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A novel fibrinolytic enzyme, from the marine fungus Penicillium steckii KU1, was purified to electrophoretic homogeneity. The fibrinolytic protease was purified to 13.56 times with a specific activity of 57.64 U/mg and final yield of 13.93 %. It was found to be a monomeric protein of 12.6 kDa, having optimum activity at 30 °C and pH 8.0. It is a plasmin-like enzyme, showing resemblance to ATP-dependent zinc metalloprotease with isoelectric point (pI) 8.0. Its activity is enhanced by Zn 2+ , and inhibited by ethylenediaminetetraacetic acid (EDTA), Co 2+ and Fe 2+ . The enzyme interaction with substrate azocasein was endothermic and with inhibitor EDTA exothermic. The K m , V max , K cat and catalytic efficiency of the enzyme for azocasein were determined to be 142.71 μg mL -1 , 285.71 μg min -1  mL -1 , 6.35 S -1 and 4.45 × 10 -2  S -1  μg -1  mL respectively. It hydrolyzed all three chains of fibrinogen within 9 h, and dissolved fibrin completely within 24 h. 2 mg/mL enzyme could dissolve blood clot completely within 30 min, with negligible hemolysis (2.60 %). Lowering the immunogenicity by the application of natural or engineered small proteins is a strategy to enhance the safety and efficacy of thrombolytic therapy. Hence, the present 12.6 kDa, plasmin-like fibrinolytic enzyme appears worthy of further investigations towards a thrombolytic therapeutic., Competing Interests: Declaration of competing interest The authors have no relevant financial or non-financial interests to disclose., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective  To compare the functional outcomes of anterior cruciate ligament (ACL) reconstruction with hamstring autograft (HA) through the all-inside (AI) technique with adjustable-loop cortical Endobutton (Smith & Nephew, Watford, Hertfordshire, England) on the sides of the femur and tibia and through the outside-in (OI) technique using an interference screw on the tibial side and a cortical Endobutton on the femoral side. Materials and Methods  The present is a double-blinded randomized controlled trial (RCT) of 44 patients undergoing arthroscopic ACL reconstruction from February 2019 to February 2022 in a tertiary care hospital. As per computer-based randomization, the patients were distributed into two groups: the AI and OI groups. Both groups were evaluated for 12 months using the Visual Analog Scale (VAS), the Lysholm Knee Scoring Scale, and part I (pain score) and part II (function score) of the Knee Society Score (KSS). Results  On postoperative day 2, the VAS score was significantly higher in the OI group ( p  = 0.0001), but insignificant ( p  = 0.807) at 6 weeks. At 3, 6, and 12 months of follow-up, the score on the Lysholm Knee Scoring Scale was significantly higher ( p  = 0.001) in the AI group. At 6 months, both parts of the KSS showed a significant difference, with the AI group presenting a better outcome ( p  = 0.04). However, at 12 months, the AI group presented a better score on part I of the KSS, but no differences were observed regarding part II. Conclusion  In a follow-up of 12 months, the patients submitted to the AI technique presented better outcome scores and pain relief than those submitted to the OI technique., Competing Interests: Conflito de Interesses Os autores não têm conflito de interesses a declarar., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution 4.0 International License, permitting copying and reproduction so long as the original work is given appropriate credit ( https://creativecommons.org/licenses/by/4.0/ ).)

Citrus sinensis (L.) Osbeck (Rutaceae), commonly known as the sweet orange, is a popular and widely consumed fruit with several medicinal properties. The present study aimed to perform the in silico screening of 18 flavonoids and eight volatile components from the peel of C. sinensis against apoptotic and inflammatory proteins, metalloprotease, and tumor suppressor markers. Flavonoids obtained higher probabilities than volatile components against selected anti-cancer drug targets. Hence, the data from the binding energies against the essential apoptotic and cell proliferation proteins substantiate that they may be promising compounds in developing effective candidates to block cell growth, proliferation, and induced cell death by activating the apoptotic pathway. Further, the binding stability of the selected targets and the corresponding molecules were analyzed by 100 ns molecular dynamics (MD) simulations. Chlorogenic acid has the most binding affinity against the important anti-cancer targets iNOS, MMP-9, and p53. The congruent binding mode to different drug targets focused on cancer shown by chlorogenic acid suggests that it may be a compound with significant therapeutic potential. Moreover, the binding energy predictions indicated that the compound had stable electrostatic and van der Waal energies. Thus, our data reinforce the medicinal importance of flavonoids from C. sinensis and expand the need for more studies, seeking to optimize results and amplify the impacts of further in vitro and in vivo studies. Communicated by Ramaswamy H. Sarma.

Uterine dehiscence (partial or complete) is a rare complication of lower segment cesarean section (LSCS). Puerperal sepsis with intra-abdominal abscess following this event has been rarely reported. The delay in diagnosis and management of the condition can result in significant morbidity and mortality. We herein report three cases of puerperal sepsis along with intra-abdominal abscess associated with uterine dehiscence following LSCS. These patients in the current case series presented with complaints of fever and abdominal pain. Early recognition and prompt treatment with diagnostic laparoscopy and or laparotomy with drainage were effective in the management of these patients., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2021, Haridas et al.)

An overview of the biological properties of phycocyanin (PC) amply illustrates that it may not have any specific functional feature towards any system at which it may elicit a specific function, but for the molecular interactions. Nevertheless, based on existing evidences, it is hypothesized that PC has more than one functional target with the interacting systems; therefore, it has diversity of effects. The mechanism of PC action remains elusive of a comprehensive idea. The present investigation focuses on the pro inflammatory enzyme, lipoxygenase (LOX) inhibiting property of PC purified from Oscillatoria sp. Enzyme kinetics studies show that the molecular composite of PC is required for its inhibition shown on LOX. Isothermal titration calorimetric study proves that one molecule of PC interacts with two molecules of LOX. Molecular dynamics simulation study pertaining to PC-LOX interactions shows it to be appropriate as a model to give molecular mechanistic insight into the varied biological properties of PC, demonstrated elsewhere in experimental studies including animal model studies. It explains that the PC-LOX interaction is of a function-freezing, protein-protein interaction in nature. The wide spectrum of properties of PC might be due to its function as a powerful protein hub showing non-specific protein-protein interactions., Competing Interests: Declaration of competing interest The authors have declared no conflict of interests., (Copyright © 2020. Published by Elsevier B.V.)

A direct acting, extracellular, fibrinolytic enzyme, ~50 KDa from marine Pseudomonas aeruginosa KU1 (PEKU1), was purified. It was found to be a metalloprotease. 60% of the total activity of the purified PEKU1 was retained at 70 °C and the enzyme was practically denatured at 80 °C, 2 h. Metal ions, such as Na + , K + and Co 2+ , were found to enhance slightly the fibrinolytic activity, while Fe 2+ , Mn 2+ and Zn 2+ were found to be inhibiting. The enzyme showed only less than 5% haemolysis, suggesting its thrombolytic administration safe. Tryptic digestion revealed its similarity to serralysin like alkaline protease of P. aeruginosa. In silico studies showed its binding of protease substrates and fibrin D-dimer in its active site. High affinity binding of bradykinin to the active site of PEKU1, confirmed by in vitro cleaving, suggested its future use as an analgesic. The purified enzyme with Na + , K + and Co 2+ , and without Fe 2+ , Mn 2+ and Zn 2+ showed thrombolysis in vivo in carrageenan induced murine tail thrombolytic model. The enzyme PEKU1, a novel protease from marine isolate Pseudomonas aeruginosa KU1 has great potential to be developed as a therapeutic agent to combat cardiovascular diseases, as well as analgesic and anti-inflammatory drug in appropriate sites., Competing Interests: Declaration of competing interest The authors have declared conflict of interests none., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: The ethno-medical significance of Clerodendrum genus raises the interest towards the characterization of its seed lectin by inexpensive and most effective technique., Objective: The focus of this study is the purification, characterization, and evaluation of the antioxidant and antiproliferative potential of a galactose-specific lectin from Clerodendrum infortunatum L. seeds., Materials and Methods: The crude extract, homogenized in 6 volumes of the saline containing 10 mM β-mercaptoethanol was subjected to pigment removal by Toyopeal HW-55 column prior to ammonium sulfate fractionation (40-80 %). The crude protein extract was then loaded to the gel filtration column Sephadex G-200 followed by affinity chromatography using activated galactose coupled Sepharose-4B., Results: The SDS-PAGE analysis showed a single band of about 30 kDa which further determined by MALDI-TOF analysis. The MALDI-TOF spectra revealed that Clerodendrum infortunatum lectin (CIL) is a homo-tetramer of 120 kDa consisting of four identical subunits of 30 kDa. The haemagglutination inhibition assay was done with purified lectin by many sugars, among which N-acetyl-D-galactosmine (NAG), D-galactose and lactose exhibited high inhibition. NAG showed the highest inhibition amongst the tested sugars, having the minimum inhibitory concentration of about 0.97 mM. The lectin exhibited a moderate antioxidant activity with an IC 50 value of 6.1 ± 0.1 mg.mL -1 and induced cell death with IC 50 of 82.8 μg.mL -1 against human gastric cancer cell line, AGS, indicated the potential of CIL for clinical and therapeutic applications., Conclusion: The present study demonstrated the moderate ability of the CIL to inhibit the growth of human gastric cancer cells, AGS either by causing cytotoxic or anti-proliferative effects. Thus, CIL due to its remarkable properties may be considered as a potential bio-molecule in tumor research and glycobiology.

2-Deoxy-D-ribose-5-phosphate aldolase (DERA) is a class I aldolase that offers access to several building blocks for organic synthesis. It catalyzes the stereoselective C-C bond formation between acetaldehyde and numerous other aldehydes. However, the practical application of DERA as a biocatalyst is limited by its poor tolerance towards industrially relevant concentrations of aldehydes, in particular acetaldehyde. Therefore, the development of proper experimental conditions, including protein engineering and/or immobilization on appropriate supports, is required. The present review is aimed to provide a brief overview of DERA, its history, and progress made in understanding the functioning of the enzyme. Furthermore, the current understanding regarding aldehyde resistance of DERA and the various optimizations carried out to modify this property are discussed.

Phospholipase A 2 (PLA 2 ) is one of the rate limiting enzymes involved in the production of arachidonic acid, a potent inflammatory mediator. PLA 2 is widely distributed all over the animal kingdom. It is also seen in inflammatory exudation and venoms of different organisms. The studies demonstrated that PLA 2 inhibitors have broad spectrum activities that they can either be used against inflammation or envenomation. In this study, the inhibitory activity of 1-napthaleneacetic acid (NAA) against porcine pancreatic PLA 2 has been explained through isothermal titration calorimetry and enzyme kinetics studies. The atomic level of interactions of NAA with PLA 2 was also studied using X-ray crystallography. Apart from these findings, the theoretical binding affinities and mode of interactions of two naphthalene-based NSAIDs such as naproxen (NAP) and nabumetone (NAB) were studied through molecular modeling. The studies proved that the selected ligands are binding at the doorway of the active site cleft and hindering the substrate entry to the active site. The study brings out a potential scaffold for the designing of broad spectrum PLA 2 inhibitors which can be used for inflammation or envenomation. © 2018 IUBMB Life, 70(10):995-1001, 2018., (© 2018 International Union of Biochemistry and Molecular Biology.)

Therapeutic effects of gallic acid (GA) have already been extensively studied. However, its interaction with lectins has not gained much attention. It is of interest to validate the binding profile of GA with Spatholobus parviflorus seed lectin. A combination of Isothermal Titration Calorimetry (ITC), haemagglutination assay and molecular docking was applied on SPL-GA interaction. ITC results showed four binding sites, stoichiometry, n=4, irrespective of the ratio of SPL:GA taken for titration. Difference among the four binding sites of a single molecule of SPL with regard to GA binding kinetic parameters was consistently varying. Similarly, the glide scores obtained for GA in the four different binding clefts of SPL were also conformed to the ITC. The binding of GA on SPL without affecting its sugar binding property could be considered as a boon for glycobiological research. From the presented studies, it could be proposed that the SPL-GA interactions may facilitate drug delivery by specific targeting/attachment by profiling of cell-surface glycans, followed by controlled release of drugs., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Proteins may utilize complex networks of interactions to create/proceed signaling pathways of highly adaptive responses such as programmed cell death. Direct binary interactions study of proteins may help propose models for protein-protein interaction. Towards this goal we applied a combination of thermodynamic kinetics and crystal structure analyses to elucidate the complexity and diversity in such interactions. By determining the heat change on the association of two galactose-specific legume lectins from Butea monosperma (BML) and Spatholobus parviflorus (SPL) belonging to Fabaceae family helped to compute the binding equilibrium. It was extended further by X-ray structural analysis of BML-SPL binary complex. In order to chart the proteins interacting mainly through their interfaces, identification of the nature of forces which stabilized the association of the lectin-lectin complex was examined. Comprehensive analysis of the BMLSPL complex by isothermal titration calorimetry and X-ray crystal structure threw new light on the lectin-lectin interactions suggesting of their use in diverse areas of glycobiology., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A novel, poly(ethyl ethylene ether) inhibitor to trypsin was purified from marine cyanobacteria, Lyngbya confervoides from the coastal areas of Thalassery, North Kerala. The kinetics and the thermodynamic parameters of its interactions with the enzyme were also studied. It was demonstrated that the substrate binding, catalytic triad of the enzyme could be blocked by the inhibitor, as expressed by molecular simulation studies. The study also showed that the cyanobacterial group could prove to be a potential source of novel enzyme inhibitors for various applications.

Piperine is a secondary metabolite of black pepper. Its uses in medicine were already studied. However, its derivatives have not gained considerable attention. In the presented study, the Lipoxygenase (LOX) inhibitory activity of piperine and its derivatives, piperonylic acid, piperic acid, and piperonal have been assessed and compared by enzyme kinetics, ITC and molecular modeling experiments. The presented investigations expressed that all the studied compounds inhibited LOX by binding at its active site. The IC(50) values of these compounds were deduced from the kinetics data and found to be 85.79, 43.065, 45.17, and 50.78 μm for piperine, piperonylic acid, piperic acid, and piperonal, respectively. The binding free energies obtained from ITC experiments were -7.47, -8.33, -8.09, and -7.86 kcal/mol for piperine, piperonylic acid, piperic acid, and piperonal, respectively. Similarly, the glide scores obtained for piperine, piperonylic acid, piperic acid, and piperonal were -7.28, -10.32, -10.72, and -9.57 kcal/mol, respectively. The results of ITC and molecular modeling experiments suggested that piperonylic acid and piperonal exhibit stronger binding at the active site than piperine does. From the presented studies, it could be concluded that derivatives of piperine may be of higher significance than piperine for certain medicinal applications, implicating (Ayurvedic) fermented herbal drugs with piperine in them., (© 2014 John Wiley & Sons A/S.)

Background: Operative hemorrhoidectomy can result in pain and altered continence from excessive excision of anoderm or surrounding tissue. We assessed a novel low-profile slotted anoscope to determine if the device would promote safe dissection, lessen trauma, and reduce operative times for hemorrhoidectomy., Methods: Patients requiring hemorrhoidectomy (June 2008 - January 2010) underwent a prospective phase-2 trial evaluating a new operating anoscope (CAD, Ethicon Endosurgery, Cincinnati, OH). Demographics and perioperative end points including bleeding, pain, fecal incontinence, stenosis, and symptom recurrence were analyzed at 4 weeks, 3 months, 6 months, and 1 year postoperatively. We compared these to patients undergoing hemorrhoidectomy (February 2010 - November 2012) with a traditional Hill-Ferguson anoscope (THF)., Results: 40 patients (CAD, 20 vs THF, 20) were included. Presenting symptoms were similar, whereas mean duration of symptoms was longer for CAD (41.2 ± 8.4 vs 27 ± 9.5 months; P < .05). Estimated blood loss was lower for CAD [8.3 mL (range = 2-40 mL) vs 11.3 mL THF (range = 5-35 mL; P = .87)]. Mean operative times were lower for the CAD than the THF group (15.6 ± 3.4 vs 26.1 ± 4.1 minutes; P < .05). Visual analog pain scores were non-significantly increased in the THF group at 4 weeks (P = .23). At 3 months, 6 months, and 1 year, there was no difference in continence., Conclusion: The CAD anoscope reduced operative times for modified Ferguson (closed) hemorrhoidectomy when compared with traditional retractors. There was no difference in incontinence or pain between groups., (© The Author(s) 2014.)

Dibenzodioxins bearing multiple electron withdrawing groups were synthesized using a simple one-step methodology including examples of molecules possessing electron acceptor groups in both ends. As a consequence internal charge delocalization occurs and the optical spectra are found to be bathochromically shifted compared to similar examples known thus far. A theoretical analysis of the molecular orbitals reveals the origin of the peaks in the dibenzodioxin optical spectra. Select examples exhibit in vitro neuro-cytotoxicity against glioma cell line C6, a finding which enhances existing knowledge about the pharmacologically relevant structural motifs in dibenzodioxins., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Intra-abdominal infections (IAIs) are a major cause of morbidity and death. We hypothesized that the involvement of specific organisms would predict death independently., Patients and Methods: All patients with IAIs treated at an academic tertiary-care facility over eight years (June 1999-June 2007) were included. The data collected were demographics, co-morbidities, source of infection, intra-abdominal culture results, type of infection (community-acquired vs. nosocomial), type of intervention (operative vs. percutaneous drainage), and outcome. The Charlson Comorbidity Index and multiple organ dysfunction score (MODS) were used in the analysis., Results: A total of 389 patients were admitted for 452 infection episodes (IEs) during the study period. None of the 129 patients with appendiceal-related infections died, and these patients were excluded from further analysis. Thus, 323 non-appendiceal IEs were evaluated. The overall mortality rate was 8.7%. The mean age of the patients was 54 y, and 50% of them were male. Intra-abdominal cultures were obtained from 303 IEs (93.8%). The most common cause of IAI was post-operative infection (44%). There were 49 distinct species isolated. The most common were Enterococcus (105), Escherichia coli (75), Streptococcus (62), Staphylococcus (51), and Bacteroides (46). Bivariable analysis revealed multiple risk factors associated with death. Logistic regression demonstrated that independent risk factors for death were age ≥65 years (odds ratio [OR] 3.92), cardiac event (OR=8.17), catheter-related blood stream infection (OR=6.16), and growth of Clostridium (OR=13.03). The growth of Streptococcus was predictive of survival. The C statistic was 0.89., Conclusions: In addition to age and intrinsic patient factors, the presence of specific bacterial organisms independently predicts death in patients with non-appendiceal IAI.

Spatholobus parviflorus seed lectin (SPL) is a heterotetrameric lectin, with two α and two β monomers. In the crystal structure of SPL α monomer, two residues at positions 240 and 241 are missing. This region was modeled based on the positional and sequence similarities. The role of metal ions in SPL structure was analyzed by 10 ns molecular dynamics simulation. MD simulations were performed in the presence and absence of metal ions to explain the loss of haemagglutinating property of the lectin due to demetallization. Demetallized structure was found to deviate drastically at the metal binding loop region. Affinity of different sugars like N-acetyl galactosamine (GalNAc), D-galactose and lactose towards the native and demetallized protein was calculated by molecular docking studies. It was found that the sugar binding site got severely distorted in demetallized lectin. Consequently, sugar binding ability of lectin might be decreasing in the demetallized condition. Isothermal titration calorimetric (ITC) analysis of the sugars in the presence of native and demetallized protein confirmed the in silico results. It was observed after molecular dynamics simulations, that significant structural deviations were not caused in the quaternary structure of demetallized lectin. It was confirmed that the structural changes modified the sugar binding ability, as well as sugar specificity of the present lectin. The role of metal ions in sugar binding is described based on the in silico studies and ITC analysis. A comprehensive analysis of the ITC data suggests that the sugar specificity of the metal bound lectin and the loss of sugar specificity due to metal chelation are not linear.

Phospholipase A2 (PLA₂) is one of the key enzymes involved in the formation of inflammatory mediators. Inhibition of PLA₂ is considered to be one of the efficient methods to control inflammation. In silico docking studies of 160 selected indole derivatives performed against porcine pancreatic PLA₂ (ppsPLA2) suggested that, CID2324681, CID8617 (indolebutyric acid or IBA), CID22097771 and CID802 (indoleacetic acid or IAA) exhibited highest binding energies. In silico analysis was carried out to predict some of the ADME properties. The binding potential of these compounds with human non pancreatic secretory PLA₂ (hnpsPLA₂) was determined using molecular docking studies. In order to corroborate the in silico results, enzyme kinetics and isothermal titration calorimetric analysis of the two selected compounds, IAA and IBA were performed against ppsPLA₂. From the analysis, it was concluded that IAA and IBA can act as competitive inhibitors to the enzyme and may be used as anti inflammatory agents.

The anti inflammatory potential of the human and microbial biotransformed derivatives of berberine were determined by molecular docking. It was revealed that almost all derivatives formed as a result of biotransformation showed increase in phospholipase A(2) binding affinity compared to berberine. The newly introduced -OH group/groups establish stronger hydrogen bonding interactions and more number of van der Waals contacts with the protein. As phospholipase A(2) is a target of anti inflammatory drugs, it might be concluded that certain biotransformed derivatives of berberine could be better anti inflammatory agents compared to berberine.

Ester bond hydrolysis of membrane phospholipids by Phospholipase A(2) and consequent release of fatty acids are the initiating steps of inflammation. It is proposed in this study that the inhibition of phospholipase A(2) is one of the ways to control inflammation. Investigations are carried out to identify the mode of inhibition of phospholipase A(2) by the n-hexadecanoic acid. It may help in designing of specific inhibitors of phospholipase A(2) as anti-inflammatory agents. The enzyme kinetics study proved that n-hexadecanoic acid inhibits phospholipase A(2) in a competitive manner. It was identified from the crystal structure at 2.5 Å resolution that the position of n-hexadecanoic acid is in the active site of the phospholipase A(2). The binding constant and binding energy have also been calculated using Isothermal Titration Calorimetry. Also, the binding energy of n-hexadecanoic acid to phospholipase A(2) was calculated by in silico method and compared with known inhibitors. It may be concluded from the structural and kinetics studies that the fatty acid, n-hexadecanoic acid, is an inhibitor of phospholipase A(2), hence, an anti-inflammatory compound. The inferences from the present study validate the rigorous use of medicated oils rich in n-hexadecanoic acid for the treatment of rheumatic symptoms in the traditional medical system of India, Ayurveda., (© 2012 John Wiley & Sons A/S.)

Biotransformation of berberine by Rhizopus oryzae leads to its demethylation, producing hydroxyl derivatives, as revealed by Fourier Transform Infra Red spectroscopy, Nuclear Magnetic Resonance and Electro Spray Ionization-Mass Spectrometric analyses. Surface Plasmon Resonance and enzyme kinetic studies showed that biotransformed derivatives of berberine had a higher inhibitory potential than berberine towards phospholipase A(2). X-ray crystal structures demonstrated that biotransformed berberine binds to PLA(2) in an entirely different, inverted orientation with respect to the binding of berberine. This study brings out the significance of biotransformation in generation of better drug-lead compounds., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A galactose-specific seed lectin from Spatholobous parviflorus (SPL) has been purified, crystallized and its X-ray structure solved. It is the first lectin purified and crystallized from the genus Spatholobus (family: Fabaceae). The crystals belong to the space group P1, with a=60.792 Å, b=60.998 Å, c=78.179 Å, α=78.68°, β=88.62°, γ=104.32°. The data were collected at 2.04 Å resolution under cryocondition, on a MAR image-plate detector system, mounted on a rotating anode X-ray generator. The coordinates of Dolichos biflorus lectin (1lu1) were successfully used for the structure solution by molecular replacement method. The primary structure of the SPL was not known earlier and it was unambiguously visible in the electron density. S. parviflorus lectin is a hetero-dimeric-tetramer with two alpha and two beta chains of 251 and 239 residues respectively. SPL has two metal ions, Ca(2+) and Mn(2+), bound to a loop region of each chain. The SPL monomers are in jelly roll form., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A galactose-specific seed lectin was purified from the legume Spatholobus parviflorus and crystallized using the hanging-drop vapour-diffusion technique. The crystals belonged to space group P1, with unit-cell parameters a = 60.998, b = 60.792, c = 78.179 Å, α = 101.32, β = 91.38, γ = 104.32°. X-ray diffraction data were collected under cryoconditions (100 K) to a resolution of 2.04 Å using a MAR image-plate detector system mounted on a rotating-anode X-ray (Cu Kα) generator. Molecular replacement using legume-lectin coordinates as a search model gave a tetrameric structure.

Antimicrobials derived from plants have been receiving increasing attention in recent years. Antimicrobial activities of a number of phytochemicals have been reported. Many present day antibiotics are ineffective against several pathogenic organisms. About 90% of Staphylococcus aureus isolates from clinical specimens is reported to have resistance against beta-lactam antibiotics. In the present study, the effect of hexane, diethyl ether, acetone and water extracts of leaves of a medicinal plant Holoptelea integrifolia has been tested against beta-lactam resistant strain of S. aureus in presence of antibiotics such as ampicillin, amoxicillin, cefotaxime and ceftriaxone. The diethyl ether extract has shown the maximum antibacterial activity and the active principle is found to be 1,4-naphthalenedione which is characterized by GC-MS and FTIR spectroscopy. The minimum inhibitory concentration (MIC) of the compound is found to be 4 mg/ml. Structural similarity of this compound with a functional group of a beta-lactamase-resistant antibiotic indicates that 1,4-naphthlenedione may be acting as an inhibitor to beta-lactamase.

Background: Intra-abdominal infections (IAIs) are an important cause of mortality and morbidity. Nosocomial IAIs (NIAIs) have been associated with higher mortality than community-acquired IAIs (CIAIs). We hypothesized that intrinsic risk factors were a better predictor of mortality than the type of infection., Methods: Patients with IAI treated at a single urban academic hospital over 8 years (June 1999-June 2007) were retrospectively reviewed. Data collected included demographics, comorbidities, source of infection, type of infection (community vs nosocomial), type of intervention (operation versus percutaneous drainage), and postoperative complications. Charlson Comorbidity Index and multiple organ dysfunction (MOD) scores were evaluated at admission and on postoperative day 7 (POD-7)., Results: There were 452 patients; 234 (51.8%) had CIAI and 218 (48.2%) had NIAI. The mean age was 51.3 +/- 0.8. The most common source of CIAI was the appendix (n = 129, 28.5%); 137 patients with NIAI had postoperative infections (30.3%). When patients with appendicitis were excluded, there was no difference in mortality or complications between patients with CIAI and NIAI. Logistic regression analysis demonstrated catheter-related bloodstream infection (P < .001; OR 7.3, 95% CI, 2.5-22.2), cardiac event (P < .001; OR 6.0, 95% CI, 2.3-16.1), and age > or = 65 (P = .009; OR 3.8, 95% CI, 1.4-8.8) to be independent risk factors for mortality. Among patients who failed initial therapy, a non-appendiceal source of infection (P < .001; OR 4.7, 95% CI, 2.3-9.8) and a Charlson score > or =2 (P = .033; OR 1.6, 95% CI, 1.0-2.6) were determined to be independent risk factors. Non-appendiceal source of infection (P = .001, OR 3.3, 95% CI, 1.6-7.0) and POD-7 MOD score > or =4 (P < .001; OR 3.4, 95% CI, 1.9-6.0) were found to be independent predictors for re-intervention., Conclusion: These results suggest mortality from IAI is strongly related to age and organ dysfunction; however, catheter-related bloodstream infection and postoperative cardiac events have a greater effect on outcome.

Background: Global parameters, such as wound class, the American Society of Anesthesiologists' physical classification score, and prolonged operative time, have been associated with the risk of surgical site infection (SSI). We hypothesized that additional risk factors for SSI would be identified by controlling for these parameters and that deep and organ/space SSI may have different risk factors for occurrence., Methods: A retrospective review was performed on general and vascular surgical patients who underwent an operation between June 2000 and June 2006 at a single institution. Patients with SSI were matched with a case-control cohort of patients without infection (no SSI) according to age, sex, ASA score, wound class, and type of operative procedure. Data were analyzed using bivariate and regression analyses., Results: Overall, 10,253 general surgical procedures were performed during the 6-year period; 316 patients (3.1%) developed SSI. In all, 300 patients with 251 superficial (83.6%), 22 deep (7.3%), and 27 organ/space (9%) SSIs were matched for comparison. Multivariate logistic regression analysis identified previous operation (odds ratio [OR], 2.4; 95% confidence interval [CI] = 1.6-3.7), duration of operation >or=75th percentile (OR, 1.8; 95% CI = 1.2-2.8), hypoalbuminemia (OR, 1.8; 95% CI = 1.1-2.8), and a history of chronic obstructive pulmonary disease (OR, 1.7; 95% CI = 1.0-2.8) as independent risk factors for SSI. Only hypoalbuminemia (OR, 2.9; 95% CI = 1.4-6.3) and a previous operation (OR, 2.0; 95% CI = 1.0-4.4) were significantly associated with deep or organ/space infections., Conclusions: These results demonstrate additional factors that increase the risk of developing SSI. Deep and organ/space infections have a different risk profile. This information should guide clinicians in their assessment of SSI risk and should identify targets for intervention to decrease the incidence of SSI.

Background: Observation following thyroidectomy and parathyroidectomy has been progressively shortened. The challenge has been to reduce the duration of postoperative observation without jeopardizing patient safety., Methods: A retrospective review of patients who underwent thyroidectomy and/or parathyroidectomy between July 1990 and March 2007 was completed to determine the frequency of life-threatening hematoma and hospital readmission and their impact on postoperative observation., Results: Of 1,050 patients, life-threatening hematoma developed in 6 (.6%) patients, 5 following bilateral and 1 following unilateral thyroidectomy. Hematoma developed 10 minutes to 7 days postoperatively, four within 4 hours, one at 21 hours, and one at 7 days. Twelve patients were readmitted an average of 5 days postoperatively for hypocalcemia, hematoma, infection, or respiratory distress., Conclusion: Without factors contributing to bleeding, unilateral thyroidectomy and parathyroidectomy can be performed as an ambulatory procedure. To maximize safety, we recommend 4-hour and 23-hour observation following unilateral and bilateral thyroidectomy, respectively.

Background: Previous studies evaluating predictive factors for conversion from laparoscopic to open cholecystectomy have drawn conflicting conclusions. We evaluated objective preoperative variables to create an accurate, accessible risk score to predict conversion., Methods: A retrospective review was performed of laparoscopic cholecystectomy patients at an urban tertiary care center. Seventy characteristics were subjected to bivariate and multivariate logistic regression analysis to identify parameters that independently predict conversion to open cholecystectomy. A model was created based on this analysis., Results: Laparoscopic cholecystectomy was performed on 1377 patients for benign gallbladder disease over a 71-month period. There were 112 (8.1%) conversions to open cholecystectomy. The correlation between the preoperative clinical diagnosis and pathologic diagnosis for acute and chronic cholecystitis was 48.6% and 94.6%, respectively. Multivariate analysis identified male gender, elevated white blood cell count, low serum albumin, ultrasound finding of pericholecystic fluid, diabetes mellitus, and elevated total bilirubin as independent predictors of conversion. These 6 factors were also associated with the pathologic diagnosis of acute cholecystitis. A model to calculate the risk for conversion was created with an area under the receiver operator curve of 0.83. The risk for conversion also can be estimated based on the number of factors identified present and ranged from 2% when 1 factor was present to 89% with 6 factors., Conclusions: These results demonstrate that conversion to open cholecystectomy can be predicted based on parameters available preoperatively. Conversion is more likely in patients who have acute cholecystitis; however, the correlation between its clinical and pathologic diagnosis is poor. Improvements in the ability to determine the risk for conversion have important implications for surgical care.

Kaliocin-1, a 31-residue synthetic peptide (FFSASCVPGADKGQFPNLCRLCA GTGENKCA), which has shown the antimicrobial activity forms the 152-182 fragment of human lactoferrin (HLf). As the octapeptide FSASCVPG forms the 2-9 fragment of kaliocin-1, in the present study, its conformation in dimethyl sulfoxide-d6 (DMSO-d6) has been determined using two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy as well as restrained molecular dynamics. Sequence specific assignments of all the 1H resonances have been carried out using 2D correlation experiments (2D DQF-COSY, TOCSY and ROESY). In dimethyl sulfoxide-d6 at 25 degrees C, the octapeptide adopts a predominantly extended backbone conformation. The calculated structure resembles closely with the reported structure of the corresponding fragment of HLf. The peptide also has sequence and structural similarity with the corresponding fragments of lactoferrins from other organisms.

The tetrapeptide Pro-Glu-Leu-Leu forms the 94-97 fragment of C globin in sea cucumber. 2% Butanediol dimethacrylate-cross linked polystyrene (2% BDDMA-PS), which had been optimized, was used for the synthesis of the tetrapeptide Pro-Glu-Leu-Leu. The peptide was synthesized by using Boc-amino acid strategy. The peptide purity was checked by RP-HPLC and the peptide was characterized by (1)H NMR spectroscopy and amino acid analysis. Conformation of the peptide was studied by 1D- and 2D- homonuclear (1)H NMR, in DMSO-d6 at 300K. The conformation of the synthetic tetrapeptide (extended backbone conformation) is not in agreement with that in C globin.

Goat milk lactoperoxidase was purified using CM-Sephadex-C-50 ion exchange chromatography and Sephadex G-100 gel filtration. The purified protein was found to have high antifungal and antibacterial activity in a thiocyanate-H2O2 medium. The results are relevant enough to suggest the exploitation of LP-thiocyanate-H2O2 system as an effective agent against many of the disease causing organisms in plants and animals.

The goat milk lactoperoxidase was purified using CM sephadex C-50 and sephadex G 100. The purity of protein was confirmed by SDS-PAGE. The purified protein was found to have antibacterial action against most of the disease causing bacteria.

The three-dimensional structures of various forms of lactoferrin, determined by high resolution crystallographic studies, have been compared in order to determine the relationship between structure and biological function. These comparisons include human apo and diferric lactoferrins, metal and anion substituted lactoferrins, the N-terminal half molecule of human lactoferrin, and bovine diferric lactoferrin. The structures themselves define the nature and location of the iron binding sites and allow anti-bacterial and putative receptor-binding regions to be mapped on to the molecular surface. The structural comparisons show that small internal adjustments can allow the accommodation of different metals and anions without altering the overall molecular structure, whereas large-scale conformational changes are associated with metal binding and release, and smaller, but significant, movements accompany species variations. The results also focus on differences in flexibility between the two lobes, and on the importance of interactions in the inter-lobe region in modulating iron release from the N-lobe and in possibly enabling binding at one site to be signalled to the other.