procedure due to the complex interactions with wheat constituents. Development of the dough starts with the addition of water and continues with mixing process. Water absorption is the amount of water that flour absorbs to maintain the optimal consistency and to produce a better dough for bread preparation. It is the appropriate amount of water you can apply to a dough before the mixture is too sticky. The farinographic results of treatments  were shown in Table 2 gas released during the fermentation. Dough stability is the stage where the gluten breaks down and the dough over mixes. In all the dough systems, mixing is a critical step, that is influenced by the speed of the mixer, dough temperature, water absorption of the flour, and shortening amount in the dough recipe. The dough stability time results were 3.98 ± 0.13, 4.17 ± 0.11, 4.32 ± 0.12, and 4.54 ± 0.6 in WF, MB5, MB10, and MB15 treatments, respectively. Mean values showed that the addition of maize bran caused a nonsignificant increase in dough stability time. This means that bran has a high potential to absorb water, and this makes the dough less stable. The mean values for the effect of maize bran on the mixing tolerance index of wheat flour and increasing pattern for mixing time were observed by applying treatments, that is, WF (control), MB5, MB10, and MB15. The mixing tolerance index was 63.83 ± 0.02, 64.07 ± 0.03, 64.67 ± 0.01, and 65.32 ± 0.02% in WF, MB5, MB10, and MB15, respectively, in wheat flour. It is obvious from these results that the mixing tolerance index of wheat flour is increased with the addition of MB.Maize bran had higher swelling strength and water retaining potential than wheat flour, and improved water absorption could minimize the negative impact of adding maize bran on the gluten network development and the quality of bread. The hydration mechanism is accomplished by forming hydrogen bonds and hydrophilic interactions with the water molecules in protein and starch molecules. Dough mixing is a procedure in which flour and water are stirred until gluten is formed as a consequence of the increased interaction among dispersed and hydrated gluten‐forming proteins. The dough development period was high in MB15 8.25 ± 0.6 min followed by WF, MB5, and MB10 were 6.75 ± 0.25, 7.66 ± 0.38, and 7.58 ± 0.38b, respectively. The development time for dough WF (6.75 ± 0.25 min) was lower than the bran‐enriched maize dough, and this could be due to increased maize bran surface area. In the process of developing dough especially bread dough, the objective was to introduce some physical changes in the dough properties, to enhance its ability to retain the COFurthermore, a decreasing trend was observed in the softness of wheat flour dough by adding maize bran with varying concentrations. The results revealed that the softness of dough was 139.4 ± 0.02, 138.5 ± 0.02, 137.7 ± 0.03, and 132.3 ± 0.06% in WF (control), MB5, MB10, and MB15, respectively.3.4.2Results regarding means values of the peak height variation of wheat flour owing to maize bran addition showed a significant increase in peak height. In MB15 treatment, the highest peak height (67.72 ± 0.11 BU) was observed in flour with 15% maize bran. The peak height results were significantly increased with the addition of maize bran  as shown in Table 3.4.3The bread dough height and after baking bread height mean values result were depicted in Figure After baking, the results for height after baking showed the maximum values. The bread height is greater than the dough height due to the baking fermentation process. Heat speed up the fermentation mechanism which illustrates when bread keeps rising in the first few minutes of baking in the oven. The bread height value was observed in samples WF, MB5, MB10, and MB15 followed by 4.96 ± 0.7, 4.38 ± 0.2, 4.06 ± 0.1, and 3.91 ± 0.06 cm, respectively. The particle of the maize bran causes the gas cell more resilient and sensitive to breakage, and that they develop a physical barrier around the gas cells, pressuring them to expand in a particular dimension . The volume of bread decreased after 4‐day duration, and values were 516.33 ± 8.02, 514.32 ± 4.58, 512.33 ± 13.79, and 489.33 ± 15.37 cm3 in WF, MB5, MB10, and MB15 treatments, respectively. WF treatment bread volume was high than other bran‐enriched treatments. The incorporation of bran to flour normally results in undesirable effects on the properties of the bread dough like. Schmiele et al.  explicated high score of sensorial overall acceptability.The authors declare no conflict of interest.5