The prevalence of neovascularity in rotator cuff tendinopathy: comparing conventional Doppler with superb microvascular imaging.

AIMS: To determine the prevalence of neovascularity in the supraspinatus tendon of patients presenting with clinically painful unilateral rotator cuff tendinopathy (RCT) using conventional colour Doppler ultrasound (CDU), power Doppler ultrasound (PDU), and superb microvascular imaging (SMI). The association between Doppler findings and clinical scores was also assessed. METHODS: The bilateral supraspinatus of consecutive patients presented with unilateral RCT clinically were evaluated with grey-scale ultrasound (tendon thickening, heterogeneous echotexture, and hypoechogenicity), CDU, PDU, and SMI. The prevalence of neovascularity and grey-scale changes on duplex imaging techniques were analysed. The relationship between neovascularity on CDU, PDU, SMI, and pain/disability as determined using a visual analogue scale (VAS) and the Oxford Shoulder Score (OSS) were assessed. RESULTS: Fifty-nine patients (mean age 53 years, 39 women) were recruited. Of the symptomatic supraspinatus tendons, 42.4% (25/59) demonstrated neovascularity on SMI, compared to 6.8% (4/59) on PDU and 5.1% (3/59) on CDU. Of the asymptomatic supraspinatus tendons, 5.1% (3/59) depicted neovascularity on SMI but not on conventional Doppler techniques. SMI showed a significant correlation with the VAS (r2 = 0.560, p<0.001) and OSS (r2 = 0.62, p<0.001). PDU weakly correlated with the VAS and OSS (r2 = 0.312, p=0.016; r2 = 0.260, p=0.047, respectively) while CDU did not show a significant relationship. CONCLUSION: SMI is superior in demonstrating neovascularity and shows better correlation with pain and functional deficit compared to conventional Doppler in patients with painful RCT. SMI also showed vascularity to a lesser degree in the asymptomatic tendon. Further large-scale studies are needed to prove the diagnostic value of SMI in the assessment of RCT.

Enhanced resonances by waveguide wrapping of a bulbed microring resonator.

A high Q-factor novel bulb design, to the best of our knowledge, inspired by the well-studied pulley microring resonator (MRR), is demonstrated and compared with circle and racetrack MRRs at wavelengths of 1.45-1.65 µm. A wrapping of 60% around the ring showed a three times Q-factor increment (Q=828) compared to a standard circle MRR (Q=206) at a telecom wavelength of 1.55 µm with a fixed ring radius of 2 µm, while further increasing the wrapping at 80% shows a decrease in its Q-factor (Q=454). A comparison of the wavelength dependence of the bulb design with other typical MRR designs suggests that bulb designs are highly sensitive to wavelength and have very high Q-factors at lower wavelengths.

The molecular mechanisms of probiotic strains in improving ageing bone and muscle of d-galactose-induced ageing rats.

AIM: The aim of this study was to evaluate the molecular mechanisms of Lactobacillus strains in improving ageing of the musculoskeletal system. METHODS AND RESULTS: The anti-ageing mechanism of three probiotics strains Lactobacillus fermentum DR9, Lactobacillus paracasei OFS 0291 and L. helveticus OFS 1515 were evaluated on gastrocnemius muscle and tibia of d-galactose-induced ageing rats. Upon senescence induction, aged rats demonstrated reduced antioxidative genes CAT and SOD expression in both bone and muscle compared to the young rats (P < 0·05). Strain L. fermentum DR9 demonstrated improved expression of SOD in bone and muscle compared to the aged rats (P < 0·05). In the evaluation of myogenesis-related genes, L. paracasei OFS 0291 and L. fermentum DR9 increased the mRNA expression of IGF-1; L. helveticus OFS 1515 and L. fermentum DR9 reduced the expression of MyoD, in contrast to the aged controls (P < 0·05). Protective effects of L. fermentum DR9 on ageing muscle were believed to be contributed by increased AMPK-α2 expression. Among the osteoclastogenesis genes studied, TNF-α expression was highly elevated in tibia of aged rats, while all three probiotics strains ameliorated the expression. Lactobacillus fermentum DR9 also reduced the expression of IL-6 and TRAP in tibia when compared to the aged rats (P < 0·05). All probiotics treatment resulted in declined proinflammatory cytokines IL-1ß in muscle and bone. CONCLUSIONS: Lactobacillus fermentum DR9 appeared to be the strongest strain in modulation of musculoskeletal health during ageing. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrated the protective effects of the bacteria on muscle and bone through antioxidative and anti-inflammatory actions. Therefore, L. fermentum DR9 may serve as a promising targeted anti-ageing therapy.

Light absorption by interacting atomic gas in quantum optical regime.

Quantum optical theory of absorption properties of interacting atoms is developed. The concept of local absorptance is introduced as a derivative of the logarithm of intensity with respect to the distance in the vicinity of a given spatial point and a moment of time. The intensity is represented by the quantum and statistically averaged normal product of creation and annihilation operators of the electromagnetic field. The development of an analytical method of the estimation for the kinetic and optical parameters for the system is proposed here. The calculation method of the absorption coefficient includes thermal atomic motion, Doppler effect, and the short-range interaction between atoms. The absorption coefficient explicitly takes into account the quantum nature of the optical field. The ability of the system to absorb or emit quanta is quantitatively expressed through the special form of interaction integrals. The specific form of integrals results from the structure of the quantum brackets. The interplay between the collective (virtual photon exchange) and binary (optically induced inter-particle bonding) processes determines the system behavior. The spectral profile of the local absorption coefficient for different atomic densities and time intervals is simulated for realistic parameters.

Phase I study of emactuzumab single agent or in combination with paclitaxel in patients with advanced/metastatic solid tumors reveals depletion of immunosuppressive M2-like macrophages.

BACKGROUND: Emactuzumab is a monoclonal antibody against the colony-stimulating factor-1 receptor and targets tumor-associated macrophages (TAMs). This study assessed the safety, clinical activity, pharmacokinetics (PK) and pharmacodynamics (PD) of emactuzumab, as monotherapy and in combination with paclitaxel, in patients with advanced solid tumors. PATIENTS AND METHODS: This open-label, phase Ia/b study comprised two parts (dose escalation and dose expansion), each containing two arms (emactuzumab, every 2 or 3 weeks, as monotherapy or in combination with paclitaxel 80 mg/m2 weekly). The dose-escalation part explored the maximum tolerated dose and optimal biological dose (OBD). The dose-expansion part extended the safety assessment and investigated the objective response rate. A PK/PD analysis of serial blood, skin and tumor biopsies was used to explore proof of mechanism and confirm the OBD. RESULTS: No maximum tolerated dose was reached in either study arm, and the safety profile of emactuzumab alone and in combination does not appear to preclude its use. No patients receiving emactuzumab monotherapy showed an objective response; the objective response rate for emactuzumab in combination with paclitaxel was 7% across all doses. Skin macrophages rather than peripheral blood monocytes or circulating colony-stimulating factor-1 were identified as an optimal surrogate PD marker to select the OBD. Emactuzumab treatment alone and in combination with paclitaxel resulted in a plateau of immunosuppressive TAM reduction at the OBD of 1000 mg administered every 2 weeks. CONCLUSIONS: Emactuzumab showed specific reduction of immunosuppressive TAMs at the OBD in both treatment arms but did not result in clinically relevant antitumor activity alone or in combination with paclitaxel. (ClinicalTrials.gov Identifier: NCT01494688).

The barriers to linkage and retention in care for women living with HIV in an high income setting where they comprise a minority group.

Women comprise a minority population of individuals living with HIV in Australia, and are often poorly represented in research and clinical trials so their needs remain largely unknown. Data suggests that they are diagnosed later than men and start antiretroviral therapy at a lower CD4 cell count. This raises the question whether there are sex specific barriers to linkage and retention in care. This study analyzed 484 surveys received from clinicians collecting demographic, virological, and reproductive health data along with perceived barriers to linkage and retention in care. Most women (67%) were estimated to have been linked into care within 28 days of diagnosis. For women who were not linked into care for more than 28 days, the most commonly reason cited was fear of disclosure to others, followed by fear of disclosure to their partner. The main reasons given for non-retention in care were related to transport, carer responsibilities, financial pressure, health beliefs and concern about stigma or disclosure.

Determinants of antibiotic prescribing for upper respiratory tract infections in an emergency department with good primary care access: a qualitative analysis.

Upper respiratory tract infections (URTIs) account for substantial attendances at emergency departments (EDs). There is a need to elucidate determinants of antibiotic prescribing in time-strapped EDs - popular choices for primary care despite highly accessible primary care clinics. Semi-structured in-depth interviews were conducted with purposively sampled physicians (n = 9) in an adult ED in Singapore. All interviews were analysed using thematic analysis and further interpreted using the Social Ecological Model to explain prescribing determinants. Themes included: (1) reliance on clinical knowledge and judgement, (2) patient-related factors, (3) patient-physician relationship factors, (4) perceived practice norms, (5) policies and treatment guidelines and (6) patient education and awareness. The physicians relied strongly on their clinical knowledge and judgement in managing URTI cases and seldom interfered with their peers' clinical decisions. Despite departmental norms of not prescribing antibiotics for URTIs, physicians would prescribe antibiotics when faced with uncertainty in patients' diagnoses, treating immunocompromised or older patients with comorbidities, and for patients demanding antibiotics, especially under time constraints. Participants had a preference for antibiotic prescribing guidelines based on local epidemiology, but viewed hospital policies on prescribing as a hindrance to clinical judgement. Participants highlighted the need for more public education and awareness on the appropriate use of antibiotics and management of URTIs. Organisational practice norms strongly influenced antibiotic prescribing decisions by physicians, who can be swayed by time pressures and patient demands. Clinical decision support tools, hospital guidelines and patient education targeting at individual, interpersonal and community levels could reduce unnecessary antibiotic use.

Probing infinity in bounded two-dimensional electrostatic systems.

The total electrostatic energy of systems of identical particles of equal charge is studied in configurations bounded in space, but divergent in the number of charges. This approach shall guide us to unveil a non-linear, functional form specifying the divergent nature of system energy. We consider fractals to be physical entities, with charges located in their vertices or nodes. This description is interesting since features, such as the corresponding fractal dimension, can characterize the total energy EN. Finally, at local length scales, we describe how energy diverges at charge accumulation points in the fractal, that is, almost everywhere by definition.

Prevalence of morphological and mechanical stiffness alterations of mid Achilles tendons in asymptomatic marathon runners before and after a competition.

OBJECTIVE: To determine the prevalence of morphological and mechanical stiffness alterations at the mid Achilles tendon in asymptomatic marathon runners before and after a competition. To assess the relationship between pre-existing Achilles tendon alterations and pain after running. MATERIALS AND METHODS: All marathon runners from a local running club who were participating in the Melbourne Marathon 2013 (full marathon category) were invited for conventional ultrasound and sonoelastography 1 week leading up to the marathon and again within 3 days post-marathon. Another group of active, healthy individuals not involved in running activities were recruited as controls. Intratendinous morphological (tendon thickness, hypoechogenicities), Doppler as well as stiffness properties of the Achilles were recorded. Achilles tendon pain was evaluated using the visual analogue scale (VAS) and Victorian Institute of Sports Assessment-Achilles (VISA-A). RESULTS: Twenty-one asymptomatic runners (42 Achilles tendons) and 20 healthy controls (40 Achilles tendons) were examined. On the pre-marathon evaluation, runners showed significantly more morphological changes on B-mode ultrasound compared to the controls (p < 0.001). Marathon running induced a significant reduction in tendon stiffness (p = 0.049) and an increase in Doppler signals (p = 0.036). Four runners (4/21, 19%) reported Achilles tendon pain after the race [VAS 4.0 (±1.9), VISA 74.2 (±10.1)]. Reduced tendon stiffness at baseline was associated with post-marathon Achilles tendon pain (p = 0.016). CONCLUSION: Marathon runners demonstrate a higher prevalence of morphological alterations compared to non-runners. Marathon running caused a significant change in Achilles tendon stiffness and Doppler signals. Pre-existing soft Achilles tendon properties on sonoelastography may be a predisposing risk for development of symptoms post-running.

Colesevelam for Type 2 diabetes mellitus: an abridged Cochrane review.

AIM: Colesevelam, a second-generation bile acid sequestrant, may be beneficial in controlling both glycaemia and lipids simultaneously. Our goal was to evaluate the systemic effects of colesevelam on Type 2 diabetes mellitus. METHOD: The original Cochrane review was conducted using the methodology for the systematic review of interventions of the Cochrane Collaboration in RevMan 5.2. We comprehensively searched the literature in several databases up to January 2012. Two reviewing authors independently selected and extracted the data, and then evaluated the quality of the randomized controlled trials that met the inclusion criteria. RESULTS: Six randomized controlled trials were selected, which ranged from 8 to 26 weeks in duration. A total of 1450 participants were divided into two groups: those treated with colesevelam and no other anti-diabetic drug treatments/placebo, or with colesevelam added on to anti-diabetic drug treatments. The colesevelam added on to anti-diabetic agents demonstrated a statistically significant reduction in the fasting blood glucose (mean difference of -0.82 mmol/l, 95% CI -1.2 to -0.44), HbA1c (mean difference -0.5%, 95% CI -0.6 to -0.4) and LDL cholesterol (mean difference -0.34 mmol/l, 95% CI -0.44 to -0.23). There were no reported data on weight. Non-severe hypoglycaemic episodes were infrequently observed. CONCLUSION: The limited number of studies concerning the treatment with colesevelam added to anti-diabetic agents showed significant effects on glycaemic control; however, more research on the reduction of cardiovascular risks is required. Furthermore, long-term data on the health-related quality of life and all-cause mortality also need to be investigated.

"Soft, hard, or just right?" Applications and limitations of axial-strain sonoelastography and shear-wave elastography in the assessment of tendon injuries.

Injury to a tendon leads to alterations in the mechanical properties of the tendon. Axial-strain sonoelastography and shear-wave elastography are relatively new, real-time imaging techniques that evaluate the mechanical properties of tendons in addition to the existing morphological and vascular information that is obtained with traditional imaging tools. Axial-strain sonoelastography displays the subjective distribution of strain data on an elastogram caused by tissue compression, whereas shear-wave elastography provides a more objective, quantitative measure of the intrinsic tissue elasticity using the acoustic push-pulse. Recent studies suggest that axial-strain sonoelastography is able to distinguish between asymptomatic and diseased tendons, and is potentially more sensitive than conventional ultrasound in detecting early tendinopathy. Shear-wave elastography seems to be a feasible tool for depicting elasticity and functional recovery of tendons after surgical management. While initial results have been promising, axial-strain sonoelastography and shear-wave elastography have not yet found routine use in wider clinical practice. Possible barriers to the dissemination of axial-strain sonoelastography technique include operator dependency, technical limitations such as artefacts and lack of reproducibility and quantification of sonoelastography data. Shear-wave elastography may improve the reproducibility of elastography data, although there is only one published study on the topic to date. Large-scale longitudinal studies are needed to further elucidate the clinical relevance and potential applications of axial-strain sonoelastography and shear-wave elastography in diagnosing, predicting, and monitoring the progress of tendon healing before they can be widely adopted into routine clinical practice.

ECFS standards of care on CFTR-related disorders: Identification and care of the disorders.

This is the third paper in the series providing updated information and recommendations for people with cystic fibrosis transmembrane conductance regulator (CFTR)-related disorder (CFTR-RD). This paper covers the individual disorders, including the established conditions - congenital absence of the vas deferens (CAVD), diffuse bronchiectasis and chronic or acute recurrent pancreatitis - and also other conditions which might be considered a CFTR-RD, including allergic bronchopulmonary aspergillosis, chronic rhinosinusitis, primary sclerosing cholangitis and aquagenic wrinkling. The CFTR functional and genetic evidence in support of the condition being a CFTR-RD are discussed and guidance for reaching the diagnosis, including alternative conditions to consider and management recommendations, is provided. Gaps in our knowledge, particularly of the emerging conditions, and future areas of research, including the role of CFTR modulators, are highlighted.

Speed of facial affect intensity recognition as an endophenotype of first-episode psychosis and associated limbic-cortical grey matter systems.

BACKGROUND: Psychotic disorders are highly heritable such that the unaffected relatives of patients may manifest characteristics, or endophenotypes, that are more closely related to risk genes than the overt clinical condition. Facial affect processing is dependent on a distributed cortico-limbic network that is disrupted in psychosis. This study assessed facial affect processing and related brain structure as a candidate endophenotype of first-episode psychosis (FEP). METHOD: Three samples comprising 30 FEP patients, 30 of their first-degree relatives and 31 unrelated healthy controls underwent assessment of facial affect processing and structural magnetic resonance imaging (sMRI) data. Multivariate analysis (partial least squares, PLS) was used to identify a grey matter (GM) system in which anatomical variation was associated with variation in facial affect processing speed. RESULTS: The groups did not differ in their accuracy of facial affect intensity rating but differed significantly in speed of response, with controls responding faster than relatives, who responded faster than patients. Within the control group, variation in speed of affect processing was significantly associated with variation of GM density in amygdala, lateral temporal cortex, frontal cortex and cerebellum. However, this association between cortico-limbic GM density and speed of facial affect processing was absent in patients and their relatives. CONCLUSIONS: Speed of facial affect processing presents as a candidate endophenotype of FEP. The normal association between speed of facial affect processing and cortico-limbic GM variation was disrupted in FEP patients and their relatives.

Unified master equation for molecules in phonon and radiation baths.

We have developed a unified quantum optical master equation that includes the dissipative mechanisms of an impurity molecule in crystals. Our theory applies generally to polyatomic molecules where several vibrational modes give rise to intramolecular vibrational redistributions. The usual assumption on identical shapes of the nuclear potentials in ground and excited electronic states and the rotating wave approximation have been relaxed, i.e. the vibrational coordinates are different in the ground and excited states, with counter-rotating terms included for generality. Linear vibrational coupling to the lattice phonons accounts for dissipations via non-radiative transitions. The interaction of a molecule with photons includes Herzberg-Teller coupling as the first order non-Condon interaction where the transition dipole matrix elements depend linearly on vibrational coordinates. We obtain new cross terms as the result of mixing the terms from the zeroth-order (Condon) and first-order (non-Condon) approximations. The corresponding Lamb shifts for all Liouvilleans are derived explicitly including the contributions of counter-rotating terms. The computed absorption and emission spectra for carbon monoxide is in good agreement with experimental data. We use our unified model to obtain the spectra for nitrogen dioxide, demonstrating the capability of our theory to incorporate all typical dissipative relaxation and decoherence mechanisms for polyatomic molecules. The molecular quantum master equation is a promising theory for studying molecular quantum memory.

The Peripheral Neuropathy Prevalence and Characteristics Are Comparable in People with Obesity and Long-Duration Type 1 Diabetes.

INTRODUCTION: Peripheral neuropathy is reported in obesity even in the absence of hyperglycaemia. OBJECTIVE: To compare the prevalence and characterise the phenotype of peripheral neuropathy in people living with obesity (OB) and long-duration type 1 diabetes (T1D). PATIENTS AND METHODS: We performed a prospective cross-sectional study of 130 participants including healthy volunteers (HV) (n = 28), people with T1D (n = 51), and OB (BMI 30-50 kg/m2) (n = 51). Participants underwent assessment of neuropathic symptoms (Neuropathy Symptom Profile, NSP), neurological deficits (Neuropathy Disability Score, NDS), vibration perception threshold (VPT) and evaluation of sural nerve conduction velocity and amplitude. RESULTS: Peripheral neuropathy was present in 43.1% of people with T1D (age 49.9 ± 12.9 years; duration of diabetes 23.4 ± 13.5 years) and 33.3% of OB (age 48.2 ± 10.8 years). VPT for high risk of neuropathic foot ulceration (VPT ≥ 25 V) was present in 31.4% of T1D and 19.6% of OB. Participants living with OB were heavier (BMI 42.9 ± 3.5 kg/m2) and had greater centripetal adiposity with an increased body fat percentage (FM%) (P < 0.001) and waist circumference (WC) (P < 0.001) compared to T1D. The OB group had a higher NDS (P < 0.001), VAS for pain (P < 0.001), NSP (P < 0.001), VPT (P < 0.001) and reduced sural nerve conduction velocity (P < 0.001) and amplitude (P < 0.001) compared to HV, but these parameters were comparable in T1D. VPT was positively associated with increased WC (P = 0.011), FM% (P = 0.001) and HbA1c (P < 0.001) after adjusting for age (R2 = 0.547). Subgroup analysis of respiratory quotient (RQ) measured in the OB group did not correlate with VPT (P = 0.788), nerve conduction velocity (P = 0.743) or amplitude (P = 0.677). CONCLUSION: The characteristics of peripheral neuropathy were comparable between normoglycaemic people living with obesity and people with long-duration T1D, suggesting that metabolic factors linked to obesity play a pivotal role in the development of peripheral neuropathy. Further studies are needed to investigate the mechanistic link between visceral adiposity and neuropathy.

Genetic diversity of secreted protein with an altered thrombospondin repeat (SPATR) of Plasmodium knowlesi clinical isolates from Malaysia.

The Plasmodium knowlesi secreted protein with an altered thrombospondin repeat (PkSPATR) is an important protein that helps in the parasite's invasion into the host cell. This protein has been regarded as one of the potential vaccine candidates against P. knowlesi infection. This study investigates the genetic diversity and natural selection of PkSPATR gene of P. knowlesi clinical isolates from Malaysia. PCR amplification of the full length PkSPATR gene was performed on 60 blood samples of infected P. knowlesi patients from Peninsular Malaysia and Malaysian Borneo. The amplified PCR products were cloned and sequenced. Sequence analysis of PkSPATR from Malaysia showed higher nucleotide diversity (CDS p: 0.01462) than previously reported Plasmodium vivax PvSPATR (p = 0.0003). PkSPATR from Peninsular Malaysia was observed to have slightly higher diversity (CDS p: 0.01307) than those from Malaysian Borneo (CDS p: 0.01212). Natural selection analysis on PkSPATR indicated significant purifying selection. Multiple amino acid sequence alignment revealed 69 polymorphic sites. The phylogenetic tree and haplotype network did not show any distinct clustering of PkSPATR. The low genetic diversity level, natural selection and absence of clustering implied functional constrains of the PkSPATR protein.

Cross-sectional study of older adults with type 2 diabetes mellitus in two rural public primary healthcare facilities in Malaysia.

Knowledge of the characteristics of older adults with type 2 diabetes mellitus (T2DM) is indispensible for improvement of their care. A cross-sectional study in two rural public primary healthcare centres in Malaysia identified 170 actively engaged older patients with T2DM, with suboptimal glycaemic control and frequent hypoglycaemia. The prevalence of multiple co-morbidities, complications of T2DM, high cardiovascular risk, neurological, musculoskeletal and visual deficits suggested high risk of disabilities and dependency but not yet disabled. This short window for interventions presents as an opportunity for development of a more comprehensive approach extending beyond glycaemia control to risk management, preventing functional loss and continuity of social participation.

High incidence of Plasmodium knowlesi malaria compared to other human malaria species in several hospitals in Malaysia.

Through the regional control programme, Malaysia has been successfully reducing the incidence of Plasmodium falciparum and Plasmodium vivax infections. However, the incidence of zoonotic malaria Plasmodium knowlesi infection is increasing and now has been the major cause of malaria in Malaysia especially Malaysian Borneo. The emergence of knowlesi infection has threatened the malaria elimination programme which the government aims to reduce the overall malaria infections by 2020. Unlike other benign human Plasmodium spp., P. knowlesi can cause fatal infections. The aim of this study was to determine the incidence and distribution of five human malaria parasites including P. knowlesi in Peninsular Malaysia and Malaysian Borneo. A total of 112 blood samples were collected from seven states and district hospitals in Peninsular Malaysia and Malaysian Borneo from year 2015 to 2016. The samples were examined by microscopy and further confirmed by nested PCR assay targeting 18S rRNA gene of Plasmodium spp. Following the nested PCR assays, a total of 54 (48.2%) samples were positive for P. knowlesi infections, 12 (10.7%) cases were positive for P. vivax infections, followed by 7 (6.3%) cases of P. falciparum and 4 (3.5%) cases of P. malariae. There were 3 cases (2.7%) of mixed infections (P. knowlesi/P. vivax). However, no cases were identified as P. ovale. A total of 32 (28.6%) cases were found as negative infections. LoopMediated Isothermal Amplification Assay (LAMP) was performed to confirm inconclusive results produced by microscopy and nested PCR. P. knowlesi showed the highest prevalence in Sarawak (n= 30), Sabah (n=13), Pulau Pinang (n=5) and Pahang (n=6). PCR and LAMP was not able to detect a large number of microscopy positive samples due to DNA degradation during storage and shipping. Among all the states involved in this study, the highest prevalence of P. knowlesi infection was found in Sabah and Sarawak.

Endotoxin-neutralizing properties of the 25 kD N-terminal fragment and a newly isolated 30 kD C-terminal fragment of the 55-60 kD bactericidal/permeability-increasing protein of human neutrophils.

The bactericidal/permeability-increasing protein (BPI) of polymorphonuclear leukocytes (PMN) is a potent cytotoxin, specific for Gram-negative bacteria, that also inhibits endotoxin activity by neutralizing isolated bacterial lipopolysaccharides (LPS). We have previously shown that an isolated 25 kD N-terminal fragment of human BPI carries all the antibacterial activities of the parent 55-60 kD molecule. In this study we have compared the LPS-neutralizing activities of human holo-BPI, the N-terminal fragment and a 30 kD C-terminal fragment that we have now isolated. We show that the N-terminal fragment also has LPS-neutralizing activity as detected by inhibition (up to 95%) of (a) activation by LPS of procoagulant proteases in Limulus amebocyte lysates, (b) LPS "priming" of PMN, and (c) LPS-mediated production of tumor necrosis factor in whole human blood. Holo-BPI and the 25 kD fragment have similar neutralizing potency (in nanomolar range) in all assays toward "smooth" LPS from Escherichia coli O111:B4 and O55:B5 (possessing long chain polysaccharide or O-antigen), and "deep rough" LPS from Salmonella minnesota Re595 mutant (possessing no O-antigen). The C-terminal fragment of BPI is devoid of antibacterial activity when tested against BPI-sensitive E. coli J5, but does have endotoxin-neutralizing activity. This activity is weak relative to holo-BPI and the 25 kD N-terminal fragment in the Limulus and PMN-priming assay, but is comparable for inhibition of TNF production in whole blood. We conclude that the principal determinants for LPS recognition and neutralization, like those for antibacterial action, reside in the N-terminal half of the BPI molecule, but that sites within the C-terminal half can also contribute to BPI-LPS interaction once LPS is detached from the bacterial envelope.