Muhammad Asif Qureshi

Differential immune cell densities in ductal carcinoma In-Situ and invasive breast cancer: Possible role of leukocytes in early stages of carcinogenesis.

Objectives: To investigate immune cell densities in pre-neoplastic (DCIS), cancer (IDC) and control breast tissues. Methods: A total of four preneoplastic, 104 cancer and 104 control samples were analyzed. Morphological classification and prognostic scoring along with quantification of immune cells/mm2 was performed. Data were entered and analyzed using SPSS version 16. Correlation of immune cell densities with various tumour sub-types was investigated using paired t-test and ANOVA. A p-value of <0.05 was considered as significant. Results: Our data show increased infiltration of lymphocytes (mean lymphocytes = 287.6cells/mm2) as well as myelocytes (mean lymphocytes = 117.1cells/mm2) in pre-neoplastic tissues. This infiltration was significantly high compared to cancer (p-value<0.001) as well as control tissues (p-value <0.001). Moreover, we report increased infiltration of lymphocytes in cancer tissues compared to controls (p-value<0.001). There was no difference in lymphocyte densities within various tumour sub-types (all p-values >0.05). Conclusion: Leukocytes may play a role in early stages of breast carcinogenesis.

Increased Tumour Infiltration of CD4+ and CD8+ T-Lymphocytes in Patients with Triple Negative Breast Cancer Suggests Susceptibility to Immune Therapy

Background: Patients with triple negative breast cancer (TNBC) have limited therapeutic options, largely because the complex tumour environment is not well-characterized. These patients are potential, but largely un-fathomed, candidates for immunotherapy. It is therefore highly relevant to characterize leukocyte complexity in TNBCs. Objective: To investigate leukocyte complexity in tumour environment of patients with TNBCs. Materials and methods: A total of 104 consecutive breast cancer patients undergoing mastectomy were recruited in the study after ethical approval. Clinico-pathological parameters were recorded and H and E staining was performed to investigate tumour morphology. Receptor status was investigated using antibodies against ER, PgR and Her-2, and patients were classified as having TNBC or non-TNBC tumours (including Luminal A, Luminal B and Her2 overexpressing tumours). Immune-cell infiltration was investigated using special stains and antibodies: α-CD3 (T-lymphocytes), α-CD20 (B-lymphocytes), α-CD4 (helper T-lymphocytes) and α-CD8 (cytotoxic T-lymphocytes). Immune cell densities were quantified as cell/mm2 using the CAP guidelines. Results: Of the 104 breast cancer patients investigated, a total of 27 (26%) had TNBC and 77(74%) non-TNBC. Patients with TNBC showed significantly increased tumour infiltration of lymphocytes (T and B-lymphocytes) compared to the patients with non-TNBC, while myelocytic infiltration was not significantly different in the two groups. Within the TNBC group, infiltration of T-lymphocytes (equal densities of CD4+ and CD8+ T-lymphocytes) was significantly higher compared to B-lymphocytes. Conclusion: Patients with TNBC show increased lymphocytic infiltration (more T-lymphocytes compared to B-lymphocytes). This suggests higher immunogenicity of TNBCs and may indicate a higher responsiveness of these cancers to immunotherapy.

HIV/AIDS registration in Pakistan

www.thelancet.com/infection Vol 15 June 2015 635 Khyberpakhtoonkhwa. This number of treatment centres is not enough to cover a population of more than 185 million. As a result, a large number of patients with HIV/AIDS remain unregistered in Pakistan. Moreover, no concrete data are available with respect to the number of people living with HIV, number of new HIV/AIDS registrations, and mortality status of HIV-infected patients in Pakistan. According to the NACP, the estimated number of people living with HIV in Pakistan is 97 400, whereas a 2014 Joint UN Program on HIV/AIDS report claims that 83 468 people were living with HIV at the end of 2013. However, these conflicting statistics are only estimates and are possibly far away from the true number. Appropriate and urgent actions must be undertaken by al l stakeholders—both Pakistani and inter national authorities—to establish an HIV/AIDS registry in Pakistan and increase public awareness. Regular public awareness campaigns, through seminars and print and electronic media could be an important initial step in this respect. Appropriate funding is needed to revolutionise the existing operations of the NACP to render it more functional and meaningful. Moreover, because Pakistan is an Islamic state in which a large proportion of people practice religion, engagement of Islamic scholars (priests, or Maulvi) could substantially increase public awareness via sermons in mosques.

Cancer registration in Pakistan: A dilemma that needs to be resolved

Dear Sir, National level cancer registration is nonexistent in Pakistan. Cancer statistics for Pakistan, as reported in the Globocan 2012 report published by the International Agency for Research on Cancer (IARC), are largely provided by the Punjab Cancer Registry (PCR)—the only provincial cancer registry in Pakistan. However, cancer statistics generated by the PCR are regional and not true representative of the whole country. Moreover, quality of these data has been stratified as category E (regional data rates) by the Globocan 2012 report, suggesting that the PCR data do not truly cover/represent cancer statistics in Pakistan. It is, therefore, extremely important to highlight the fact that a noticeable proportion of cancers in Pakistan are unregistered. This is an alarming situation requiring urgent response from all the stakeholders including Pakistani as well as international authorities. It is important to note that establishment of national level cancer registries demands huge logistical support including appropriate funds, human resource and political will amongst others. Nevertheless, and at least to begin with, several hospital-based registries could be initiated that can ultimately pool data to contribute toward provincial and subsequently national level cancer statistics. Indeed, there are several hospitals within the country that maintain their own records without contributing toward provincial and national statistics, probably because such forums are lacking in Pakistan. A successful model in Pakistan is the hospital-based cancer registry of the Shaukat Khanum Memorial Cancer and Research Centre, a major contributor of the PCR. Another example is the “currently-non-functional” Karachi Cancer Registry (KCR) that was founded by Dr. Yasmin Bhurguri. However, and after her death a couple of years ago, the KCR could not generate any further data. Several international examples also exist including the Surveillance, Epidemiology and End Results Program that is amongst the top research and funding priorities of the National Cancer Institute. In summary, there is a dire need that the government of Pakistan pays serious attention and allocates appropriate funding to establish hospital based, provincial as well as national level cancer registries. Moreover, international organizations such as the IARC and the Union for International Cancer Control should support Pakistan in this regard. Establishment of cancer statistics will not only provide incidence, prevalence, and mortality data, but will also allow the health care policy makers to devise appropriate screening and therapeutic strategies according to cancer burden in the country.

HIV-1 genetic diversity, geographical linkages and antiretroviral drug resistance among individuals from Pakistan

Worldwide antiretroviral therapy (ART) has reduced the mortality and morbidity rates in individuals with HIV infection. However, the increasing occurrence of drug resistance is limiting treatment options. In recent years, Pakistan has witnessed a concentrated epidemic of HIV. It is very important to identify geographical linkages and mutations that generate selective pressure and drive resistance of HIV in our population. The aim of this work was to identify genetic diversity and drug resistance patterns of HIV in Pakistan, using available sequences and bioinformatics tools, which may help in selecting effective combination of available drugs. A total of 755 Pakistani HIV gag, pol and env sequences were retrieved from the Los Alamos HIV database. Sequences were aligned with reference sequences of different subtypes. For geographical linkages, sequences of predominant subtypes were aligned with sequences of the same subtypes from different countries. Phylogenetic trees were constructed using the maximum-likelihood method in MEGA 7 software. For drug resistance analysis, sequences were entered into the Stanford University HIV Drug Resistance Database. Phylogenetic trees for studying genetic diversity showed that 82% of the sequences were of subtype A, while the rest of the sequences were of subtypes B (9.5%), K (2%), D (2%) and AE (1%). Moreover, trees that were constructed to examine geographical linkages showed close clustering of strains with those of the neighboring countries Afghanistan and India, as well as some African countries. A search for drug resistance mutations showed that 93% of the sequences had no major or minor mutations. The remaining 7% of the sequences contained a major mutation, Y115F, which causes the virus to exhibit low to intermediate resistance against lamivudine and emtricitabine. Our data indicate that HIV subtype A is the major subtype, while subtypes K, D and AE are also present in our country, suggesting gradual viral evolution and possible entry of different subtypes from neighboring countries. These data suggest that HIV is still sensitive to most of the antiretroviral drugs used in Pakistan.

Correction to: HIV-1 genetic diversity, geographical linkages and antiretroviral drug resistance among individuals from Pakistan

Unfortunately, the co-author’s name Nouman Mughal was incorrectly published in the original article and the same is corrected here in this erratum.

Frequency of Her2/neu expression in colorectal adenocarcinoma: A study from developing South Asian country

Results: Majority of our study population was aged over 35 years (74.3%) with a predominance of males (58.6%). 10% were overweight; 20% had a moderate alcohol consumption; 1.4% had diabetes; 38.5% were clinical stage 3 of the WHO and 8.7% in stage 4. The median CD4 count was 124 (19-618) cells / microliter. 62.9% of patients were treated with AZT+3TC+NVP; 10% in TNF+3TC+EFV; 7.1% for EFV+TNF+EMTRIC. The prevalence of hepatic steatosis was 59.2% and 14.2% of patients had advanced liver fibrosis. Factors associated with steatosis were an albumin level of less than 3.5 g/ dL OR=10.8 (1.5 to 264.6) p=0.02; elevation of alanine aminotransferase (ALT) OR=3.2 (1.1 to 9.8) p=0.02.

Cytogenetic and Molecular Analyses of Philadelphia Chromosome Variants in CML (chronic myeloid leukemia) Patients from Sindh using Karyotyping and RT-PCR.

Objective: To determine the frequency of Philadelphia chromosome (Ph) and its variants in chronic myeloid leukemia (CML) cases at a tertiary care hospital of Sindh. Methods: The study was conducted at the Department of Pathology, Liaquat University of Medical and Health Sciences, Jamshoro and Isra University Hospital, Hyderabad during May-to-September 2014. Bone marrow and peripheral blood samples from a total of 145 diagnosed cases of CML were collected. Cytogenetic analyses were performed using karyotyping as per the International System for Human Cytogenetic Nomenclature guidelines. All karyotypic images were analyzed using the Cytovision software. In order to identify BCR-ABL transcripts, RT-PCR was performed. Statistical analysis of the data was done using SPSS-version-21.0. Results: Of the 145 samples, a total of 133 (91.7%) were positive for the Ph (Ph+) while 12 (8.3%) were negative for the Ph (Ph-). Of the 133 Ph+ samples, standard karyotypes were noted in 121 (91%), simple variants in 9 (6.7%) and complex variants in 3 (2.3%) of the samples. All the Ph+ samples (n=133) showed BCR-ABL positivity. Of the 12 Ph- samples, a total of 7 (58.3%) were BCR-ABL-positive and 5 (41.6%) were BCR-ABL-negative. Conclusion: Frequency of the Ph was found to be of 90.9% in CML patients using a highly sensitive technique, the RT-PCR. Cytogenetic abnormalities were at a lower frequency. Cytogenetic and molecular studies must be conducted for better management of CML cases. These findings could be very useful in guiding the appropriate therapeutic options for CML patients.