Relation of Neutrophil-Lymphocyte Ratio with Clinical Severity in Patients with Guillain-Barre Syndrome.

Guillain-Barre syndrome (GBS) is an acute autoimmune polyradiculoneuropathy. Neutrophil-lymphocyte ratio (NLR) is a novel prognostic and inflammatory marker in patients with neurological diseases. The study was designed to observe the relation between neutrophil lymphocyte ratio and the clinical severity in patients with Guillain-Barre Syndrome (GBS). This cross-sectional descriptive study was conducted at the department of Neurology and Medicine, Mymensingh Medical College and Hospital from April 2019 to September 2020. Total 58 patients with GBS were enrolled according to the inclusion and exclusion criteria within 7 days of development of symptoms. The clinical diagnosis of GBS was done according to Ausbury and Cornblath's diagnostic criteria; moreover clinical severity was done according to Hughes and Rees scale, Medical Research Council (MRC) grade, cranial nerve involvement and autonomic involvement. After measuring complete blood count, NLR was calculated by dividing neutrophil count to lymphocyte count. Data analysis was done on SPSS 23.0. Mean age of the GBS patients was 36.21±11.55 years. Among 58 respondents 70.69% (41) were male and 29.31% (17) were female. Most of the patients had a GBS severity score of 4(62.07%) followed by 3(27.59%) and 5(10.34%). The mean NLR of the respondents was 3.22±2.25. Acute motor axonal neuropathy (AMAN) was in 48.28% respondents and their mean NLR was 3.89±0.31, 31.03% had AIDP and mean NLR was 3.28±0.46 and 20.69% had Acute motor sensory axonal neuropathy (AMSAN) and mean NLR was 4.5±0.52. The mean NLR of the MRC grade 0, 1, 2, 3 patients was 6.61±1.78, 3.39±0.92, 2.71±1.39 and 1.45±0.40 respectively. Hughes score had a positive correlation (r=0.5333 and p<0.01) and MRC grade had a negative correlation (r= - 0.76805 and p<0.01) with NLR. Severity of GBS was significantly associated with raised NLR. Increase Hughes and Rees scale and decrease MRC grade also related to increased NLR.

Combination of cell culture and quantitative PCR (cc-qPCR) to assess disinfectants efficacy on Cryptosporidium oocysts under standardized conditions.

Oocysts of Cryptosporidium parvum are resistant to environmental conditions and many disinfectants. A combination of cell culture and quantitative real time PCR (cc-qPCR) is established for evaluation of anticoccidial disinfectants against C. parvum. C. parvum oocysts were treated with disinfectants, washed and oocysts were incubated with HCT-8 cell monolayers in the presence of excystation medium for 3h. Subsequently, unbound parasites were removed by washing with growing medium and the infected monolayers were further maintained in fresh growing medium for 48h. Genomic DNA was extracted from each sample and qPCR performed targeting a specific sequence of the 70kDa heat shock protein gene in order to quantify development. Treatment of oocysts with cresolic disinfectants demonstrated dose dependent reduction of viability of oocysts. More than 98% inactivations were recorded with at least 2% concentration of cresolic disinfectants after 2h of treatment. Bleach (sodium hypochlorite) at 6% solution induced 92.7% inactivation of C. parvum oocysts after 2h. Thermally treated oocysts (56 and 70 degrees C for 20min) demonstrated complete inactivation, whereas at 38 degrees C no inactivation was observed. Application of Neopredisan((R)) 135-1 and Aldecoc((R)) TGE (4% for 2h) as recommended according to the current guidelines stipulated by DVG (German Veterinary Society) consistently inactivated more than 99.5% of oocysts. The suggested cc-qPCR method appeared to be suited for standardized testing of inactivation measures, particularly for evaluation of chemical disinfectants and thus cc-qPCR is proposed as an alternative to the established chicken infectivity model for Eimeria tenella for testing anticoccidial disinfectants. A minimum inactivation of 99.5% in cc-qPCR model is claimed as a suitable threshold for certification of chemical products for disinfection of coccidia oocysts.

Effects of curcumin on Cryptosporidium parvum in vitro.

Cryptosporidium parvum is a zoonotic protozoan parasite having peculiarities among the apicomplexa that could be responsible for its resistance to some drugs and disinfectants against coccidia. The awareness of Cryptosporidium as a health problem in man and animal is increasing and potent drugs are urgently needed. Curcumin, a natural polyphenolic compound, has been found to be active against a variety of diseases including anticarcinogenic, antimicrobial, and antiprotozoal effects. We investigated the effects of curcumin on infectivity and development of C. parvum in a recently established in vitro system combining infection of human ileocecal adenocarcinoma cell cultures with quantification of intracellular parasites by quantitative polymerase chain reaction. Curcumin was found to be effective (>95% inhibition of parasite growth) at 50 microM for 24 h when infected cultures were exposed for more than 12 h. Withdrawal of curcumin after 24 h of exposure did not result in a significant resumption of C. parvum growth. The invasion of host cells by sporozoites (infectivity) was found to be inhibited at least 65% in the presence of 200 microM curcumin. No significant reduction of viability of C. parvum oocysts after incubation with curcumin was recorded. Altogether, curcumin showed promising anticryptosporidial effects under in vitro conditions and deserves further exploration.

Combination of cell culture and quantitative PCR for screening of drugs against Cryptosporidium parvum.

Cryptosporidium parvum is a zoonotic pathogen causing self-limiting diarrhea in immunocompetent patients. An assay combining cell culture and real time quantitative PCR (qPCR) is reported here to verify drug efficacy against C. parvum in vitro. The monolayers of Human ileocecal adenocarcinoma cells (HCT-8) were infected by sporozoites excysted directly on the cells and were incubated with monensin, halofuginone bromide and hexadecylphosphocholine until 45h post infection (p.i.). The genomic DNA was extracted at 3, 27 and 45h p.i. and subjected to qPCR targeting the 70kDa heat shock protein gene to quantify the development of C. parvum. The reliability of the method was validated by testing of monensin and halofuginone bromide, which are well known to be effective in vitro. With the dose dependency monensin and halofuginone showed a maximum inhibition of 98.15% and 98.05% at 0.144 and 25microM, respectively, compared with non-treated controls at the endpoint incubation, confirming previous reports. The reduction of the parasite DNA reproduction over 27h p.i. compared with 3h p.i. was found to be as 97-99% in 0.144microM monensin and 99% in 25microM halofuginone treated cells. The new antileishmanial compound hexadecylphosphocholine (24.5microM, Miltefosine) showed 78-98% inhibition at 45h p.i., however, the reproduction of parasite DNA was reduced to 96-98% over 27h p.i. The method has the potential to easily and reliably assess anticryptosporidial compounds in adequately equipped routine laboratories.

Quality of health care from patient perspectives.

The quality of health care is the consequence of strong link between service providers and user of the services. Perceived quality is one of the principal determinant of utilisation and non-utilisation of health services, a major issue in developing countries. Considering this, the present study was aimed to assess the quality of care in in-patient and outpatient departments of rural and urban government hospitals in Bangladesh. A total of 2420 patients were interviewed. The patients were selected by using systematic random sampling technique. Results revealed that age, waiting time, time spent for patient examination, place of treatment, income, years of schooling and male sex appeared to be independent predictors of patient satisfaction (p<0.001). Age, waiting time and years of schooling were negatively related with level of satisfaction indicating younger patients, less waiting time and patients with less education were more satisfied, whereas time spent for examination, income were positively related with patient's satisfaction. Patients attending at the urban hospitals and male sex were also significantly associated with patient's satisfaction. The study recommends that both short and long-term policy action should be adopted for quality assurance of the existing health care facilities in Bangladesh.