# Antibacterial and Antiproliferative Potential of Green, Roasted, and Spent Coffee Extracts
## 2 Results and Discussion
### Antibacterial Activity
\begin{table}
\begin{tabular}{c c c c c c c c c c c c} \hline \hline  & & & & **MIC (mg\(\cdot\)mL\({}^{-1}\))** & & & & & & & \\ \hline \multicolumn{1}{c}{} & \multicolumn{1}{c}{} & \multicolumn{1}{c}{} & **ATCC** & & & & & & & \\ \hline \multicolumn{1}{c}{**Extract**} & **Sample** & **EC1** & **EClo** & **Sd** & **Sa1** & **Ef** & **Kp** & **EC2** & **Sh** & **Sho** & **Sa2** \\ \hline \multirow{3}{*}{Ethanolic} & GB & 8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & 8 & \textgreater{}8 & \textgreater{}8 & 8 \\  & RC & 8 & \textgreater{}8 & 8 & 2 & 8 & 4 & 4 & 4 & 2 & 4 \\  & SCG & 8 & 8 & 8 & 8 & 8 & 2 & 2 & 2 & 2 & 2 \\ \cline{2-11}  & GB & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 \\  & RC & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & \textgreater{}8 & 4 & 4 & 4 & 4 \\  & SCG & \textgreater{}8 & 8 & 8 & 4 & 8 & 2 & 4 & 4 & 8 & 4 \\ \cline{2-11}  & C\(-\) & \(+\) & \(+\) & \(+\) & \(+\) & \(+\) & \(+\) & \(+\) & \(+\) & \(+\) & \(+\) \\  & C\(+\) & \(-\) & \(-\) & \(-\) & \(-\) & \(-\) & \(-\) & \(-\) & \(-\) & \(-\) & \(-\) \\ \hline \hline \end{tabular}
\begin{tabular}{c c c c c c c c c c} \hline \hline  & & & & & & & & & & \\  & EClo, _Enterobacter_ & _clone_ & 700323; & Sd _Salmonella dublin_ & 9676; & Sa1, _Staphylococcus aureus_ & 25,923; Ef, _Enterococcus faecalis_ & 29212. Clinically isolated methicillin-resistant strains: Kp, _Klebsiella pneumoniae_; EC, _Escherichia coli_; Sh, _Staphylococcus haemolyticus_; Sho, _Staphylococcus homitis_; and Sa2, _Staphylococcus aureus_. C\(-\): negative control (+, bacterial growth); C+: positive control, amikacin at 100 mg\(\cdot\)mL\({}^{-1}\) (\(-\), no bacterial growth).  
\end{table} 
Table 2: Minimal inhibitory concentration (MIC) of coffee extracts. Analysis of In Vitro Antiproliferative Activity, Morphological Changes, Apoptosis, and Cell-Cycle Arrest  
Cancers are characterized by uncontrolled cell proliferation and increased oxidative stress. Evidence has shown that some coffee metabolites can reduce viability and cell proliferation in transformed cells. We compared the antiproliferative effects of the aqueous and ethanolic extracts of GB, RC, and SCG. Our results show that the aqueous and ethanolic extracts from GB, RC, and SCG affected cell viability differently in A549 and C33A cell lines. In the antiproliferative activity assays, the SCG and RC extracts led to a significant decrease in viability in C33A cells (\(p<0.001\)). The most significant effect in C33A was observed with ethanolic extracts from GB and RC; these extracts diminished cell viability by up to 58.5% and 57.6%, respectively. The most efficacious concentrations were 100, 125, and 150 \(\upmu\)g\(\cdot\)mL\({}^{-1}\). Notably, the ethanolic extract of SCG diminished cell proliferation in 30.5% of cells at 50 \(\upmu\)g\(\cdot\)mL\({}^{-1}\), and in 53.9% of cells at 25 \(\upmu\)g\(\cdot\)mL\({}^{-1}\) (Figure 1A). The aqueous extract of SCG showed a reduction in cell viability by 56.7% and by 27.6% at 50 and 150 \(\upmu\)g\(\cdot\)mL\({}^{-1}\), respectively (Figure 1C). Morphological changes associated with apoptosis include cellular size reduction, cytoplasmic inclusions, and cellular fragmentation [38]. We observed morphological changes in both cell lines treated for 24 h with the extracts; however, the changes were more evident in C33A cells. Ethanolic extracts caused cellular debris, as occurs with paclitaxel treatment, for most concentrations in C33A cells (Figure 1B). Furthermore, changes in cell morphology and reductions in the number of cells per field were evident, mainly at concentrations of 50 and 150 \(\upmu\)g\(\cdot\)mL\({}^{-1}\) in all aqueous extracts, but mainly in extracts from SCG (Figure 1D).