Predictors of elevated C-reactive protein among pre-treatment, newly diagnosed breast cancer patients: A cross-sectional study.

BACKGROUND & AIMS: Accumulating evidence showed that inflammation contributes markedly to cancer progression, with C-reactive protein (CRP) being one of the lengthily studied inflammation marker. For breast cancer (BCa), pre-treatment elevated CRP upon diagnosis was linked with increased mortality. This study aimed to identify factors predictive of elevated CRP in pre-treatment BCa population that can serve as potential therapeutic targets to reduce inflammation. METHODS: This is a cross-sectional study using multiple logistic regression to identify predictors of elevated CRP among pre-treatment, newly diagnosed BCa patients. Studied variables were socio-demographic and medical characteristics, anthropometric measurements [body weight, Body Mass Index, body fat percentage, fat mass/fat free mass ratio, muscle mass, visceral fat], biochemical parameters [albumin, hemoglobin, white blood cell (WBC), neutrophil, lymphocyte], energy-adjusted Dietary Inflammatory Index, handgrip strength (HGS), scored Patient Generated-Subjective Global Assessment, physical activity level and perceived stress scale (PSS). RESULTS: A total of 105 participants took part in this study. Significant predictors of elevated CRP were body fat percentage (OR 1.222; 95 % CI 1.099-1.358; p < 0.001), PSS (OR 1.120; 95 % CI 1.026-1.223; p = 0.011), low vs normal HGS (OR 41.928; 95 % CI 2.155-815.728; p = 0.014), albumin (OR 0.779; 95 % CI 0.632-0.960; p = 0.019), and WBC (OR 1.418; 95% CI 1.024-1.963; p = 0.036). CONCLUSION: Overall, predictors of elevated CRP in pre-treatment, newly diagnosed BCa population were body fat percentage, PSS, HGS category, albumin and WBC.

Sequence analysis of LipL41 and LipL21: Prospective Outer Membrane Proteins (OMPs) in early diagnosing leptospirosis.

Leptospirosis is a zoonotic disease mostly occurring in tropical climate countries. The etiology of the disease is due to microbes from the genus Leptospira. Higher number of cases reported worldwide indicated the disease is not easily eradicated. Leptospirosis shares the most common febrile symptoms such as dengue, Zika and yellow fever thus making it difficult to differentiate the disease at an early stage. The widely used current detection via PCR, uses the bacterial outer membrane protein (OMP) as their target region. However, the heterogeneity and variation of the genome cause false negative results. Lipoprotein LipL41 is the third most abundant outer membrane lipoprotein among pathogenic species and it is surface exposed and expressed during infection thus making it a suitable candidate in identifying pathogenic Leptospira. LipL21 on the other hand is a potential candidate in identifying the intermediate species. The study aimed in designing suitable PCR primers in identifying pathogenic and intermediate species of Leptospira through bioinformatics analysis on the bacterial OMPs. LipL41 and LipL21 were chosen as the suitable target sequence to be used as PCR primers in detecting the pathogenic and intermediate species, respectively. The designed primers indicated positive feedback upon tested with their respective bacterial DNA extract. These lipoproteins may serve as potential PCR primers to be used with clinical samples in diagnosing leptospirosis.•The etiology of the illness is due to bacteria from the genus Leptospira.•PCR utilizes the bacterial external membrane protein (OMP) thus the heterogeneity and variety of the genome cause bogus adverse outcomes.•The suitable candidates are LipL41, the third most abundant outer membrane lipoprotein, whereas LipL21 is a potential candidate in identifying the intermediate species.

Development of nanoemulsion for efficient brain parenteral delivery of cefuroxime: designs, characterizations, and pharmacokinetics.

BACKGROUND AND AIM: Drugs that are effective against diseases in the central nervous system and reach the brain via blood must pass through the blood-brain barrier (BBB), a unique interface that protects against potential harmful molecules. This presents a major challenge in neuro-drug delivery. This study attempts to fabricate the cefuroxime-loaded nanoemulsion (CLN) to increase drug penetration into the brain when parenterally administered. METHODS: The nanoemulsions were formulated using a high-pressure homogenization technique and were characterized for their physicochemical properties. RESULTS: The characterizations revealed a particle size of 100.32±0.75 nm, polydispersity index of 0.18±0.01, zeta potential of -46.9±1.39 mV, viscosity of 1.24±0.34 cps, and osmolality of 285.33±0.58 mOsm/kg, indicating that the nanoemulsion has compatibility for parenteral application. CLN was physicochemically stable within 6 months of storage at 4°C, and the transmission electron microscopy revealed that the CLN droplets were almost spherical in shape. The in vitro release of CLN profile followed a sustained release pattern. The pharmacokinetic profile of CLN showed a significantly higher Cmax, area under the curve (AUC)0-t , prolonged half-life, and lower total plasma clearance, indicating that the systemic concentration of cefuroxime was higher in CLN-treated rats as compared to cefuroxime-free treated rats. A similar profile was obtained for the biodistribution of cefuroxime in the brain, in which CLN showed a significantly higher Cmax, AUC0-t , prolonged half-life, and lower clearance as compared to free cefuroxime solution. CONCLUSION: Overall, CLN showed excellent physicochemical properties, fulfilled the requirements for parenteral administration, and presented improved in vivo pharmacokinetic profile, which reflected its practical approach to enhance cefuroxime delivery to the brain.

Characterization of resistance to selected antibiotics and Panton-Valentine leukocidin-positive Staphylococcus aureus in a healthy student population at a Malaysian University.

INTRODUCTION: This study aims to assess the antimicrobial susceptibility profiles of Staphylococcus aureus strains isolated from university students and to determine the prevalence of constitutive and inducible clindamycin resistance, the latter being able to cause therapeutic failure due to false in vitro clindamycin susceptibility. METHODS: S. aureus strains were isolated from the nasal swabs of 200 health sciences students of a Malaysian university. Twelve classes of antibiotics were used to evaluate the antimicrobial susceptibility profiles with the macrolide-lincosamide-streptogramin B (MLSB) phenotype for inducible clindamycin resistance determined by the double-diffusion test (D-test). Carriage of resistance and virulence genes was performed by PCR on S. aureus isolates that were methicillin resistant, erythromycin resistant and/or positive for the leukocidin gene, pvl (n=15). RESULTS: Forty-nine isolates were viable and identified as S. aureus with four of the isolates characterized as methicillin-resistant S. aureus (MRSA; 2.0%). All isolates were susceptible to the antibiotics tested except for penicillin (resistance rate of 49%), erythromycin (16%), oxacillin (8%), cefoxitin (8%) and clindamycin (4%). Of the eight erythromycin-resistant isolates, iMLSB was identified in five isolates (three of which were also MRSA). The majority of the erythromycin-resistant isolates harbored the msrA gene (four iMLSB) with the remaining iMLSB isolate harboring the ermC gene. CONCLUSION: The presence of MRSA isolates which are also iMLSB in healthy individuals suggests that nasal carriage may play a role as a potential reservoir for the transmission of these pathogens.

Genetic diversity of hepatitis B co-infection with hepatitis C, D and E viruses among Malaysian chronic hepatitis B patients.

BACKGROUND: Hepatitis B virus co-infection with other strains of viral hepatitis is associated with increased risk of liver cirrhosis and hepatic decompensation. OBJECTIVES: This is a prevalence study that assessed the genetic diversity of chronic hepatitis B patients and coinfection. METHODS: Chronic hepatitis B patients enrolled in this study were tested for antibodies of other hepatitis viruses using ELISA kits. Patient clinical profiles were collected and partial genes of HBV, HCV, and HEV were amplified, sequenced, and analyzed using phylogenetic analysis. The associations between variables were determined using the chi-squared test. RESULTS: Of the 82 patients recruited for this study, 53.7% were non-cirrhotic, 22.0% cirrhotic, 20.7% acute flare and 3.7% hepatocellular carcinoma. Majority (58%) of patients had a high level of ALT (≥34 U/L). Sequence analysis showed HBV (63.9%) belonged to genotype B, HEV belonged to genotype 4 while HCV belonged to genotype 3a and the genotypes were found to be significantly associated with the clinical stage of the patients (χ2=56.632; p<0.01). Similarly, Hepatitis B e antigen was also found to be significantly associated with the clinical stage of infection (χ2=51.952; p<0.01). CONCLUSION: This study revealed that genetic diversity was found to have a significant impact on the severity of infection.

Expression of Human Cytokine Genes Associated with Chronic Hepatitis B Disease Progression.

BACKGROUND: Hepatitis viruses are non-cytopathic viruses that lead to the infection and pathogenesis of liver diseases as a result of immunologically mediated events. OBJECTIVE: To investigate the expression of human inflammatory cytokines in chronic hepatitis B patients according to the severity of the infection. METHODS: We recruited a total of 120 patients, 40 of whom from cirrhotic, 40 non-cirrhotic, and 40 acute flare chronic hepatitis B and 40 healthy controls. For all groups total cellular RNA was extracted from whole blood samples, genomic DNA was eliminated, and cDNA was synthesized using the RT2 first strand kit, as instructed by the manufacturer. The real-time profiler PCR array was performed on a master cycler ep realplex and the data were analyzed using an online data analysis software. RESULTS: Non-cirrhotic chronic hepatitis B patients were found to significantly upregulate interleukin 10 receptors that regulate the balance between T helpers 1 and 2. On the other hand, patients with cirrhosis were found to have significant upregulated interleukin 3 gene expression. CONCLUSION: Our finding suggests that upregulation of anti-inflammatory and downregulation of pro-inflammatory cytokines may play a role in the progression of non-cirrhotic chronic hepatitis B patients to cirrhotic and acute flare. However, a multi-center study with a larger sample size is needed to confirm our findings.

Leptospirosis: Molecular trial path and immunopathogenesis correlated with dengue, malaria and mimetic hemorrhagic infections.

Immuno-pathogenesis of leptospirosis can be recounted well by following its trail path from entry to exit, while inducing disastrous damages in various tissues of the host. Dysregulated, inappropriate and excessive immune responses are unanimously blamed in fatal leptospirosis. The inherent abilities of the pathogen and inabilities of the host were debated targeting the severity of the disease. Hemorrhagic manifestation through various mechanisms leading to a fatal end is observed when this disease is unattended. The similar vascular destructions and hemorrhage manifestations are noted in infections with different microbes in endemic areas. The simultaneous infection in a host with more than one pathogen or parasite is referred as the coinfection. Notably, common endemic infections such as leptospirosis, dengue, chikungunya, and malaria, harbor favorable environments to flourish in similar climates, which is aggregated with stagnated water and aggravated with the poor personal and environmental hygiene of the inhabitants. These factors aid the spread of pathogens and parasites to humans and potential vectors, eventually leading to outbreaks of public health relevance. Malaria, dengue and chikungunya need mosquitoes as vectors, in contrast with leptospirosis, which directly invades human, although the environmental bacterial load is maintained through other mammals, such as rodents. The more complicating issue is that infections by different pathogens exhibiting similar symptoms but require different treatment management. The current review explores different pathogens expressing specific surface proteins and their ability to bind with array of host proteins with or without immune response to enter into the host tissues and their ability to evade the host immune responses to invade and their affinity to certain tissues leading to the common squeal of hemorrhage. Furthermore, at the host level, the increased susceptibility and inability of the host to arrest the pathogens' and parasites' spread in different tissues, various cytokines accumulated to eradicate the microorganisms and their cellular interactions, the antibody dependent defense and the susceptibility of individual organs bringing the manifestation of the diseases were explored. Lastly, we provided a discussion on the immune trail path of pathogenesis from entry to exit to narrate the similarities and dissimilarities among various hemorrhagic fevers mentioned above, in order to outline future possibilities of prevention, diagnosis, and treatment of coinfections, with special reference to endemic areas.

Prevalence and molecular characterisation of Schistosoma haematobium among primary school children in Kebbi State, Nigeria

Schistosomiasis is the major source of morbidity in Sub-Saharan Africa and Asia. It is estimated that 207 million people are infected, of which 97% are in Africa. The aim of this study was the determining of prevalence as well as the phylogeny of S. haematobium among school children in Argungu Emirate, Kebbi State Nigeria. A total of 325 urine samples was collected from school children between 7 to 14 years. S. heamatobium eggs was examined under dissecting microscope and DNA was extracted from urine sample and COX1 gene was amplified by nested PCR. The PCR products were purified, sequenced and analysed. This study showed a prevalence of 32.09%, with male pupils having the highest prevalence. S. haematobium infections in children who fetch water in the river have 24 times higher risk of being infected while those who bath in the river have 158 times higher risk of being infected. Our sequences were phylogenetically related to S. haematobium isolate U82266 from Kenya and consistence with the predominant species in Africa. This was the first S. haematobium and S. mansoni co-infection reported in Nigeria. S. haematobium infection is prevalent among school age and significantly associated with water contact.

Genotypic characterization of Malaysian human isolates of Streptococcus pneumoniae from carriage and clinical sources.

This study characterized carriage and clinical pneumococcal isolates for serotypes, penicillin susceptibility, virulence genes and restriction fragment length polymorphism (RFLP) pattern of penicillin binding protein (PBP) genes. DNA fingerprint of isolates was generated by BOX-PCR. Majority of serotypes were 23F followed by 19F, 19A and 6A. Twenty-four percent of isolates were penicillin non-susceptible (PNSP). All of the targeted virulence genes were detected in all isolates with the exception of pili; 20.6% (n=22) for PI-1 and 14.0% (n=15) for PI-2. Of the 13 isolates which carried both PI-1 and PI-2, 10 were of clinical origin. Digested pbp-DNA produced three PBP-RFLP profiles for pbp1a (A1 to A3), six profiles for pbp2b (B1 to B6) and seven for pbp2x (X1 to X7) mostly in PNSPs. Based on BOX-PCR analysis, the majority of isolates were genetically diverse with a small number of potentially related isolates carrying pili genes. No obvious genotypic association was observed pertaining to carriage and clinical origin of isolates.

Determination of phenotypes and pneumococcal surface protein A family types of Streptococcus pneumoniae from Malaysian healthy children.

BACKGROUND: There is limited information about pneumococcal carriage among healthy children in Malaysia. Therefore, this study was conducted to determine the prevalence rate, serotype distribution, susceptibility pattern, and pneumococcal surface protein A (PspA) family types of Streptococcus pneumoniae isolates in the nasal carriage of children 5 years old or younger in three day care centers in Kuala Lumpur, Malaysia. METHODS: Nasal swabs were collected from 195 healthy children, age 5 years or younger, from June to December 2010. S pneumoniae was identified by phenotypic and genotypic methods. The serotyping was performed using Pneumotest kit (Statens Serum Institut, Copenhagen, Denmark) and the susceptibility pattern was determined by using the E-test method (AB Biodisk, Solna, Sweden). PspA family typing was done using polymerase chain reaction. RESULTS: S pneumoniae was found in the nasal carriage of 35.4% of children (69 of 195) and penicillin resistance was found in 23.2% (16 of 69). Among the 69 isolates, multidrug-resistant S pneumoniae (MDRSP) was present in 20.3%. All 16 penicillin-resistant S pneumoniae (PRSP) isolates were resistant to erythromycin and 14 PRSPs (87.5%) were resistant to co-trimoxazole. The six most common serotypes were 6A, 23F, 19A, 6B, 19F, and 15C, which were found in 87% of all isolates. Of the 69 isolates, 24.6% belonged to PspA family 1, 71.0% to PspA family 2, and 4.3% to PspA family 3. CONCLUSION: Twenty-eight of the isolates (40.6%) belonged to serotypes included in the pneumococcal polysaccharide vaccines (PCV) 7 and 10, whereas 48 (69.5%) were included in PCV13. The high rate of PRSP and MDRSP supports the need for continuing surveillance of pneumococcal carriage. The major PspA families were 1 and 2 (95.7%), thus making them suitable candidates for future vaccines.

Modified silver staining in 2DE improves protein detection even at extremely low sample concentration.

The typical concentration of protein loaded varies from 0.13 to 1.40 µg/µL for a classical silver staining method in 2DE gel. Here, we present a simple modified classical silver staining method by modifying the silver impregnation and development reaction steps. This modified method detects the protein spots at extremely low loaded concentrations, ranging from 0.0048 to 0.0480 µg/µL. We recommend this modified silver staining as an excellent method for the limited biological samples used for silver-stained 2DE analysis. Altogether, the protocol takes close to two days from first dimension separation to second dimension separation, followed by silver staining, scanning, and analysis.