Cryo-EM structures of wild-type and E138K/M184I mutant HIV-1 RT/DNA complexed with inhibitors doravirine and rilpivirine.

Structures trapping a variety of functional and conformational states of HIV-1 reverse transcriptase (RT) have been determined by X-ray crystallography. These structures have played important roles in explaining the mechanisms of catalysis, inhibition, and drug resistance and in driving drug design. However, structures of several desired complexes of RT could not be obtained even after many crystallization or crystal soaking experiments. The ternary complexes of doravirine and rilpivirine with RT/DNA are such examples. Structural study of HIV-1 RT by single-particle cryo-electron microscopy (cryo-EM) has been challenging due to the enzyme's relatively smaller size and higher flexibility. We optimized a protocol for rapid structure determination of RT complexes by cryo-EM and determined six structures of wild-type and E138K/M184I mutant RT/DNA in complexes with the nonnucleoside inhibitors rilpivirine, doravirine, and nevirapine. RT/DNA/rilpivirine and RT/DNA/doravirine complexes have structural differences between them and differ from the typical conformation of nonnucleoside RT inhibitor (NNRTI)-bound RT/double-stranded DNA (dsDNA), RT/RNA-DNA, and RT/dsRNA complexes; the primer grip in RT/DNA/doravirine and the YMDD motif in RT/DNA/rilpivirine have large shifts. The DNA primer 3'-end in the doravirine-bound structure is positioned at the active site, but the complex is in a nonproductive state. In the mutant RT/DNA/rilpivirine structure, I184 is stacked with the DNA such that their relative positioning can influence rilpivirine in the pocket. Simultaneously, E138K mutation opens the NNRTI-binding pocket entrance, potentially contributing to a faster rate of rilpivirine dissociation by E138K/M184I mutant RT, as reported by an earlier kinetic study. These structural differences have implications for understanding molecular mechanisms of drug resistance and for drug design.

Impact of surgical timing and type of operative procedure on outcomes in periprosthetic hip fractures: an observational study at an NHS trust centre in the UK.

PURPOSE: There is a global trend of increased periprosthetic fractures due to the growing number of arthroplasty procedures. The present study assessed the impact of factors such as time to surgery and type of surgery on the outcomes, which have been seldom evaluated for periprosthetic fractures. METHODS: An observational study was conducted on consecutive 87 patients within an NHS district hospital trust in the UK. Patients who underwent a complete hip replacement prior to the fracture, received fixation therapy, or underwent revision surgery within the specified time were screened. Patients were grouped in two ways: based on time to surgery and based on surgery type. Logistic regression models were performed to assess for statistically significant differences in post-operative complication, 30-day, and 1-year mortality rates between groups, whilst adjusting for age, gender, and ASA grade. RESULTS: Forty-one patients underwent open reduction and internal fixation (ORIF), 29 patients underwent revision arthroplasty, and 17 patients were subjected to both, ORIF and revision arthroplasty. Sixty of the 87 patients were operated on > 48 h of injury. The median hospital stay was significantly lower in the ORIF plus revision arthroplasty group, versus other surgical groups (p < 0.05) whilst it was significantly higher in the group of patients who underwent surgery after 48 h of injury (p < 0.05). Numerically higher mortality was noted in the revision arthroplasty group (31.03%, p > 0.05). The group that was operated after 48 h of injury showed greater mortality but was comparable to the other group (25% vs. 14.81%, p > 0.05). For post-operative complications, none of the variables were significantly predictive (p > 0.05). However, for 30-day mortality, ASA grade (p = 0.04) and intra-operative complications (p = 0.0001) were significantly predictive. Additionally, for 1-year mortality, ASA grade (p = 0.004) was noted to be significantly predictive. CONCLUSION: Revision and delayed periprosthetic fracture management (> 48 h after injury) group showed a numerically greater mortality risk; however, this finding was not statistically significant. ASA grading at baseline is predictive of mortality for periprosthetic fractures.

Investigation of efficient photoconduction and enhanced luminescence characteristics of Ce-doped ZnO nanophosphors for UV sensors.

This study involves the single-step, mass-scale productive synthesis, photoconduction, and luminescence characteristics of pure and cerium rare-earth-ion-doped ZnO (CZO) nanophosphors with different Ce concentrations (Ce: 0, 2, 4, 6, and 8 wt.%) synthesized using the solid-state reaction method. The synthesized nanophosphors were characterized for their structural, morphological, optical, and photoconductivity (PC) properties using X-ray diffraction (XRD), field-effect scanning electron microscopy (FE-SEM), energy dispersive spectroscopy, Fourier-transform infrared (FT-IR), photoluminescence (PL), and PC measurements. The sharp diffraction peaks of XRD results exhibit the formation of crystalline hexagonal wurtzite ZnO nanostructures. The decrease in diffraction peak intensities of CZO with an increase in Ce concentrations signifies the deterioration of the ZnO crystal. FE-SEM images exhibit the good crystalline quality of nanophosphors composed of spherical- and elongated-shaped nanoparticles that are distributed consistently on the surface. The energy dispersive X-ray pattern of the 4 wt.% Ce-doped ZnO (CZO4 ) sample confirms the doping of Ce in ZnO. The presence of chemical bonds and functional groups corresponds to transmittance peaks established using FT-IR spectroscopy. Deconvoluted PL spectra show two major emission peaks, one in the UV region, which is near-band-edge, and the other in the visible region ranging from ~456 to 561 nm. In PC studies, current-voltage (I-V) and current-time (I-T) characteristics, that is, rise/decayin current under dark as well as UV light conditions, are also investigated. Efficient photoconduction is observed in CZO samples. The obtained results indicate the suitability to luminescent and photosensor applications.

Valgus arthritic knees can be classified into nine phenotypes.

PURPOSE: It is not shown whether anatomical variations exist in valgus arthritic limbs as to support individualized component and limb alignment. The null hypothesis was that there was no phenotypic variation of coronal femoro-tibial morphology in valgus knees. The aim was to determine whether distinct phenotypes of valgus knees could be identified to help surgical planning and classifying valgus knees for outcome studies. METHODS: Full-leg weight-bearing radiographs of 233 knees (182 preoperative; 51 of contralateral arthritic knee) were measured for HKA (hip-knee-ankle angle), VCA (valgus correction angle), mLDFA (lateral mechanical distal femoral angle), aLDFA (lateral anatomical distal femoral angle), MPTA (medial proximal tibial angle), MNSA (medial neck shaft angle), TAMA (angle between tibial mechanical and anatomical axes), and TBA (tibial bowing angle). RESULTS: Nine phenotypes were identified encompassing all 233 knees which could be clubbed into 4 broad types. Type 1 Neutral knees (12.5%) had almost normal values (mean VCA 5.3°, mLDFA 86.9°, aLDFA 81.1°). Type 2 'Intra-articular valgus' (22.7%) showed lateral compartment bone loss (mean mLDFA 83.9°; MPTA 90.2°). Type 3 'Extra-articular valgus' (35.2%) had extra-articular deformity: 3a showed valgus femoral bowing (mean VCA 2.7°); 3b valgus tibial bowing; 3c showed valgus tibial bowing with lateral femoral condyle wear (mean mLDFA 84.3°). Type 4 'Varus' type (29.6%) had features of varus knees: 4a had varus femoral bowing (VCA 8.3°); distal femur in 4b was akin to varus knees (mean mLDFA 89.3°) with lateral tibial bone loss (mean MPTA 91.2°). 4c had varus tibial bowing and deficient lateral femoral condyle (mLDFA 83.7°). 4d had varus tibial bowing and lateral tibial bone loss (mean MPTA 89.8°). CONCLUSIONS: The study identified four broad groups of valgus arthritic knees with nine phenotypes based on coronal plane variations in femoro-tibial morphology. This study may be of value in planning and performing corrective osteotomies, and planning the optimal position of femoral and tibial components in unicompartmental and total knee arthroplasty. The classification presented in this study may aid in categorizing valgus knees for outcome studies. LEVEL OF EVIDENCE: IV.

Arthritic knees with more than 10° valgus can have soft-tissue imbalance in flexion.

PURPOSE: Valgus knees have inferior outcomes compared to varus knees. There is little data regarding soft-tissue balance in flexion which may influence outcome in valgus knees undergoing TKA. The purpose of this study was to evaluate whether there is imbalance between medial and lateral flexion gaps in valgus deformity. A secondary aim was to compare soft-tissue balance in knees with valgus deformity less than 10° with those exceeding 10°. The null hypothesis was that there was no soft-tissue imbalance in 90° of flexion irrespective of magnitude of deformity. METHODS: 64 valgus knees (52 female and 12 male) with deformity from 0.5 to 27.5° (mean 188.77, SD 6.21) were studied in 54 patients (mean age 67.81 y, SD 8.69) undergoing navigated TKA. Medial and lateral gaps in extension and at 90° of flexion were compared (using Independent-samples t test) between knees with valgus < 10° with those > 10° using a validated dynamic method after resection of cruciates, menisci and osteophytes, and then after final trialling. RESULTS: Mean initial medial-lateral (ML) gap difference in extension was 2.63 mm (SD 2.63) and 2.09 mm (SD 3.78) in flexion, being tighter laterally. Initial ML gap differences in extension and flexion correlated with valgus deformity (R = - 0.514; p = 0.00001; R = - 0.325; p = 0.01, respectively). Initial ML gap differences in extension correlated with those in flexion (R = 0.42; p = 0.0005). Mean ML flexion and extension gap differences were 1.30 mm (SD 3.67) and 1.26 mm (SD 1.92) in knees with < 10° valgus, and 3.17 mm (SD 3.71) and 4.29 mm (SD 2.45) in those > 10° valgus; p values were 0.026 and < 0.001 respectively. CONCLUSION: The lateral flexion gap in valgus knees may be narrower than the medial flexion gap, especially in knees with > 10° deformity. This contrasts with native and varus knees, in which it exceeds the medial gap. This novel study indicates the need to identify valgus knees with lateral flexion gap tightness by distracting the posterior femoral condyles from the proximal tibia by dynamic stressing of the soft-tissues after resection of cruciates, menisci, and osteophytes, with the knee flexed to 90°. These findings, highlighting the need for restoring flexion gap balance, may improve the inferior outcomes in valgus knees. LEVEL OF EVIDENCE: IV.

Seven phenotypes of varus osteoarthritic knees can be identified in the coronal plane.

PURPOSE: Recommendations for resecting distal femur and proximal tibia in mechanical and anatomical alignment techniques are standardized. Kinematic alignment propagates individualizing resection planes. Whether significant variation exists, to warrant departure from standardized resection planes, has not been shown thus far in a large cohort of knees and with a wide range of varus deformity. The null hypothesis of this study was that there was no phenotypic variation in varus osteoarthritic knees. The aim of this paper was to determine whether distinct phenotypes could be identified, based on variations in coronal femoral and tibial morphology, which could aid in surgical planning and categorizing varus knees for future studies. METHODS: 2129 full-leg weightbearing radiographs were analyzed (1704 preoperative; 425 of contralateral arthritic knee). Measurements made were of HKA (hip-knee-ankle angle), VCA (valgus correction angle), mLDFA (lateral mechanical distal femoral angle), aLDFA (lateral anatomical distal femoral angle), MPTA (medial proximal tibial angle), MNSA (medial neck shaft angle), TAMA (angle between tibial mechanical and anatomical axes), and TPDR (percentage length of tibia proximal to extra-articular deformity). RESULTS: Seven distinct types were identified covering 2021 knees, reducible to 4 broad phenotypes: 11% were Type 1 'Neutral' knees showing values close to reported normal knees (mean VCA 5.5°, mLDFA 87°, aLDFA 81°). 38% were Type 2 'Intra-articular varus' with medial intra-articular bone loss (mean mLDFA 90.9°, MPTA 85.4°, VCA of 5.7°). 41% were Type 3 'Extra-articular varus' with extra-articular deformity (EAD). Type 3a had proximal tibial EAD; Type 3b had tibial diaphyseal EAD; Type 3c had femoral EAD (mean VCA 8.7°, HKA 166°), and severe medial bone loss (mean mLDFA 92°, MPTA 83°). 9% were Type 4 'Valgoid type' with features of valgus knees: Type 4a had medial femoral bowing (mean VCA 2.9°); Type 4b had significant distal femoral valgus (mean mLDFA 85.3°, aLDFA 78.6°). CONCLUSIONS: The null hypothesis that there was no phenotypic variation in varus osteoarthritic knees was rejected as considerable variation was found in coronal morphology of femur and tibia. Four broad phenotypic groups could be identified. Plane of the knee joint articular surface was quite variable. This has relevance to planning and performance of corrective osteotomies, unicompartmental and total knee arthroplasty. LEVEL OF EVIDENCE: III, retrospective cohort study.

Role of Etophylline and Theophylline Prolonged Release Tablet in COVID-19 Associated Sinus Node Dysfunction.

During COVID-19 pandemic, one of the most common arrythmia reported with this illness is sinus bradycardia. Treatment for COVID-19 and associated cardiac dysfunction is still evolving. Temporary pacemaker insertion is difficult due to pandemic and risk of spread of infection to the additional staff involved. Orciprenaline stimulates the sino-atrial and atrioventricular nodes and accelerates atrioventricular conduction. Theophylline improves sinus node function in subjects with sinus bradycardia and enhances atrioventricular nodal conduction We report a case series of 10 patients admitted in dedicated COVID-19 ICUs and developed sinus node dysfunction. All of these patients were started on etophylline and theophylline prolonged release tablet (150mg) once a day. On subsequent follow up after 72 hours, all patients reported heart rate well within normal range. COVID-19 virus directly involves the myocardium by entering the cardiac myocytes resulting in inflammation and injury. As the sinus bradycardia due to COVID-19 is usually transient and respond well this drug, short course of this drug could be added to treat this arrythmia in future.

Alkynamide phthalazinones as a new class of TbrPDEB1 inhibitors (Part 2).

Inhibitors against Trypanosoma brucei phosphodiesterase B1 (TbrPDEB1) and B2 (TbrPDEB2) have gained interest as new treatments for human African trypanosomiasis. The recently reported alkynamide tetrahydrophthalazinones, which show submicromolar activities against TbrPDEB1 and anti-T. brucei activity, have been used as starting point for the discovery of new TbrPDEB1 inhibitors. Structure-based design indicated that the alkynamide-nitrogen atom can be readily decorated, leading to the discovery of 37, a potent TbrPDEB1 inhibitor with submicromolar activities against T. brucei parasites. Furthermore, 37 is more potent against TbrPDEB1 than hPDE4 and shows no cytotoxicity on human MRC-5 cells. The crystal structures of the catalytic domain of TbrPDEB1 co-crystalized with several different alkynamides show a bidentate interaction with key-residue Gln874, but no interaction with the parasite-specific P-pocket, despite being (uniquely) a more potent inhibitor for the parasite PDE. Incubation of blood stream form trypanosomes by 37 increases intracellular cAMP levels and results in the distortion of the cell cycle and cell death, validating phosphodiesterase inhibition as mode of action.

Alkynamide phthalazinones as a new class of TbrPDEB1 inhibitors.

Several 3',5'-cyclic nucleotide phosphodiesterases (PDEs) have been validated as good drug targets for a large variety of diseases. Trypanosoma brucei PDEB1 (TbrPDEB1) has been designated as a promising drug target for the treatment of human African trypanosomiasis. Recently, the first class of selective nanomolar TbrPDEB1 inhibitors was obtained by targeting the parasite specific P-pocket. However, these biphenyl-substituted tetrahydrophthalazinone-based inhibitors did not show potent cellular activity against Trypanosoma brucei (T. brucei) parasites, leaving room for further optimization. Herein, we report the discovery of a new class of potent TbrPDEB1 inhibitors that display improved activities against T. brucei parasites. Exploring different linkers between the reported tetrahydrophthalazinone core scaffold and the amide tail group resulted in the discovery of alkynamide phthalazinones as new TbrPDEB1 inhibitors, which exhibit submicromolar activities versus T. brucei parasites and no cytotoxicity to human MRC-5 cells. Elucidation of the crystal structure of alkynamide 8b (NPD-048) bound to the catalytic domain of TbrPDEB1 shows a bidentate interaction with the key-residue Gln874 and good directionality towards the P-pocket. Incubation of trypanosomes with alkynamide 8b results in an increase of intracellular cAMP, validating a PDE-mediated effect in vitro and providing a new interesting compound series for further studies towards selective TbrPDEB1 inhibitors with potent phenotypic activity.

Liquid assisted pulsed laser ablation synthesized copper oxide nanoparticles (CuO-NPs) and their differential impact on rice seedlings.

Hydroponic experiments were conducted to investigate impact of laser ablated copper oxide nanoparticles (CuO-NPs) on rice seedlings. The present work demonstrates that exposure of lower concentrations (5, 10, 20, and 50 µM) of CuO-NPs enhance growth (in terms of fresh and dry weight and length), of rice seedlings. However, at higher concentrations (100, 200, and 500 µM) of CuO-NPs, growth (in terms of length, fresh weight and dry weight) decreased significantly (P < 0.05). Further, photosynthetic pigments (total chlorophyll and carotenoids) and protein contents were also found to be in accordance with the results of growth. This had occurred due to enhanced level of CuO-NPs accumulation at higher doses which also enhanced the level of oxidative stress markers such as hydrogen peroxide (H2O2) and malondialdehyde (MDA). Chlorophyll a fluorescence parameters (Fv/Fm and qP and except NPQ) and amount of some minerals (Ca, Mg, Na, and K) increased at lower concentrations of CuO-NPs. In contrast, the levels of Fv/Fm and qP were significantly (P < 0.05) reduced at higher concentration of CuO-NPs, which might be due to enhanced accumulation of Cu and oxidative stresses markers. Our results showed that lower dosages of pulsed laser ablated CuO-NPs (5, 10, 20, and 50 µM) might be beneficial for growth and development of rice seedlings.

Target highlights from the first post-PSI CASP experiment (CASP12, May-August 2016).

The functional and biological significance of the selected CASP12 targets are described by the authors of the structures. The crystallographers discuss the most interesting structural features of the target proteins and assess whether these features were correctly reproduced in the predictions submitted to the CASP12 experiment.

Structure and N-acetylglucosamine binding of the distal domain of mouse adenovirus 2 fibre.

Murine adenovirus 2 (MAdV-2) infects cells of the mouse gastrointestinal tract. Like human adenoviruses, it is a member of the genus Mastadenovirus, family Adenoviridae. The MAdV-2 genome has a single fibre gene that expresses a 787 residue-long protein. Through analogy to other adenovirus fibre proteins, it is expected that the carboxy-terminal virus-distal head domain of the fibre is responsible for binding to the host cell, although the natural receptor is unknown. The putative head domain has little sequence identity to adenovirus fibres of known structure. In this report, we present high-resolution crystal structures of the carboxy-terminal part of the MAdV-2 fibre. The structures reveal a domain with the typical adenovirus fibre head topology and a domain containing two triple ß-spiral repeats of the shaft domain. Through glycan microarray profiling, saturation transfer difference nuclear magnetic resonance spectroscopy, isothermal titration calorimetry and site-directed mutagenesis, we show that the fibre specifically binds to the monosaccharide N-acetylglucosamine (GlcNAc). The crystal structure of the complex reveals that GlcNAc binds between the AB and CD loops at the top of each of the three monomers of the MAdV-2 fibre head. However, infection competition assays show that soluble GlcNAc monosaccharide and natural GlcNAc-containing polymers do not inhibit infection by MAdV-2. Furthermore, site-directed mutation of the GlcNAc-binding residues does not prevent the inhibition of infection by soluble fibre protein. On the other hand, we show that the MAdV-2 fibre protein binds GlcNAc-containing mucin glycans, which suggests that the MAdV-2 fibre protein may play a role in viral mucin penetration in the mouse gut.

Some of the most interesting CASP11 targets through the eyes of their authors.

The Critical Assessment of protein Structure Prediction (CASP) experiment would not have been possible without the prediction targets provided by the experimental structural biology community. In this article, selected crystallographers providing targets for the CASP11 experiment discuss the functional and biological significance of the target proteins, highlight their most interesting structural features, and assess whether these features were correctly reproduced in the predictions submitted to CASP11. Proteins 2016; 84(Suppl 1):34-50. © 2015 The Authors. Proteins: Structure, Function, and Bioinformatics Published by Wiley Periodicals, Inc.

Crystal structure of the fibre head domain of bovine adenovirus 4, a ruminant atadenovirus.

BACKGROUND: In adenoviruses, primary host cell recognition is generally performed by the head domains of their homo-trimeric fibre proteins. This first interaction is reversible. A secondary, irreversible interaction subsequently takes place via other adenovirus capsid proteins and leads to a productive infection. Although many fibre head structures are known for human mastadenoviruses, not many animal adenovirus fibre head structures have been determined, especially not from those belonging to adenovirus genera other than Mastadenovirus. METHODS: We constructed an expression vector for the fibre head domain from a ruminant atadenovirus, bovine adenovirus 4 (BAdV-4), consisting of amino acids 414-535, expressed the protein in Escherichia coli, purified it by metal affinity and cation exchange chromatography and crystallized it. The structure was solved using single isomorphous replacement plus anomalous dispersion of a mercury derivative and refined against native data that extended to 1.2 Å resolution. RESULTS: Like in other adenoviruses, the BAdV-4 fibre head monomer contains a beta-sandwich consisting of ABCJ and GHID sheets. The topology is identical to the fibre head of the other studied atadenovirus, snake adenovirus 1 (SnAdV-1), including the alpha-helix in the DG-loop, despite of them having a sequence identity of only 15 %. There are also differences which may have implications for ligand binding. Beta-strands G and H are longer and differences in several surface-loops and surface charge are observed. CONCLUSIONS: Chimeric adenovirus fibres have been used to retarget adenovirus-based anti-cancer and gene therapy vectors. Ovine adenovirus 7 (OAdV-7), another ruminant atadenovirus, is intensively tested as a basis for such a vector. Here, we present the high-resolution atomic structure of the BAdV-4 fibre head domain, the second atadenovirus fibre head structure known and the first of an atadenovirus that infects a mammalian host. Future research should focus on the receptor-binding properties of these fibre head domains.

Extensive Metallosis in a Primary Knee Arthroplasty as a Result of Polyethylene Wear: Is It Avoidable?

Metallosis is known to occur in metal-on-metal arthroplasty and has been of concern to orthopaedic surgeons worldwide. It is a rare, late complication of total knee arthroplasty (TKA), in which metal-on-metal contact leads to metal debris deposition in the surrounding tissue. Reasons for metal-on-metal contact could range from wear of the polyethylene insert to abnormal joint biomechanics. Many components can affect the development of metallosis, with polyethylene wear being the most common cause of metallosis. This paper discusses the case of an 85-year-old man who developed metallosis, attributed to polyethylene wear, 24 years after undergoing TKA. It also highlights the different components of knee prostheses, evaluates the efficacy of different types of polyethylene, and explores whether ceramic coating can improve TKA outcomes and reduce complications such as metallosis.

Use of a Trephine to Extract a Fractured Corail Femoral Stem During Revision Total Hip Arthroplasty: Tips From Our Case Report.

Despite the significant advancements in the field of total hip arthroplasty (THA) and prosthesis designs, fracture of the modular femoral stem after THA can occur rarely and needs attention. Orthopaedic surgeons face a daunting task when it comes to the removal of a broken stem. The use of a trephine reamer has been evaluated for extracting the distal femoral stem, and this case report tries to address some key tips for the same. A 67-year-old obese male, without any major medical comorbidities, presented to the Royal Lancaster Infirmary orthopaedic outpatient department with a complaint of acute-on-chronic right anterior thigh pain that worsened over a few weeks. He had a history of bilateral staged uncemented THA done around 12 years ago. The plain radiological images confirmed the presence of a fracture of the Corail femoral stem. A posterior approach was used to dislocate the hip and the distal broken part of the stem was removed using trephines. Reamers were used and care was taken to prevent thermal necrosis by using intermittent saline lavage. After the removal of the fractured femoral stem, a cemented femoral revision THA was performed, which was uneventful. The patient walked without any aid or thigh pain postoperatively during his last follow-up. Using trephines to remove broken femoral stems is an effective and safe method. Intraoperative measures can help in avoiding heat necrosis while using a trephine reamer for extracting the fractured femoral stem. Regular follow-up and counselling are important after THA, to avoid a delayed diagnosis of non-traumatic femoral stem fractures.

Crystallization of the C-terminal head domain of the fibre protein from a siadenovirus, turkey adenovirus 3.

Turkey adenovirus 3 belongs to the genus Siadenovirus. Its predicted fibre protein consists of an N-terminal virus-attachment domain, a central shaft domain and a head domain at the C-terminus. The head domain has little sequence identity to known adenovirus fibre head structures. Crystals of the fibre head domain consisting of amino acids 304-454 with an N-terminal purification tag were produced. Crystals of native and selenomethionine-derivatized protein belonged to space group I23 (unit-cell parameter 99 Å). They diffracted synchrotron radiation to 2.0 and 2.14 Šresolution, respectively, and are expected to contain one monomer in the asymmetric unit.

Crystallization of the C-terminal domain of the fibre protein from snake adenovirus 1, an atadenovirus.

Adenovirus fibre proteins play an important role in determining viral tropism. The C-terminal domain of the fibre protein from snake adenovirus type 1, a member of the Atadenovirus genus, has been expressed, purified and crystallized. Crystals were obtained belonging to space groups P2(1)2(1)2(1) (two different forms), I2(1)3 and F23. The best of these diffracted synchrotron radiation to a resolution of 1.4 Å. As the protein lacks methionines or cysteines, site-directed mutagenesis was performed to change two leucine residues to methionines. Crystals of selenomethionine-derivatized crystals of the I2(1)3 form were also obtained and a multi-wavelength anomalous dispersion data set was collected.

Controlled synthesis, characterization, and application of iron oxide nanoparticles for oral delivery of insulin.

Impurity-free, controlled synthesis of iron oxide nanoparticle, in ultrapure water and chitosan, using laser ablation technique and its application for type II diabetes management through oral delivery of insulin-loaded iron oxide-chitosan nanocomposite is presented. The purity of the nanoparticle is monitored by laser-induced breakdown spectroscopy technique. The synthesized iron oxide nanoparticle was characterized by UV/Vis absorption spectroscopy, and morphological study was performed by scanning electron microscope. The intensity of absorption peak and wavelength corresponding to peak of the nanoparticle prepared in water and chitosan is dependent on the laser energy used for ablation purpose. Red shift in the absorption peak wavelength was observed by increasing laser energy. In addition to red shift, an increase in intensity of absorption peak was also seen when ablating laser energy was increased. The appearance of a weak peak around 295 nm was observed in iron oxide-chitosan nanocomposite. The spherical shape of the nanoparticle synthesized at the lower laser energy has gradually changed to triangular and irregular shaped structures as ablating laser energy was increased. The spherical nanoparticles loaded with insulin were used for oral delivery for diabetic management. The iron oxide-chitosan nanocomposite loaded with insulin has resulted in reduction in blood glucose level in mild diabetic, subdiabetic, and severely diabetic rats; more than 51 % reduction in blood glucose level, compared to the control group, has been achieved in the present work.