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Aims The aim of this study was to construct the improved pMAL expression vector to increase the efficacy of purification of small native peptides and their clear-cut separation from MBP tag. The modifications we introduced can be applied to many expression vectors. Methods and Results To improve the pMAL expression vector, we introduced the His6 tag and the enterokinase cleavage site (Ek) downstream from the MBP tag and Xa cleavage site on the original vector. For cloning of a desired peptide DNA, the enterokinase site contains a unique BsaBI restriction site adjacent to the original multi-cloning site. This redesigned pMAL vector was optimized for the purification of cytoplasmic (pMALc5HisEk) and periplasmic (pMALp5HisEk) peptides. The purification of native and active peptide (P) was obtained following two-step affinity chromatography. In the first step, the entire MBP-His6-Ek-P fusion protein is purified using the Ni-NTA agarose column. This fusion protein was cleaved with active His6 tagged enterokinase. In the second step, the further purification was performed by column containing the mixture of amylose and Ni-NTA agarose resins. This removes both the MBP-His6 and His6-enterokinase leaving pure native protein in solution. These new vectors and the two-step purification protocol were successfully applied in purification of active native small antimicrobial peptides (AMPs), lactococcin A and human β-defensin. Conclusions We constructed the improved pMAL expression vectors and established the pipeline and optimal conditions for their use in efficient purification of large amounts of active native small peptides. Significance and Impact of the Study Choice of expression vector impacts on the efficiency of expression and purification of desired proteins. The idea of redesigning pMAL vector was driven by the need for rapid purification of larger amounts of active native AMPs. This newly improved pMAL vector, the cloning strategy, expression conditions and two-step purification protocol represent a unique simple approach which can be applied in every laboratory.

Lactic acid bacterium Lactococcus lactis BGBU1-4 produces 43 amino acids (aa) long bacteriocin, lactolisterin BU (LBU), a 5.161 kDa peptide with potent antibacterial activity against many Gram-positive pathogens. In addition, BGBU1-4 produces an additional unknown product of 3.642 kDa with antibacterial activity. Here, we determined that the significant amount of naturally produced LBU breaks down to create a 3.642 kDa truncated form of LBU bacteriocin consisting of 31 N-terminal aa (LBU1-31) that exhibits 12.5% the antibacterial activity of the full-length LBU. We showed that chemically synthesized LBU is stable and 50% less active than native LBU, and so we used the synthetic peptides of LBU and its variants to further study their activities and antibacterial potential. Deletion analysis of LBU revealed that the 24 N-terminal aa of LBU (LBU1-24) are responsible for antibacterial activity, while downstream aa (25–43) determine the species-specific effectiveness of LBU. Although LBU1-31 contains aa 1–24, the truncation at position 31 is predicted to change the structure within aa 15–31 and might impact on antibacterial activity. Intriguingly, whole genome sequencing and genome mining established that BGBU1-4 is abundant in genes that encode potential antibacterials, but produces LBU and its breakdown product LBU1-31 exclusively.

The central element of the integrative traffic planning is Sustainable urban mobility plan (SUMP). It is based on EU documents and guidelines set by European commission. Integrated traffic planning does not reject, but upgrades current planning practices and it has a long-term and strategic vision striving towards sustainable mobility. SUMP of the Municipality of Veles is aiming towards an attractive public passenger transport, branched network of safe cycling routes and good conditions for pedestrians. It focuses on city center as a regulated, attractive, accessible and safe urban space. This paper summarized the sustainable measures and projects that are planned for implementation and have impact on the city and its inhabitants.

The issue of Climate Change has raised so much attention for decades, specifically in recent years. Modern trends in urban architecture which tried to create living machines similar to each other in any context, have now been proven to be insufficient and unless we decide to respect nature and make our buildings cooperate with their contexts as soon as possible we cannot address further threads regarding our future life. If we take a look at traditional architecture existing in old towns and rural area, we can see how people managed to bond a new construction with its surroundings. In this article we study different architectural patterns which exists in old architectures of towns and villages in Iran; the country which can be a good case study due to its rich diversity of nature, climate zones and architectural background. In Iran we have a range of climate of cities from hot and dry to humid and very cold ones. So, depending on the city climate, water supplies and energy sufficiency people tried to create best residential buildings. Here we try to check out some examples.

Introduction: After marsupialisation of large jaw cysts, there comes to the formation of bone defects which do not guarantee the stability of the blood clot and therefore are filled with iodine gauze. Any replacement of iodine gauze, toilet of the wound, causes minor or major bleeding, which slows wound epithelialisation and organization. Therefore, making the partial acrylic denture obturator is recommended to be done a few weeks after the surgery. Case report: This work presents two patients who had partial acrylic denture obturator made on the tenth day after the surgery. This allowed significantly easier toilet of the wounds, faster epithelialization and organization, and thus faster bone healing. Conclusion: Restored function of chewing, swallowing and speech, as well as the aesthetic appearance suggest that early use of acrylic obturator prosthesis is very significant.

Numerous applications in wireless sensor networks (WSNs) require collecting data without loss during transmission, long-term sensing, and long system lifetime. Achieving reliable data transfer and long system lifetime is difficult because, from one hand, the wireless transfer is error prone, and, on the other, sensor node (SN), as battery powered device, is energy limited. By using some power-aware techniques, such as duty-cycling and power-gating, it is possible to reduce the energy consumption at an acceptable level. The requirements for higher level of reliability during wireless data transfer have increased the use of error correcting codes (ECCs). Codes represent an effective means of providing protection against injection of single-/double-/multiple bit errors over noisy communication channel. The two basic mechanisms to recover erroneous packets in any network are Automatic Repeat Request (ARQ), and Forward Error Correction (FEC). As energy consumption is a major issue in concern in WSN, packet retransmission is not an option and FEC would be preferred over ARQ. In this paper, an efficient scheme, based on two-dimensional (rectangular) block ECC code, referred as Two Dimensional Single Error Correction and Double Error Detection (2D SEC-DED), has been developed. By using 2D SEC-DED encoding all single-bit and 99.9% of double −/multiple-bit errors, within transferred packets, are recovered. In this way, the number of retransmissions, when WSN operates in harsh environmental (bit error rate (BER), BER > 10− 4) is decreased, what means that not only energy saving but also extension of the transmit range (transmission distance between the transmitter and receiver), is achieved. As illustration, for indoor environment (the path loss exponent, also known as propagation constant or space loss factor, α = 4) at the target BER of 5 · 10− 4, the proposed encoding scheme is able to improve the transmission distance by about 18 m or the received signal strength (RSSI) by about 8.5dBm compared to WSN without error correction (WSN which use Cycle Redundancy Check (CRC) encoding as error detection mechanism).

The national economic and political conditions of a country inevitably affect the development of its social housing sector. However, in order to prevent social exclusion, the quality of social housing has to at least be similar to the national average quality. As the practice of social housing in Serbia is still in its early stages, and the norms for this type of housing have only applied since 2013, this research strives to influence the formation of a more complete basis for the development of contemporary social housing models in Serbia, with the aim of improving the quality of social housing and achieving the prevailing housing quality in the country. A comparative analysis of the norms defined for social housing and those defined for market housing, as well as the results from the last census concerning the average housing area per member in Serbia, were used to define the position of social housing in comparison to other types of housing in the country. The analysis pointed out that the spatial comfort of social housing units is lower than other forms of housing in the country. In addition to which the unit area and the individual rooms are inadequately defined by the current social housing standard. The second part of the research, which is based on relevant architectural recommendations regarding the functional minimum regarding spatial comfort of the apartment units, aims to point out the necessary minimum of the housing space. The results of this research define recommendations for the minimum spatial comfort of social housing units: 1) the minimum floor area per member; and 2) the minimum floor area and width of the individual rooms.

The complex of the Pasini konaci in Vranje, was built in the second half of the 18th century, and it is composed of two buildings. The pasha used to reside in the Selamluk with his entourage, while the Haremluk was a residence of the female part of the family. Due to the continuing use of these two buildings, their architectonic characteristics and historical environment were preserved to a great extent. Nowadays, the Selamluk and Haremluk have been declared the cultural property of great importance for Serbia. The first part of the paper about the Selamluk treated the genesis of spatial organization and architectonic form of the building on the basis of historical sources. This part analyzes the structural and decorative elements of the Selamluka in Vranje as a typical representative of the Balkan Oriental housing architecture. Cultural, historical, artistic and craftsmanship values, based on the material acquired during the field work, were discussed. The detailed study and comparative analysis of the Selamluk characteristics provided a contribution to understanding of specific values in the architectonic evaluation of town structures in this part of the Balkan peninsula.

In this paper, we present one approach in design of self-tuning all-pass, band-pass, low-pass and notch filters based on phase control loops with voltage-controlled active components and analyze their stability as well. The main idea is to vary signal delay of the filter and in this way to achieve phase correction. The filter phase characteristics are tuned by varying the transconductance of the operational transconductance amplifier or capacitance of an MOS varicap element, which are the constituents of filters. This approach allows us to implement active filters with capacitance values of order of pF, making the complete filter circuit to be amenable for realization in CMOS technology. The phase control loops are characterized by good controllable delay over the full range of phase and frequency regulation, high stability, and short settling (locking) time. The proposed circuits are suitable for implementation as a basic building RF function block, used in phase and frequency regulation, frequency synthesis, clock generation recovery, filtering, selective amplifying etc.

Pasini konaci (Pasha?s residence), Selamluk and Haremluk in Vranje, are typical representatives of Balkan Oriental architecture of 18th century. In spite of the buildings becoming very decrepit and prone to dilapidation due to their structural characteristics, in terms of their function and construction technique, they represent an evidence of the living standard of a social class and the social relationships of the historical age they belong to. With the aid of an analysis of historical data, the part 1 of this paper presented the results of the research of the origins and development of the Selamluk in Vranje and its surrounding area with an attempt to determine architectonic genesis from the original structure, through the destruction phases and reconstruction to the present day status. In addition to the information published in the relevant literature, the data were collected by analyzing old plans, photographs and documents archived in various cultural institutions. In the part 2 of this paper, the structural and decorative elements of the Selamluk are discussed for the first time in literature.

The circuit-level simulation analysis of the single event transient response of an on-chip single event latchup protection switch (SPS cell), previously designed and developed in the IHP 250 nm CMOS process, is presented. The SPS cell provides the latchup protection for standard cells on the principle of power domain control. It is based on a sensing/driving PMOS transistor which acts both as a latchup sensor and a driver for the standard cells, and includes additional PMOS and NMOS transistors for controlling the sensing/driving transistor and interfacing to the external control logic. The previous work has confirmed the SPS cell's functionality in the case of single event latchup, and the experimental investigation has proven that the SPS cell is immune to single event latchup for LET values up to 74.8 MeV?cm2/mg. This case study extends the previous research by introducing the circuit-level simulation of the SPS cell's response to the single event transients (SETs). Using the square pulse current as a SET model, the amplitude and duration of the SET-induced voltage pulses at the SPS cell's outputs have been analyzed with respect to the injected charge, operating temperature, supply voltage, load (number of standard cells connected to the SPS cell) and sensing/driving transistor's channel width. The results have shown that the immunity of the SPS cell to SETs can be significantly enhanced by connecting a larger number of standard cells to the SPS cell and increasing the sensing/driving transistor's size, without any area overhead. An analytical expression for calculating the critical charge in terms of the transistor size and the number of standard cells connected to the SPS cell has been derived. Moreover, the SPS cell can be used as a SET sensor for detecting the levels of injected charge which cannot be mitigated by the increase of transistor size and load, and in conjunction with the external control logic it could be possible to measure the SET duration and perform online corrections of the SET-induced faults in the standard cells supplied by the SPS cell.

The bacterial cell envelope retains a highly dense cytoplasm. The properties of the cytoplasm change with the metabolic state of the cell, the logarithmic phase (log) being highly active and the stationary phase metabolically much slower. Under the differing growth phases, many different types of stress mechanisms are activated in order to maintain cellular integrity. One such response in enterobacteria is the phage shock protein (Psp) response that enables adaptation to the inner membrane (IM) stress. The Psp system consists of a transcriptional activator PspF, negative regulator PspA, signal sensors PspBC, with PspA and PspG acting as effectors. The single molecule imaging of the PspF showed the existence of dynamic communication between the nucleoid-bound states of PspF and membrane via negative regulator PspA and PspBC sensors. The movement of proteins in the cytoplasm of bacterial cells is often by passive diffusion. It is plausible that the dynamics of the biomolecules differs with the state of the cytoplasm depending on the growth phase. Therefore, the Psp response proteins might encounter the densely packed glass-like properties of the cytoplasm in the stationary phase, which can influence their cellular dynamics and function. By comparing the properties of the log and stationary phases, we find that the dynamics of PspF are influenced by the growth phase and may be controlled by the changes in the cytoplasmic fluidity.

Wound healing is a complex process that includes hemostasis, inflammation, proliferation and tissue remodeling. Growth factors are natural biological mediators that regulate crucial cellular processes involved in the tissue repair, such as DNA synthesis, angiogenesis, metabolic activity, migration, chemotaxis, proliferation, differentiation and matrix synthesis. The most important growth factors that play a part in the extraction-wound healing process and bone tissue regeneration are: platelet - derived growth factor-PDGF, transforming growth factor Beta-TGF β, insulin-like growth factor-lGF, bone morfogenetic protein-BMP-2, BMP- 7 ,vascular endothelial growth factor- VEGF, fibroblast growth factor-FGF. Growth factors appear at various concentrations at different times, so that the wound age may be estimated by their age. Except wound healing growth factors may be used in better oseointegration of imlants, alveolar ridge augmentation, alveolit etc. Studies of physiological processes in which growth factors have a regulatory role indicate that these molecules rarely act in biological isolation. The study of the interaction between the growth factors in the alveolar bone can explain tissue ability to heal even under adverse conditions, such as infection and radiation.

Although single-family housing in the city is considered higher quality and preferred type of housing, housing crisis, as a permanent actual problem in the world, requires adequate solutions. In this sense, housing in multi-family housing buildings can be considered as a necessity (social, moral, economic, etc.), but in fact, now and in the future, it is the main form of housing construction which can give the solution for housing problems. However, to make this housing type more attractive and acceptable to the occupants, it is necessary to improve it by the implementation of individualization modalities, and in that way make its qualities much closer to the preferred single-family housing.

It is generally known that a man causes changes in the environment and thus many of these changes affect the relationship between man and his environment. Additionally, population increase and urban concentration directly contribute to pollution and degradation of the environment. Considering that building construction alone represents the largest segment of energy consumption in EU (over 40%), it is logical that it has aroused the interest of scientific and professional community in terms of finding new technologies for increasing building energy efficiency and applying renewable energy sources. Satisfaction of energy demand of a country depends not only on the available energy and economic resources, but also on economic-political and energy conditions in general environment.

Wireless sensor networks (WSNs) include a large number of distributed sensor nodes that consist of sensor, microcontroller and transceiver block. As the main source of energy sensor nodes usually use non-rechargeable batteries, so that the stored energy is a very important resource of a WSN. The main goal of this paper is estimating the battery lifetime of a sensor node in respect to the usage pattern. Calculating the battery remaining capacity is performed in real application conditions by using information from datasheets for all its blocks. Simulation results show that the design and operation modes of the sensor node affect the battery lifetime.

Background/purpose Tooth extraction is often followed by a number of different complications that demand additional treatment. In order to accelerate healing processes and decrease the complication occurrence various agents, growth factors, natural and synthetic antioxidants (e.g coenzyme Q10-CoQ10), are applied. Due to the partially known health-promoting effects of CoQ10 we decided to assess potential of it's encapsulated in nanoliposomes form on wound healing process following tooth extraction. Materials and methods Effects of free and encapsulated form of CoQ10 on wound healing processes after tooth extraction in rats, 3 and 7 days following surgical procedure, was studied by means of tissue biochemical (myeloperoxidase activity and nitric oxide (NO) concentrations) and pathohistological analysis. Results The obtained results indicate that the encapsulated form of CoQ10 compared to control and CoQ10 treated animals statistically significantly decreases inflammatory process estimated through myeloperoxidase activity and NO concentrations, as well as based on histopathological analysis 3 and 7 days following surgery. Conclusion The results of this study unequivocally prove that the encapsulation of CoQ10 in nanoliposomes enhances CoQ10 activity by accelerating wound healing process after tooth extraction.

The phage-shock protein (Psp) system is believed to manage membrane stress in all Enterobacteriaceae and has recently emerged as being important for virulence in several pathogenic species of this phylum. The core of the Psp system consists of the pspA–D operon and the distantly located pspG gene. In Salmonella enterica serovar Typhimurium (S. Typhimurium), it has recently been reported that PspA is essential for systemic infection of mice, but only in NRAMP1+ mice, signifying that attenuation is related to coping with divalent cation starvation in the intracellular environment. In the present study, we investigated the contribution of individual psp genes to virulence of S. Typhimurium. Interestingly, deletion of the whole pspA–D set of genes caused attenuation in both NRAMP1+ and NRAMP1− mice, indicating that one or more of the psp genes contribute to virulence independently of NRAMP1 expression in the host. Investigations of single gene mutants showed that knock out of pspB reduced virulence in both types of mice, while deletion of pspA only caused attenuation in NRAMP1+ mice, and deletion of pspD had a minor effect in NRAMP1− mice, while deletions of either pspC or pspG did not affect virulence. Experiments addressed at elucidating the role of PspB in virulence revealed that PspB is dispensable for uptake to and intracellular replication in cultured macrophages and resistance to complement-induced killing. Furthermore, the Psp system of S. Typhimurium was dispensable during pIV-induced secretin stress. In conclusion, our results demonstrate that removal of PspB reduces virulence in S. Typhimurium independently of host NRAMP1 expression, demonstrating that PspB has roles in intra-host survival distinct from the reported contributions of PspA.

Introduction: Soft laser could be used for the treatment of different painful diseases and conditions in the oral cavity. One of them is Herpes labialis. The aim of the paper was to study the efficiency of soft laser in the treatment of Herpes labialis. Material and method: The study included 43 patients. Laser therapy was applied in 23 patients, whereas 20 patients were treated by Ung. Acycklovir. Results: The level of statistical significance of differences in mean values of pain between the groups was maximum (p

Experiences of the countries with developed social housing sector indicate that the role of social housing should not be treated in a limited manner, considered to be just a roof over one’s head, whereby housing quality will be neglected. Contemporary approaches to the development of social housing models are focused on users themselves, with a special concern for the housing quality. Because the units of social housing have the minimal size, it is necessary to provide such amenities so as to overcome the shortage of usable housing space. It is also desirable that the design of social housing dwellings should have a simulative effect on the tenants within this areas. This paper discusses the spatial and physical parameters which could affect the housing quality of this type of dwellings. Key words: housing quality, housing standard, housing unit, social housing

Recognizing the uncertainty of future city development is the basic starting point of sustainable urban planning. Flexibility in design enables the development of “age-friendly cities”, reducing the risks of functional failures caused by changing living circumstances. Planning strategies that enable adaptation of the system to changing environmental, functional, social and technological conditions are essential to the long service life and the resilience of buildings and hence to sustainable city development. This paper considers the implementation of flexible design strategies within the concept of sustainability, defined as a key tool of urban renewal and resilient city development. Value-enhancing flexibility in urban and architectural design is an essential resource for all participants of integrative city development: designers, managers, financial analysts, investors, regulators and academics. Considering this, it is essential to support the flexibility in both conceptual and legislative framework of sustainable urban strategies. Key words : sustainability, flexibility, adaptability, urban resilience

The clock is one of the most critical signals in any synchronous system. As CMOS technology has scaled, supply voltages have dropped chip power consumption has increased and the effects of jitter due to clock frequency increase have become critical and jitter budget has become tighter. This article describes design and development of low-cost mixed-signal programmable jitter generator with high resolution. The digital technique is used for coarse-grain and an analogue technique for fine-grain clock phase shifting. Its structure allows injection of various random and deterministic jitter components in a controllable and programmable fashion. Each jitter component can be switched on or off. The jitter generator can be used in jitter tolerance test and jitter transfer function measurement of high-speed synchronous digital circuits. At operating system clock frequency of 220 MHz, a jitter with 4 ps resolution can be injected.

The description of residential buildings in the past is made possible by studying the physical structure of the preserved architectural and movable archaeological finds, and also by the analysis of the written (epigraphic and numismatic) material findings, as well as the analysis of the written literary works which, through the author?s subjectivity, documented the social life. That is why the knowledge of the classical antiquity construction handbooks can only contribute to the understanding of the architecture, housing conditions, and thus facilitate the reconstruction of the ancient houses. Works about residential buildings by Columella, Vitruvius and Faventinus, three authors from different periods of the mature antiquity, are fully preserved. This paper presents the results of a comparative analysis of their works, which indicate that the recommendations for determining the properties of various functional units within the residential building were similar over a longer period of time observed here.

The phage shock protein (Psp) response maintains integrity of the inner membrane (IM) in response to extracytoplasmic stress conditions and is widely distributed amongst enterobacteria. Its central component PspA, a member of the IM30 peripheral membrane protein family, acts as a major effector of the system through its direct association with the IM. Under non-stress conditions PspA also negatively regulates its own expression via direct interaction with the AAA+ ATPase PspF. PspA has a counterpart in cyanobacteria called Vipp1, which is implicated in protection of the thylakoid membranes. PspA's and Vipp1's conserved N-terminal regions contain a putative amphipathic helix a (AHa) required for membrane binding. An adjacent amphipathic helix b (AHb) in PspA is required for imposing negative control upon PspF. Here, purified peptides derived from the putative AH regions of PspA and Vipp1 were used to directly probe their effector and regulatory functions. We observed direct membrane-binding of AHa derived peptides and an accompanying change in secondary structure from unstructured to alpha-helical establishing them as bona fide membrane-sensing AH's. The peptide-binding specificities and their effects on membrane stability depend on membrane anionic lipid content and stored curvature elastic stress, in agreement with full length PspA and Vipp1 protein functionalities. AHb of PspA inhibited the ATPase activity of PspF demonstrating its direct regulatory role. These findings provide new insight into the membrane binding and function of PspA and Vipp1 and establish that synthetic peptides can be used to probe the structure-function of the IM30 protein family.

Transcription regulation in a temporal and conditional manner underpins the lifecycle of enterobacterial pathogens. Upon exposure to a wide array of environmental cues, these pathogens modulate their gene expression via the RNA polymerase and associated sigma factors. Different sigma factors, either involved in general 'house-keeping' or specific responses, guide the RNA polymerase to their cognate promoter DNAs. The major alternative sigma54 factor when activated helps pathogens manage stresses and proliferate in their ecological niches. In this chapter, we review the function and regulation of the sigma54-dependent Phage shock protein (Psp) system-a major stress response when Gram-negative pathogens encounter damages to their inner membranes. We discuss the recent development on mechanisms of gene regulation, signal transduction and stress mitigation in light of different biophysical and biochemical approaches.

Diabetes mellitus (DM) and chronic periodontitis are common chronic diseases in adults in the world population. DM has a strong influence on the oral cavity and represents a risk factor for gingivitis and periodontitis. Low-level laser therapy (LLLT) has proven effective in the reduction of inflammation and swelling. The aim of the present study was to evaluate the efficacy of LLLT in diabetic periodontitis through histological analysis. A total of 300 diabetics with chronic periodontal disease and teeth indicated for extraction were assigned into six equal groups. In the groups 1 and 4, indicated teeth were extracted before treatment, and in the rest of the groups upon completion of the entire treatment. All patients received oral hygiene instructions and full-mouth conservative periodontal treatment. In groups 3 and 6, LLLT was applied (670 nm, 5 mW, 2 J/cm(2), 16 min, 5 days). Histologic findings of gingival tissue treated with LLLT showed expressed healing, as is evident by the absence of inflammatory cells. Tissue edema could not be seen, and the number of blood vessels was reduced. In the gingival lamina, propria pronounced collagenization and homogenization were present. It can be concluded that LLLT has shown efficacy in the treatment of periodontitis in diabetics. Because of more pronounced alterations of periodontium in diabetics, the use of LLLT is of particular importance.

Background/Aim. Ropivacaine is a relatively new longacting local anesthetic. The aim of this study was to compare the postoperative analgesic effect of topical anesthetics ropivacaine 0.75% and lidocaine 2% with adrenaline in the postoperative treatment of periapical lesions in the maxilla. Methods. The study was conducted on 60 subjects, divided into two groups. The study-group received 0.75% ropivacaine without a vasoconstrictor, while the control group was treated with 2% lidocaine with adrenaline (1 : 80.000). Block anesthesia for n. infraorbitalis was used and local anesthetics were applied also on the palatine side for the end branches of n. nasopalatinus. The following parameters were observed: time elapsed from the application of an anesthetic until the first occurrence of pain after the surgery and first intake of an analgesic, the intensity of initial pain, pain intensity 6 h after the application of anesthetics and the total number of analgesics taken within 24 h after the completion of surgery. Results. The pain appeared statistically significantly earlier in the patients who had been given lidocaine with adrenaline (p < 0.001), while statistically significantly higher mean values of initial postoperative pain (p < 0.05) and pain intensity 6 h after the intervention (p < 0.01) were also registered in the same group of patients. In the period of 24 h upon the intervention, the study-group patients were taking less analgesics as compared to the control-group subjects (46.6% vs 73.3%), who were given analgesics earlier, although no statistically significant differences were observed related to the number of analgesic doses taken. Conclusion. The results of our study indicate a better postoperative analgesic effect of ropivacaine as compared to lidocaine with adrenaline.

Phage shock proteins (Psp) and their homologues are found in species from the three domains of life: Bacteria, Archaea and Eukarya (e.g. higher plants). In enterobacteria, the Psp response helps to maintain the proton motive force (PMF) of the cell when the inner membrane integrity is impaired. The presumed ability of ArcB to sense redox changes in the cellular quinone pool and the strong decrease of psp induction in ΔubiG or ΔarcAB backgrounds suggest a link between the Psp response and the quinone pool. The authors now provide evidence indicating that the physiological signal for inducing psp by secretin-induced stress is neither the quinone redox state nor a drop in PMF. Neither the loss of the H+-gradient nor the dissipation of the electrical potential alone is sufficient to induce the Psp response. A set of electron transport mutants differing in their redox states due to the lack of a NADH dehydrogenase and a quinol oxidase, but retaining a normal PMF displayed low levels of psp induction inversely related to oxidised ubiquinone levels under microaerobic growth and independent of PMF. In contrast, cells displaying higher secretin induced psp expression showed increased levels of ubiquinone. Taken together, this study suggests that not a single but likely multiple signals are needed to be integrated to induce the Psp response. Electronic supplementary material The online version of this article (doi:10.1007/s00284-011-9869-5) contains supplementary material, which is available to authorized users.

Residence of the elderly is analyzed in this work. Experiences of the developed world in this field can be extremely beneficial to the housing policy in Serbia. The elderly are facing serious facility shortages for living compared to the actual demand. The subject-matter of this work is to carry out research into the institutional forms of taking up residence of the elderly in Serbia and France, since France is a welfare state. By analyzing French pattern, some useful recommendations can be found for the development of the Serbian pattern.

Batteries are the main source of energy for low-power electronics such as micro-electro mechanical systems (MEMS), wireless sensor networks, embedded devices for remote sensing and control, etc. With the limited capacity of finite power sources and the need for supplying energy for the lifetime of a system/device there is a requirement for self-powered devices. Using conventional batteries is not always good design solution because batteries require human intervention to replace them (very often in hard-accessible and harsh-environmental conditions). Therefore, acquiring the electrical power, by using an alternative source of energy that is needed to operate these devices is a major concern. The process of extracting energy from the surrounding environment and converting it into consumable electrical energy is known as energy harvesting or power scavenging. The energy harvesting sources can be used to increase the lifetime and capability of the devices by either replacing or augmenting the battery usage. There are various forms of energy that can be scavenged, like solar, mechanical, thermal, and electromagnetic. Nowadays, there is a big interest in the field of research related to energy harvesting. This paper represents a survey for identifying the sources of energy harvesting and describes the basic operation of principles of the most common energy harvester. As first, we present, in short, the conversion principles of single energy source harvesting systems and point to their benefits and limitations in their usage. After that, hybrid structures of energy harvesters which simultaneously combine scavenged power from different ambient sources (solar, thermoelectric, electromagnetic), with aim to support higher load at the output, are considered.

Sensing the environment and responding appropriately to it are key capabilities for the survival of an organism. All extant organisms must have evolved suitable sensors, signaling systems, and response mechanisms allowing them to survive under the conditions they are likely to encounter. Here, we investigate in detail the evolutionary history of one such system: The phage shock protein (Psp) stress response system is an important part of the stress response machinery in many bacteria, including Escherichia coli K12. Here, we use a systematic analysis of the genes that make up and regulate the Psp system in E. coli in order to elucidate the evolutionary history of the system. We compare gene sharing, sequence evolution, and conservation of protein-coding as well as noncoding DNA sequences and link these to comparative analyses of genome/operon organization across 698 bacterial genomes. Finally, we evaluate experimentally the biological advantage/disadvantage of a simplified version of the Psp system under different oxygen-related environments. Our results suggest that the Psp system evolved around a core response mechanism by gradually co-opting genes into the system to provide more nuanced sensory, signaling, and effector functionalities. We find that recruitment of new genes into the response machinery is closely linked to incorporation of these genes into a psp operon as is seen in E. coli, which contains the bulk of genes involved in the response. The organization of this operon allows for surprising levels of additional transcriptional control and flexibility. The results discussed here suggest that the components of such signaling systems will only be evolutionarily conserved if the overall functionality of the system can be maintained.

The bacterial phage shock protein (Psp) response functions to help cells manage the impacts of agents impairing cell membrane function. The system has relevance to biotechnology and to medicine. Originally discovered in Escherichia coli, Psp proteins and homologues are found in Gram-positive and Gram-negative bacteria, in archaea and in plants. Study of the E. coli and Yersinia enterocolitica Psp systems provides insights into how membrane-associated sensory Psp proteins might perceive membrane stress, signal to the transcription apparatus and use an ATP-hydrolysing transcription activator to produce effector proteins to overcome the stress. Progress in understanding the mechanism of signal transduction by the membrane-bound Psp proteins, regulation of the psp gene-specific transcription activator and the cell biology of the system is presented and discussed. Many features of the action of the Psp system appear to be dominated by states of self-association of the master effector, PspA, and the transcription activator, PspF, alongside a signalling pathway that displays strong conditionality in its requirement.

Single family housing is for many reasons considered a more favorable form of housing than the multi-family one. Hence, designing of housing in a multi-family housing structure is a special challenge for designers, because it is expected that the dwelling comfort offered by the multi-family structure is as similar to one of living in a house as possible, that is to seek analogies with the family house when designing a multi-family building. There is a number of possible ways to individualize a multi-family building, regarding the apartments, architectonic composition or urban composition, whose realization would contribute to enhancement of multi-family housing quality.

Introduction. Reconstruction of perinasal area is still a challenge for the surgeon who is involved in excisional tumor surgery. The authors report their experience in reconstruction of hemi-nose defects with the free microvascular submental island flap prelaminated with oral mucosa. Material and methods. A 73-year-old male patient with recurrent extensive basal cell carcinoma of the right hemi-nose (nasal ala and lateral nasal part) and portion of the upper lip underwent to excisional surgery and creation of lateral nasal defect 3x2cm in size. Six months after the first surgery the reconstruction of postoperative defect was done by means of free microvascular submental flap prelaminated with oral mucosa. Results. After necrosis of flap 3mm x 4mm in size around the tip zone of the nose on the 7th postoperative days, the rest of microvascular submental flap prelaminated with oral mucosa survived completely with good cosmetic and nasal respiratory results. Conclusion. This flap can be used successfully in reconstruction of the hemi-nose area. Color and texture of the flap match with adjacent tissue and adequate diameter of submental vessels and reasonably long vascular pedicle enables successful microvascular anastomosis.

Introduction The oroantral communication is a pathologic communication between the oral cavity and maxillary sinus. It originates with extraction of the upper lateral teeth. Primary plastics communication, which is more extensive than 5 mm has been unsuccessful in 16%. Small power lasers have positive reaction on wounds healing. The aim of this work was to determine the lasers effects on slice epithelization after the plastics more extensive than 5 mm. Material and methods The experimental research was done on dogs with extraction of the upper second premolars on both sides, and formed oroantral communication having diameter of 10 mm. After the plastics of sinus, the left side slices were exposed to radiation for 7 days, and the opposite slices healed spontaneously. Eight points (8x1J) were treated for 100s by GaAlAs laser, power 10 mW and wavelength 670 nm. Biopses of the slices connections were taken on the 14th day to be laboratory treated and examined microscopially. The study included 36 examinees with communication diameter of 10 mm and performed plastics of sinuses. In half of the examinees wounds were exposed to radiation for 7 days, and in other examinees they healed spontaneously. The results were analyzed on the fourteenth day according to the scale: 1 - complete healing, without dehiscention; 2 - incomplete healing, with minimal dehiscention; 3 - communication did not heal, with partial dehiscention; 4 - open communication, with significant dehiscention. Results and discussion The microscopic analysis shows that no wounds exposed to radiation were overcast with mucous membrane which had smooth sub epithelia chronic inflammation and inflammable infiltrate, and strong fibroplasias and granulations. Wounds exposed to radiation had mucous membrane without any signals of inflammation. Laser radiation causes anti-inflammatory reaction, i.e. it provokes reduction of exudation, alteration and proliferation, it blocks cyclo- and lipo-oxygenation by delaying the synthesis of prostaglandin, stimulates neutrophyll, macrophage and lissome activity and it activates the function of immune complex T and B lymphocytes, so this difference could be primary referred to the action of laser. Our clinical study shows that complete healing of oroantral communication was recorded in 88,8% of the examinees who were exposed to radiation in relation to 50%, of those who did not receive radiation therapy which is statistically much higher percentage (?2 test < 0, 05). The surgery was repeated in 5,6% of those who had received radiation therapy and in 16,7% of those who had not been exposed to radiation. Laser radiation stimulates changing of ADP in ATP and it accelerates cells metabolism, it increases microcirculation and accelerates substance exchange of cells, it increases DNK and RNK synthesis and stimulates cells division, which cause quicker regeneration of epithelia, i.e. it accelerates the process of wound healing. Conclusion It can be concluded that small power laser can be used successfully as additional method of treatment, after closing of oroantral communication surgically.

Introduction. Ropivacaine is used in orthopedcs, gyneacology, surgery, ophtamology, whereas experience about its usage in dentistry is still limited. The aim of this research was to compare the anesthetic effect between local anesthetics ropivacaine and bupivacaine, in surgical disposals of chronical periapical lessions in maxilla. Material and methods. The study included the patients that had indications for surgical removal of chronical periapical lessions at one of the frontal teeth of upper jaw. The total total number of examinees was 60, and they were devided in two groups. Ropivacaine chloride (0.75%) was used as a local anesthetic in one group (Naropin® 0,75%; Astra Zeneca), and in the other one bupivacaine chloride (0.5%) (Vexelit® 0,5%; Zdravlje). The autors applied 1,8 ml of block anesthesia for the n. infraorbitalis (intraoral approach) as well as 0,2 ml of the local anesthetic from the palatine side for the final branches of n. nasopalatinus in order to observe the folloving anesthetic parameters. I. Beginning of anesthesia was followed by the appearence of upper lip numbness. II. Pain rating scale according to Sisk was used for the objective measurement of the anesthesia quality. III. The pain intensity during the intervention was measured by visual analogous scale, on which the patient denoted the intensity of pain he had felt during the intervention. IV. Duration of anesthetic effect - it is followed by soft tissues numbness. Results and discussion. After the ropivacaine application anesthesia effect started in 1.57 min. and after the usage of bupivacaine in 1.67 min. The mean duration of soft tissue numbness after the application of ropivacaine was 321 minutes. Bupivacaine had a shorter anesthetic effect - 296.5 minutes. The quality of anesthesia after the usage of ropivacaine was assessed by the surgeons with average mark - 1.76. Interventions in which this anesthetic was used were performed with minimal pain and without additional anesthesia. The quality of anesthesia after the usage of bupivacaine (3.03) was statistically much worse (p

To survive and colonise their various environments, including those used during infection, bacteria have developed a variety of adaptive systems. Amongst these is phage shock protein (Psp) response, which can be induced in Escherichia coli upon filamentous phage infection (specifically phage secretin pIV) and by other membrane-damaging agents. The E. coli Psp system comprises seven proteins, of which PspA is the central component. PspA is a bifunctional protein that is directly involved in (i) the negative regulation of the psp-specific transcriptional activator PspF and (ii) the maintenance of membrane integrity in a mechanism proposed to involve the formation of a 36-mer ring complex. Here we established that the PspA negative regulation of PspF ATPase activity is the result of a cooperative inhibition. We present biochemical evidence showing that an inhibitory PspA–PspF regulatory complex, which has significantly reduced PspF ATPase activity, is composed of around six PspF subunits and six PspA subunits, suggesting that PspA exists in at least two different oligomeric assemblies. We now establish that all four putative helical domains of PspA are critical for the formation of the 36-mer. In contrast, not all four helical domains are required for the formation of the inhibitory PspA–PspF complex. Since a range of initial PspF oligomeric states permit formation of the apparent PspA–PspF dodecameric assembly, we conclude that PspA and PspF demonstrate a strong propensity to self-assemble into a single defined heteromeric regulatory complex.

Schwannoma is a benign nerve tumour that originates from Schwann cells, which cover peripheral nerves. Intraosseous schwannoma of the jaw is rare. Primary schwannoma of the mandible presenting as a periapical radiolucency on a non-vital endodontically treated tooth has rarely been referred to in the English medical literature. A rare case of intraosseous schwannoma is reported in a 23-year-old woman. The patient presented clinically with a painful swelling on the right side of the mandible and with a radiologically ill-defined unilocular periapical lesion around the mesial root of the endodontically treated first molar. Surgical treatment involved complete excision of the lesion with a mesial and distal root resection. Histological and immunohistochemical examination showed that the lesion was a primary intraosseous schwannoma of the mandible. Immunohistochemical examination showed a positive and strong reaction for S-100 protein, characteristic of neural tumours. 1 year following surgery, there were no regional recurrences. Radiographically, mandibular intraosseous schwannoma is difficult to differentiate from bone tumours such as ameloblastoma, myxoma, fibrous dysplasia, neurofibroma, central giant cell lesion or periapical lesion, so it should be included in the list of possible periapical pathoses.

In CMOS multistage clock buffer design, the duty-cycle of clock is liable to be changed when the clock passes through several buffer stages. The pulse-width may be changed due to unbalance of the p- and n-OS transistors in the long buffer. This paper describes a delay locked loop with double edge synchronization for use in a clock alignment process. Results of its SPICE simulation, that relate to 1.2 μm CMOS technology, shown that the duty-cycle of the multistage output pulses can be precisely adjusted to (50 ± 1)% within the operating frequency range, from 55 MHz up to 166 MHz.

Transcription regulation in a temporal and conditional manner underpins the lifecycle of enterobacterial pathogens. Upon exposure to a wide array of environmental cues, these pathogens modulate their gene expression via the RNA polymerase and associated sigma factors. Different sigma factors, either involved in general ‘house-keeping’ or specific responses, guide the RNA polymerase to their cognate promoter DNAs. The major alternative sigma54 factor when activated helps pathogens manage stresses and proliferate in their ecological niches. In this chapter, we review the function and regulation of the sigma54-dependent Phage shock protein (Psp) system—a major stress response when Gram-negative pathogens encounter damages to their inner membranes. We discuss the recent development on mechanisms of gene regulation, signal transduction and stress mitigation in light of different biophysical and biochemical approaches.

The paper presents the application of the principle of flexibility of architectonic space on the concrete example of flat composition. This example demonstrates that with a proper choice of structural spans dimensions and regular arrangement of sanitary and ventilation vertical assemblies, a structure with high flexibility in terms of flat space organization and floor space organization, can be achieved.

A single event latchup protection switch (SPS) has been developed in the IHP 250 nm bulk CMOS technology. The SPS has been designed as a standard library cell intended for implementation in the radiation-tolerant application specific integrated circuits (ASICs). It provides detection of single event latchup and subsequent shut-down of power supply to critical elements within the chip to restore the normal operation. However, the SPS cell might be also susceptible to single event transients. In that regard, this work presents the simulation-based analysis of the response of SPS cell in the case of single event transients. The dependence of the single event transient response with respect to the injected charge, supply voltage, load and sensing transistor's size has been analyzed.

Two-component systems play a central part in bacterial signal transduction. Phosphorelay mechanisms have been linked to more robust and ultra-sensitive signalling dynamics. The molecular machinery that facilitates such a signalling is, however, only understood in outline. In particular the functional relevance of the dimerization of a non-orthodox or hybrid histidine kinase along which the phosphorelay takes place has been a subject of debate. We use a combination of molecular and genetic approaches, coupled to mathematical and statistical modelling, to demonstrate that the different possible intra- and inter-molecular mechanisms of phosphotransfer are formally non-identifiable in Escherichia coli expressing the ArcB non-orthodox histidine kinase used in anoxic redox control. In order to resolve this issue we further analyse the mathematical model in order to identify discriminatory experiments, which are then performed to address cis- and trans-phosphorelay mechanisms. The results suggest that exclusive cis- and trans-mechanisms will not be operating, instead the functional phosphorelay is likely to build around a sequence of allosteric interactions among the domain pairs in the histidine kinase. This is the first detailed mechanistic analysis of the molecular processes involved in non-orthodox two-component signalling and our results suggest strongly that dimerization facilitates more discriminatory proof-reading of external signals, via these allosteric reactions, prior to them being further processed.

The region of the prtR gene coding for the active site of PrtR proteinase was detected in natural isolates of lactobacilli, previously determined as Lactobacillus rhamnosus. This region was present in all L. rhamnosus strains with proteolytic activity. The PCR primers used were constructed on the basis of the sequence of the catalytic domain of the prtR proteinase gene. These primers generated in colony-PCR procedure specific 611 1-bp product with DNA from natural isolates of L. rhamnosus. No PCR amplifications using these primers were obtained for closely related bacteria of genus Lactobacillus, regardless of their proteolytic activity. In addition, these primers could be used singly or in multiplex PCR together with the Lactobacillus genus-specific primers. Compared with the other proteinases within the genus Lactobacillus (PrtP, PrtB and PrtH) which retained the activity in cell-free proteinase extracts, PrtR proteinase showed proteolytic activity only under in vivo conditions (whole cells of the producing strains).

The phage shock protein (Psp) F regulon response in Escherichia coli is thought to be induced by impaired inner membrane integrity and an associated decrease in proton motive force (pmf). Mechanisms by which the Psp system detects the stress signal and responds have so far remained undetermined. Here we demonstrate that PspA and PspG directly confront a variety of inducing stimuli by switching the cell to anaerobic respiration and fermentation and by down-regulating motility, thereby subtly adjusting and maintaining energy usage and pmf. Additionally, PspG controls iron usage. We show that the Psp-inducing protein IV secretin stress, in the absence of Psp proteins, decreases the pmf in an ArcB-dependent manner and that ArcB is required for amplifying and transducing the stress signal to the PspF regulon. The requirement of the ArcB signal transduction protein for induction of psp provides clear evidence for a direct link between the physiological redox state of the cell, the electron transport chain, and induction of the Psp response. Under normal growth conditions PspA and PspD control the level of activity of ArcB/ArcA system that senses the redox/metabolic state of the cell, whereas under stress conditions PspA, PspD, and PspG deliver their effector functions at least in part by activating ArcB/ArcA through positive feedback.

Variable delay elements are often used in different types of high-speed integrated circuits, mainly intended for delay compensation, skew equalization, etc. These circuits are normally realized as hybrid, composed of digital and analog controlled parts. The digital part is used for coarse-grain, while the analog for fine-grain delay variation. Efficient analog delay element architecture is proposed in this paper. The proposal is based on modification of the standard current starved delay element solution. An analytical equation that corresponds to the delay of the circuit is given also. In terms of control voltage, the proposed circuit has a linear delay transfer function in the whole range of regulation. Improvement is achieved at a cost of small hardware overhead in respect to the standard solution. Delay linearity error is less than 1% and the agreement between analytical model and simulation results is good, i.e. the error is less than 5%.