Detection of carbapenemases blaOXA48-blaKPC-blaNDM-blaVIM and extended-spectrum-ß-lactamase blaOXA1-blaSHV-blaTEM genes in Gram-negative bacterial isolates from ICU burns patients.

BACKGROUND AND OBJECTIVES: Burn patients are highly susceptible to invasion by multidrug-resistant Gram-negative bacteria (MDR-GNB) through post-burn damage. The prevalence of MDR-GNB isolated from burns patients has increased dramatically in the last decade, representing a serious risk to patients admitted to burns units worldwide. The challenges of managing infected burns patients are exacerbated in poor resource settings. This study was designed to develop a pathway for the rapid diagnosis of multidrug-resistant (MDR) Gram-negative infections and identify the bacterial genes including blaOXA1, blaTEM, and blaSHV encoding ESBLs and blaOXA48, blaKPC, blaNDM, and blaVIM encoding carbapenemases from the patient of post burns infection.  METHODS: Clinical isolates were collected (August 2017 to August 2018) from Intensive care unit (ICU) of Burn Centre. Antibiotic susceptibility testing and phenotypic detection of ESBLs and carbapenemases was performed by disk diffusion, double disk synergy test (DDST), combination disk test (CDT), and Imipenem + EDTA combined disk test (IMP + EDTA CDT). Polymerase chain reaction (PCR) detection was performed for ESBLs blaOXA1-blaSHV-blaTEM and carbapenemases genes blaOXA48-blaKPC-blaNDM-blaVIM RESULTS: In total, of 170 Gram-negative isolates, 104 (61.2%) were confirmed as multidrug-resistant (MDR); Pseudomonas aeruginosa was found to be the most prevalent 43/104 (41.4%), followed by Klebsiella pneumoniae 17/104 (16.4%), Acinetobacter baumannii12/104 (11.5%), and 6/104 Proteus mirabilis (5.8%). All isolates (100%) were resistant to cefotaxime and ceftazidime, while the meropenem resistance was 58.7%. ESBL and carbapenemase genotypes were found to be associated with higher MAR index (0.65-0.88) and MIC (> 32 µg/ml) values P. aeruginosa was the major ESBL and carbapenemase producer as determined by phenotypic testing and PCR. blaTEM positive isolates among ESBLs producers were predominant 81.8% (27/33), followed by 27.3% blaOXA1 and blaSHV, respectively. blaVIM positive isolates among carbapenemase producers were predominant 47.7% (21/44), followed by 27.3% blaKPC, 20.5% blaOXA48, and 11.4% blaNDM positive isolates. CONCLUSIONS: The predominant organism causing burn infections was ESBL and carbapenemase-producing Pseudomonas aeruginosa. There are only limited effective antibiotics against such strains. blaVIM and blaTEM individually and in co-existence with blaKPC, blaOXA48, blaSHV, and blaOXA1 confer antimicrobial resistance in burns patients. Rapid detection of ESBL and carbapenemase genes will inform treatment strategies improving the outcome for post-burn patients in ICU.

A quasi experimental study to compare thermo-regulator blanket with conventional method (incubator) for temperature regulations in preterm, low birth weight neonates landing at emergency department of a tertiary care paediatric facility.

Objective: To compare incubator and thermal blanket for thermoregulation in preterm and low birth weight neonates. METHODS: The pilot study was conducted from January to March 2018 at the emergency department of the National Institute of Child Health, Karachi, and comprised preterm and low birth weight neonates without any co-morbidity who were randomised to control and interventional groups. Data was collected using a proforma about demographics, weight, gestational age and brief clinical history. Temperature, blood pressure and pulse were monitored along with other routine care after every 15 minutes in the incubator, and the same procedure was adopted for thermal blankets. Data was analysed using Statistical Package for Social Sciences (SPSS) Version 23. RESULTS: Of the 38 subjects, 19(50%) each were cases and controls. The mean gestational age between the groups was not significantly different (p>0.05). Temperature for the first 60 minutes remained similar in the two groups, but at 75 minutes and onwards, the incubator showed a significantly better results (p<0.05). Conclusion: Thermal blankets can sufficiently provide a hyperthermic environment to the neonates, but an incubator is a more reliable option to provide sustained temperature.

Antioxidant properties of Milk and dairy products: a comprehensive review of the current knowledge.

Milk and dairy products are integral part of human nutrition and they are considered as the carriers of higher biological value proteins, calcium, essential fatty acids, amino acids, fat, water soluble vitamins and several bioactive compounds that are highly significant for several biochemical and physiological functions. In recent years, foods containing natural antioxidants are becoming popular all over the world as antioxidants can neutralize and scavenge the free radicals and their harmful effects, which are continuously produced in the biological body. Uncontrolled free radicals activity can lead to oxidative stresses, which have been implicated in breakdown of vital biochemical compounds such as lipids, protein, DNA which may lead to diabetes, accelerated ageing, carcinogenesis and cardiovascular diseases. Antioxidant capacity of milk and milk products is mainly due to sulfur containing amino acids, such as cysteine, phosphate, vitamins A, E, carotenoids, zinc, selenium, enzyme systems, superoxide dismutase, catalase, glutathione peroxidase, milk oligosaccharides and peptides that are produced during fermentation and cheese ripening. Antioxidant activity of milk and dairy products can be enhanced by phytochemicals supplementation while fermented dairy products have been reported contained higher antioxidant capacity as compared to the non-fermented dairy products. Literature review has shown that milk and dairy products have antioxidant capacity, however, information regarding the antioxidant capacity of milk and dairy products has not been previously compiled. This review briefly describes the nutritional and antioxidant capacity of milk and dairy products.

Connective tissue growth factor contributes to joint homeostasis and osteoarthritis severity by controlling the matrix sequestration and activation of latent TGFß.

OBJECTIVES: One mechanism by which cartilage responds to mechanical load is by releasing heparin-bound growth factors from the pericellular matrix (PCM). By proteomic analysis of the PCM, we identified connective tissue growth factor (CTGF) and here investigate its function and mechanism of action. METHODS: Recombinant CTGF (rCTGF) was used to stimulate human chondrocytes for microarray analysis. Endogenous CTGF was investigated by in vitro binding assays and confocal microscopy. Its release from cut cartilage (injury CM) was analysed by Western blot under reducing and non-reducing conditions. A postnatal, conditional CtgfcKO mouse was generated for cartilage injury experiments and to explore the course of osteoarthritis (OA) by destabilisation of the medial meniscus. siRNA knockdown was performed on isolated human chondrocytes. RESULTS: The biological responses of rCTGF were TGFß dependent. CTGF displaced latent TGFß from cartilage and both were released on cartilage injury. CTGF and latent TGFß migrated as a single high molecular weight band under non-reducing conditions, suggesting that they were in a covalent (disulfide) complex. This was confirmed by immunoprecipitation. Using CtgfcKO mice, CTGF was required for sequestration of latent TGFß in the matrix and activation of the latent complex at the cell surface through TGFßR3. In vivo deletion of CTGF increased the thickness of the articular cartilage and protected mice from OA. CONCLUSIONS: CTGF is a latent TGFß binding protein that controls the matrix sequestration and activation of TGFß in cartilage. Deletion of CTGF in vivo caused a paradoxical increase in Smad2 phosphorylation resulting in thicker cartilage that was protected from OA.

A discoidin domain receptor 1 knock-out mouse as a novel model for osteoarthritis of the temporomandibular joint.

Discoidin domain receptor 1 (DDR-1)-deficient mice exhibited a high incidence of osteoarthritis (OA) in the temporomandibular joint (TMJ) as early as 9 weeks of age. They showed typical histological signs of OA, including surface fissures, loss of proteoglycans, chondrocyte cluster formation, collagen type I upregulation, and atypical collagen fibril arrangements. Chondrocytes isolated from the TMJs of DDR-1-deficient mice maintained their osteoarthritic characteristics when placed in culture. They expressed high levels of runx-2 and collagen type I, as well as low levels of sox-9 and aggrecan. The expression of DDR-2, a key factor in OA, was increased. DDR-1-deficient chondrocytes from the TMJ were positively influenced towards chondrogenesis by a three-dimensional matrix combined with a runx-2 knockdown or stimulation with extracellular matrix components, such as nidogen-2. Therefore, the DDR-1 knock-out mouse can serve as a novel model for temporomandibular disorders, such as OA of the TMJ, and will help to develop new treatment options, particularly those involving tissue regeneration.

Influence of flaxseed-derived diglyceride-based high internal phase Pickering emulsions on the rheological and physicochemical properties of myofibrillar protein gels.

The study aimed to investigate the influence of flaxseed-derived diglyceride-based high internal phase Pickering emulsions (HIPPE) at different levels (0%, 10%, 20%, 30%, 40%, and 50%) on the rheological and physicochemical properties of myofibrillar protein (MPs) gels. The study indicated that with increasing HIPPE levels, there was a significant increase in whiteness while a decrease in water-holding capacity. The gels with 10% HIPPE levels had higher ionic bonds, while those with 40% and 50% HIPPE levels showed higher hydrogen bonds. By increasing HIPPE levels in the formation of MP gels, the T2 relaxation time was found to decrease. Additionally, in all MP gels, G' values were significantly higher than G" values over time. Adding lower contents of HIPPE levels resulted in a more compact microstructure. These findings indicate that flaxseed-derived diglyceride-based HIPPEs could be utilized as fat substitutes in meat products to enhance their nutritional quality.

Effect of flaxseed-derived diglyceride-based high internal phase Pickering emulsion on the quality characteristics of reformulated beef burgers.

The study aimed to fabricate healthier beef burgers using high internal phase Pickering emulsion (HIPPE) as animal fat substitute. In this context, HIPPE stabilized by modified soy protein isolates was produced with flaxseed-derived diglycerides (DAGs). Beef burgers were prepared by substituting beef backfat with HIPPE at varying levels (0%, 25%, 50%, 75%, and 100%). Reformulated burgers showed a significant decrease in WHC (from 89.75 to 77.38%), pH (from 5.73 to 5.58), L* values (from 53.5 to 44.5), and b* values (22.9 to 21.8), while a significant increase in a* values (from 24.4 to 6.7), cooking loss (from 20.25 to 30.62), and cooking shrinkage (from 11.27 to 13.05). Texture attributes, including hardness, chewiness, and gumminess, decreased up to 50% fat substitution and increased with increasing levels of fat substitution. Moreover, the rheological properties (G' and G'') and T2 relaxation time were increased with increasing fat replacement. The reformulation with HIPPE resulted in a decrease in SFA (from 3896 to 1712 mg/100 g), ω-6/ω-3 ratio (from 5.29 to 0.47), atherogenic index (from 0.57 to 0.13), and thrombogenic index (from 1.46 to 0.15) and increase in PUFA/SFA ratio (from 0.20 to 2.79). Notably, burgers with 50% fat substitution were more preferred regarding tenderness, while those with 100% fat substitution obtained higher scores for color and flavor than all other treatments. In conclusion, 50% fat replacement using flaxseed-derived diglyceride-based HIPPE improved beef burgers' textural profile and fatty acid composition without compromising the sensory characteristics.

Predictors of Preterm Neonatal Mortality in India and Pakistan: A Secondary Analysis of Data from PURPOSe Study.

Objective. To create a prediction model for preterm neonatal mortality. Methods. A secondary analysis was conducted using data from a prospective cohort study, the Project to Understand and Research Preterm Pregnancy Outcome South Asia. The Cox proportional hazard model was used and adjusted hazard ratios (AHR) with 95% confidence intervals (95% CI) were reported. Results. Overall, 3446 preterm neonates were included. The mean age of preterm neonates was 0.65 (1.25) hours and 52% were female. The preterm neonatal mortality rate was 23.3%. The maternal factors predicting preterm neonatal death was any antepartum hemorrhage, AHR 1.99 (1.60-2.47), while neonatal predictors were preterm who received positive pressure ventilation AHR 1.30 (1.08-1.57), temperature <35.5°C AHR 1.18 (1.00-1.39), and congenital malformations AHR 3.31 (2.64-4.16). Conclusion. This study identified key maternal and neonatal predictors of preterm neonatal mortality, emphasizing the need for targeted interventions and collaborative public health efforts to address disparities and regional variations.

The primary cilium as a dual sensor of mechanochemical signals in chondrocytes.

The primary cilium is an immotile, solitary, and microtubule-based structure that projects from cell surfaces into the extracellular environment. The primary cilium functions as a dual sensor, as mechanosensors and chemosensors. The primary cilia coordinate several essential cell signaling pathways that are mainly involved in cell division and differentiation. A primary cilium malfunction can result in several human diseases. Mechanical loading is sense by mechanosensitive cells in nearly all tissues and organs. With this sensation, the mechanical signal is further transduced into biochemical signals involving pathways such as Akt, PKA, FAK, ERK, and MAPK. In this review, we focus on the fundamental functional and structural features of primary cilia in chondrocytes and chondrogenic cells.

Effect of Different Packaging Atmosphere on Microbiological Shelf Life, Physicochemical Attributes, and Sensory Characteristics of Chilled Poultry Fillets.

This trial was conducted to evaluate the effect of overwrap, vacuum, and modified atmosphere packaging (MAP) on poultry breast fillets' microbiological, biochemical shelf life and sensory attributes. The fillets were divided into 4 groups, and each of the treatments was replicated 3 times with 60 breast fillets. The first group was a control group with overwrap packaging; the second group was vacuum packed (VP); the third and fourth groups were MAP-1: 0% O2, 40% CO2, 60% N2, and MAP-2: 20% O2, 40% CO2, 40% N2. The microbiological and biochemical analyses were performed for the total viable count, coliform count, Pseudomonas count, Salmonella count, total volatile basic nitrogen (TVB-N), pH, cooking loss, color, lipid oxidation, tenderness, and sensory analysis. The data were analysed through two-way ANOVA by Minitab (Minitab 17.3.1). Meat treated with understudy MAP compositions and vacuum packaging reduced total viable count, Pseudomonas count, and total coliform count than control (p<0.05). TVB-N remained below the recommended limit throughout storage except aerobic packaging (p<0.05). Cooking loss (%) was lowered and showed non-significant results (p>0.05) between vacuum packaging and both MAP concentrations. The meat stored in MAP-2 was characterised by higher (p<0.05) visual scores. Whilst MAP-1 showed higher (p<0.05) L* values and overall acceptability. Sample packaged under aerobic packaging showed significant (p<0.05) results for b* and thiobarbituric acid reactive substances (TBARS). Meat stored in aerobic packaging showed higher (p<0.05) shear force values. The outcome of this trial may help to promote the application of understudy MAP compositions and rapid detection of microbes by biochemical analysis under local conditions.

Variants in ALDH1A2 reveal an anti-inflammatory role for retinoic acid and a new class of disease-modifying drugs in osteoarthritis.

More than 40% of individuals will develop osteoarthritis (OA) during their lifetime, yet there are currently no licensed disease-modifying treatments for this disabling condition. Common polymorphic variants in ALDH1A2, which encodes the key enzyme for synthesis of all-trans retinoic acid (atRA), are associated with severe hand OA. Here, we sought to elucidate the biological significance of this association. We first confirmed that ALDH1A2 risk variants were associated with hand OA in the U.K. Biobank. Articular cartilage was acquired from 33 individuals with hand OA at the time of routine hand OA surgery. After stratification by genotype, RNA sequencing was performed. A reciprocal relationship between ALDH1A2 mRNA and inflammatory genes was observed. Articular cartilage injury up-regulated similar inflammatory genes by a process that we have previously termed mechanoflammation, which we believe is a primary driver of OA. Cartilage injury was also associated with a concomitant drop in atRA-inducible genes, which were used as a surrogate measure of cellular atRA concentration. Both responses to injury were reversed using talarozole, a retinoic acid metabolism blocking agent (RAMBA). Suppression of mechanoflammation by talarozole was mediated by a peroxisome proliferator-activated receptor gamma (PPARγ)-dependent mechanism. Talarozole was able to suppress mechano-inflammatory genes in articular cartilage in vivo 6 hours after mouse knee joint destabilization and reduced cartilage degradation and osteophyte formation after 26 days. These data show that boosting atRA suppresses mechanoflammation in the articular cartilage in vitro and in vivo and identifies RAMBAs as potential disease-modifying drugs for OA.

Effect of Packaging Type and Aging on the Meat Quality Characteristics of Water Buffalo Bulls.

The present study determined the effect of the packaging type and aging time on the meat quality of water buffalo (Bubalus bubalis) bulls. A total of n = 36 longissimus lumborum (LL) muscles from n = 18 buffalo bulls were obtained. Half LL muscles were packed in modified atmosphere packaging (Hi-O2 MAP), vacuum packaging (VP), and oxygen-permeable packaging (OP) on day 1, while the other half were aged for 7 days. Meat instrumental color, cooking loss, Warner-Bratzler shear force (WBSF), thiobarbituric acid reactive substances (TBARS), and total volatile basic nitrogen (TVB-N) of the LL steaks were analyzed, both on unaged and aged buffalo meat. Color CIE L* and C* values on all display days and a* on the first 4 days of the simulated retail display under Hi-O2 MAP packaging were significantly higher than those of the VP and OP. WBSF and TBARS values were also higher under Hi-O2 MAP as compared to the other packaging. Steaks under OP exhibited lower cooking loss but higher TVB-N values than the MAP and VP. The 7-day-aged buffalo meat indicated higher instrumental color (L*, a* and C*), cooking loss, and lower WBSF values than fresh meat. This study concluded that Hi-O2 MAP improved the color; however, it negatively influenced the buffalo meat's WBSF and TBAR values. Furthermore, VP and aging were the most effective in decreasing the WBSF values of buffalo meat.

Effect of Animal Age, Postmortem Calcium Chloride Marination, and Storage Time on Meat Quality Characteristics of M. longissimus thoracis et lumborum of Buffalo Bulls.

This study investigated the effect of animal age, calcium chloride marination, and storage time on meat quality characteristics of buffalo bulls to suggest a cost-effective method of improving buffalo meat quality. The current study was designed considering the importance of buffalo meat and the usage of meat from spent buffalo animals in local markets of South Asian countries. A total of 36 animals comprised of 18 young and 18 spent buffalo bulls were selected. After slaughtering and 24 h of postmortem chilling, striploins were separated and cut into 16 steaks and equally divided into two groups, i.e., either marinated with calcium chloride or not. Meat quality characteristics were recorded on 0, 2, 4, 6, 8, and 10 days of storage. The results showed that the pH value of young animals was higher than the value of spent animals and pH was increased over the storage time. Color b*, C*, and h* values were higher in spent animals as compared with the young animals; however, values of colors L* and h* were higher and a* was lower in marinated samples than the values of non-marinated samples. Color a* and C* values were increased and h* was decreased with lengthening the storage time. The meat cooking loss was higher in marinated and the water-holding capacity was higher in non-marinated meat samples. Shear force values were lower in young animals and marinated samples than the values of spent animals and non-marinated meat samples, respectively. Sensory characteristic scores of marinated samples were better than the non-marinated samples. In conclusion, calcium chloride marination can be used to improve the quality characteristics of buffalo meat.

Effect of Carcass Electrical Stimulation and Suspension Methods on Meat Quality Characteristics of Longissimus lumborum of Young Buffalo (Bubalus bubalis) Bulls.

Buffalo animals are slaughtered at their early age and carcasses are chilled rapidly which deteriorates its meat quality and decreases the consumer likeliness of buffalo meat. This study investigated the appropriate methods to prevent the quality deterioration of buffalo meat during chilling. Twenty four 18-mon-old buffalo bulls were slaughtered, electrically stimulated and suspended either by hip or achilles tendon. After 24 h postmortem, meat quality characteristics were recorded. Results showed that electrical stimulation (ES) led to rapid decline of carcass pH compared to non-ES method (p<0.05). Furthermore, electrically stimulated meat presented lower shear force accompanied with the higher CIE L*, a*, and b* values (p<0.05). Suspension methods only affect the meat shear values and were lowered in hip suspended samples. It can be concluded that ES combined with hip suspension can be adopted to prevent the meat quality deterioration of young buffalo bulls during postmortem storage.

Effect of Wet Aging on Color Stability, Tenderness, and Sensory Attributes of Longissimus lumborum and Gluteus medius Muscles from Water Buffalo Bulls.

The present study aimed to investigate the effect of wet aging on meat quality characteristics of Longissimus lumborum (LL) and Gluteus medius (GM) muscles of buffalo bulls. Meat samples from six aging periods, i.e., 0 day (d) = control, 7 d, 14 d, 21 d, 28 d, and 35 d, were evaluated for pH, color, metmyoglobin content (MetMb%), cooking loss, water holding capacity (WHC), myofibrillar fragmentation index (MFI), Warner-Bratzler shear force (WBSF), and sensory evaluation. The pH, instrumental color redness (a *), yellowness (b *), chroma (C *), and MetMb% values were increased, while the lightness (L *) and hue angle (h *) values showed non-significant (p > 0.05) differences in both LL and GM muscles in all aging periods. The cooking loss increased while WHC decreased till 35 days of aging. MFI values significantly (p < 0.05) increased, while WBSF values decreased; in addition, sensory characteristics were improved with the increase in the aging period. Overall, the color, tenderness, and sensory characteristics were improved in LL and GM muscles until 28 and 21 days of aging, respectively. Based on the evaluated meat characteristics, 28 days of aging is required to improve the meat quality characteristics of LL, whereas 21 days of aging is suitable for GM muscle.

Meat Production and Supply Chain Under COVID-19 Scenario: Current Trends and Future Prospects.

The COVID-19 pandemic impacted meat production, supply chain, and meat prices that caused a severe socio-economic crisis worldwide. Initially, meat and meat products' prices increased due to less production and increased demand because of panic buying. Whereas, later on, both meat production and demand were significantly decreased due to lockdown restrictions and lower purchasing power of the consumers that results in a decrease in meat prices. In early April 2020, meat packing facilities started to shut down due to the rapid spread of the COVID-19 virus among workers. Furthermore, meat producers and processors faced difficulty in harvesting and shipment of the products due to lockdown situations, decrease in labor force, restrictions in movement of animals within and across the country and change in legislation of local and international export market. These conditions adversely impacted the meat industry due to decrease in meat production, processing and distribution facilities. It is suggested that the integration among all the meat industry stakeholders is quite essential for the sustainability of the industry's supply chain to cope with such devastating conditions the future may hold. This review aimed to discuss different aspects of the meat industry and supply chain during the COVID-19 pandemic and proposed some future directions.

Effect of lactic acid spray on microbial and quality parameters of buffalo meat.

We evaluated the effect of lactic acid spray on micro-flora, instrumental color, shelf-life and sensory attributes of buffalo meat displayed under Modified Atmosphere Packaging. Buffalo calf carcasses (n = 12) were sliced into equal sagittal halves, n = 6 halves were randomly assigned to each of four treatments i.e. 2% LA, 4% LA, 6% LA and control. Afterwards, sirloin and tenderloin were vacuum packed and aged for 7 days. Later, steaks were packed in high-oxygen MAP. Microbial load, instrumental color, shelf-life and sensory attributes were evaluated at different days. Aerobic plate count of sprayed carcass and steaks was significantly lower than un-sprayed control. Similarly, though non-significant, redness and chroma value of sprayed carcass meat was found better than un-sprayed control. Lactic acid sprayed meat did not differ in terms of sensory attributes. It is concluded that spraying buffalo carcasses with 2-4% lactic acid after slaughter not only enhances microbial quality but it may also improve its instrumental color.

Radicalization Leading to Violence: A Test of the 3N Model.

The present research examines the social cognitive processes underlying ideologically-based violence through the lens of the 3N model of radicalization. To test this theory, we introduce two new psychometric instruments-a social alienation and a support for political violence scale-developed in collaboration with 13 subject matter experts on terrorism. Using these instruments, we test the theory's hypotheses in four different cultural settings. In Study 1, Canadians reporting high levels of social alienation (Need) expressed greater support for political violence (Narrative), which in turn positively predicted wanting to join a radical group (Network), controlling for other measures related to political violence. Study 2a and 2b replicated these findings in Pakistan and in Spain, respectively. Using an experimental manipulation of social alienation, Study 3 extended these findings with an American sample and demonstrated that moral justification is one of the psychological mechanisms linking social alienation to supporting political violence. Implications and future directions for the psychology of terrorism are discussed.

High frequency and molecular epidemiology of metallo-ß-lactamase-producing gram-negative bacilli in a tertiary care hospital in Lahore, Pakistan.

Background: Metallo-ß-lactamase (MBL)-producing isolates have a strong impact on diagnostic and therapeutic decisions. A high frequency of MBL-producing gram-negative bacilli has been reported worldwide. The current study was based on determining the incidence of MBL-producing imipenem-resistant clinical isolates and investigating the ß-lactamase gene variants in strains conferring resistance to a carbapenem drug (imipenem). Methods: A total of 924 gram negative isolates were recovered from a tertiary care hospital in Lahore, Pakistan, during a two-year period (July 2015 to February 2017). The initial selection of bacterial isolates was based on antibiotic susceptibility testing. Strains resistant to imipenem were processed for the molecular screening of ß-lactamase genes. Statistical analysis for risk factor determination was based on age, gender, clinical specimen and type of infection. Results: The rate of imipenem resistance was calculated to be 56.51%. Among the 142 strains processed, the phenotypic tests revealed that the incidence of MBLs was 63.38% and 86.61% based on the combination disc test and the modified Hodge test, respectively. The frequencies of bla TEM, bla SHV, bla OXA, bla IMP-1, and bla VIM genes were calculated to be 46%, 34%, 24%, 12.5% and 7%, respectively. The co-expression of bla MBL (bla IMP and bla VIM) and bla ESBL (bla TEM, bla SHV, bla OXA) was also detected through multiplex and singleplex PCR. bla OXA, bla TEM and bla SHV coexisted in 82% of the isolates. Co-expression of ESBL and MBL genes was found in 7% of the isolates. Conclusion: To our knowledge, this is the first report from Pakistan presenting the concomitant expression of bla OXA, bla TEM and bla SHV with bla IMP-1 and bla VIM in MBL-producing gram-negative bacilli.