Cytomorphologic differentiation of benign and malignant mammary tumors in fine needle aspirate specimens from irradiated female Sprague–Dawley rats

Background: Fine needle aspiration (FNA) offers a rapid and minimally invasive means to distinguish malignant from benign neoplasms. However, few studies have been published regarding the cytopathology of mammary tumors in rats despite widespread use of the rat model for breast cancer formation and inhibition.
Objective: The purpose of this study was to determine the diagnostic accuracy of FNA cytology and to develop distinguishing cytologic criteria for the diagnosis of radiation-induced benign and malignant mammary tumors in rats.
Methods: In a study of radiation-induced mammary carcinogenesis, 100 Sprague–Dawley rats with cutaneous masses were randomly chosen for FNA. The aspirates were smeared, fixed, and stained with a modified Papanicolaou procedure for diagnostic evaluation. Cytologic and histologic diagnoses (benign vs malignant) were compared, and diagnostic accuracy was calculated using the histologic diagnosis as the criterion standard. FNA smears were scored semiquantitatively on a scale of 1–4 for cellularity, atypia, nuclear size, chromatin pattern, nuclear membrane thickness, nucleoli, and mitoses. The background was evaluated for necrosis, hemorrhage, inflammation, and mucosecretory material. Cytomorphologic features were compared statistically between benign and malignant tumors, based on the histologic diagnosis.
Results: The sensitivity of FNA was 92.3% and specificity was 89.4% for the detection of malignancy. However, 14% of specimens, all fibroadenomas by histology, had insufficient cells for cytologic evaluation, for an overall accuracy rate of 78.0%. Malignant tumors had significantly higher scores for all cytomorphologic features, and were significantly more likely to contain cell clusters and necrotic debris.
Conclusions: FNA is an accurate method for differentiating benign and malignant rat mammary tumors.     

Physiological response of crossbred sheep under nutritional scarcity

Nutritional scarcity during summer and winter is a major constraint for sheep rearing in the semi-arid tropical region. In this region, a new crossbred sheep has been developed by the crossing of a native sheep breed Malpura with a breed of hot-humid coastal region breed Garole. A study was conducted for 9 weeks to assess the physiological response of crossbred (Garole x Malpura x Malpura, GMM) rams under nutritional scarcity in the semi-arid tropical region. Eighteen adult crossbred rams were randomly allocated into three groups of six animals each. The groups are G1 (control, ad libitum feeding), G2 (20% less than ad libitum feeding), and G3 (30% less than ad libitum feeding). The animals were stall-fed with a diet consisting of 70% roughage (Cenchrus ciliaris hay) and 30% concentrate feed. Body weight gain, average daily gain, and feed intake were significantly (P?<?0.05) lower in G2 and G3 as compared to G1 rams. Water intake was significantly (P?<?0.05) higher in G2 and G3. Physiological responses, blood biochemical and endocrine profile did not differ between the groups. The results indicate that the crossbred rams evolved through the crossing of the hot semi-arid tropical region breed with hot humid coastal region breed are able to maintain their body homeostasis despite nutritional scarcity although their body weight decreased.

     

Heritability estimates and genetic correlations of various production and reproductive traits of different grades of dairy cattle reared under subtropical condition

Inheritance of economic traits and their genetic correlation was studied in different breeds and their various grades (crossbred’s cattle) under the subtropical condition. Data on principal parameters comprising 10 years, that is 2004–2014, were utilized for the present study. Different grades of dairy cows comprising Holstein Friesian (HF), Sahiwal x Friesian (SxHF), Jersey (J), Jersey x Achai (JxAC) and Achai (AC) were included in the study. Heritability estimates and genetic correlations of some important productive and reproductive traits were worked out. Heritability estimates of certain economic traits were found to be: birthweight, 0.32 ± 0.181; age at maturity, 0.11 ± 0.136; age at first calving, 0.19 ± 0.162; days open, 0.09 ± 0.121; calving interval, 0.14 ± 0.211; dry period, 0.11 ± 0.124; lactation length, 0.04 ± 0.212 and lactation yield, 0.46 ± 0.206. Genetic correlation showed that body weight was significantly and positively correlated with lactation length (0.30) and lactation yield (0.81), while negatively correlated with age at maturity, age at first calving, days open, calving interval and dry period (?0.09, ?0.07, ?0.16, ?0.34 and ?0.002, respectively). Calving interval was positively and significantly correlated with the dry period (0.56), lactation length (0.72) and lactation yield (0.37). Moderate to higher heritability estimates with adequate genetic variance was found for body weight and lactation yield. The moderate to higher heritability estimates of birthweight in the present study revealed that this important trait might be considered in selection criteria.     

Localization of the motor neuron somata of geniohyoid muscle in rat: A horseradish peroxidase study

The aim of the study was to investigate the location of motor neuron somata of geniohyoid muscle in rat. Nine Sprague‐Dawley rats were used in this study. Operations were performed under general anaesthesia. Nembutal sodium, 40 mg per kg intraperitoneally was used for anaesthesia. 0.02 to 0.05 ml of 30% horseradish peroxidase (Sigma Type VI) solution in normal saline was injected into the exposed right geniohyoid muscle. After 48 hr, the animals were fixed by perfusion through left ventricle of heart, first by 100 ml normal saline and then with 500 ml of 1.25% glutaraldehyde and 1% paraformaldehyde in 0.1 M phosphate buffer, pH 7.4, at room temperature, and finally with 500 ml of 10% sucrose in the same buffer at 4°C. The medulla oblongata and first cervical segment of spinal cord were removed, kept in 10% sucrose in above phosphate buffer at 4°C for 24 hr. Thereafter, their serial transverse sections were cut in a cryostat at a thickness of 60 μm. The sections were treated according to tetramethyl benzidine (TMB)‐horseradish peroxidase (HRP) method. HRP‐labelled neuron somata were observed at the following sites: (a) In ventral part of right main hypoglossal nucleus in upper two‐thirds of the closed part of medulla oblongata. (b) In ventrolateral subnucleus of hypoglossal nucleus in lower third of closed part of medulla oblongata. (c) At spinomedullary junction, they were located in dorsomedial part of right ventral grey column; a few were also seen here scattered on right side of central canal and among corticospinal fibres.     

Modulation of seminal plasma content in extended semen improves the quality attributes of ram spermatozoa following liquid preservation at 3–5°C

Seminal plasma (SP) is known to induce motility and capacitation in spermatozoa curtailing their lifespan when preserved. Hence, this study was conducted to examine the effects of removal of SP from sperm surface prior to liquid preservation either by high dilution (1/15) or by washing and the poststorage treatment with SP (15% and 25%, v/v) on the quality attributes of liquid‐preserved ram semen. Over the period of storage, the rapid motility (66.0% and 71.1% vs. 58.3%), straightness (87.1% and 82.1% vs. 79.4%), average path velocity (152.3 and 152.0 µm/s vs. 133.3 µm/s) and the straight‐line velocity (131.3 and 127.8 µm/s vs. 108.5 µm/s) were significantly (p < 0.05) higher in both the high‐dilution and wash groups as compared to the control (1/3 dilution). The functional membrane integrity (82.3% vs. 77.2%) and noncapacitated sperm count (65.0% vs. 58.7%) were also significantly (p < 0.05) higher in the high‐dilution and wash groups, respectively, as compared to the control. The poststorage treatment of sperm with SP significantly (p < 0.05) increased the functional membrane integrity (70.1% vs. 53.8%) and most of the motility attributes as compared to the control (without SP). In conclusion, both the removal of SP prior to liquid preservation and poststorage treatment with SP significantly improved the quality attributes of ram spermatozoa.     

Enzymatic surface modification of polyacrylonitrile and its copolymers: Effects of polymer surface area and protein adsorption

Polyacrylonitrile (PAN) is a widely used polymer in the textile industry. PAN contains cyano groups on the surface due to which it possess low hydrophilicity and limits its application. Thus, there is a need to modify the functional groups on the surface of PAN for its industrial demand to improve moisture uptake, dyeability with ionic dyes, without affecting mechanical properties. A number of strategies such as chemical treatment, plasma treatment, enzymatic treatment etc. have been applied for the surface modification of polymer but enzymatic treatment are advantageous over plasma treatment and chemical treatment. In enzymatic treatment, reaction is limited to polymer surface only, and provides milder condition with less damage to polymer. In present study, it was found that enzyme system of Amycolatopsis sp.IITR215 was effective enzyme system for modification of surface nitrile groups of polyacrylonitrile. PAN powder was treated with the cell free extract of Amycolatopsis sp.IITR215 and it was found that the nitrile metabolizing enzymes of this strain were efficiently able to transform -CN to -COOH groups present on the surfaces of PAN powder. The formation of carboxyl group was quantified by ammonia released and dye binding assay. Further, confirmation of carboxyl group on polymer was done by FTIR and XPS. This study indicates that, specific adsorption of enzyme probably plays an important role in the enzymatic surface modification of polymer.     

Simultaneous HPLC determination of gamma amino butyric acid (GABA) and lysine in selected Pakistani rice varieties by pre-column derivatization with 2-Hydroxynaphthaldehyde

A selective and sensitive HPLC method for the simultaneous determination of gamma amino butyric acid (GABA) and lysine in Pakistani rice varieties was developed. Both analytes were detected in rice varieties as Schiff base derivatives with 2-hydroxynaphthaldehyde. The derivatives were analyzed on a reverse phase C-18 column with Diode Array Detector (DAD) at 254 nm. The calibration curves were found linear over a concentration range of 3.83–34.58 μg/mL for GABA and 5.16–48.68 μg/mL for lysine with a correlation coefficient of 0.998 for both standards. GABA and lysine contents were found higher in brown rice varieties (4.1–6.58 mg/100 g for GABA and 15.1–27.6 mg/100 g for lysine) than the polished varieties (0.32–0.47 mg/100 g GABA and 13.1–19.8 mg/100 g lysine). This method could be used for separation and quantification of GABA and lysine simultaneously in food samples, especially in cereal seeds.     

Spatio-temporal variability of drought characteristics across Pakistan

Climate change in Pakistan has a great impact on the spatial and temporal variation of precipitation and ultimately alters the frequency and duration of droughts. In this study, spatial and temporal trend analyses of precipitation and droughts were observed at 58 meteorological stations across Pakistan from 1981 to 2018. The existing trend analysis methods were evaluated to address the issue of serial correlation in the climatic data. Results of precipitation analysis showed significant decreasing trends in winter (November, December) and significant increasing trends were observed in summer (June and September) at a confidence level of 95 percent. The magnitude of the precipitation trends showed the highest variation during summer season and the least variation in winter season. Rotated Principal Component (RPC) analysis showed the severe droughts (high positive loading) in southeastern side (Sindh province) of Pakistan due to lack of summer rains. Furthermore, variance correction approaches are identified as the most suitable in coping with the effect of serial correlation. The highest drought frequencies were observed in the southern areas of Pakistan and the drought events are expected to occur more frequently in the late winter, early spring, and early autumn, while droughts were expected to occur least frequently in summer.

     

Isolation and identification of bovine Brucella isolates from Pakistan by biochemical tests and PCR

Brucellosis is endemic in bovines in Pakistan. The Brucella species and biovars involved, however, are unknown. The objectives of the present study were to isolate and characterize brucellae from seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes which had recently aborted. The seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes were collected from the Potohar Plateau, Pakistan. Isolation of brucellae was done on modified Farrell’s serum dextrose agar. Isolates were characterized by conventional biotyping methods, while molecular typing was done by genus (B4/B5) and species-specific (Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis) polymerase chain reaction (PCR). A total of 30 isolates were recovered from milk (n?=?5), aborted fetuses (n?=?13), and vaginal swabs (n?=?12). Most isolates were from cattle (56.7 %). All of them were identified as B. abortus biovar 1 based on conventional biotyping methods and genus and species-specific PCR. This preliminary study provides the first report on the prevalence of B. abortus biovar 1 in cattle and buffaloes in Pakistan.