Development of Data-driven Machine Learning Models and their Potential Role in Predicting Dengue outbreak.

ABSTRACT: Dengue fever is one of the most widespread vector-borne viral infections in the world, resulting in increased socio-economic burdens. The WHO has reported that 2.5 billion people are infected with dengue fever across the world, resulting in high mortalities in tropical and subtropical regions. The current article endeavors to present an overview of predicting dengue outbreaks through data-based machine-learning models. This artificial intelligence model uses real-world data such as dengue surveillance, climatic variables, and epidemiological data and combines big data with machine learning algorithms to forecast dengue. Monitoring and predicting dengue incidences have been significantly enhanced through innovative approaches. This involves gathering data on various climatic factors, including temperature, rainfall, relative humidity, and wind speed, along with monthly records of dengue cases. The study functions as an efficient warning system, enabling the anticipation of dengue outbreaks. This early warning system not only alerts communities but also aids relevant authorities in implementing crucial preventive measures.

Expression of pro-inflammatory cytokines (IL-6 & IL-18) exacerbate the risk of diabetic nephropathy in the Pakistani population.

BACKGROUND: Diabetic nephropathy (DN) is a micro-chronic diabetic consequence induced by metabolic and hemodynamic abnormalities. Free radicals react with other critical cellular components, causing progression of aberrant renal function. OBJECTIVE: This case control study was aimed to determine the role of IL-6 and IL-18 in diabetic nephropathy in Pakistani population. METHODS AND MATERIALS: The study's subjects (n = 180 from Lahore, Gujranwala, and Karachi) were divided into control, diabetes mellitus (DM) and diabetic nephropathy (DN) groups. The serum concentration of IL-6 & IL-18 were determined by enzyme-linked immunosorbent assay (ELISA). The expression analysis of IL-6 & IL-18 were performed by Real Time PCR. RESULTS: The significant increase in serum levels of IL-6 were observed among the control, DM and DN groups (15.3 ± 24.1 pg/ml, 34.7 ± 24.0 pg/ml, 52.6 ± 33.2 pg/ml) whereas no significant difference was observed in serum levels of IL-18. The expression analysis of IL-6 was increased by more than forty three fold in DN group (n-fold = ~43.6) as compared to DM & control whereas the expression profile of IL-18 decreased in DN group (n-fold = ~0.89). In DN group the correlation analysis revealed direct association of GFR with serum IL-6 (r = 0.1114) & inverse relationship with serum IL-18 (r = - 0.097). In multiple regression analysis using GFR as the dependent variable, BMI and expression of IL-18 were determinants in DM subjects, but only uric acid in DN subjects. CONCLUSION: The present study implicates that increased expression of IL-6 and decreased of IL-18 was associated with development of DN in Pakistani population.

Current status of insecticide resistance and its underlying mechanisms in Aedes aegypti (L.) in Punjab, Pakistan.

Aedes aegypti (Linnaeus) plays an important role as a vector of different deadly diseases particularly dengue fever. Insecticides are used as a primary tool to control Ae. aegypti. However, due to the excessive use of insecticides on agricultural, public health, and industrial levels, mosquitoes have developed resistance. In this study, the current susceptibility status of Ae. aegypti mosquitoes against different insecticides (Temephos, DDT, dieldrin, Malathion, Bendiocarb, Permethrin, Cypermethrin, and Lambda-cyhalothrin) was evaluated in district Lahore and district Muzaffargarh of Punjab, Pakistan. For this purpose, WHO bioassays and biochemical assays were performed on Ae. aegypti population from Lahore (APLa) and Aedes population from Muzaffargarh (APMg). Results of APLa and APMg showed high levels of resistance against the larvicide Temephos. Resistance against all adulticides was also observed in APLa and APMg (% mortality < 98%). The biochemical assays indicated statistically significant elevated levels of detoxification enzymes in APLa and APMg. APLa showed slightly higher levels as compared to APMg. Mosquitoes were also screened for the presence of kdr mutations. The results revealed no mutation in domain II while the presence of mutation F1534C in domain III was found in both field populations. The results showed the presence of moderate to high grade resistance against all insecticides in Ae. aegypti in district Lahore and district Muzaffargarh of Punjab, Pakistan.

Nanoformulations and their mode of action in insects: a review of biological interactions.

While nanoparticles (NPs) can be used as insecticides by themselves, they can also be carriers for insecticidal chemicals. Existing literature suggests that the smaller the NP size, the greater the toxicity and penetration into the insect's body. Nonetheless, there is a lack of literature pertaining to the mode of action within insects. This review article summarizes the currently available entomological studies on the mechanisms of NP-insect interactions. Externally, NPs affect pigmentation and integrity of the cuticle, while internally they induce immune responses and alter gene expression leading to altered protein, lipid, and carbohydrate metabolism along with cellular toxicity that impairs development and reproduction of the insect. Consequently, insects are incapacitated due to the disruption of the nutrient intake, production of reactive oxygen species and altered biochemical activity while some NPs can promote growth and development as well as diminish the effects of nontarget toxicity.

Association of Interleukin 10 (IL-10) Gene with Type 2 Diabetes Mellitus by Single Nucleotide Polymorphism of Its Promotor Region G/A 1082.

Interleukin 10 (IL-10) is an anti-inflammatory cytokine with lower circulating levels in patients with type 2 diabetes mellitus (T2DM). Cytokines such as IL-10 downregulate the production of pro-inflammatory cytokines, which impair proper function of insulin. So any mutation in the IL-10 gene results in increased production of proinflammatory cytokines, which in turn affect insulin action and cause T2DM. In this study, a polymorphism (rs1800896) in the gene (IL-10) and its association with T2DM was determined with the amplification-refractory mutation system with PCR (ARMS-PCR). Study subjects were divided in two groups, control and T2DM. DNA was extracted from blood, and ARMS-PCR was performed by using specific primers for the promoter region. An amplified product was obtained at 258 base pairs (bp). In the control group, heterozygous bands with both alleles (A/G) were present, whereas AA and GG homozygosity was observed in the T2DM group. This polymorphism (rs1800896) in the IL-10 gene is associated with T2DM. Physical measurements were also obtained, and significant differences between the groups were determined with an independent t-test. Significance was based on a p-value level of 0.05 or less, and highly significant was based on a p-value of 0.01 or less at 95% confidence interval.

Assessment of abnormalities in lipid profiles of patients with chronic kidney disease from different hospitals of Lahore, Pakistan: A case control study.

The current study aimed to evaluate lipid profile parameters in patients with chronic kidney disease (CKD). This case-control study was conducted at the Zoology Department of Lahore College for Women University, Lahore, Pakistan, from September 2016 to June 2017 and analysed lipid profiles of patients with CKD attending different hospitals of Lahore. Of the randomly collected 100 blood samples, 50 were healthy, while the remaining 50 were patients with CKD. Lipid profile parameters were analysed using Hitachi 704 Analyser. For data analysis Chi-square test was used by means of SSPS 17. Results showed that all the parameters, including high density lipoprotein, triglycerides and low density lipoprotein were considerably altered in patients with CKD as compared to the control group. The values of triglycerides and LDL parameters were higher, while HDL cholesterol was lower in patients with CKD as compared to controls.

Expression of Gstt1 in Type 2 Diabetic Retinopathy Patients.

Type 2 diabetes mellitus (T2DM) is the most common metabolic and multifactorial disease in which both genetic and environmental factors are involved. In diabetes, the defects in the cellular metabolism result in higher levels of free radicals. These radicals react with other vital cellular molecules, which are responsible in diabetes side effects and known as diabetic retinopathy (DR), a disease of the retina that results in impairment or loss of vision. In the present study, we investigated the expression of glutathione S-transferases class theta 1 (GSTT1) in type 2 DR subjects. Highly significant differences (P ≤ 0.05) were observed in GSST1 expression in DR patients compared with diabetic and control groups.

Evaluation of Fish Health Status and Histopathology in Gills and Liver Due to Metal Contaminated Sediments Exposure.

Health status of freshwater fish, Cirrhina mrigala was studied by qualitative and quantitative histopathological analysis, alterations in frequency/prevalence percentages (%) and histological alteration indices (HAI) in response to metal contaminated sediments of the River Ravi aquatic ecosystem. Histo-structures of gill and liver samples of fish were analyzed and comparison between the degree of damage of the alterations in fish organs was performed after exposure to metal contaminated sediments for 7, 14 and 28 days under semi-static water renewal bioassays. Histopathological studies revealed marked histological alterations in the gills and liver of exposed fish as compared to normal tissue structure observed in control fish. The frequency and prevalence percentages observed in 28-day exposed fish were significantly higher as compared to 7- and 14-day exposed fish. The order of frequency and prevalence percentage for gills and liver of exposed fish was as: 28-day > 14-day > 7-day. The highest prevalence percentages recorded were 83 and 80% as focal area of necrosis in gill and liver, respectively, after 28-day exposure. The lowest prevalence percentage observed in 7-day exposed Cirrhina mrigala was dilation of sinusoids (17%).

Assessment of infection control practices in teaching hospitals of Quetta.

OBJECTIVE: To identify the gaps in infection control and prevention practices in teaching hospitals. METHODS: This cross-sectional study was conducted at Bolan Medical Complex and Sandeman Medical College Hospital, Quetta, from August 2012 to January 2013.The study comprised members (n=7) of infection control committee who were interviewed through a self-developed, closed-ended questionnaire and their perception regarding infection control and prevention was recorded. Data was analysed using SPSS 16. RESULTS: Only 3(42.9%) of the committee members believed that the administrative factors for causing hospital-acquired infections were nurse-patient ratio. On the patient care side, 1(14.3%) participants at one of the hospitals attributed infections to antibiotic use, 5(71.4%) to invasive medical device and 1(14.3%) to other factors. CONCLUSIONS: Poor perception held by the members of infection control committee was the basic cause of bad outcome. Capacity-building of all the stakeholders is required.

Analysis of Insect toxicity and repellent activity of Phytochemicals from "Skimmia laureola, Nair" against "Black garden ant, Lasius niger" of Pakistan.

A study was conducted to evaluate the toxicity and repellency of essential oils from root, stem and leaves of Nazar panra, Skimmia laureola (DC.) Zucc. Ex Walp. of family (Sapindales: Rutaceae) ver. Nair of Pakistan. The oils were tested at three concentrations i.e. 1, 5 and 10%. Black garden ant, Lasius niger L. (Hymenoptera: Formicidae) of Pakistan were selected and exposed to essential oils at room temperature. All essential oils showed Insecticidal activity with LC(50)=10.15, while dose dependant effect was significant with R(2)=0.98. It can be concluded that the three Essential oils in this study have both Insecticidal as well as repellent effect.

Assessing arthropod biodiversity with DNA barcoding in Jinnah Garden, Lahore, Pakistan.

Previous difficulties in arthropod taxonomy (such as limitations in conventional morphological approaches, the possibility of cryptic species and a shortage of knowledgeable taxonomists) has been overcome by the powerful tool of DNA barcoding. This study presents a thorough analysis of DNA barcoding in regards to Pakistani arthropods, which were collected from Lahore's Jinnah Garden. The 88 % (9,451) of the 10,792 specimens that were examined were able to generate DNA barcodes and 83% (8,974) of specimens were assigned 1,361 barcode index numbers (BINs). However, the success rate differed significantly between the orders of arthropods, from 77% for Thysanoptera to an astounding 93% for Diptera. Through morphological exams, DNA barcoding, and cross-referencing with the Barcode of Life Data system (BOLD), the Barcode Index Numbers (BINs) were assigned with a high degree of accuracy, both at the order (100%) and family (98%) levels. Though, identifications at the genus (37%) and species (15%) levels showed room for improvement. This underscores the ongoing need for enhancing and expanding the DNA barcode reference library. This study identified 324 genera and 191 species, underscoring the advantages of DNA barcoding over traditional morphological identification methods. Among the 17 arthropod orders identified, Coleoptera, Diptera, Hemiptera, Hymenoptera, and Lepidoptera from the class Insecta dominated, collectively constituting 94% of BINs. Expected malaise trap Arthropod fauna in Jinnah Garden could contain approximately 2,785 BINs according to Preston log-normal species distribution, yet the Chao-1 Index predicts 2,389.74 BINs. The Simpson Index of Diversity (1-D) is 0.989, signaling high species diversity, while the Shannon Index is 5.77, indicating significant species richness and evenness. These results demonstrated that in Pakistani arthropods, DNA barcoding and BOLD are an invaluable tool for improving taxonomic understanding and biodiversity assessment, opening the door for further eDNA and metabarcoding research.

Evaluation of cyto-genotoxicity biomarkers, changes in histology and antioxidant defense system of Oreochromis niloticus induced by the industrial effluents.

Aquatic pollution mainly by industrial effluents has been a major concern since a few decades. The current study evaluated cyto-genotoxicity of industrial effluents on Oreochromis niloticus exposed to sublethal levels by hematotoxicity, blood biochemistry analysis, micronucleus assay, antioxidants and cerebral toxicity. The significant elevation in differential leukocytes of exposed fish was indicative of infections and compromised immune system. The acute and chronic industrial effluent exposure caused significant decline in aspartame transaminase (AST) and alanine transaminase (ALT) and renal function enzymes. Necrosis, hyperplastic growth, hypertrophy and toxicant accumulation exhibited cerebral toxicity potential of industrial toxicants. A significant decrease in antioxidants, GSH, SOD and catalase (0.14, 0.66 and 1549 unit/mg protein) in chronic exposure group in comparison to 0.18, 2.83, 7680 and 6200.8 values of GSH, SOD, GPx and CAT, respectively. Results showed that acute and chronic industrial effluent exposure caused genotoxicity with higher frequencies of formation of micronuclei and cytokaryotic fusion.

Association of adiponectin gene expression with atrial fibrillation in a Pakistani populace.

Adiponectin, an adipocytokine produced and secreted by adipose tissue, has anti-diabetic, anti-atherogenic, and anti-inflammatory properties. This case-control study was aimed to assess the expression and serum levels of adiponectin in subject suffereing from atrial fibrillation (AF). The study's subjects (n = 690) were enrolled from the Punjab Institute of Cardiology, Lahore and were grouped into control, AF without Metabolic syndrome (MetS), and AF with MetS groups. Along with the collection of demographic data, an analysis of adiponectin and biochemical parameters were performed. A highly significant difference in serum levels of adiponectin was observed among the control, AF without MetS, and AF with MetS groups (61.61 ± 45.30 ng/ml, 37.20 ± 19.46 ng/ml, 63.78 ± 61.69 ng/ml). The expression analysis of adiponectin was decreased (n-fold = Ì´ 0.30) in AF without MetS group as compared to control group (n-fold = ~ 1.16) but increased in AF with MetS group (n-fold = Ì´ 6.26). The correlation analysis revealed a highly significant positive relationship between the expression of the adiponectin gene with waist-to-hip ratio (WHR) in AF without MetS group. Whereas, serum adiponectin was negatively related to serum triglycerides (TG) in AF with MetS group. In multiple regression analysis using adiponectin expression as the dependent variable, WHR was a determinant in AF without MetS. Whereas, when serum adiponectin was used as the dependent variable, serum TG was the determinant in group AF with MetS. The present study implicates that decreased expression and serum levels of adiponectin were associated with the development of AF in which WHR and serum TG also contributed towards the onset of atrial fibrillation.

Heavy metal toxicity arising from the industrial effluents repercussions on oxidative stress, liver enzymes and antioxidant activity in brain homogenates of Oreochromis niloticus.

Industrial effluents reaching to the aquatic ecosystem is one of the major causes of environmental pollution and exposure to industrial effluents containing harmful substances may be a serious threat to human health. Therefore, the present study aimed to determine the sub-lethal (1/5th of predetermined LC50) impact of industrial effluents from Sundar Industrial Estate on Oreochromis niloticus with proper negative control. The physicochemical analysis of industrial effluents showed enormous loads of inorganic pollutants and exhibited high mean levels of heavy metals, Mn, Fe, Pb, Ni, Cr, Hg, As, Zn and Fe with statistically significant differences at p < 0.05. Highest level of Mn and Fe was detected in effluent's samples as 147.36 ± 80.91 mg/L and 90.52 ± 32.08 mg/L, respectively. Exposure led to increase in serum biochemical parameters alanine aminotransferase + 25%, aspartate aminotransferase + 20% and alkaline phosphatase + 7% over control although superoxide dismutase, catalase, glutathione peroxidase and reduced glutathione significantly increased as 3.42, 2.44, 4.8 and 8 folds, respectively in metabolically active tissue brain which indicated stress caused by industrial effluents. The results concluded that industrial effluent has potent oxidative stress inducers on one hand whereas histoarchitectural and physiological toxicity causing contaminants on the other. This condition may adversely affect the health of aquatic organisms, the fish and ultimately the human beings.

Correction: Mutational analysis in sodium-borate cotransporter SLC4A11 in consanguineous families from Punjab, Pakistan.

[This corrects the article DOI: 10.1371/journal.pone.0273685.].

Mutational analysis in sodium-borate cotransporter SLC4A11 in consanguineous families from Punjab, Pakistan.

AIM: To identify the molecular basis of Congenital Hereditary Endothelial Dystrophy CHED caused by mutations in SLC4A11, in the consanguineous Pakistani families. METHODS: A total of 7 consanguineous families affected with Congenital Hereditary Endothelial Dystrophy were diagnosed and registered with the help of ophthalmologists. Blood samples were collected from affected and unaffected members of the enrolled families. Mutational analysis was carried out by DNA sequencing using both Sanger and Whole Exome Sequencing (WES). Probands of each pedigree from the 7 families were used for WES. Results were analyzed with the help of different bioinformatics tools. RESULTS: The sequencing results demonstrated three known homozygous mutations in gene SLC4A11 in probands of 7 families. These mutations p.Glu675Ala, p.Val824Met, and p.Arg158fs include 2 missense and 1 frameshift mutation. The mutations result in amino acids that were highly conserved in SLC4A11 across different species. The mutations were segregated with the disease phenotype in the families. CONCLUSION: This study reports 3 mutations in 7 families. One of the pathogenic mutations (p.R158fs) was identified for the first time in the Pakistani population. However, two mutations (p.Glu675Ala, p.Val824Met) were previously reported in two and one Pakistani family respectively. As these mutations segregate with the disease phenotype and bioinformatics tool also liable them as pathogenic, they are deemed as probable cause of underlying disease.

A DNA barcode survey of insect biodiversity in Pakistan.

Although Pakistan has rich biodiversity, many groups are poorly known, particularly insects. To address this gap, we employed DNA barcoding to survey its insect diversity. Specimens obtained through diverse collecting methods at 1,858 sites across Pakistan from 2010-2019 were examined for sequence variation in the 658 bp barcode region of the cytochrome c oxidase 1 (COI) gene. Sequences from nearly 49,000 specimens were assigned to 6,590 Barcode Index Numbers (BINs), a proxy for species, and most (88%) also possessed a representative image on the Barcode of Life Data System (BOLD). By coupling morphological inspections with barcode matches on BOLD, every BIN was assigned to an order (19) and most (99.8%) were placed to a family (362). However, just 40% of the BINs were assigned to a genus (1,375) and 21% to a species (1,364). Five orders (Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera) accounted for 92% of the specimens and BINs. More than half of the BINs (59%) are so far only known from Pakistan, but others have also been reported from Bangladesh (13%), India (12%), and China (8%). Representing the first DNA barcode survey of the insect fauna in any South Asian country, this study provides the foundation for a complete inventory of the insect fauna in Pakistan while also contributing to the global DNA barcode reference library.

Biomarkers of Metabolic Syndrome: Role in Pathogenesis and Pathophysiology Of Atrial Fibrillation.

The relationship between Metabolic syndrome and Atrial Fibrillation is confirmed by many studies. The components of Metabolic syndrome cause remodeling of the atrial. Metabolic syndrome and metabolic derangements of the syndrome could be the cause of the pathogenesis of AF. This review article discusses the major biomarkers of Metabolic syndrome and their role in the pathogenesis of AF. The biomarkers are adiponectin, leptin, Leptin/ Adiponectin ratio, TNF-α, Interleukin-6, Interleukin-10, PTX3, ghrelin, uric acid, and OxLDL.The elevated plasma levels of adiponectin were linked to the presence of persistent AF. Leptin signaling contributes to angiotensin-II evoked AF and atrial fibrosis. Tumor necrosis factor-alpha involvement has been shown in the pathogenesis of chronic AF. Similarly, Valvular AF patients showed high levels of TNF-α. Increased left atrial size was associated with the interleukin-6 because it is a well-known risk factor for AF. Interleukin-10 as well as TNF-α were linked to AF recurrence after catheter ablation. PTX3 could be superior to other inflammatory markers that were reported to be elevated in AF. The serum ghrelin concentration in AF patients was reduced and significantly increased after treatment. Elevated levels of uric acid could be related to the burden of AF. Increased OxLDL was found in AF as compared to sinus rhythm control.

Recovery of E. coli From Liver and Spleen of Broiler Birds and the Effects of Induced High Ammonia Level on Haematobiochemical Parameters and Its Amelioration by Different Modifiers.

The poultry sector is one of the most vibrant segments of the agriculture industry of Pakistan. In addition to different infections, ammonia (NH3) production from litter material of broiler is the most harmful pollutant and causes serious threats for the environment. To overcome this problem, different methods are proposed assuring poultry bird's health and production. This study was carried out to evaluate the effect of toxic levels of NH3 on the haematology and serum proteins of broiler birds and its amelioration by using different modifiers. The recovery of Escherichia coli (E. coli) from liver and spleen of broiler birds was also carried out. A total of 100 birds were divided into 5 separate groups (groups A-D). The groups C, D and E were treated with potassium aluminium sulphate, aluminium silicate and Yucca schidigera plant extract, respectively. Blood and tissue samples were collected after slaughtering the birds at 42 days of age. This study revealed increased RBC, total leucocyte count, Hb and heterophils percentage. Serum proteins were decreased in Yucca-treated and potassium aluminium sulphate-treated groups. This study concluded that NH3 production was reduced by the application of different modifiers, and these modifiers also neutralized the changes in blood parameters induced by NH3.

Analysis of the growth performance, stress, profile of fatty acids and amino acids and cortisol in Tilapia (Oreochromis niloticus), cultured at high stocking density using in-pond raceway system.

In-pond raceway system technology (IPRS) was introduced in Pakistan in 2019 as solution for sustainable aquaculture approach by effectively increasing production, reducing pollution and facilitating feed and pond management. Fingerlings of GIFT Tilapia (Oreochromis niloticus) (n = 16,500 in each raceway, initial weight = 32.00 ± 1.26 g) were stocked in June 2019 in two IPRS raceways (area of each raceway = 220 m3) for 171 days until harvested on November 30, 2019. Fingerlings stocked in traditional earthen ponds (area of each pond = 6167 m3) were studied as control (n = 3000 in each pond, initial weight = 32.00 ± 1.26 g). Average harvested biomass from raceways was 57.33 kg/m3 with an average daily weight gain per fish of 4.47 g per day. On the other hand, average harvested biomass from control ponds was 0.38 kg/m3 with an average daily weight gain per fish of 4.60 g per day. Average feed conversion ratio (FCR) in both raceways and control ponds was recorded as 1.25 and 1.24, respectively. Overall survival rate in both raceways and control ponds was above 99%. No sign of any disease was noted at any stage in both study groups. Crude protein and fats contents did not reduce in any raceway despite of high stocking density and sharp seasonal changes. Profile of essential and non-essential amino acids were found to be upto nutritional requirements of adult human. High levels of n-3 and n-6 fatty acids in fish collected from raceways as compared to those in traditional earthen pond show that muscle quality was not compromised due to high stocking density in small area. Return on investment excluding capital cost was 47.05 which implies that IPRS technology can be economically feasible with further modifications.