Medical Laboratory Journal

Analysis of broncho alveolar lavage adenosine deaminase assay in patients with clinically diagnosed pulmonary tuberculosis

Pankaj Garg

Background: The sputum smear-negative pulmonary tuberculosis (PTB) is a diagnostic challenge for physicians. It has been shown that adenosine deaminase (ADA) activity increases in various body fluids of patients with tuberculosis (TB). A prospective clinical trial was conducted to determine the effectiveness of ADA activity in bronchoalveolar lavage (BAL) in subjects who have sputum smear-negative PTB.
Methods: A total of 29 patients (M/F: 15/14), mean age (36.8 years), were enrolled in our study from October 2021 to August 2022 after providing written consent. The mean duration of symptoms was 41.66 days. Out of 29 patients, 21 patients had BAL ADA 4.81±1.68 unit??, for whom AKT treatment was started and cured, while four patients with BAL ADA 4.50±2.86 unit? did not improve, and four patients with BAL ADA 6.52±1.16 whose AKT treatment is ongoing at present but clinically improved. The sensitivity of BAL ADA with the outcome of 29 patients is 75%, while for BAL CBNAAT, it is 80%. When we apply a formula for combined sensitivity for the parallel test, then it comes to 95%, which indicates a great number of patients gets the benefit when we apply both tests simultaneously.
Results: We conclude that ADA activity was significantly increased in BAL.
Conclusion: BAL ADA is a useful and effective investigation for the diagnosis of PTB.

Clinical and microbiological study of vancomycin-resistant enterococci isolated from colonized and infected patients with special reference to risk factors

Deepa Devhare

Background: Vancomycin-resistant enterococci (VRE) has become a growing concern in healthcare settings as a major cause of many nosocomial infections worldwide. Risk factors associated with VRE are important to study. High-risk patients need to be screened and isolated to prevent the spread of infection and colonization. The present study aims to investigate the clinical spectrum, risk factors, and source of transmission of VRE in infected and colonized patients.
Methods: A prospective observational study was carried out for 1 year. A total of 200 Enterococcus species isolated from clinical samples such as urine, pus, blood, sterile body fluids, and stool from 200 patients without infection were included in the study. Stool samples were screened to measure the prevalence of VRE colonization. All samples were screened for vancomycin resistance using the Kirby-Bauer disc diffusion method. Vancomycin MIC was detected using the macrobroth dilution method. Demographic and clinical history of the patients were recorded.
Results: Vancomycin resistance was detected in 7 (3.5%) of 200 enterococci isolates from clinical samples. Urinary tract infection (n = 5, 71.4%) was the most common clinical illness caused by VRE. Gut colonization was found in 12 (6%) out of 200 patients screened for VRE. A history of previous antibiotic exposure was a significant risk factor in the current study and was associated with VRE infection and colonization. Endogenous bloodstream infection caused by VRE was found in one patient with VRE colonization.
Conclusion: The findings of this study highlight the significant burden of VRE on patients, both those infected and colonized. The emergence of multidrug-resistant bacteria in healthcare settings, a consequence of inappropriate antibiotic use, is a serious concern that warrants further research and our continued attention.

An update on the increasing prevalence of multidrug-resistant pathogens found in mechanically ventilated patients in central India

Sonakshi Dwivedi

Background: Rampant and irrational use of antibiotics led to antimicrobial resistance in intensive care units, directly influencing the clinical outcome. The prior introduction of antibiotics, especially broad-spectrum antibiotics, has been identified as a leading cause of hospital-acquired pneumonia. The present study aims to examine the existing scenario of antibiotic resistance due to multidrug-resistant organisms that are detected in mechanically ventilated patients.
Methods: This cross-sectional study was conducted in the department of Microbiology of a tertiary care hospital in Central India. A total of 410 endotracheal secretions were collected. The endotracheal aspirate of adult patients admitted to the medicine intensive care unit and on mechanical ventilation was received at the microbiology laboratory for processing by standard bacteriological techniques. Drug susceptibility testing was done using the Kirby-Bauer disc diffusion method according to the indications mentioned in Clinical and Laboratory Standards Institute 2021.
Results: Out of 410 collected endotracheal secretion samples, 332 (81 %) samples demonstrated bacterial growth. A total of 265 (80%) cases fulfilled the inclusion criteria. From 265 samples, 92 (34.7 %) patients were clinically and microbiologically confirmed as cases of ventilator-associated pneumonia. Over eighty percent of gram-negative bacilli were multidrug-resistant strains (Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa).
Conclusion: Real understanding of multidrug-resistant pathogens, early isolation as well as avoiding long-term antibiotic intake can reduce mortality levels currently linked with late-onset ventilator-associated pneumonia.

Chemiluminescence vs hemagglutination assay in the detection of Anti-Treponema pallidum antibodies

Toukam Michel

Background: Syphilis has serious consequences for human health and poses challenges for biological diagnosis. Techniques such as chemiluminescence are still poorly known in our context. Our objective was to compare chemiluminescence and hemagglutination assays with immunoblot as a reference test for detecting anti-Treponema pallidum antibodies.
Methods: Anti-Treponema pallidum antibodies were detected in serum samples. Comparison of chemiluminescence and hemagglutination assays was carried out with immunoblot as the reference test, based on sensitivity, specificity, positive and negative predictive values, and kappa statistic, using IBM-SPSS version 24 software. The significance level was set at 5%.
Results: A total of 600 participants were recruited, with a prevalence of syphilis infection of 3%. Sensitivity and specificity were 100% and 96.2% for chemiluminescence, and 93.8% and 100% for hemagglutination, respectively. Cohen's kappa statistic between the two tests showed satisfactory agreement (0.768). Cohen's kappa agreements between chemiluminescence/immunoblot and plate hemagglutination assay/immunoblot were greater than 0.8, indicating excellent agreement with the reference test (p<0.0001).
Conclusion: Chemiluminescence is more sensitive but less specific than hemagglutination. The use of chemiluminescence as a first-line test for the diagnosis of syphilis should be considered.

An overview of the production of AmpC and Metallo-β-Lactamase enzymes in Escherichia coli

Kirandeep Kaur

Escherichia coli is a Gram-negative, rod-shaped bacterium, responsible for 90% of all community-acquired infections and 50% of hospital-acquired infections, with opportunistic infections found in intensive care unit (ICU) patients. The β-lactam antibiotics, which inhibit cell wall synthesis, are known for their high efficacy and broad-spectrum activity. They also have low toxicity and provide long-term effects, making them widely used drugs against Gram-negative bacteria. Bacteria develop resistance to β-lactams primarily through the expression of hydrolytic enzymes, called β-lactamases, which are divided into serine β-lactamases (Classes A, C, and D) and metallo-β-lactamases (Class B), based on their molecular mechanism. This study aimed to clarify the mechanism of action of β-lactams against Gram-negative bacilli and to emphasize the multidrug resistance of cephalosporins and carbapenems to E. coli.

In vitro anti-leishmanial effects of garlic extract on Leishmania major: A Systematic Review

Roghiyeh Faridnia

Background: Leishmaniasis is a vector-borne parasitic disease caused by an obligate intracellular protozoan. Despite the significant frequency of cutaneous leishmaniasis, there is still no appropriate prevention, control and treatment. In recent years, a few studies have been performed on the effects of garlic extract for the treatment of cutaneous leishmaniasis. Current drugs, such as glucantime, have several side effects and lead to direct cellular damage. If an herbal-based drug reduces Leishmania by augmenting the immune system, it has the potential to possess a wider margin of safety. This study aimed to investigate the effects of in vitro systematic review of the effects of garlic (Allium sativum) and its compounds on Leishmania major.
Methods: Five English databases (PubMed, Google Scholar, Embase, Scopus, and Web of Science) were searched until the end of December 2022. The syntax and specific tags have been used for each database. The studies with poor methodology, inadequate information, inappropriate analysis, and confusing presentation were excluded from the current study. The quality of articles was assessed by eleven questions developed by the authors and the data were extracted from the selected articles and discussed.
Results: In this study, 198 articles were selected in the search step, of which five eligible articles were included for examination. The most commonly used solvent for the preparation of garlic extract was distilled water (60%), followed by methanol (40%). In these studies, the bulb of garlic is mostly used (80%). Cell type of J774 was used in only one study (20%), and murine peritoneal macrophages were used in the other studies (80%). The effect of garlic on Leishmania major was strong (80%). Studies have shown that garlic extract or its active compounds can increase cellular immune responses, which play an important role in inhibiting the Leishmania parasite. This is associated with the activation of macrophages and increased IFN-γ levels and NO production.
Conclusion: According to the studies, the effect of different combinations of garlic on the Leishmania parasite has been shown. However, the exact mechanism of the anti-Leishmanial effect of garlic has not been determined. Thus, this issue needs further investigation.

Evaluation of different decalcifying agents on histochemical and immunohistochemical staining properties of canine osseous tissue

Esmail Ranjbar

Background: The purpose of tissue processing is to strengthen the tissue and place it in a suitable environment for slicing without causing damage. In routine histopathology, decalcification of calcified tissues is a principal step before tissue processing. Thus, the aim of this study is to evaluate the effect of several decalcifying agents on morphological and antigenic preservation in canine mandibular tissue.
Methods: Four different decalcifying solutions, including 5% nitric acid (NA), 10% and 20% formic acid (FA), and 10% ethylene diamine tetraacetic acid (EDTA), were employed to remove mineral salts from specimens harvested from mandibular osseous tissues of four healthy Mongrel dogs (32 samples). The solutions were compared regarding the decalcification time, ease of tissue slicing, morphological organization after Alizarin red S staining, and osteocalcin antigenic preservation.
Results: Considering the short decalcification time and ease of sectioning, 5% NA) yielded better results than 10% and 20% FA and 10% EDTA (pH 7.4). In terms of the preservation of morphology and antigenicity of the tissue samples, 10% EDTA was found to be the most optimal solution, followed by 10% and 20% FA and 5% NA.
Conclusion: Our findings support EDTA as a highly preferred choice for the decalcification of canine osseous tissue when aiming for immunohistochemistry, despite its time-consuming nature. However, for general histological staining procedures, 20% FA and 5% NA are preferred.

Evaluation prevalence of Rh system major antigens (D, C, c, E, e) and their phenotypes in blood donors of Golestan province, Iran

Alireza Ahmadi

Background: The understanding of blood group phenotypes is currently limited to the ABO and Rh blood group systems. This study aimed to determine the frequency of Rh system antigens (D, C, c, E, e) and the phenotypes of the system in blood donors. Identifying the blood group phenotypes of donors in any population is important for improving healthcare services and better serving patients.
Methods: This descriptive study was carried out on 575 donors (Turkmen and Fars) in blood transfusion centers in Golestan Province, Iran. A cell suspension (3-5%) from each sample was prepared in normal saline and exposed to Rh system antisera using the haemagglutination technique. The Rh phenotype was then determined based on the most common genotype.
Results: For the Rh system, the antigen frequencies of D, C, c, E, and e were 87.76%, 73.6%, 72.1%, 30.83%, and 93.59%, respectively. The most common phenotypes among the Turkmen and Fars donors were R1R1, R1r, and rr, respectively, while the least common phenotypes were R2Rz and ryry. The phenotypes r'r' and ryry were not detected in the Turkmen donors, and the phenotype r''r' was not identified in the Fars donors.
Conclusion: Identifying the prevalence of blood group antigens in donors from each region is crucial for organizing negative antigen blood units, preparing compatible blood for multitransfused patients, and preventing the development of alloantibodies in these patients.

Effect of combined training on gene expression of nuclear factor-κB and Sirtuin 1 in fast and slow twitch muscles in elderly male rats

Mehdi Mogharnasi

Background: The aim of this study is to investigate the effects of ten weeks of combined training on the gene expression of nuclear factor-κB and sirtuin 1 in fast and slow twitch muscles of aged male rats.
Methods: Sixteen rats, each 24 months old, were randomly divided into two groups: combined training and control (Eight rats per group). Combined exercises were performed four sessions per week, including two days of endurance and two days of resistance. The exercises took place in a container measuring 50x50x100 cm, filled with water maintained at 30±1°C. On the first day, the animals swam for five minutes in water at a height equal to 100% of their body length, without weights. On the second and third days, the rats swam for 10 minutes with the water height equal to 120% of their body length. On the fourth and fifth days, they swam for 15 minutes with the water height at 140% of their body length, which remained constant during the study period. Data were analyzed using two-way analysis of variance with SPSS version 22 software, with the significance level at P≤0.05.
Results: After ten weeks of combined training, a significant difference was observed in the gene expression of nuclear factor-κB and sirtuin 1 between the training and control groups (P=0.001 for both).
Conclusion: According to the results of this research, performing combined exercises in water with appropriate intensity and duration can regulate inflammatory and anti-inflammatory pathways, thereby strengthening muscles and reducing muscle wasting and atrophy in the elderly.

Assessment of parasitic contamination in the soil of public parks Gorgan city

Farideh Tohidi

Background: Soil contamination with parasites poses a significant and concerning risk for the transmission of parasitic infections to humans and animals. Considering the crucial role of soil as a potential source of parasite transmission, this study aimed to investigate the presence of helminth and protozoan parasites in the soil of public parks in Gorgan City.
Methods: In this study, a total of 80 soil samples were collected from 16 public parks in Gorgan City in 2022. The soil samples were collected from various areas within the parks, including flower-making, children's play areas, walking paths, sitting areas, and garbage dumps. To isolate the parasites from the soil samples, a saturated solution of sodium nitrate was utilized. The samples were carefully examined under a microscope, and all isolates were stained and subjected to further investigation using special acid-fast stain.
Results: Toxocara eggs were the most commonly detected parasite, present in 40% of the samples (n=32). Coccidia oocysts were found in 12.5% of the samples (n=10), which could potentially be Sarcocystis or Toxoplasma oocysts. Mites were also detected in 12.5% of the samples (n=10). Additionally, Cryptosporidium oocysts were identified using Ziehl-Neelsen's modified staining method, and they were found in 10% of the samples (n=8).
Conclusion: Health education plays a vital role in raising awareness about preventive measures. Collecting reservoirs of parasites (Such as dogs and cats) and planning for the proper disposal of human waste also play an important role in prevention of this disease.