Hollow-fiber liquid-phase microextraction (HF-LPME) and electromembrane extraction (EME) are miniaturized extraction techniques, and have been coupled with various analytical instruments for trace analysis of heavy metals, drugs and other organic compounds, in recent years. HF-LPME and EME provide high selectivity, efficient sample cleanup and enrichment, and reduce the consumption of organic sol-vents to a few micro-liters per sample. HF-LPME and EME are compatible with different analytical in-struments for chromatography, electrophoresis, atomic spectroscopy, mass spectrometry, and electrochemical detection. HF-LPME and EME have gained significant popularity during the recent years. This review focuses on hollow fiber based techniques (especially HF-LPME and EME) of heavy metals and pharmaceuticals (published 2017 to May 2019), and their combinations with atomic spectroscopy, UV-VIS spectrophotometry, high performance liquid chromatography, gas chromatography, capillary elec-trophoresis, and voltammetry.

AIM: To study the pattern of presentation and to highlight the common causes of primary benign orbital lesions.of Jinnah Postgraduate Medical Centre Karachi from July 1997 to August 2001 and then from September 2001 to date (Continued) at the Department of Ophthalmology of Chandka Medical College & Hospital Larkana. Only patients with primary benign orbital lesions were included in this study. All the patients were admitted in eye ward from the out patients department. The diagnosis of the disease was based on the presentation, clinical examination, investigations and histopathology of excised mass. A total of 68 patients were included in this study. The age range was from 2 months to 60 years. Out of 68, 27 (39.7%) patients were male and 41 (60.3%) were female. The left orbit was involved in 35 (51.5%) and right orbit was involved in 33 (48.5%).revealed that superficial capillary hemangiomas of the eye lid were the most common lesion 26 (38.2%) followed by deep orbital cavernous hemangiomas 5 (7.4%), lymphangiomas 5 (7.4%), orbital varices 4 (5.9%), gliomas 7 (10.3%), meningiomas 5 (7.4%), neurofibromas 5 (7.4%), neurofibromatosis 4 (5.9%), schwannomas 2 (2.9%), and pleomorphic adenoma (benign mixed cell tumor) of lacrimal gland 5 (7.4%) cases.treatment can prevent the patient from visual and life threatening complications.

AIM:To evaluate the site, size,histology,and recurrence of the tumor.METHODS: We prospectively evaluated 53 patients with histological diagnosis of retinoblastoma involving the eye, orbit, intra cranial cavity and other tissues, at Department of Ophthalmology, Chandka Medical College Hospital, Larkana. To assess the extent of the tumor investigations, including X-ray-orbits, ocular B-scan, CT-scan/MRI-Oorbits-brain, whole body bone scanning, X-ray chest PA view, ultrasound abdomen, lumbar punc-ture, complete blood cell count, blood chemistry as-sessment, including renal and hepatic function para-meters, were performed. The selection of mode of treatment like surgical procedures, local radiotherapy and systemic chemotherapy used depend on the site, size, recurrence and metastasis of the tumor. The patients with local and systemic metastases were referred to the oncologist at the Larkana Institute of the nuclear medicine and radiotherapy for local radiation and systemic chemotherapy. Follow up examinations were carried out at 1, 3 , 6 and 12 months after the operation, and then annually for a further 5 years or longer. The tumor's site, size, histological type, and recurrence were evaluated.RESULTS: Of these 53 patients the age range was from 8 months to 8 years. 25 (47%) were male and 28 (53%) were female. The laterality of the tumor was in left eye 25 (47%) cases, right eye 19 (36%) cases, and both eyes 9 (17%) cases. The presentation of the patient at 1st visit was strabismus 3 (6%) cases, pseudohypopyon 4 (8%) cases, orbital pseudo orbital inflammation 6 cases (11%), mass with metastases 7 (13%) cases, fungating mass 8 (15%) cases, leu-kocoria 12(23%) cases and orbital invasion 13(25%) cases. The surgical procedures performed were enucleation alone in 19 (36%) cases, enucleation with modified exentration followed by local radiotherapy and systemic chemotherapy in 19 (36%) cases and enucleation with total exentration followed by local radiotherapy and systemic chemotherapy in 15 (28%) cases. The histological types of the tumor were well- differentiated retinoblastoma with abundant rosettes in 25 (47%) cases and highly undifferentiated retino- blastoma 28 (53%) cases . After primary surgery, the recu-rrences and metastases of the tumors were noticed in 37 (70%) cases due to optic nerve involvement beyond the point of surgical transaction, orbital invasion and sy-stemic metastasis.CONCLUSION: Any child with red eye or white pupillary reflex should be suspected of having retinoblastoma, until proved otherwise, and should be promptly referred to an ophthalmologist, because early presentation of patient in the initial stage of the tumor will result in decreased risk of tumor extension, recurrence and mortality.

The severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),which caused the coronavirus disease 2019(COVID-19)pandemic,has affected more than 400 million people worldwide.With the recent rise of new Delta and Omicron variants,the efficacy of the vaccines has become an important question.The goal of various studies has been to limit the spread of the virus by utilizing wireless sensing technologies to prevent human-to-human interactions,particularly for healthcare workers.In this paper,we discuss the current literature on invasive/contact and non-invasive/non-contact technologies(including Wi-Fi,radar,and software-defined radio)that have been effectively used to detect,diagnose,and monitor human activities and COVID-19 related symptoms,such as irregular respiration.In addition,we focused on cutting-edge machine learning algorithms(such as generative adversarial networks,random forest,multilayer perceptron,support vector machine,extremely randomized trees,and k-nearest neighbors)and their essential role in intelligent healthcare systems.Furthermore,this study highlights the limitations related to non-invasive techniques and prospective research directions.

PURPOSE: The promoter methylation status of cell cycle regulatory genes plays a crucial role in the regulation of the eukaryotic cell cycle. CpG cytosines are actively subjected to methylation during tumorigenesis, resulting in gain/loss of function. E2F5 gene has growth repressive activities; various studies suggest its involvement in tumorigenesis. This study aims to investigate the epigenetic regulation of E2F5 in breast cancer to better understand tumor biology. METHODS: The promoter methylation status of 50 breast tumor tissues and adjacent normal control tissues was analyzed. mRNA expression was determined using SYBR® green quantitative polymerase chain reaction (PCR), and methylation-specific PCR was performed for bisulfite-modified genomic DNA using E2F5-specific primers to assess promoter methylation. Data was statistically analyzed. RESULTS: Significant (p<0.001) upregulation was observed in E2F5 expression among tumor tissues, relative to the control group. These samples were hypo-methylated at the E2F5 promoter region in the tumor tissues, compared to the control. Change in the methylation status (Δmeth) was significantly lower (p=0.022) in the tumor samples, indicating possible involvement in tumorigenesis. Patients at the postmenopausal stage showed higher methylation (75%) than those at the premenopausal stage (23.1%). Interestingly, methylation levels gradually increased from the early to the advanced stages of the disease (p<0.001), which suggests a putative role of E2F5 methylation in disease progression that can significantly modulate tumor biology at more advanced stage and at postmenopausal age (Pearson's r=0.99 and 0.86, respectively). Among tissues with different histological status, methylation frequency was higher in invasive lobular carcinoma (80.0%), followed by invasive ductal carcinoma (46.7%) and ductal carcinoma in situ (20.0%). CONCLUSION: Methylation is an important epigenetic factor that might be involved in the upregulation of E2F5 gene in tumor tissues, which can be used as a prognostic marker for breast cancer.
Biology ; Breast Neoplasms* ; Breast* ; Carcinogenesis ; Carcinoma, Ductal ; Carcinoma, Intraductal, Noninfiltrating ; Carcinoma, Lobular ; Cell Cycle ; Disease Progression ; DNA ; E2F5 Transcription Factor ; Epigenomics ; Eukaryotic Cells ; Genes, Regulator ; Humans ; Methylation* ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; RNA, Messenger ; Up-Regulation*

Limited studies on prevalence and risk factors of Premature Coronary Artery Disease (PCAD) were done in Malaysia, primarily on lipid profile. This cross sectional study aims to identify any changing patterns in prevalence and risk factors of Premature CAD between 2000 and 2012. From 2000 to 2012 we included 21862 patients who underwent the first Percutaneous Coronary Intervention (PCI). Analysis of risk factors was done to 1660 and 2098 patients from year 2007 and 2012 respectively. Age of less than 45 years was taken as PCAD. Data was collected from PCI database of National Heart Institute (NHI), NHI TrakCare System, and patients’ medical records. PCAD significantly decreased from 18.8% (2000) to 11.6% (2012). Malay ethnicity showed increasing trend over the years from 55.1% to 66.9%. Multiple logistic regression analysis in 2007 showed that smoking had the higher risk (AOR=2.52), followed by male gender (AOR=2.06), family history of PCAD (AOR=1.96), Indian ethnicity, (AOR=1.65), triglycerides level (AOR=1.20) and BMI (AOR=1.06). In 2012, family history of PCAD had the highest risk (AOR=2.00) followed by smoking (AOR=1.91) and BMI (AOR=1.11). There are changes in risk factors patterns of premature CAD between 2007 and 2012. Most of them are preventable at earlier stage.

Background: Malignant catarrhal fever (MCF) is a highly fatal lymphoproliferative disease of cattle, deer, bison, water buffalo, and pigs caused by the gamma-herpesviruses alcelaphine herpesvirus-1 (AlHV-1) and ovine herpesvirus-2 (OvHV-2). Objectives: This study aimed to determine the prevalence of OvHV-2 in sheep, goats, cattle, and buffalo in Rawalpindi and Islamabad, Pakistan, by applying molecular and phylogenetic methods. Methods: Blood samples were aspirated from sheep (n = 54), goat (n = 50), cattle (n = 46) and buffalo (n= 50) at a slaughterhouse and several farms. The samples were subjected to heminested polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis of the OvHV-2 POL gene and the OvHV-2 ORF75 tegument protein gene. Results: The highest percentage of MCF positive samples was in sheep (13%), whereas goat, cattle, and buffalo had lower positive percentages, 11%, 9%, and 6.5%, respectively. Four OvHV-2-positive PCR products obtained from sheep samples were sequenced. The sequences obtained were submitted to the NCBI GenBank database (MK852173 for the POL gene;MK840962, MK852171, and MK852172 for the ORF75 tegument protein gene). Phylogenetic analysis revealed a close similarity of study sequences with those of worldwide samples. Conclusions This study is the first cross-sectional study on the prevalence and molecular detection of OvHV-2 in apparently healthy cattle and buffalo that could be carrying OvHV-2 acquired from OvHV-2-positive sheep and goats. The results indicate that OvHV-2 is circulating in Pakistan. Further studies are needed to characterize OvHV-2 and elucidate further its prevalence.

Background: Malignant catarrhal fever (MCF) is a highly fatal lymphoproliferative disease of cattle, deer, bison, water buffalo, and pigs caused by the gamma-herpesviruses alcelaphine herpesvirus-1 (AlHV-1) and ovine herpesvirus-2 (OvHV-2). Objectives: This study aimed to determine the prevalence of OvHV-2 in sheep, goats, cattle, and buffalo in Rawalpindi and Islamabad, Pakistan, by applying molecular and phylogenetic methods. Methods: Blood samples were aspirated from sheep (n = 54), goat (n = 50), cattle (n = 46) and buffalo (n= 50) at a slaughterhouse and several farms. The samples were subjected to heminested polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis of the OvHV-2 POL gene and the OvHV-2 ORF75 tegument protein gene. Results: The highest percentage of MCF positive samples was in sheep (13%), whereas goat, cattle, and buffalo had lower positive percentages, 11%, 9%, and 6.5%, respectively. Four OvHV-2-positive PCR products obtained from sheep samples were sequenced. The sequences obtained were submitted to the NCBI GenBank database (MK852173 for the POL gene;MK840962, MK852171, and MK852172 for the ORF75 tegument protein gene). Phylogenetic analysis revealed a close similarity of study sequences with those of worldwide samples. Conclusions This study is the first cross-sectional study on the prevalence and molecular detection of OvHV-2 in apparently healthy cattle and buffalo that could be carrying OvHV-2 acquired from OvHV-2-positive sheep and goats. The results indicate that OvHV-2 is circulating in Pakistan. Further studies are needed to characterize OvHV-2 and elucidate further its prevalence.

Scabies occurs in human due to the burrowing ectoparasite Sarcoptes scabiei var. hominis resulting in intense itching and inflammation, and manifesting as a skin allergy. Limited information is available about the genetic diversity of S. scabiei in human. In this study, we characterized S. scabiei var. hominis using nuclear marker ITS-2, mitochondrial marker 16S and microsatellite markers. To examine the extent of the genetic variation, individual mite gDNA was first amplified using ITS-2, 16S and microsatellite primers by PCR, later amplicons were sequenced directly and analysed by MEGA 7. Sequence analysis of ITS- 2 showed no host segregation and geographical isolation, whereas 16S indicated presence of both hosts adapted and geographically segregated populations of S. scabiei. Results of microsatellites revealed polymorphism as allelic variability between and within the populations studied. The different varieties of Sarcoptes mites belong to different host species and geographic regions recommended that Sarcoptes mites are genetically isolated. This is the first report on the molecular characterization of S. scabiei var. hominis from Pakistan. Additionally, genetic studies including a greater number of specimens are required to comprehend the molecular epidemiology of Sarcoptes mite in Pakistan.

Purpose: Laparoscopic approach to colonic tumor requires skill set and resources to be established as routine standard of care in most centers around the world. It presents particular challenge in country like Pakistan due to economic constrain and lack of teaching and training opportunities available for surgeons to be trained to deliver such service. The aim of this study is to look into changing practice of our institution from conventional approach of open to laparoscopic surgery for right colon cancer. Methods: Consecutive patients between January 2010 to December 2018 who presented to Shaukat Khanum Memorial Cancer Hospital and Research Centre with diagnosis of right colon (cecum, ascending and transverse colon) adenocarcinoma and underwent surgical resections were included in this study. Results: A total of 230 patients with adenocarcinoma of the right colon underwent curative resections during the study period. Of these, 141 patients (61.3%) underwent laparoscopic surgery while open resection was performed in 89 patients (38.7%). Five-year disease-free survival (DFS) of patients with American Joint Committee on Cancer (AJCC) stage III (80.9% vs. 54.8%, P = 0.021) was significantly better if these patients underwent laparoscopic surgery while a trend toward better DFS (96.7% vs. 84.1%, P = 0.111) was also observed in AJCC stage II patients, although this difference was not significant. Conclusion This study demonstrates the adoption of a laparoscopic approach for right colon cancer over 10 years. With a standardized approach and using the principle of oncological surgery, we incorporated this in our minimally invasive surgery practice at our institution.