Development of collagenous scaffolds for wound healing: characterization and in vivo analysis.

The development of wound dressings from biomaterials has been the subject of research due to their unique structural and functional characteristics. Proteins from animal origin, such as collagen and chitosan, act as promising materials for applications in injuries and chronic wounds, functioning as a repairing agent. This study aims to evaluate in vitro effects of scaffolds with different formulations containing bioactive compounds such as collagen, chitosan, N-acetylcysteine (NAC) and ε-poly-lysine (ε-PL). We manufactured a scaffold made of a collagen hydrogel bioconjugated with chitosan by crosslinking and addition of NAC and ε-PL. Cell viability was verified by resazurin and live/dead assays and the ultrastructure of biomaterials was evaluated by SEM. Antimicrobial sensitivity was assessed by antibiogram. The healing potential of the biomaterial was evaluated in vivo, in a model of healing of excisional wounds in mice. On the 7th day after the injury, the wounds and surrounding skin were processed for evaluation of biochemical and histological parameters associated with the inflammatory process. The results showed great cell viability and increase in porosity after crosslinking while antimicrobial action was observed in scaffolds containing NAC and ε-PL. Chitosan scaffolds bioconjugated with NAC/ε-PL showed improvement in tissue healing, with reduced lesion size and reduced inflammation. It is concluded that scaffolds crosslinked with chitosan-NAC-ε-PL have the desirable characteristics for tissue repair at low cost and could be considered promising biomaterials in the practice of regenerative medicine.

Spatiotemporal variation in phenolic levels in galls of calophyids on Schinus polygama (Anacardiaceae).

The expression of plant secondary metabolism is strongly controlled by plant both in time and space. Although the variation of secondary metabolites, such as soluble and structural phenolics (e.g., lignins), has been largely observed in gall-inducing insects, and compared to their non-galled host organs, only a few datasets recording such variation are available. Accordingly, the relative importance of spatiotemporal variability in phenolic contents, and the influence of gall developmental stages on the original composition of host organs are poorly discussed. To address this knowledge gap, we histochemically determined the sites of polyphenol and lignin accumulation, and the polyphenol contents in three developmental stages of two calophyid galls and their correspondent host organs. Current results indicate that the compartmentalization of phenolics and lignins on Schinus polygama (Cav.) Cabrera follows a similar pattern in the two-calophyid galls, accumulating in the outer (the external tissue layers) and in the inner tissue compartments (the cell layers in contact with the gall chamber). The non-accumulation in the median compartment (median parenchyma layers of gall wall with vascular bundles, where gall inducer feeds) is important for the inducer, because its mouth apparatus enter in contact with the cells of this compartment. Also, the concentration of phenolics has opposite dynamics, decreasing in leaf galls and increasing in stem galls, in temporal scale, i.e., from maturation toward senescence. The concentration of phenolics in non-galled host organs, and in both galls indicated the extended phenotype of Calophya rubra (Blanchard) and C. mammifex Burckhardt & Basset (Hemiptera: Sternorrhyncha: Psylloidea: Calophyidae) over the same host plant metabolic potentiality.

Older adults' dependence in activities of daily living: Implications for oral health.

OBJECTIVE: To assess the relationship between ADL and oral health status in older adults. DESIGN AND SAMPLE: Quantitative, analytical, cross-sectional research with 280 people aged ≥60 in public primary health care centers in the city of Fortaleza, Northeastern Brazil. MEASURES: Collection of sociodemographic data, oral discomfort data, and general health data and application of the Community Oral Health Indicator-COHI and the activities of daily living scale. RESULTS: Significant differences in functional dependence were found between age groups (p < .001), incomes (p = .034), and levels of education (p = .003). Tooth loss (OR = 2.13; p = 0.025), having no teeth (OR = 2.97), difficulties in chewing and swallowing food (OR = 2.56; p = 0.003), problems with taste of food (OR = 2.06; p = .044), poor self-rated general health (OR = 3.7; p = .001), hearing problems (OR = 2.23; p = .009) and speech problems (OR = 3.15; p = .002) increased chances for functional dependence. Dependent individuals were less likely to use toothpaste, thereby increasing the potential for caries (p = .013). Age 80 or older (p < .001), ages 70-79 (p < .001), three or more visible dental cavities (p = .030) and difficulty chewing and swallowing food (p = .027) remained associated with dependence in ADL in the logistic regression model. CONCLUSION: Functional dependence in activities of daily living has implications for the oral health status of older people.

Stomatognathic function in Duchenne muscular dystrophy: a case-control study.

AIM: This study aimed to analyse electromyographic activity, masticatory efficiency, muscle thickness, and bite force of individuals with Duchenne muscular dystrophy (DMD). METHOD: Forty males aged 4-15 years, 20 with DMD and 20 healthy age-, height-, and weight-matched controls, underwent electromyography and ultrasonography of temporalis, masseter, and sternocleidomastoid muscles during postural control of the jaw, mastication, and maximal molar bite force. RESULTS: The normalized electromyography signals showed higher activity in masseter and temporal muscles at rest, during protrusion, left and right laterality, and fatigue condition in the group with DMD than in the comparison group (p≤0.05). For masticatory efficiency of cycles, in analysis of non-habitual chewing of flavourless gum, and habitual chewing of peanuts and raisins, the group with DMD presented lower averages (p≤0.05). For the muscle thickness, the results showed that there was a lower muscle thickness in the group with DMD for all muscles during the rest and maximal voluntary contraction, except for masseter and sternocleidomastoid in the maximal voluntary contraction. In the maximal molar bite force, the group with DMD presented higher values for both sides than the comparison group (p≤0.05). INTERPRETATION: Patients with DMD show muscle changes related to the stomatognathic system, in their activity, bite force, and muscle thickness.

Preparation and characterization of PEGylated amylin.

Amylin is a pancreatic hormone that plays important roles in overall metabolism and in glucose homeostasis. The therapeutic restoration of postprandial and basal amylin levels is highly desirable for patients with diabetes who need to avoid glucose excursions. Protein conjugation with polyethylene glycol (PEG) has long been known to be a convenient approach for extending the biological effects of biopharmaceuticals. We have investigated the reactivity of amylin with methoxy polyethylene glycol succinimidyl carbonate and methoxy polyethylene glycol succinimidyl propionate, which have an average molecular weight of 5 kDa. The reaction, which was conducted in both aqueous and organic (dimethyl sulfoxide) solvents, occurred within a few minutes and resulted in at least four detectable products with distinct kinetic phases. These results suggest a kinetic selectivity for PEGylation by succinimidyl derivatives; these derivatives exhibit enhanced reactivity with primary amine groups, as indicated by an evaluation of the remaining amino groups using fluorescamine. The analysis of tryptic fragments from mono- and diPEGylated amylin revealed that conjugation occurred within the 1-11 amino acid region, most likely at the two amine groups of Lys(1). The reaction products were efficiently separated by C-18 reversed phase chromatography. Binding assays confirmed the ability of mono- and diPEGylated amylin to interact with the amylin co-receptor receptor activity-modifying protein 2. Subcutaneous administration in mice revealed the effectiveness of monoPEG-amylin and diPEG-amylin in reducing glycemia; both compounds exhibited prolonged action compared to unmodified amylin. These features suggest the potential use of PEGylated amylin to restore basal amylin levels.

Waste-derived volatile fatty acids as carbon source for added-value fermentation approaches.

The establishment of a sustainable circular bioeconomy requires the effective material recycling from biomass and biowaste beyond composting/fertilizer or anaerobic digestion/bioenergy. Recently, volatile fatty acids attracted much attention due to their potential application as carbon source for the microbial production of high added-value products. Their low-cost production from different types of wastes through dark fermentation is a key aspect, which will potentially lead to the sustainable production of fuels, materials or chemicals, while diminishing the waste volume. This article reviews the utilization of a volatile fatty acid platform for the microbial production of polyhydroxyalkanoates, single cell oil and omega-3 fatty acids, giving emphasis on the fermentation challenges for the efficient implementation of the bioprocess and how they were addressed. These challenges were addressed through a research project funded by the European Commission under the Horizon 2020 programme entitled 'VOLATILE-Biowaste derived volatile fatty acid platform for biopolymers, bioactive compounds and chemical building blocks'.

Electroacupuncture Reduces Inflammation but Not Bone Loss on Periodontitis in Arthritic Rats.

Periodontitis and rheumatoid arthritis (RA) are inflammatory diseases characterized by chronic inflammation and bone erosion. Electroacupuncture (EA) shows anti-inflammatory and anti-resorptive effects in experimental periodontitis (EP) and in RA. It is important to investigate whether EA shows these effects in periodontal tissues in the presence of these two inflammatory diseases or not. For this, Wistar rats were divided into six groups: control (C); experimental rheumatoid arthritis (RA; bovine type II collagen-induced (CII)); experimental periodontitis (EP); RA/EP (RA + EP); EP/EA (EP treated with EA); RA/EP/EA (RA + EP treated with EA). EP was induced 21 days after RA induction and EA was performed previously and during the EP induction period, every 3 days until the 36th experimental day. The rats were euthanized on day 39. RA was evaluated by edema and the withdrawal threshold of hind paws. The maxillae were removed, and alveolar bone loss (ABL) and bone radiographic density (BRD) were evaluated. Immunohistochemical analyses for interleukins (IL)-6 and -17 and nuclear factor (NF)-κB were performed. Our results showed that EA reduced only the pain intensity in arthritic rats. Histomorphometric, macroscopic, and radiographic analyses did not show differences between the control and EP/EA groups. EA caused a reduction in ABL and BRD only in the presence of EP. EA caused a reduction in IL-6 and -17 in all groups, but NF-κB was only reduced in the arthritic rats with EP. In conclusion, EA reduced the inflammation related to periodontitis in arthritic rats but did not prevent ABL.

An innovative proposal for anophthalmic cavity reconstruction surgery with bone cement: a case report.

We present a patient who underwent evisceration surgery after spontaneous rupture of the ocular globe due to long-data uncontrolled glaucoma, with posterior placement of an orbital implant made of a bone cement compound based on polymethylmethacrylate as alternative materials were not available. Such a compound is characterized by excellent biocompatibility and low cost, which makes it an interesting alternative for treatment. The anophthalmic socket was successfully filled, providing proper esthetic results and favorable conditions for the posterior scleral prosthesis implantation. No complications were observed during 10 months of follow-up. We believe that, in the absence of alternative materials, low-cost materials may be used in emergency settings to repair anophthalmic cavities and provide satisfactory functional and esthetic outcomes.

Elemental concentration and migratability in bioplastics derived from organic waste.

In line with the Circular Economy approach, the production of polyhydroxyalkanoate (PHA) with organic waste as the feedstock may a biotechnological application to reduce waste and recover high-value materials. The potential contaminants that could transfer from bio-waste to a PHA include inorganic elements, such as heavy metals. Hence, the total content and migratability of certain elements were evaluated in several PHA samples produced from different origins and following different methods. The total content of certain elements in PHA ranged between 0.0001 (Be) and 49,500 mg kg-1 (Na). The concentrations of some alkaline (Na and K) and alkaline earth (Ca and Mg) metals were highest, which are of little environmental concern. The feedstock type and PHA stabilisation and extraction procedures affected the element contents. Several sets of experiments were conducted to evaluate the migration of elements from the PHA samples under different storage times, temperatures, and pH levels. The total contents of some heavy metals (As, Cd, Fe, Hg, Ni, Pb, and Zn) in PHA produced from fruit waste or crops (commercial PHA) were lower than those in the PHA samples produced from the mixture of the organic fraction of municipal waste and sludge from wastewater treatment. Both the PHA obtained by extraction from wet biomass (acid storage) with aqueous phase extraction reagents and commercial PHA were below the migration limits stipulated by the current Toy Safety Directive and by Commission Regulation (EU) October 2011 on plastic materials and articles intended to come into contact with food under frozen and refrigerated conditions.

Preventing False Negatives for Histochemical Detection of Phenolics and Lignins in PEG-Embedded Plant Tissues.

Polyethylene glycol (PEG) is a low-cost and advantageous embedding medium, which maintains the majority of cell contents unaltered during the embedding process. Some hard or complex plant materials are better embedded in PEG than in other usual embedding media. However, the histochemical tests for phenolics and lignins in PEG-embedded plant tissues commonly result in false negatives. We hypothesize that these false negatives should be prevented by the use of distinct fixatives, which should avoid the bonds between PEG and phenols. Novel protocols for phenolics and flavanols detection are efficiently tested, with fixation of the samples in ferrous sulfate and formalin or in caffeine and sodium benzoate, respectively. The differentiation of lignin types is possible in safranin-stained sections observed under fluorescence. The Maule's test faultlessly distinguishes syringyl-rich from guaiacyl- and hydroxyphenyl-rich lignins in PEG-embedded material under light microscopy. Current hypothesis is corroborated, that is, the adequate fixation solves the false-negative results, and the new proposed protocols fill up some gaps on the detection of phenolics and lignins.

Reconstrução microcirúrgica de mandíbula com retalho osteocutâneo de fíbula, com auxílio de modelo 3D, em paciente vítima de trauma: relato de caso / Mandible microsurgical reconstruction with osteocutaneous fibular flap, with the assistance of a 3D model, in a trauma patient: case report

Trata-se de um relato de caso de reconstrução microcirúrgica de mandíbula com retalho osteocutâneo de fíbula em um paciente de 16 anos, vítima de agressão com projétil de arma de fogo e trauma contuso em face, acarretando em fratura cominutiva de mandíbula, com perda de elementos dentários e lesões extensas de partes moles. Após tratamento da fase aguda, optou-se pela reconstrução microcirúrgica com retalho livre de fíbula, devido à extensão do defeito mandibular e possibilidade de reabilitação dentária. O uso de modelos tridimensionais da mandíbula e fíbula foram excepcionalmente úteis para tornar possível o melhor planejamento cirúrgico das áreas de osteotomia, com diminuição do tempo de isquemia do retalho da cirurgia.

This is a case report of the mandible microsurgical reconstruction with osteocutaneous fibular flap in a 16-year-old patient. A victim of an assault with a firearm projectile and blunt trauma to the face, resulting in a comminuted mandible fracture, loss of dental elements, and extensive soft tissue injuries. After treatment of the acute phase, we chose microsurgical reconstruction with a free fibula flap due to the mandibular defect's extension and the possibility of dental rehabilitation. The use of three-dimensional models of the mandible and fibula was exceptionally helpful in making the best surgical planning of the osteotomy areas, decreasing the surgery flap's ischemia time.

Spatial distribution of floating marine debris in offshore continental Portuguese waters.

This study presents data on abundance and density of macro-floating marine debris (FMD), including their composition, spatial distribution and potential sources off continental Portugal. FMD were assessed by shipboard visual surveys covering ±252,833 km(2) until the 220 nm limit. The FMD average density was 2.98 items/km(2) and abundance amounted to 752,740 items. Unidentified plastics constitute the major bulk of FMD (density=0.46 items/km(2); abundance=117,390 items), followed by styrofoam, derelict or lost materials from fisheries, paper/cardboard and wood material. The North sector of the area presents higher FMD diversity and abundances, probably as a result of the high number of navigation corridors and fisheries operating in that sector. Most FMD originate from local sources, namely discharges from vessels and derelict material from fisheries. Considering the identifiable items, cables and fishing lines were the only fishing related items among the top ten FMD items in Portuguese offshore waters.

Monoconjugation of Human Amylin with Methylpolyethyleneglycol.

Amylin is a pancreatic hormone cosecreted with insulin that exerts unique roles in metabolism and glucose homeostasis. The therapeutic restoration of postprandial and basal amylin levels is highly desirable in diabetes mellitus. Protein conjugation with the biocompatible polymer polyethylene glycol (PEG) has been shown to extend the biological effects of biopharmaceuticals. We have designed a PEGylated human amylin by using the aminoreactive compound methoxylpolyethylene glycol succinimidyl carbonate (mPEGsc). The synthesis in organic solvent resulted in high yields of monoPEGylated human amylin, which showed large stability against aggregation, an 8 times increase in half-life in vivo compared to the non-conjugated amylin, and pharmacological activity as shown by modulation of cAMP production in MCF-7 cell line, decrease in glucagon and modulation of glycemia following subcutaneous administration in mice. Altogether these data reveal the potential use of PEGylated human amylin for the restoration of fasting amylin levels.

An innovative proposal for anophthalmic cavity reconstruction surgery with bone cement: a case report / Proposta inovadora para reconstrução de cavidades anoftálmicas utilizando cimento ósseo: um relato de caso

ABSTRACT We present a patient who underwent evisceration surgery after spontaneous rupture of the ocular globe due to long-data uncontrolled glaucoma, with posterior placement of an orbital implant made of a bone cement compound based on polymethylmethacrylate as alternative materials were not available. Such a compound is characterized by excellent biocompatibility and low cost, which makes it an interesting alternative for treatment. The anophthalmic socket was successfully filled, providing proper esthetic results and favorable conditions for the posterior scleral prosthesis implantation. No complications were observed during 10 months of follow-up. We believe that, in the absence of alternative materials, low-cost materials may be used in emergency settings to repair anophthalmic cavities and provide satisfactory functional and esthetic outcomes.

RESUMO Apresentamos um paciente que foi submetido à cirurgia de evisceração após ruptura espontânea do globo ocular devido a glaucoma não controlado de longa data, com posterior colocação de implante orbital feito de cimento ósseo, composto à base de polimetilmetacrilato, diante da indisponibilidade de materiais alternativos. Tal composto se caracteriza pela excelente biocompatibilidade e baixo custo, o que o torna uma alternativa interessante para o tratamento. A cavidade anoftálmica foi preenchida com sucesso, fornecendo resultados estéticos adequados e condições favoráveis para o implante posterior de prótese escleral. Nenhuma complicação foi observada durante os 10 meses de seguimento. Acredi­tamos que, na ausência de materiais alternativos, materiais de baixo custo podem ser usados em situações emergenciais para preencher cavidades anoftálmicas e prover resultados funcionais e estéticos satisfatórios.

Functional analysis of the stomatognathic system in individuals infected with human immunodeficiency virus.

PURPOSE: To understand the effects of HIV type 1 on the function of the masticatory muscles. METHODS: Sixty individuals were selected from both genders, aged between 22 and 57years (mean 36.77±9.33years), and divided into two groups: Group HIVG, 30 individuals with HIV subtype 1, and Group CG, 30 healthy individuals (control group). The individuals were submitted to assessments of their masticatory muscle activity, biting strength and thickness of the masticatory muscles by means of electromyography, maximal molar bite force and ultrasound imaging, respectively. The resultant data were statistically analyzed by t-tests (p<0.05). RESULTS: The HIVG normalized EMG data from a mandibular rest position, protrusion, right and left laterality movement of the jaw showed that HIVG presented a relative increase in EMG activity compared to the CG. The HIVG had a lower masticatory cycle ability while chewing Parafilm M®, Raisins and Peanuts when compared to CG. During rest conditions, the ultrasound images showed a greater average muscular thickness in the right and left temporal region compared to CG. Upon maximal voluntary contraction, an increased average thickness was seen in the temporalis muscles and left sternocleidomastoid muscle when compared to the CG. CONCLUSION: Based on the results of this research, it can be concluded that individuals with acquired immunodeficiency syndrome showed muscular changes related to the stomatognathic system, especially concerning EMG activity and muscle thickness.

Efficiency of the Polyethylene-Glycol (PEG) Embedding Medium for Plant Histochemistry.

Histochemical analyses in plants are commonly performed on hand-made sections of fresh materials. The disadvantages of embedding in historesin, paraffin or paraplast® are the alterations to cellular contents, the high costs and few evident results, depending on the test. Polyethylene-glycol (PEG), as a low cost, hydrophilic medium that maintains most of the cellular features similar to fresh conditions, may be useful for obtaining good histochemical results in thinner and homogeneous sections. The current study aimed to compare the efficiency of PEG as an embedding medium for histochemical analyses of primary and secondary metabolites accumulation. Using hand-made sections of fresh samples (T1) as a comparison, we tested the influence of the use of Karnovsky's solution as a fixative (T2) versus embedding in PEG (T3). The samples herein analyzed comprise leaves, stems, seeds and insect galls of different plant species. Neither the Karnovsky's fixative nor the embedding in PEG altered the histochemical results for starch, lipids, terpenoids, proteins and reducing sugars in T1, T2, and T3. However, PEG binds to phenols, such as tannins, flavonoids and lignins, thereby presenting false negatives in T3.

Enhanced bioproduction of poly-3-hydroxybutyrate from wheat straw lignocellulosic hydrolysates.

Polyhydroxyalkanoates (PHAs) are bioplastics that can replace conventional petroleum-derived products in various applications. One of the major barriers for their widespread introduction in the market is the higher production costs compared with their petrochemical counterparts. In this work, a process was successfully implemented with high productivity based on wheat straw, a cheap and readily available agricultural residue, as raw material. The strain Burkholderia sacchari DSM 17165 which is able to metabolise glucose, xylose and arabinose, the main sugars present in wheat straw hydrolysates (WSHs), was used. Results in shake flask showed that B. sacchari cells accumulated about 70%gpoly(3-hydroxybutyrate)(P(3HB))/g cell dry weight (CDW) with a yield of polymer on sugars (YP/S) of 0.18g/g when grown on a mixture of commercial C6 and C5 sugars (control), while these values reached about 60%gP(3HB)/g CDW and 0.19g/g, respectively, when WSHs were used as carbon source. In fed-batch cultures carried out in 2L stirred-tank reactors (STRs) on WSH, a maximum polymer concentration of 105 g/L was reached after 61 hours of cultivation corresponding to an accumulation of 72% of CDW. Polymer yield and productivity were 0.22 gP(3HB)/g total sugar consumed and 1.6g/L hour, respectively. The selected feeding strategy successfully overcame the carbon catabolite repression (CCR) phenomenon observed with sugar mixtures containing hexoses and pentoses. This is the first work describing fed-batch cultivations aiming at PHA production using real lignocellulosic hydrolysates. Additionally, the P(3HB) volumetric productivities attained are by far the highest ever achieved on agricultural waste hydrolysates.

Production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) by Burkholderia sacchari using wheat straw hydrolysates and gamma-butyrolactone.

Burkholderia sacchari DSM 17165 is able to grow and produce poly(3-hydroxybutyrate) both on hexoses and pentoses. In a previous study, wheat straw lignocellulosic hydrolysates (WSH) containing high C6 and C5 sugar concentrations were shown to be excellent carbon sources for P(3HB) production. Using a similar feeding strategy developed for P(3HB) production based on WSH, fed-batch cultures were developed aiming at the production of the copolymer P(3HB-co-4HB) (poly(3-hydroxybutyrate-co-4-hydroxybutyrate)) by B. sacchari. The ability of this strain to synthesize P(3HB-co-4HB) was first shown in shake flasks using gamma-butyrolactone (GBL) as precursor of the 4HB units. Fed-batch cultures using glucose as carbon source (control) and GBL were developed to achieve high copolymer productivities and 4HB incorporations. The attained P(3HB-co-4HB) productivity and 4HB molar% were 0.7g/(Lh) and 4.7molar%, respectively. The 4HB incorporation was improved to 6.3 and 11.8molar% by addition of 2g/L propionic and acetic acid, respectively. When WSH were used as carbon source under the same feeding conditions, the values achieved were 0.5g/(Lh) and 5.0molar%, respectively. Burkholderia sacchari, a strain able to produce biopolymers based on xylose-rich lignocellulosic hydrolysates, is for the first time reported to produce P(3HB-co-4HB) using gamma butyrolactone as precursor.

Analysis of endodontist posture utilizing cinemetry, surface electromyography and ergonomic checklists.

The postural risk factors for dentists include the ease of vision in the workplace, cold, vibration and mechanical pressure in tissues, incorrect posture, functional fixity, cognitive requirements and work-related organizational and psychosocial factors. The objective was to analyze the posture of endodontists at the workplace. Eighteen right-handed endodontists aged 25 to 60 years (34±3) participated in the study. Electromyography, kinemetry, ergonomic scales (RULA and Couto's checklist) and biophotogrammetry were used to analyze the posture of endodontists during root canal treatment of the maxillary right first and second molars using rotary and manual instrumentation. The variations observed in the electromyographic activities during the performance of rotary and manual techniques suggest that the fibers of the longissimus region, anterior and medium deltoid, medium trapezium, biceps, triceps brachii, brachioradialis and short thumb abductor muscles underwent adaptations to provide more accurate functional movements. Computerized kinemetry and biophotogrammetry showed that, as far as posture is concerned, rotary technique was more demanding than the manual technique. In conclusion, the group of endodontists evaluated in this study exhibited posture disorders regardless of whether the rotary or manual technique was used.

Amylin conjugation with methoxyl polyethyleneglycol.

We modified amylin chemically by conjugating methoxyl polyethyleneglycol succinimidyl carbonate (mPEGSC) of varying molecular weights (1 kDa, 2 kDa and 5 kDa). The reaction occurred within a few minutes, resulting in at least four distinct PEGylated products. The reaction products were separated by reversed-phase chromatography and identified by mass-spectrometry. The monoPEGylated and diPEGylated amylin products were generated rapidly through conjugation to the two amino groups of the N-terminal lysine residue. Both PEGylated amylin products bound to the receptor activity-modifying protein 1 (RAMP1). Pharmacological evaluation by subcutaneous administration in mice of monoPEGylated and diPEGylated amylin obtained with mPEG-SC 5 kDa revealed that both compounds modulated glycemia for longer times than unmodified amylin. Collectively, these data demonstrate the potential of bioconjugation with mPEG for the design of amylin therapeutics with sustained action.