Leukocytes Classification for Leukemia Detection Using Quantum Inspired Deep Feature Selection

Leukocytes, also referred to as white blood cells (WBCs), are a crucial component of the human immune system. Abnormal proliferation of leukocytes in the bone marrow leads to leukemia, a fatal blood cancer. Classification of various subtypes of WBCs is an important step in the diagnosis of leukemia. The method of automated classification of WBCs using deep convolutional neural networks is promising to achieve a significant level of accuracy, but suffers from high computational costs due to very large feature sets. Dimensionality reduction through intelligent feature selection is essential to improve the model performance with reduced computational complexity. This work proposed an improved pipeline for subtype classification of WBCs that relies on transfer learning for feature extraction using deep neural networks, followed by a wrapper feature selection approach based on a customized quantum-inspired evolutionary algorithm (QIEA). This algorithm, inspired by the principles of quantum physics, outperforms classical evolutionary algorithms in the exploration of search space. The reduced feature vector obtained from QIEA was then classified with multiple baseline classifiers. In order to validate the proposed methodology, a public dataset of 5000 images of five subtypes of WBCs was used. The proposed system achieves a classification accuracy of about 99% with a reduction of 90% in the size of the feature vector. The proposed feature selection method also shows a better convergence performance as compared to the classical genetic algorithm and a comparable performance to several existing works

New Onset Pancytopenia in Adults: Clinico-etiological Profile at a Tertiary Care Hospital

Objective: To evaluate and compare common disorders leading to new onset pancytopenia among both genders. Methods: It was a cross-sectional study conducted at the Department of Pathology at Watim Medical College and Watim General Hospital, Rawat, from November 2020 to November 2021.A total of 98 adult patients with persistent pancytopenia for one week who gave consent for bone marrow aspiration and a trephine biopsy were selected. A blood-complete picture with peripheral smears along with bone marrow aspiration biopsy were done. Trephine needle biopsy and cytochemical staining were performed when required. Results: There were 44 males and 54 female patients. The mean age was 43.49 ±19 years. Common presenting complaints were pallor (82.6%), weakness (63.2%), petechial haemorrhages (45.9%) and fever (23.5%). The hemoglobin value, white blood cell count and platelet count were 6.5± 1.65 g/dl, 3.20 ±1.43 x 109/L and 55.21 ±28.01 x 109/L respectively. Major causes were megaloblastic anemia with female predominance (S.D: 0.003) in 43.8% patients, aplastic anemia with male predominance (S.D:0.001) in 17.3%, portal hypertension induced splenomegaly in 13.2%, infectious diseases in 7%, leukemias in 5.1% and Myelodysplastic syndromes in 3.1%. Conclusion: Megaloblastic anaemia is the major cause of pancytopenia in females and aplastic anemia in male pancytopenic patients

Molecular Genetic Studies and Delineation of the Oculocutaneous Albinism Phenotype in the Pakistani Population

Background: Oculocutaneous albinism (OCA) is caused by a group of genetically heterogeneous inherited defects that result in the loss of pigmentation in the eyes, skin and hair. Mutations in the TYR, OCA2, TYRP1 and SLC45A2 genes have been shown to cause isolated OCA. No comprehensive analysis has been conducted to study the spectrum of OCA alleles prevailing in Pakistani albino populations. Methods: We enrolled 40 large Pakistani families and screened them for OCA genes and a candidate gene, SLC24A5. Protein function effects were evaluated using in silico prediction algorithms and ex vivo studies in human melanocytes. The effects of splice-site mutations were determined using an exon-trapping assay. Results: Screening of the TYR gene revealed four known (p.Arg299His, p.Pro406Leu, p.Gly419Arg, p.Arg278*) and three novel mutations (p.Pro21Leu, p.Cys35Arg, p.Tyr411His) in ten families. Ex vivo studies revealed the retention of an EGFP-tagged mutant (p.Pro21Leu, p.Cys35Arg or p.Tyr411His) tyrosinase in the endoplasmic reticulum (ER) at 37°C, but a significant fraction of p.Cys35Arg and p.Tyr411His left the ER in cells grown at a permissive temperature (31°C). Three novel (p.Asp486Tyr, p.Leu527Arg, c.1045-15 T \u3e G) and two known mutations (p.Pro743Leu, p. Ala787Thr) of OCA2 were found in fourteen families. Exon-trapping assays with a construct containing a novel c.1045-15 T \u3e G mutation revealed an error in splicing. No mutation in TYRP1, SLC45A2, and SLC24A5 was found in the remaining 16 families. Clinical evaluation of the families segregating either TYR or OCA2 mutations showed nystagmus, photophobia, and loss of pigmentation in the skin or hair follicles. Most of the affected individuals had grayish-blue colored eyes. Conclusions: Our results show that ten and fourteen families harbored mutations in the TYR and OCA2 genes, respectively. Our findings, along with the results of previous studies, indicate that the p.Cys35Arg, p.Arg278* and p.Gly419Arg alleles of TYR and the p.Asp486Tyr and c.1045-15 T \u3e G alleles of OCA2 are the most common causes of OCA in Pakistani families. To the best of our knowledge, this study represents the first documentation of OCA2 alleles in the Pakistani population. A significant proportion of our cohort did not have mutations in known OCA genes. Overall, our study contributes to the development of genetic testing protocols and genetic counseling for OCA in Pakistani families

Delineating the Molecular and Phenotypic Spectrum of the <i>CNGA3</i>-Related Cone Photoreceptor Disorder in Pakistani Families

Cone photoreceptor dysfunction represents a clinically heterogenous group of disorders characterized by nystagmus, photophobia, reduced central or color vision, and macular dystrophy. Here, we described the molecular findings and clinical manifestations of achromatopsia, a partial or total absence of color vision, co-segregating with three known missense variants of CNGA3 in three large consanguineous Pakistani families. Fundus examination and optical coherence tomography (OCT) imaging revealed myopia, thin retina, retinal pigment epithelial cells loss at fovea/perifovea, and macular atrophy. Combination of Sanger and whole exome sequencing revealed three known homozygous missense variants (c.827A>G, p.(Asn276Ser); c.847C>T, p.(Arg283Trp); c.1279C>T, p.(Arg427Cys)) in CNGA3, the α-subunit of the cyclic nucleotide-gated cation channel in cone photoreceptor cells. All three variants are predicted to replace evolutionary conserved amino acids, and to be pathogenic by specific in silico programs, consistent with the observed altered membrane targeting of CNGA3 in heterologous cells. Insights from our study will facilitate counseling regarding the molecular and phenotypic landscape of CNGA3-related cone dystrophies

Delineating the Molecular and Phenotypic Spectrum of the CNGA3-Related Cone Photoreceptor Disorder in Pakistani Families

Cone photoreceptor dysfunction represents a clinically heterogenous group of disorders characterized by nystagmus, photophobia, reduced central or color vision, and macular dystrophy. Here, we described the molecular findings and clinical manifestations of achromatopsia, a partial or total absence of color vision, co-segregating with three known missense variants of CNGA3 in three large consanguineous Pakistani families. Fundus examination and optical coherence tomography (OCT) imaging revealed myopia, thin retina, retinal pigment epithelial cells loss at fovea/perifovea, and macular atrophy. Combination of Sanger and whole exome sequencing revealed three known homozygous missense variants (c.827A&gt;G, p.(Asn276Ser); c.847C&gt;T, p.(Arg283Trp); c.1279C&gt;T, p.(Arg427Cys)) in CNGA3, the &alpha;-subunit of the cyclic nucleotide-gated cation channel in cone photoreceptor cells. All three variants are predicted to replace evolutionary conserved amino acids, and to be pathogenic by specific in silico programs, consistent with the observed altered membrane targeting of CNGA3 in heterologous cells. Insights from our study will facilitate counseling regarding the molecular and phenotypic landscape of CNGA3-related cone dystrophies

Molecular Genetic Studies and Delineation of the Oculocutaneous Albinism Phenotype in the Pakistani Population

Background: Oculocutaneous albinism (OCA) is caused by a group of genetically heterogeneous inherited defects that result in the loss of pigmentation in the eyes, skin and hair. Mutations in the TYR, OCA2, TYRP1 and SLC45A2 genes have been shown to cause isolated OCA. No comprehensive analysis has been conducted to study the spectrum of OCA alleles prevailing in Pakistani albino populations. Methods: We enrolled 40 large Pakistani families and screened them for OCA genes and a candidate gene, SLC24A5. Protein function effects were evaluated using in silico prediction algorithms and ex vivo studies in human melanocytes. The effects of splice-site mutations were determined using an exon-trapping assay. Results: Screening of the TYR gene revealed four known (p.Arg299His, p.Pro406Leu, p.Gly419Arg, p.Arg278*) and three novel mutations (p.Pro21Leu, p.Cys35Arg, p.Tyr411His) in ten families. Ex vivo studies revealed the retention of an EGFP-tagged mutant (p.Pro21Leu, p.Cys35Arg or p.Tyr411His) tyrosinase in the endoplasmic reticulum (ER) at 37°C, but a significant fraction of p.Cys35Arg and p.Tyr411His left the ER in cells grown at a permissive temperature (31°C). Three novel (p.Asp486Tyr, p.Leu527Arg, c.1045-15 T \u3e G) and two known mutations (p.Pro743Leu, p. Ala787Thr) of OCA2 were found in fourteen families. Exon-trapping assays with a construct containing a novel c.1045-15 T \u3e G mutation revealed an error in splicing. No mutation in TYRP1, SLC45A2, and SLC24A5 was found in the remaining 16 families. Clinical evaluation of the families segregating either TYR or OCA2 mutations showed nystagmus, photophobia, and loss of pigmentation in the skin or hair follicles. Most of the affected individuals had grayish-blue colored eyes. Conclusions: Our results show that ten and fourteen families harbored mutations in the TYR and OCA2 genes, respectively. Our findings, along with the results of previous studies, indicate that the p.Cys35Arg, p.Arg278* and p.Gly419Arg alleles of TYR and the p.Asp486Tyr and c.1045-15 T \u3e G alleles of OCA2 are the most common causes of OCA in Pakistani families. To the best of our knowledge, this study represents the first documentation of OCA2 alleles in the Pakistani population. A significant proportion of our cohort did not have mutations in known OCA genes. Overall, our study contributes to the development of genetic testing protocols and genetic counseling for OCA in Pakistani families

Molecular genetic studies and delineation of the oculocutaneous albinism phenotype in the Pakistani population

<p>Abstract</p> <p>Background</p> <p>Oculocutaneous albinism (OCA) is caused by a group of genetically heterogeneous inherited defects that result in the loss of pigmentation in the eyes, skin and hair. Mutations in the <it>TYR</it>, <it>OCA2</it>, <it>TYRP1</it> and <it>SLC45A2</it> genes have been shown to cause isolated OCA. No comprehensive analysis has been conducted to study the spectrum of <it>OCA</it> alleles prevailing in Pakistani albino populations.</p> <p>Methods</p> <p>We enrolled 40 large Pakistani families and screened them for <it>OCA</it> genes and a candidate gene, <it>SLC24A5</it>. Protein function effects were evaluated using <it>in silico</it> prediction algorithms and <it>ex vivo</it> studies in human melanocytes. The effects of splice-site mutations were determined using an exon-trapping assay.</p> <p>Results</p> <p>Screening of the <it>TYR</it> gene revealed four known (p.Arg299His, p.Pro406Leu, p.Gly419Arg, p.Arg278*) and three novel mutations (p.Pro21Leu, p.Cys35Arg, p.Tyr411His) in ten families. <it>Ex vivo</it> studies revealed the retention of an EGFP-tagged mutant (p.Pro21Leu, p.Cys35Arg or p.Tyr411His) tyrosinase in the endoplasmic reticulum (ER) at 37°C, but a significant fraction of p.Cys35Arg and p.Tyr411His left the ER in cells grown at a permissive temperature (31°C). Three novel (p.Asp486Tyr, p.Leu527Arg, c.1045-15 T > G) and two known mutations (p.Pro743Leu, p.Ala787Thr) of <it>OCA2</it> were found in fourteen families. Exon-trapping assays with a construct containing a novel c.1045-15 T > G mutation revealed an error in splicing. No mutation in <it>TYRP1</it>, <it>SLC45A2,</it> and <it>SLC24A5</it> was found in the remaining 16 families. Clinical evaluation of the families segregating either <it>TYR</it> or <it>OCA2</it> mutations showed nystagmus, photophobia, and loss of pigmentation in the skin or hair follicles. Most of the affected individuals had grayish-blue colored eyes.</p> <p>Conclusions</p> <p>Our results show that ten and fourteen families harbored mutations in the <it>TYR</it> and <it>OCA2</it> genes, respectively. Our findings, along with the results of previous studies, indicate that the p.Cys35Arg, p.Arg278* and p.Gly419Arg alleles of <it>TYR</it> and the p.Asp486Tyr and c.1045-15 T > G alleles of <it>OCA2</it> are the most common causes of OCA in Pakistani families. To the best of our knowledge, this study represents the first documentation of <it>OCA2</it> alleles in the Pakistani population. A significant proportion of our cohort did not have mutations in known <it>OCA</it> genes. Overall, our study contributes to the development of genetic testing protocols and genetic counseling for OCA in Pakistani families.</p

Identities and frequencies of variants in CYP1B1 causing primary congenital glaucoma in Pakistan

Purpose: Primary congenital glaucoma (PCG) is a clinically and genetically heterogeneous disease. The present study was undertaken to find the genetic causes of PCG segregating in 36 large consanguineous Pakistani families. Methods: Ophthalmic examination including fundoscopy, or slit-lamp microscopy was performed to clinically characterize the PCG phenotype. Genomic nucleotide sequences of the CYP1B1 and LTBP2 genes were analyzed with either Sanger or whole exome sequencing. In silico prediction programs were used to assess the pathogenicity of identified alleles. ClustalW alignments were performed to determine evolutionary conservation, and three-dimensional (3D) modeling was performed using HOPE and Phyre2 software. Results: Among the known loci, mutations in CYP1B1 and LTBP2 are the common causes of PCG. Therefore, we analyzed the genomic nucleotide sequences of CYP1B1 and LTBP2, and detected probable pathogenic variants cosegregating with PCG in 14 families. These included the three novel (c.542T>A, c.1436A>G, and c.1325delC) and five known (c.868dupC, c.1168C>T, c.1169G>A, c.1209InsTCATGCCACC, and c.1310C>T) variants in CYP1B1. Two of the novel variants are missense substitutions [p.(Leu181Gln), p.(Gln479Arg)], which replaced evolutionary conserved amino acids, and are predicted to be pathogenic by various in silico programs, while the third variant (c.1325delC) is predicted to cause reading frameshift and premature truncation of the protein. A single mutation, p.(Arg390His), causes PCG in six (~43%) of the 14 CYP1B1 mutations harboring families, and thus, is the most common variant in this cohort. Surprisingly, we did not find any LTBP2 pathogenic variants in the families, which further supports the genetic heterogeneity of PCG in the Pakistani population. Conclusions: In conclusion, results of the present study enhance our understanding of the genetic basis of PCG, support the notion of a genetic modifier of CYP1B1, and contribute to the development of genetic testing protocols and genetic counseling for PCG in Pakistani families