Medical Laboratory Journal

Utility of immunofixation in complementing and empowering serum protein electrophoresis in the diagnosis of paraproteinemia: Experience at a tertiary care center

Imran Ahmed Siddiqui

Background: ‘M’ proteins or paraproteins refer to immunoglobulins that are produced by clonal plasma cells and are a characteristic feature of monoclonal gammopathies. Routine electrophoresis on agarose gel and immunofixation can be used to detect immunoglobulin paraprotein (M-protein). We aimed to evaluate the performance of agarose gel electrophoresis alone and in combination with immunofixation for detecting serum M-proteins.
Methods: One hundred and twenty-three patients suspected of paraproteinemia were evaluated. Routine serum protein electrophoresis (SPE) and immunofixation electrophoresis (IFE) protocols were performed. Data from SPE, and SPE-IFE (gel images and electrophoretograms) were collected and reviewed.
Results: 21% cases were confirmed using the SPE-IFE combination, and among them, 33% had positive light chain (λ) only on IFE. Similarly, nine cases with biclonal gammopathy on SPE were characterized by IFE.
Conclusion: IFE can be a confirmatory test in cases where SPE results are not reliable and it can be a complementary test when characterization of the M protein detected on SPE is required.

New insight into antibiotic resistance in urinary tract infections: Interplay between community and hospital acquired UTI

Sangita Bhalavi

Background: Urinary tract infections (UTIs) are among the most common types of infections affecting people in community and hospital settings. Bacteria are the leading cause of UTIs, followed by fungi. 39% of all healthcare-associated infections (HAIs) affecting all age groups are UTIs, causing high morbidity and mortality rates. The antibiotic susceptibility pattern of causative organisms is changing due to improper antibiotic use. The study was conducted to determine the microbiological profile of both community and HAIs and their antimicrobial susceptibility pattern.
Methods: Clean-catch, mid-stream urine samples collected in the universal wide-mouthed sterile containers were transported to the laboratory. Samples were processed by standard conventional microbiological procedures. Antimicrobial susceptibility was done using the Kirby-Bauer disc diffusion method on Mueller-Hinton agar plates.
Results: The most common causative organisms among gram-negative bacteria were E coli (26.05%), followed by Klebsiella spp (20.37%), and Enterococcus spp (12.81%) was more common among the gram-positive bacteria. Non-albicans Candida (64.10%) were more commonly isolated than Candida albicans (35.90%). E coli was highly susceptible to nitrofurantoin and fosfomycin, and Klebsiella spp and Enterococcus spp were similarly highly susceptible. Antibiotic resistance was more common among bacteria isolated in HAIs.
Conclusion: In both settings, E coli was the most common causative organism. The incidence of non-albicans Candida species has increased in comparison to Candida albicans. Antimicrobial susceptibility to empirical 3rd-generation cephalosporins and fluoroquinolones has drastically decreased. Hospital-acquired UTIs are a rising threat to the healthcare system and community. Based on hospitals’ antimicrobial policy formulated by studying antimicrobial susceptibility patterns, empirical treatment should be chosen.

Creatine kinase as a supportive test in thyroid disorders

Jincy W

Background: Musculoskeletal disorders are common in patients with hypothyroidism, and are also observed in thyrotoxicosis. Creatine kinase (CK) is present in the muscles and is involved in energy metabolism. This study aimed to estimate the serum CK levels in patients with hypothyroidism, hyperthyroidism, and healthy individuals. Moreover, the correlation of CK levels with markers of thyroid function is assessed.
Methods: A total of 120 patients with hypothyroid and 120 with hyperthyroid were compared with 120 healthy individuals aged 20-60 years. The thyroid status was assessed by determining the serum thyroid-stimulating hormone (TSH) and free thyroxine (fT4) using chemiluminescent immunoassay. Serum CK was measured the by kinetic method. Statistical analysis was performed by analysis of variance and Pearson’s correlation to investigate the correlations between CK and thyroid hormones.
Results: A significant increase (P<0.0001) and a significant decrease (P<0.0001) in serum CK were observed in hypothyroid patients (253.98±129.04 IU/L) and in hyperthyroid patients (34.68±13.15 IU/L), respectively, compared to the control group (72.9±29.01 IU/L). A negative correlation was found between fT4 and CK (r: -0.4253, P<0.0005).
Conclusion: It could be concluded that CK activity in serum may be a useful additional test in thyroid disorders.

The potential of mononuclear cells as a predictive marker for the level of stem cells in autologous peripheral blood stem cell transplantation in Multiple Myeloma: A Review Article

Darshana Kottahachchi

Multiple myeloma (MM) is a plasma cell neoplasm that is characterized by the clonal proliferation of malignant plasma cells in the bone marrow. It is considered the second most common hematological malignancy which accounts for approximately 1% - 2% of all cancers and among 10% of hematological malignancies. Autologous peripheral blood stem cell Transplantation (PBSCT) is the best treatment for MM. Since the optimum harvested stem cell yield is a crucial factor for sufficient engraftment, the enumeration of Mononuclear cell (MNC) count in peripheral blood and harvested CD 34+ stem cell count can be considered as the best predictive markers for the best timing of apheresis which positively correlates with engraftment outcome of PBSCT.
MNC count can be obtained using either a hematological analyzer or peripheral blood smear while flow cytometry is the advanced technology that can be used to enumerate CD 34+ stem cell count other than peripheral blood smear. The unavailability of a flow cytometer, the expensiveness of this method, and the lack of trained personnel regarding this new technology, especially in lower-middle-income countries cause disturbance in the enumeration of stem cells. In such a situation, this review describes the importance of establishing an association between peripheral blood MNCs and harvested CD 34+ cells. Furthermore, this association facilitates conducting effective PBSCT for MM patients even in the absence of a flow cytometer and eventually, it focuses on decentralizing the treatment of PBSCT.

Bacteriological profile of pyogenic infections and their antimicrobial susceptibility in a tertiary care hospital in central India

Bhawana Bajare

Background: Pyogenic infection is a significant cause of morbidity. Infection with multidrug-resistant strains poses a major difficulty in the treatment. The study was conducted to know the bacteriological profile of pyogenic infections and their antibiotic susceptibility.
Methods: A cross-sectional study was conducted in a tertiary care hospital from October 2021 to March 2022. Isolates from pus specimens were subjected to an antibiotic sensitivity test using the Kirby-Bauer method as per CLSI 2021.
Results: Out of 752 samples, etiology could be revealed in 510 (68.4%) specimens. Enterobacterales dominated the profile, with K pneumoniae isolated in the maximum number of specimens. Staphylococcus aureus was the culprit in 14% of the cases. Non-fermenters were isolated in 17% of the cases. Methicillin resistance in S aureus was 67%. Gram-positive cocci showed high sensitivity to linezolid. For both Piptaz and carbapenem, Enterobacterales and non-fermenters showed around 50% and 60% susceptibility, respectively.
Conclusion: Continuous surveillance of the aetiologic agents of pyogenic infections and their antibiotic sensitivity pattern needs to be done to design and implement the antibiotic policy for the infection in our set-up.

The correlation between serum leptin levels and lipid profile among non-obese type 2 diabetes mellitus patients and non-diabetic individuals

Chandralekshmy Chandrika

Background: Type 2 DM is a heterogeneous group of disorders characterized by insulin resistance, impaired insulin secretion, increased glucose production, and abnormal fat metabolism. Diabetes mortality primarily results from microvascular complications such as diabetic retinopathy, nephropathy, and neuropathy, as well as macrovascular complications like coronary artery, peripheral vascular, and cerebrovascular diseases. Patients with diabetes usually have changes in adipose tissue metabolism and abnormalities in the secretion of adipokines such as leptin. The present study aims to study the relationship between serum leptin levels and lipid profile parameters among non-obese type 2 diabetes mellitus patients and non-diabetic individuals.
Methods: This hospital-based cross-sectional study was conducted among 41 type 2 diabetic patients and 41 non-diabetic individuals of both sexes between the ages of 40 and 70. Fasting blood glucose (FBS), serum leptin, total cholesterol (TC), triglyceride (TG), and high-density lipoprotein (HDL) levels were assayed.
Results: The mean serum leptin levels among diabetic patients were lower than those of non-diabetic individuals, and this difference in mean was statistically significant. The study showed a significant negative correlation between serum leptin, TC, TG, and low-density lipoprotein (LDL), and a positive correlation with HDL. In the final regression model, serum leptin showed a statistically significant association with FBS and HDL.
Conclusion: This study demonstrated that serum leptin levels can be a strong predictor of low HDL levels in diabetic patients. It can also contribute to raised levels of total cholesterol, triglyceride, and LDL, which are responsible for macrovascular complications in diabetics.

Phylogenetic group determination and genetic diversity of Escherichia coli isolated from domestic animals’ stool specimens and human clinical samples

maryam adabi

Background: Escherichia coli consists of a wide range of strains with huge diversity in their genome, distributed in nature and the alimentary tracts of animals and humans. This study analyzed the phylogenetic group determination and genetic diversity of E. coli strains isolated from domestic animals and human clinical samples.
Methods: Twenty E. coli isolates from domestic animals were analyzed for phylogenetic grouping. Also, 100 clinical samples and 20 animal samples were evaluated by the enterobacterial repetitive intergenic consensus–polymerase chain reaction (ERIC-PCR) technique. The results and the similarity between the strains were determined based on the Dice similarity coefficient in the SAHN program of the NTSYS-pc software.
Results: The frequency of phylogroups among animal samples were A = 5%, B1 = 65%, B2 = 20%, and D = 10%. Based on the ERIC-PCR results, the clinical strains were allocated into 19 clusters. Most strains were in the E7 cluster. Fifty percent of the E. coli isolated from animal specimens belonged to the E4 group, and the lowest number of strains was in the E3 and E5 (1 strain) groups.
Conclusion: The results confirmed the efficiency and usefulness of the ERIC-PCR tool for the identification and classification of bacteria. Also, we demonstrated the most phylogroup among animal samples.

Detection and identification of Prevotella Melaninogenica in saliva samples of patients with ulcerative colitis

Nazanin Mortazavi

Background: Ulcerative colitis (UC) is a long-term inflammatory bowel disease (IBD) caused by abnormal immune responses, leading to inflammation and scarring in the large intestine. The bacteria Prevotella melaninogenica, found in the intestine and mouth, may contribute to UC. This study focuses on the detection of P. melaninogenica in the saliva of UC patients and compares them with the healthy control (HC) group.
Methods: The present study was a case-control study including 40 UC patients and 40 healthy controls (HCs) with an average age of 43.0 3± 10.3. This study used a real-time PCR test to investigate the frequency and average number of P. melaninogenica from the 16S rRNA gene sequence of P. melaninogenica in both groups.
Results: P. melaninogenica was more frequent in UC patients (77.5%) than HCs (45%) (p = 0.003). The patient group had more bacteria (339.31 ± 1082.29) than HCs (61.29 ± 154.03) (p = 0.005). Women in the UC group had more P. melaninogenica (492.35 ± 1427.61) than the control group (56.98 ± 123.50) (p = 0.0342). Similarly, men in the UC group (262.85 ± 664.97) had more bacteria than the control group (72.62 ± 222.76) (p = 0.015).
Conclusion: The current study showed that dysbiosis in P. melaninogenica, a bacterium in human saliva, could be important in the development of UC. Further investigation is needed to evaluate its use as a potential biomarker in the UC.

Investigating the changes of hippocampal m-RNA gene expression of neurotrophin-3 and its receptor (Tropomyosin Receptor Kinase C) in the recovery of movements of rats with spinal cord injury during two types of endurance exercises

Sadegh Cheragh Birjandi

Background: This study aimed to investigate the effect of four weeks of selected endurance training on neurotrophin-3 (NT-3) and tropomyosin receptor kinase C (TrkC) gene expression in hippocampal areas of rats with spinal cord injury (SCI).
Methods: In this experimental study, the rats were separated into six equal groups. First, the animals were put under general anesthesia and had their SCI. Then, for four weeks, they were subjected to two kinds of endurance training programs. However, the control injury group received no intervention or training. Following the completion of the training regimes, molecular tests were done using the qRT-PCR technique to evaluate changes in the gene expression of NT-3 and TrkC from the animals' hippocampus.
Results: The expression of NT-3 and TrkC genes were significantly reduced in the SCI model compared to the healthy control group, but it was increased in the SCI + exercise 1 and SCI + exercise 2 groups compared to the SCI group. NT-3 levels did not vary significantly between the SCI + exercise 1 and SCI + exercise 2 groups, although alterations in TrkC levels altered.
Conclusion: In addition to enhancing locomotion in animals with SCI, the endurance training regimens in this research were effective on the expression of NT-3 and TrkC genes and may play a role in axonal development and neuronal survival in SCI recovery.

Cerium interfering effect on iron intestinal absorption in rats using the Everted Gut Sac (EGS) method

Ali Asghar Moshtaghie

Background: Certain trace elements, like cerium, have the potential to disrupt iron metabolism. This study explored the impact of cerium on intestinal iron absorption, focusing on the initial stage of iron metabolism. We employed the rat everted gut sac (EGS) segments to assess the interference caused by cerium. The primary objectives of this study were to examine the absorption of cerium in the intestines and to compare iron absorption in the presence and absence of cerium.
Methods: For the EGS experiment, segments of the rat's duodenum, ileum, or jejunum were promptly excised, cut into 5-6 cm segments, and rinsed with a physiological solution. These freshly prepared rat EGS segments were then incubated in Earle's medium containing iron (III) and/or cerium (III). We examined the impact of ascorbic acid, glucose, and different time intervals on the intestinal absorption of cerium and iron. Specifically, we investigated how glucose (5 mM) and ascorbic acid (2.8 mM) affected the absorption of cerium and iron at various concentrations (ranging from 0 to 200 mg/L). Additionally, we assessed the interfering effect of cerium on iron absorption.
Results: The results indicated that the maximum intestinal absorption of Fe (III) and Ce (III) occurred at a concentration of 200 mg/L. Furthermore, it was observed that their uptake increased following the reduction by ascorbic acid. The absorption of these elements also rose in the presence of glucose, suggesting energy-dependent transport. Additionally, a consistent cerium concentration was found to decrease iron absorption by 24.3% (P ≤ 0.05).
Conclusion: Based on the results, cerium likely reduces iron uptake by competing with iron. Cerium can also disrupt iron metabolism and lead to iron-related metabolic disorders. However, further studies at the molecular and intracellular levels are needed to gain a better understanding of this mechanism.