Neutrophil-lymphocyte and platelet-lymphocyte ratios in rheumatoid arthritis patients: Relation to disease activity

Aim of the work: To assess the neutrophil–lymphocyte ratio (NLR) and platelet–lymphocyte ratio (PLR) in rheumatoid arthritis (RA) patients and compare between active cases and those in remission. Patients and methods: The study included 50 RA patients and 20 matched control. Patients were enrolled into 2 equally divided groups; group A (active) with a disease activity score (DAS-28) ≥2.6 and group B (remission) <2.6. Laboratory investigations included the calculation of the NLR and PLR for all subjects. Results: The mean age of patients was 40.7?±?10.1?years and the mean of disease duration was 5.9?±?3.4?years. The DAS-28 was 3.9?±?0.9 in active patients and 2.1?±?0.3 in those in remission (p?=?.001). NLR was 2.8?±?2.1 in the patients and 2.1?±?0.59 in the control (p?=?.15). PLR was 1.7?±?0.9 in the patients and 1.27?±?0.46 in the control (p?=?.09). Active patients had an NLR of 3.27?±?2.81 and PLR of 1.8?±?1.2 while they were 2.3?±?0.84 and PLR 1.5?±?0.59 in patients in remission (p?=?.05 and p?=?.18 respectively). There was a significant difference regarding NLR and PLR between active patients and control (2.1?±?0.59 and 1.27?±?0.46; p?=?.03 and p?=?.04 respectively). In active patients, the NLR and PLR significantly correlated with the patients age (p?=?.02 and p?=?.006) and with the DAS-28 (p?=?.001 and p?=?.03 respectively). Conclusion: NLR and PLR are 2 emerging inflammatory biomarkers which could be used to evaluate disease activity in active RA patients. A larger scale longitudinal study is recommended to confirm the present results and further demonstrate the relation to medications received and disease outcome.     

Endothelin ETA receptor/lipid peroxides/COX-2/TGF-β1 signalling underlies aggravated nephrotoxicity caused by cyclosporine plus indomethacin in rats

Background and Purpose

Cyclosporine (CSA) and non-steroidal anti-inflammatory drugs (NSAIDs) are co-prescribed for some arthritic conditions. We tested the hypothesis that this combined regimen elicits exaggerated nephrotoxicity in rats via the up-regulation of endothelin (ET) receptor signalling.

Experimental Approach

The effects of a 10 day treatment with CSA (20 mg·kg⁻¹·day⁻¹), indomethacin (5 mg·kg⁻¹·day⁻¹) or their combination on renal biochemical, inflammatory, oxidative and structural profiles were assessed. The roles of ET_A receptor and COX-2 pathways in the interaction were evaluated.

Key Results

Oral treatment with CSA or indomethacin elevated serum urea and creatinine, caused renal tubular atrophy and interstitial fibrosis, increased renal TGF-β1, and reduced immunohistochemical expressions of ET_A receptors and COX-2. CSA, but not indomethacin, increased renal ET-1, the lipid peroxidation product malondialdehyde (MDA) and GSH activity. Compared with individual treatments, simultaneous CSA/indomethacin exposure caused: (i) greater elevations in serum creatinine and renal MDA; (ii) loss of the compensatory increase in GSH; (iii) renal infiltration of inflammatory cells and worsening of fibrotic and necrotic profiles; and (iv) increased renal ET-1 and decreased ET_A receptor and COX-2 expressions. Blockade of ET_A receptors by atrasentan ameliorated the biochemical, structural, inflammatory and oxidative abnormalities caused by the CSA/indomethacin regimen. Furthermore, atrasentan partly reversed the CSA/indomethacin-evoked reductions in the expression of ET_A receptor and COX-2 protein.

Conclusions and Implications

The exaggerated oxidative insult and associated dysregulation of the ET_A receptor/COX-2/TGF-β1 signalling might account for the aggravated nephrotoxicity caused by the CSA/indomethacin regimen. The potential renoprotective effect of ET_A receptor antagonism might be exploited therapeutically.Tables of Links

Bone-tissue engineering: complex tunable structural and biological responses to injury,drug delivery,and cell-based therapies

Bone loss and failure of proper bone healing continues to be a significant medical condition in need of solutions that can be implemented successfully both in human and veterinary medicine. This is particularly true when large segmental defects are present, the bone has failed to return to normal form or function, or the healing process is extremely prolonged. Given the inherent complexity of bone tissue – its unique structural, mechanical, and compositional properties, as well as its ability to support various cells – it is difficult to find ideal candidate materials that could be used as the foundation for tissue regeneration from technological platforms. Recently, important developments have been made in the implementation of complex structures built both at the macro- and the nano-level that have been shown to positively impact bone formation and to have the ability to deliver active biological molecules (drugs, growth factors, proteins, cells) for controlled tissue regeneration and the prevention of infection. These materials are diverse, ranging from polymers to ceramics and various composites. This review presents developments in this area with a focus on the role of scaffold structure and chemistry on the biologic processes that influence bone physiology and regeneration.     

Natural product diversity from the endophytic fungi of the genus Aspergillus

The endophytic fungus Aspergillus is considered as an enormous source of chemical leads with promising biological activities. Different Aspergillus species have proved their ability to produce plenty of secondary metabolites including butenolides, alkaloids, terpenoids, cytochalasins, phenalenones, ρ-terphenyls, xanthones, sterols, diphenyl ether and anthraquinone derivatives with diverse biological activities, such as anti-cancer, antifungal, anti-bacterial, anti-viral, anti-inflammatory, antitrypanosomal and antileishmanial activities. From January 2015 until December 2019, three hundred and sixty-one secondary metabolites were reported from different endophytic Aspergillus species. This review discusses the isolated secondary metabolites from different endophytic Aspergillus species reported from January 2015 to December 2019 along with their reported biological activities and structural aspects whenever applicable.

The endophytic fungus Aspergillus is the precious source of chemical biodiversity.     

Ki67 expression in breast cancer: Correlation with prognostic markers and clinicopathological parameters in Saudi patients

Objectives:

To evaluate Ki67 immunoexpression pattern in Saudi breast cancer (BC) patients and investigate any possible predictive or prognostic value for Ki67.

Methods:

This is a retrospective study designed to quantitatively assess the Ki67 proliferative index (PI) in retrieved paraffin blocks of 115 Saudi BC patients diagnosed between January 2005 and March 2015 at the Department of Pathology, King Fahd Hospital, Al Madinah Al Munawarah, Kingdom of Saudi Arabia. The Ki67 PI was correlated with individual and combined immunoprofile data of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2/neu) with their clinicopathological parameters.

Results:

Ki67 immunoreactivity was highly expressed (>25% of the tumor cells were positive) in 85 (73.9%) patients. The Ki67 PI was significantly associated with poor prognostic clinicopathological parameters including old age (p<0.02), high tumor grade (p<0.01), lymph node metastasis (p<0.001), and Her-2/neu positivity (p<0.009). However, the association with ER positivity, PR positivity, tumor size, and lymphovascular invasion were not statistically significant. The Ki67 PI was significantly associated with BC molecular subtypes that were Her2/neu positive (luminal B and HER-2) subtypes compared with the Her2/neu negative (luminal A) subtype (p<0.04).

Conclusion:

The Ki67 PI is significantly higher in Saudi BC patients comparing with the reported literature. Ki67 PI was highest in the HER-2 and luminal-B molecular subtypes. Along with other prognostic indicators, Ki67 PI may be useful in predicting prognosis and management of Saudi BC patients.Breast cancer (BC) is one of the most common malignancy in the world.1 Although in the Kingdom of Saudi Arabia (KSA), the incidence of BC is much lower than in the Western world; it is still the most common malignancy in the Saudi women. According to the Saudi Cancer registry,2 BC accounted for approximately 23% of all the newly diagnosed female cancers. An additional significant fact on BC in KSA is its special presentation; as it predominantly affects the younger population, frequently presents as higher histological grades and in advanced clinical stages.2-4 Apart from the problem of its being highly prevalent globally and locally; BC has also shown its divergent nature with regards to its clinical course, response to treatment, and prognostic outcomes. Thus, the new molecular classification of BC has emerged on the basis of biomarkers. In the initial stages, the hormones namely, the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (Her2) played their roles. It was only 15 years ago, that the molecular classification of BC was proposed by Californian scientists, initially there were 4 major classes: luminal-like, basal-like, normal-like, and Her2 positive.5 Consequently, a fifth class was added, dividing one of the major luminal class to luminal A and luminal B classes.6-8 Ki67 had been known to be an important proliferation biomarker since 1980s. It has recently become an essential component of routine biomarker profile for BC, along with ER, PR, and Her2, to assist the oncologists in delivering optimum treatment to BC patients. Its role as a poor prognostic biomarker is well established, and a number of studies have found a significant correlation between Ki67 positivity with that of histological parameters such as nuclear grades and mitotic figures.9 Recent studies have also proved its predictive role in both the antihormonal therapy and chemotherapy for the efficacy of the treatment. The aim of this study is to examine the Ki67 biomarker in the BC patients and the immunohistochemically on the paraffin embedded blocks. Subsequently, to correlate the Ki67 findings with individual and combined immunoprofile data of ER, PR, and Her2/neu, as well as with their clinicopathological parameters to identify any specific differences in our BC cases as compared with western cases. This is may be important in investigating any predictive or prognostic role of Ki67 in managing BC patients in KSA population.     

Endogenous signals released from necrotic cells augment inflammatory responses to bacterial endotoxin

Stressed cells undergoing necrosis release molecules that acts as endogenous danger signals to alert and activate innate immune cells. Both HMGB1 and HSP70 are induced in activated monocytes/macrophages and also are released from stressed or injured cells. We investigated whether HMGB1 and HSP70 released from necrotic monocytes/macrophages, can act as danger signals to mediate proinflammatory cytokine responses to bacterial endotoxin or lipopolysaccharide (LPS). We show that cell lysate, obtained from necrotic cells directly stimulates the proinflammatory cytokine and chemokine responses in human monocyte/macrophage cell line, THP-1, as revealed by the induction of TNF-alpha, IL-6 and IL-8 mRNA expression and protein production. In the presence of LPS, necrotic cell lysate induced a more robust increase in all three proteins. We found that HMGB1 and HSP70 were indeed present in the necrotic cell lysate and were responsible for the significant induction of the proinflammatory cytokine expression, as neutralization with antibodies against both proteins blocked the increase in the cytokine production seen after incubating LPS-stimulated cells with the necrotic cell lysate. We also found that the newly identified triggering receptor expressed on myeloid cells-1 (TREM-1) was involved in mediating the HMGB1- and HSP70-induced cytokine production. Blocking TREM-1 on THP-1 cells with a recombinant chimera prevented the increase in cytokine production, while simultaneous blocking of TLR4 and TREM-1 completely abolished the proinflammatory response, suggesting that TREM-1 synergizes with TLR4 to mediate the effects of such signals from necrotic cells. In addition, blocking HMGB1 or HSP70 simultaneously with TREM-1 did not decrease the cytokine level further, confirming the involvement of TREM-1 in mediating the effect of HMGB1 and HSP70. Although the interaction of HMGB1 and HSP70 with TREM-1 induced I kappa B alpha and p38 expression, both of which are required for the inflammatory cytokine expression, blockade of TREM-1 did not affect I kappa B alpha expression but markedly reduced p38 activation, as revealed by Western blot analysis. Together, these results demonstrate that HMGB1 and HSP70 released from necrotic cells function as endogenous danger signals to augment the proinflammatory responses in monocytes/macrophage and that TREM-1 relays such signals to the cytokine expression cascade. This mechanism may contribute to the amplification and persistence of the inflammatory response to bacterial infection.     

Frequency and features of nocturnal enuresis in Pakistani children aged 5 to 16 years based on ICCS criteria: a multi-center cross-sectional study from Karachi,Pakistan

Background

Nocturnal enuresis (NE) is a common symptom in children worldwide. International Children’s Continence Society (ICCS) defines enuresis as either mono-symptomatic, NE with lower urinary tract symptoms and NE with co-morbid conditions. The objectives of this study were to determine the frequencies and types of NE and associated symptoms and conditions in children aged 5 to 16?years based on ICCS criteria.

Methods

A multi-center cross sectional study was conducted between November 2012 and December 2013 in the primary care clinics of four hospitals in Karachi. Children aged five to fifteen years were included through consecutive sampling. Informed consent was obtained from the parents and a pre-coded semi-structured questionnaire was used to obtain the information. Data was entered on SPSS version 20.0 and multivariable logistic regression analysis was used for data analysis.

Results

Out of 429 children aged between five and sixteen years, 243(56.9%) were boys and the remaining 186(43.1%) were girls. One hundred and eighty three children (43%) had nocturnal enuresis (NE). Forty four (10.3%), had mono-symptomatic NE, 57(31.1%) had associated lower urinary tract symptoms (NE-LUTS), whereas 30 (16.3%) had NE with a co-morbid condition. Fifty two (28.4%) NE’s had at least one of both LUTS and a co-morbid condition. Out of the 246(57%) non-enuretic’s, 31(12.6%) had a LUTS, 95(38.6%) had a co-morbid condition and 57(23.2%) had at least one of both LUTS and a co-morbid condition. The remaining 63 (25.6%) were symptom free. Increased voiding frequency, urgency, dysuria, suprapubic pain and daytime incontinence were the LUTS significantly associated with NE. Co-morbid conditions significantly associated with NE included constipation, congenital defects, developmental delay, and learning and sleep problems.

Conclusion

Although NE can be an only symptom, it is often associated with lower urinary tract symptoms like dysuria, urgency, suprapubic pain, and daytime incontinence. Children presenting with NE often have co-morbid conditions like constipation, urinary tract infection, sleep disorders, and developmental delay. Many children presenting with these conditions as the primary complaint may also have NE. It should be addressed as unrecognized and untreated NE can cause additional morbidity and distress.

     

Mucosal co-delivery of ketorolac and lidocaine using polymeric wafers for dental application

The current study aimed to investigate the effectiveness of a developed sodium alginate and polyvinylpyrrolidone K-25 (PVP K-25) polymeric wafer for the co-delivery of ketorolac and lidocaine to soft tissues for healing and pain control following gingivectomy. Nine ketorolac/lidocaine lyophilized wafers were formulated and assessed for their hydration capacity, mucoadhesion ability and in vitro release profile to select the optimum system for further clinical investigation. Wafer F6 containing 2:1 sodium alginate to PVP K-25 and 10% glycerol showed optimum properties and was selected for the clinical study. Twenty patients were included in the study and the ketorolac/lidocaine wafer was assessed versus a market product. Visual pain analog was evaluated daily for the first week and wound healing index was evaluated for one week, two weeks and one month following the procedure. The developed ketorolac/lidocaine polymeric wafer proved to be an effective method of reducing pain and discomfort together with enhancing wound healing following gingivectomy.